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Sample records for tetraploid medicago sativa

  1. Molecular Diversity and Population Structure of a Worldwide Collection of Cultivated Tetraploid Alfalfa (Medicago sativa subsp. sativa L.) Germplasm as Revealed by Microsatellite Markers.

    Science.gov (United States)

    Qiang, Haiping; Chen, Zhihong; Zhang, Zhengli; Wang, Xuemin; Gao, Hongwen; Wang, Zan

    2015-01-01

    Information on genetic diversity and population structure of a tetraploid alfalfa collection might be valuable in effective use of the genetic resources. A set of 336 worldwide genotypes of tetraploid alfalfa (Medicago sativa subsp. sativa L.) was genotyped using 85 genome-wide distributed SSR markers to reveal the genetic diversity and population structure in the alfalfa. Genetic diversity analysis identified a total of 1056 alleles across 85 marker loci. The average expected heterozygosity and polymorphism information content values were 0.677 and 0.638, respectively, showing high levels of genetic diversity in the cultivated tetraploid alfalfa germplasm. Comparison of genetic characteristics across chromosomes indicated regions of chromosomes 2 and 3 had the highest genetic diversity. A higher genetic diversity was detected in alfalfa landraces than that of wild materials and cultivars. Two populations were identified by the model-based population structure, principal coordinate and neighbor-joining analyses, corresponding to China and other parts of the world. However, lack of strictly correlation between clustering and geographic origins suggested extensive germplasm exchanges of alfalfa germplasm across diverse geographic regions. The quantitative analysis of the genetic diversity and population structure in this study could be useful for genetic and genomic analysis and utilization of the genetic variation in alfalfa breeding.

  2. Molecular Diversity and Population Structure of a Worldwide Collection of Cultivated Tetraploid Alfalfa (Medicago sativa subsp. sativa L. Germplasm as Revealed by Microsatellite Markers.

    Directory of Open Access Journals (Sweden)

    Haiping Qiang

    Full Text Available Information on genetic diversity and population structure of a tetraploid alfalfa collection might be valuable in effective use of the genetic resources. A set of 336 worldwide genotypes of tetraploid alfalfa (Medicago sativa subsp. sativa L. was genotyped using 85 genome-wide distributed SSR markers to reveal the genetic diversity and population structure in the alfalfa. Genetic diversity analysis identified a total of 1056 alleles across 85 marker loci. The average expected heterozygosity and polymorphism information content values were 0.677 and 0.638, respectively, showing high levels of genetic diversity in the cultivated tetraploid alfalfa germplasm. Comparison of genetic characteristics across chromosomes indicated regions of chromosomes 2 and 3 had the highest genetic diversity. A higher genetic diversity was detected in alfalfa landraces than that of wild materials and cultivars. Two populations were identified by the model-based population structure, principal coordinate and neighbor-joining analyses, corresponding to China and other parts of the world. However, lack of strictly correlation between clustering and geographic origins suggested extensive germplasm exchanges of alfalfa germplasm across diverse geographic regions. The quantitative analysis of the genetic diversity and population structure in this study could be useful for genetic and genomic analysis and utilization of the genetic variation in alfalfa breeding.

  3. Comparative gene expression profiles between heterotic and non-heterotic hybrids of tetraploid Medicago sativa

    Directory of Open Access Journals (Sweden)

    Nettleton Dan

    2009-08-01

    Full Text Available Abstract Background Heterosis, the superior performance of hybrids relative to parents, has clear agricultural value, but its genetic control is unknown. Our objective was to test the hypotheses that hybrids expressing heterosis for biomass yield would show more gene expression levels that were different from midparental values and outside the range of parental values than hybrids that do not exhibit heterosis. Results We tested these hypotheses in three Medicago sativa (alfalfa genotypes and their three hybrids, two of which expressed heterosis for biomass yield and a third that did not, using Affymetrix M. truncatula GeneChip arrays. Alfalfa hybridized to approximately 47% of the M. truncatula probe sets. Probe set signal intensities were analyzed using MicroArray Suite v.5.0 (MAS and robust multi-array average (RMA algorithms. Based on MAS analysis, the two heterotic hybrids performed similarly, with about 27% of genes showing differential expression among the parents and their hybrid compared to 12.5% for the non-heterotic hybrid. At a false discovery rate of 0.15, 4.7% of differentially expressed genes in hybrids (~300 genes showed nonadditive expression compared to only 0.5% (16 genes in the non-heterotic hybrid. Of the nonadditively expressed genes, approximately 50% showed expression levels that fell outside the parental range in heterotic hybrids, but only one of 16 showed a similar profile in the non-heterotic hybrid. Genes whose expression differed in the parents were three times more likely to show nonadditive expression than genes whose parental transcript levels were equal. Conclusion The higher proportions of probe sets with expression level that differed from the parental midparent value and that were more extreme than either parental value in the heterotic hybrids compared to a non-heterotic hybrid were also found using RMA. We conclude that nonadditive expression of transcript levels may contribute to heterosis for biomass

  4. Construction of two genetic linkage maps in cultivated tetraploid alfalfa (Medicago sativa) using microsatellite and AFLP markers

    Science.gov (United States)

    Julier, Bernadette; Flajoulot, Sandrine; Barre, Philippe; Cardinet, Gaëlle; Santoni, Sylvain; Huguet, Thierry; Huyghe, Christian

    2003-01-01

    Background Alfalfa (Medicago sativa) is a major forage crop. The genetic progress is slow in this legume species because of its autotetraploidy and allogamy. The genetic structure of this species makes the construction of genetic maps difficult. To reach this objective, and to be able to detect QTLs in segregating populations, we used the available codominant microsatellite markers (SSRs), most of them identified in the model legume Medicago truncatula from EST database. A genetic map was constructed with AFLP and SSR markers using specific mapping procedures for autotetraploids. The tetrasomic inheritance was analysed in an alfalfa mapping population. Results We have demonstrated that 80% of primer pairs defined on each side of SSR motifs in M. truncatula EST database amplify with the alfalfa DNA. Using a F1 mapping population of 168 individuals produced from the cross of 2 heterozygous parental plants from Magali and Mercedes cultivars, we obtained 599 AFLP markers and 107 SSR loci. All but 3 SSR loci showed a clear tetrasomic inheritance. For most of the SSR loci, the double-reduction was not significant. For the other loci no specific genotypes were produced, so the significant double-reduction could arise from segregation distortion. For each parent, the genetic map contained 8 groups of four homologous chromosomes. The lengths of the maps were 2649 and 3045 cM, with an average distance of 7.6 and 9.0 cM between markers, for Magali and Mercedes parents, respectively. Using only the SSR markers, we built a composite map covering 709 cM. Conclusions Compared to diploid alfalfa genetic maps, our maps cover about 88–100% of the genome and are close to saturation. The inheritance of the codominant markers (SSR) and the pattern of linkage repulsions between markers within each homology group are consistent with the hypothesis of a tetrasomic meiosis in alfalfa. Except for 2 out of 107 SSR markers, we found a similar order of markers on the chromosomes between the

  5. High-resolution melting analysis for SNP genotyping and mapping in tetraploid alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Han, Yuanhong; Khu, Dong-Man; Monteros, Maria J

    2012-02-01

    Single nucleotide polymorphisms (SNPs) represent the most abundant type of genetic polymorphism in plant genomes. SNP markers are valuable tools for genetic analysis of complex traits of agronomic importance, linkage and association mapping, genome-wide selection, map-based cloning, and marker-assisted selection. Current challenges for SNP genotyping in polyploid outcrossing species include multiple alleles per loci and lack of high-throughput methods suitable for variant detection. In this study, we report on a high-resolution melting (HRM) analysis system for SNP genotyping and mapping in outcrossing tetraploid genotypes. The sensitivity and utility of this technology is demonstrated by identification of the parental genotypes and segregating progeny in six alfalfa populations based on unique melting curve profiles due to differences in allelic composition at one or multiple loci. HRM using a 384-well format is a fast, consistent, and efficient approach for SNP discovery and genotyping, useful in polyploid species with uncharacterized genomes. Possible applications of this method include variation discovery, analysis of candidate genes, genotyping for comparative and association mapping, and integration of genome-wide selection in breeding programs. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11032-011-9566-x) contains supplementary material, which is available to authorized users.

  6. The Medicago sativa gene index 1.2: a web-accessible gene expression atlas for investigating expression differences between Medicago sativa subspecies

    Science.gov (United States)

    Alfalfa (Medicago sativa L.) is the primary forage legume crop species in the United States and plays essential economic and ecological roles in agricultural systems across the country. Modern alfalfa is the result of hybridization between tetraploid M. sativa ssp. sativa and M. sativa ssp. falcata....

  7. Alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Fu, Chunxiang; Hernandez, Timothy; Zhou, Chuanen; Wang, Zeng-Yu

    2015-01-01

    Alfalfa (Medicago sativa L.) is a high-quality forage crop widely grown throughout the world. This chapter describes an efficient protocol that allows for the generation of large number of transgenic alfalfa plants by sonication-assisted Agrobacterium-mediated transformation. Binary vectors carrying different selectable marker genes that confer resistance to phosphinothricin (bar), kanamycin (npt II), or hygromycin (hph) were used to generate transgenic alfalfa plants. Intact trifoliates collected from clonally propagated plants in the greenhouse were sterilized with bleach and then inoculated with Agrobacterium strain EHA105. More than 80 % of infected leaf pieces could produce rooted transgenic plants in 4-5 months after Agrobacterium-mediated transformation.

  8. Lucerna (Medicago sativa L.) kraljica krmnih kultura

    OpenAIRE

    Halagić, Stevo

    2005-01-01

    Lucerna (Medicago sativa L.) je jedna od njavrijednijih, pa stoga i najraširenijih krmnih kultura u proizvodnji voluminozne stoËne hrane na oranicama i sa pravom nosi ime “kraljica krmnih kultura”.

  9. Karyotypic evolution of the Medicago complex: sativa-caerulea-falcata inferred from comparative cytogenetic analysis.

    Science.gov (United States)

    Yu, Feng; Wang, Haiqing; Zhao, Yanyan; Liu, Ruijuan; Dou, Quanwen; Dong, Jiangli; Wang, Tao

    2017-04-21

    Polyploidy plays an important role in the adaptation and speciation of plants. The alteration of karyotype is a significant event during polyploidy formation. The Medicago sativa complex includes both diploid (2n = 2× = 16) and tetraploid (2n = 2× = 32) subspecies. The tetraploid M. ssp. sativa was regarded as having a simple autopolyploid origin from diploid ssp. caerulea, whereas the autopolyploid origin of tetraploid ssp. falcata from diploid form ssp. falcata is still in doubt. In this study, detailed comparative cytogenetic analysis between diploid to tetraploid species, as well as genomic affinity across different species in the M. sativa complex, were conducted based on comparative mapping of 11 repeated DNA sequences and two rDNA sequences by a fluorescence in situ hybridization (FISH) technique. FISH patterns of the repeats in diploid subspecies caerulea were highly similar to those in tetraploid subspecies sativa. Distinctly different FISH patterns were first observed in diploid ssp. falcata, with only centromeric hybridizations using centromeric and multiple region repeats and a few subtelomeric hybridizations using subtelomeric repeats. Tetraploid subspecies falcata was unexpectedly found to possess a highly variable karyotype, which agreed with neither diploid ssp. falcata nor ssp. sativa. Reconstruction of chromosome-doubling process of diploid ssp. caerulea showed that chromosome changes have occurred during polyploidization process. The comparative cytogenetic results provide reliable evidence that diploid subspecies caerulea is the direct progenitor of tetraploid subspecies sativa. And autotetraploid ssp. sativa has been suggested to undergo a partial diploidization by the progressive accumulation of chromosome structural rearrangements during evolution. However, the tetraploid subspecies falcata is far from a simple autopolyploid from diploid subspecies falcata although no obvious morphological change was observed between these two

  10. Ploidy Effects in Isogenic Populations of Alfalfa (Medicago sativa L.) 1

    Science.gov (United States)

    Meyers, Steven P.; Brinegar, A. Chris; Schrader, Larry E.; Jordan, Douglas B.; Ogren, William L.

    1983-01-01

    Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCase) was isolated from isogenic diploid-tetraploid and tetraploid-octoploid sets of alfalfa (Medicago sativa L.) leaves. Molecular weights of RuBPCase subunits were similar across ploidy levels of both isogenic sets with subunits of 52,000 and 14,000. Apparent Km(CO2) values and substrate specificity factors (VcKo/VoKc) of RuBPCase were similar across ploidy levels of both isogenic sets. These results indicate that ploidy had no effect on the kinetic properties of RuBPCase in alfalfa. Images Fig. 1 PMID:16662937

  11. The Medicago sativa gene index 1.2: a web-accessible gene expression atlas for investigating expression differences between Medicago sativa subspecies.

    Science.gov (United States)

    O'Rourke, Jamie A; Fu, Fengli; Bucciarelli, Bruna; Yang, S Sam; Samac, Deborah A; Lamb, JoAnn F S; Monteros, Maria J; Graham, Michelle A; Gronwald, John W; Krom, Nick; Li, Jun; Dai, Xinbin; Zhao, Patrick X; Vance, Carroll P

    2015-07-07

    Alfalfa (Medicago sativa L.) is the primary forage legume crop species in the United States and plays essential economic and ecological roles in agricultural systems across the country. Modern alfalfa is the result of hybridization between tetraploid M. sativa ssp. sativa and M. sativa ssp. falcata. Due to its large and complex genome, there are few genomic resources available for alfalfa improvement. A de novo transcriptome assembly from two alfalfa subspecies, M. sativa ssp. sativa (B47) and M. sativa ssp. falcata (F56) was developed using Illumina RNA-seq technology. Transcripts from roots, nitrogen-fixing root nodules, leaves, flowers, elongating stem internodes, and post-elongation stem internodes were assembled into the Medicago sativa Gene Index 1.2 (MSGI 1.2) representing 112,626 unique transcript sequences. Nodule-specific and transcripts involved in cell wall biosynthesis were identified. Statistical analyses identified 20,447 transcripts differentially expressed between the two subspecies. Pair-wise comparisons of each tissue combination identified 58,932 sequences differentially expressed in B47 and 69,143 sequences differentially expressed in F56. Comparing transcript abundance in floral tissues of B47 and F56 identified expression differences in sequences involved in anthocyanin and carotenoid synthesis, which determine flower pigmentation. Single nucleotide polymorphisms (SNPs) unique to each M. sativa subspecies (110,241) were identified. The Medicago sativa Gene Index 1.2 increases the expressed sequence data available for alfalfa by ninefold and can be expanded as additional experiments are performed. The MSGI 1.2 transcriptome sequences, annotations, expression profiles, and SNPs were assembled into the Alfalfa Gene Index and Expression Database (AGED) at http://plantgrn.noble.org/AGED/ , a publicly available genomic resource for alfalfa improvement and legume research.

  12. Complex patterns of autopolyploid evolution in alfalfa and allies (Medicago sativa; Leguminosae).

    Science.gov (United States)

    Havananda, Tee; Brummer, E Charles; Doyle, Jeff J

    2011-10-01

    Although there is growing evidence that autopolyploidy is a widespread and important evolutionary phenomenon, it has received less attention than allopolyploidy. Medicago sativa comprises several diploid and autopolyploid taxa, including autotetraploid cultivated alfalfa, and affords an opportunity to elucidate the evolutionary history of a morphologically and genetically complex autopolyploid system. Phylogenies and haplotype networks were constructed from two chloroplast noncoding regions (rpl20-rps12 and trnS-trnG spacers) across seven diploid and polyploid infraspecific taxa of M. sativa and five additional closely related Medicago species, and genetic differentiation was estimated. The two most prominent M. sativa autopolyploids have contrasting evolutionary histories. Chloroplast data support a simple autopolyploid origin of subsp. sativa (alfalfa) from diploid subsp. caerulea, from which it is distinguishable in several quantitative characters. In contrast, morphologically identical diploid and autopolyploid cytotypes of subsp. falcata were found to possess very different chloroplast haplotypes, suggesting past introgression from M. prostrata into the polyploid. Despite the presence of hybrids between tetraploid subspecies falcata and sativa, there was little evidence of introgression of chloroplast genomes from either subspecies into the other. Autopolyploid evolution in M. sativa is complicated and has followed very different paths in different subspecific taxa. The potential exists for gene flow in virtually all combinations of subspecies both within and between ploidies, yet despite the existence of hybrids, morphologically and genetically distinctive subspecies persist.

  13. Comparative Analysis of the Symbiotic Efficiency of Medicago truncatula and Medicago sativa under Phosphorus Deficiency

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    Sulieman, Saad; Schulze, Joachim; Tran, Lam-Son

    2013-01-01

    Phosphorus (P)-deficiency is a major abiotic stress that limits legume growth in many types of soils. The relationship between Medicago and Sinorhizobium, is known to be affected by different environmental conditions. Recent reports have shown that, in combination with S. meliloti 2011, Medicago truncatula had a lower symbiotic efficiency than Medicago sativa. However, little is known about how Medicago–Sinorhizobium is affected by P-deficiency at the whole-plant level. The obj...

  14. Stress responses in alfalfa (Medicago sativa L. )

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    Kessmann, H.; Edwards, R.; Dixon, R.A. (Samuel Roberts Noble Foundation, Ardmore, OK (USA)); Geno, P.W. (Oklahoma State Univ., Stillwater (USA))

    1990-09-01

    The isoflavonoid conjugates medicarpin-3-O-glucoside-6{double prime}-O-malonate (MGM), afrormosin-7-O-glucoside (AG), and afrormosin-7-O-glucoside-6{double prime}-O-malonate (AGM) were isolated and characterized from cell suspension cultures of alfalfa (Medicago sativa L.), where they were the major constitutive secondary metabolites. They were also found in alfalfa roots but not in other parts of the plant. The phytoalexin medicarpin accumulated rapidly in suspension cultured cells treated with elicitor from Colletotrichum lindemuthianum, and this was subsequently accompanied by an increase in the levels of MGM. In contrast, net accumulation of afrormosin conjugates was not affected by elicitor treatment. Labeling studies with ({sup 14}C)phenylalanine indicated that afrormosin conjugates were the major de novo synthesized isoflavonoid products in unelicited cells. During elicitation, ({sup 14}C)phenylalanine was incorporated predominantly into medicarpin, although a significant proportion of the newly synthesized medicarpin was also conjugated. Treatment of {sup 14}C-labeled, elicited cells with L-{alpha}-aminooxy-{beta}-phenylpropionic acid, a potent inhibitor of PAL activity in vivo, resulted in the initial appearance of labeled medicarpin of very low specific activity, suggesting that the phytoalexin could be released from a preformed conjugate under these conditions. Our data draw attention to the involvement of isoflavone hydroxylases during the constitutive and elicitor-induced accumulation of isoflavonoids and their conjugates in alfalfa cell cultures.

  15. Stress Responses in Alfalfa (Medicago sativa L.)

    Science.gov (United States)

    Choudhary, Arvind D.; Lamb, Christopher J.; Dixon, Richard A.

    1990-01-01

    Protoplasts derived from cell suspensions of alfalfa (Medicago sativa L.) responded to treatment with fungal elicitor (FE) by an increase in endogenous chalcone synthase (CHS) activity but were unresponsive to reduced glutathione (GSH). Preexposure of protoplasts to polyethylene glycol and electroporation resulted in strong responsiveness to GSH but little change in responsiveness to FE. Protoplasts from suspension cultures which had been subcultured more than 12 times lost responsiveness to GSH, but not FE, as assessed by measuring expression of a chimeric gene containing a bean CHS promoter linked to a bacterial chloramphenicol acetyltransferase (CAT) reporter gene. In protoplasts in which putative cis-acting CHS promoter sequences had been coelectroporated in trans with the intact CHS promoter-CAT construct, the extent of CAT expression depended upon the elicitor used (FE or GSH), the age (number of times subcultured) of the cells from which the protoplasts were isolated, and the nature of the coelectroporated CHS promoter sequence. For example, a region of the CHS promoter from -326 to -141 behaved as a trans-activator when coelectroporated with the CAT construct into unelicited protoplasts isolated from newly initiated cell suspensions, but the same region acted as a trans-silencer in the same protoplasts in the presence of FE. This silencer activity was much reduced in GSH-treated protoplasts. The results suggest that there are differences in the signal transduction pathways for elicitation of CHS transcription by FE and GSH, which involve previously indentified cis-elements in the CHS promoter. PMID:16667919

  16. Evaluation of models for assessing Medicago sativa L. hay

    African Journals Online (AJOL)

    UFS Campus

    Abstract. A study was conducted to evaluate current proposed models for assessing Medicago sativa L. hay quality, using near infrared reflectance spectroscopy (NIRS) analyses and Cornell Nett Carbohydrate and. Protein System (CNCPS) milk production prediction as a criterion of accuracy. Application of the.

  17. Evaluation of models for assessing Medicago sativa L. hay

    African Journals Online (AJOL)

    UFS Campus

    ² Agricultural Modeling and Training Systems 418 Davis Rd Cortland, NY 13045, USA. Abstract. A study was conducted to evaluate current proposed models for assessing Medicago sativa L. hay quality, using near infrared reflectance spectroscopy (NIRS) analyses and Cornell Nett Carbohydrate and. Protein System ...

  18. Prediction of chemical composition of South African Medicago sativa ...

    African Journals Online (AJOL)

    The near infrared reflectance spectroscopy (NIRS) to predict chemical and digestibility parameters was investigated. Samples (n = 168) representing the spectral characteristics of the South African. Medicago sativa L. hay population were chemically analysed for the development of calibration equations. Values for r² and ...

  19. Productivity evaluation of Medicago sativa cultivars under irrigation ...

    African Journals Online (AJOL)

    Many Medicago sativa cultivars are available in South Africa, each with unique characteristics. This study evaluated 26 cultivars under irrigation on two ecotopes in terms of number of cuttings, dry matter (DM) production, seasonal DM production curve, DM production curve over lifespan and plant density, over nine growing ...

  20. A model for assessing Medicago Sativa L. hay quality | Scholtz ...

    African Journals Online (AJOL)

    A study was conducted to identify chemical parameters and/or models for assessing. Medicago sativa L. (L) hay quality, using near infrared reflectance spectroscopy (NIRS) analysis and Cornell Net Carbohydrate and Protein System (CNCPS) milk prediction as a criterion of accuracy. Milk yield (MY) derived from the ...

  1. Sample preparation of Medicago sativa L. hay for chemical analysis ...

    African Journals Online (AJOL)

    The objective of this study was to quantify the effect of the grinding procedure on the moisture and crude protein concentration of a ground Medicago sativa L. hay sample for quality grading. An additional aim was to investigate the accuracy of electronic moisture testers (EMT). Variance of analyses revealed significant ...

  2. Is There a Relationship between Infection by Rhizobia and Occurrence of Disomatic Nuclei in Nodules of Alfalfa (Medicago sativa L.)? 1

    Science.gov (United States)

    Shanklin, John; Schrader, Larry E.

    1986-01-01

    Cytological examination of nodules from diploid, tetraploid, and octoploid alfalfa (Medicago sativa L.) plants revealed that the proportion of nodule cells infected by rhizobia was not significantly affected by nuclear ploidy of the host plant. Flow cytometry was used to determine the influence of host plant nuclear ploidy on the nuclear ploidy of infected cells. In nodules from diploid plants, most of the nuclei were tetraploid, whereas in nodules from tetraploid plants, about half of the nodule nuclei were tetraploid and half were octoploid; in octoploid plants, most of the nodule nuclei were octoploid. The occurrence of disomatic nuclei was independent of infection of nodule cells by rhizobia, because diploid plants had mostly disomatic nodule nuclei, and octoploid plants had mostly monosomatic nodule nuclei, whereas all nodules maintained a constant proportion of infected to uninfected cells. These results do not support the earlier hypothesis that infected nodule cells contain disomatic nuclei. PMID:16664599

  3. Sexual Polyploidization in Medicago sativa L.: Impact on the Phenotype, Gene Transcription, and Genome Methylation.

    Science.gov (United States)

    Rosellini, Daniele; Ferradini, Nicoletta; Allegrucci, Stefano; Capomaccio, Stefano; Zago, Elisa Debora; Leonetti, Paola; Balech, Bachir; Aversano, Riccardo; Carputo, Domenico; Reale, Lara; Veronesi, Fabio

    2016-04-07

    Polyploidization as the consequence of 2n gamete formation is a prominent mechanism in plant evolution. Studying its effects on the genome, and on genome expression, has both basic and applied interest. We crossed two diploid (2n = 2x = 16) Medicago sativa plants, a subsp. falcata seed parent, and a coerulea × falcata pollen parent that form a mixture of n and 2n eggs and pollen, respectively. Such a cross produced full-sib diploid and tetraploid (2n = 4x = 32) hybrids, the latter being the result of bilateral sexual polyploidization (BSP). These unique materials allowed us to investigate the effects of BSP, and to separate the effect of intraspecific hybridization from those of polyploidization by comparing 2x with 4x full sib progeny plants. Simple sequence repeat marker segregation demonstrated tetrasomic inheritance for all chromosomes but one, demonstrating that these neotetraploids are true autotetraploids. BSP brought about increased biomass, earlier flowering, higher seed set and weight, and larger leaves with larger cells. Microarray analyses with M. truncatula gene chips showed that several hundred genes, related to diverse metabolic functions, changed their expression level as a consequence of polyploidization. In addition, cytosine methylation increased in 2x, but not in 4x, hybrids. Our results indicate that sexual polyploidization induces significant transcriptional novelty, possibly mediated in part by DNA methylation, and phenotypic novelty that could underpin improved adaptation and reproductive success of tetraploid M. sativa with respect to its diploid progenitor. These polyploidy-induced changes may have promoted the adoption of tetraploid alfalfa in agriculture. Copyright © 2016 Rosellini et al.

  4. Selective lignin downregulation leads to constitutive defense response expression in alfalfa (Medicago sativa L.)

    National Research Council Canada - National Science Library

    Lina Gallego-Giraldo; Yusuke Jikumaru; Yuji Kamiya; Yuhong Tang; Richard A. Dixon

    2011-01-01

    Downregulation of hydroxycinnamoyl CoA: shikimate hydroxycinnamoyl transferase (HCT) in alfalfa (Medicago sativa) reduces lignin levels and improves forage quality and saccharification efficiency for bioethanol production...

  5. [Quality standard of uygur medicine Medicago sativa seeds].

    Science.gov (United States)

    Ding, Wen-Huan; Xu, Hai-Yan; Wang, Dong-Dong; Li, Jie; Tian, Shu-Ge

    2013-11-01

    In this paper, microscopic identification method was adopted to observe the microscopic characters of ten batches of Medicago sativa seeds. And M. sativa seeds were identificated by TLC method in contrast to trigonelline and stachydrine hydrochloride. The impurities, moisture, ash, sour insoluble ash were detected based on Chinese Pharmacopoeia 2010 version (Vol I ). An HPLC method was also established for determination of trigonelline in the M. sativa seeds. The contents of impurities, moisture, ash, sour insoluble ash should not exceed 5%, 10%, 6%, and 2%, respectively. The content of trigonelline should be not less than 0.795 6 mg x g(-1). The experimental methods were accurate and reliable, and can be used as the quality control of the seeds of M. sativa.

  6. Comparative Analysis of the Symbiotic Efficiency of Medicago truncatula and Medicago sativa under Phosphorus Deficiency

    Science.gov (United States)

    Sulieman, Saad; Schulze, Joachim; Tran, Lam-Son Phan

    2013-01-01

    Phosphorus (P)-deficiency is a major abiotic stress that limits legume growth in many types of soils. The relationship between Medicago and Sinorhizobium, is known to be affected by different environmental conditions. Recent reports have shown that, in combination with S. meliloti 2011, Medicago truncatula had a lower symbiotic efficiency than Medicago sativa. However, little is known about how Medicago–Sinorhizobium is affected by P-deficiency at the whole-plant level. The objective of the present study was to compare and characterize the symbiotic efficiency of N2 fixation of M. truncatula and M. sativa grown in sand under P-limitation. Under this condition, M. truncatula exhibited a significantly higher rate of N2 fixation. The specific activity of the nodules was much higher in M. truncatula in comparison to M. sativa, partially as a result of an increase in electron allocation to N2versus H+. Although the main organic acid, succinate, exhibited a strong tendency to decrease under P-deficiency, the more efficient symbiotic ability observed in M. truncatula coincided with an apparent increase in the content of malate in its nodules. Our results indicate that the higher efficiency of the M. truncatula symbiotic system is related to the ability to increase malate content under limited P-conditions. PMID:23459233

  7. Occurrence of transgenic feral alfalfa (Medicago sativa subsp. sativa L.) in alfalfa seed production areas in the United States

    Science.gov (United States)

    Genetically-engineered glyphosate-resistant alfalfa (Medicago sativa subsp. sativa) was commercialized in 2011. The potential risk of transgene dispersal into the environment is not clearly understood for alfalfa, a perennial crop that is cross-pollinated by insects. We gathered data on feral and tr...

  8. Exploring the plant-associated bacterial communities in Medicago sativa L

    National Research Council Canada - National Science Library

    Pini, Francesco; Frascella, Arcangela; Santopolo, Luisa; Bazzicalupo, Marco; Biondi, Emanuele G; Scotti, Carla; Mengoni, Alessio

    2012-01-01

    ... in crop improvement and protection. Medicago sativa L. is a legume crop of high economic importance as forage in temperate areas and one of the most popular model plants for investigations on the symbiosis with nitrogen fixing rhizobia...

  9. Alfalfa (Medicago Sativa L As A Promising Forage In Indonesia

    Directory of Open Access Journals (Sweden)

    Sajimin

    2011-06-01

    Full Text Available Alfalfa (Medicago sativa L or Lucerne is a perennial herbaceous legume with superior forage quality. It is the most important forage crop in the world and it was the first domesticated forage crop. Alfalfa is able to fix nitrogen from the air through a symbiotic relationship with Rhizobium bacteria with N production 7.85 – 10.37 g/m2. Its rooting system can reach 4.5 m that allows it to escape drought. Forage production can reach 15.48 tons of dry matter per ha/year and containing 18.0 – 29.1 % crude protein. Plants can live 3 to 12 years depending on climatic conditions and crop varieties. However, alfalfa is not a tropical plant, thus it has not been widely cultivated in Indonesia. The problem of alfalfa cultivation are high pest attacks and competition with weeds. Therefore, alfalfa cultivation requires attention and good management to obtain optimum yield.

  10. Stress Responses in Alfalfa (Medicago sativa L.) 1

    Science.gov (United States)

    Kessmann, Helmut; Edwards, Robert; Geno, Paul W.; Dixon, Richard A.

    1990-01-01

    The isoflavonoid conjugates medicarpin-3-O-glucoside-6″-O-malonate (MGM), afrormosin-7-O-glucoside (AG), and afrormosin-7-O-glucoside-6″-O-malonate (AGM) were isolated and characterized from cell suspension cultures of alfalfa (Medicago sativa L.), where they were the major constitutive secondary metabolites. They were also found in alfalfa roots but not in other parts of the plant. The phytoalexin medicarpin accumulated rapidly in suspension cultured cells treated with elicitor from Colletotrichum lindemuthianum, and this was subsequently accompanied by an increase in the levels of MGM. In contrast, net accumulation of afrormosin conjugates was not affected by elicitor treatment. Labeling studies with [14C]phenylalanine indicated that afrormosin conjugates were the major de novo synthesized isoflavonoid products in unelicited cells. During elicitation, [14C]phenylalanine was incorporated predominantly into medicarpin, although a significant proportion of the newly synthesized medicarpin was also conjugated. Treatment of 14C-labeled, elicited cells with l-α-aminooxy-β-phenylpropionic acid, a potent inhibitor of PAL activity in vivo, resulted in the initial appearance of labeled medicarpin of very low specific activity, suggesting that the phytoalexin could be released from a preformed conjugate under these conditions. Our data draw attention to the involvement of isoflavone hydroxylases during the constitutive and elicitor-induced accumulation of isoflavonoids and their conjugates in alfalfa cell cultures. PMID:16667691

  11. Biofertilizer in the nutritional quality of alfalfa (Medicago sativa L.

    Directory of Open Access Journals (Sweden)

    Rodrigo Luis Lemes

    2016-06-01

    Full Text Available The objective of this study was to evaluate the response of alfalfa (Medicago sativa L. in the nutritional composition to the application of biofertilizers. The experiment was conducted with increasing doses of biofertilizers in a greenhouse at the Faculty of Veterinary Medicine / UNESP, Araçatuba - Sao Paulo, Brazil, from April to October 2010. The experimental design was completely randomized with six biofertilizer doses from cattle manure (0, 25, 50, 100, 200, and 400 m3 ha-1 and five replications. Cuts were performed, on average, every 27 days, 10 cm above the ground when 10% of the plants were flowering. Biofertilization had a positive significant impact on foliar nitrogen, potassium, calcium, magnesium, sulfur, and shoot iron concentrations. The values of crude protein, acid detergent fiber, and neutral detergent fiber did not differ between doses of biofertilizers. Biofertilization is a viable alternative for nutrition of this species, showing positive results in the nutritional composition of alfalfa. However, but long-term studies are necessary to assess the environmental impact of these fertilizers.

  12. Chemical characterisation of bioactive compounds in Medicago sativa growing in the desert of Oman.

    Science.gov (United States)

    Hanif, Muhammad Asif; Al-Maskari, Ahmed Yahya; Al-Sabahi, Jamal Nasser; Al-Hdhrami, Ibtisam; Khan, Muhammad Mumtaz; Al-Azkawi, Ahlam; Hussain, Abdullah Ijaz

    2015-01-01

    Medicago sativa Linn growing in Omani desert were chemically characterised using flame photometry, inductively coupled plasma, gas chromatography-mass spectrometry and high performance liquid chromatographic (HPLC) analysis. HPLC analyses were performed to determine the phenolics and flavonoids present in M. sativa. The major compounds detected in M. sativa leaves were protchaechenic acid (3.22%), hydroxyl benzoic acid (1.05%), β-Phenyl caffate (0.97%) and kaempherol (0.89%). Pterostilbene, a cholesterol-lowering compound, was detected in M. sativa.

  13. Beta-Amylases from Alfalfa (Medicago sativa L.) Roots 1

    Science.gov (United States)

    Doehlert, Douglas C.; Duke, Stanley H.; Anderson, Laurens

    1982-01-01

    Amylase was found in high activity (193 international units per milligram protein) in the tap root of alfalfa (Medicago sativa L. cv. Sonora). The activity was separated by gel filtration chromatography into two fractions with molecular weights of 65,700 (heavy amylase) and 41,700 (light amylase). Activity staining of electrophoretic gels indicated the presence of one isozyme in the heavy amylase fraction and two in the light amylase fraction. Three amylase isozymes with electrophoretic mobilities identical to those in the heavy and the light amylase fractions were the only amylases identified in crude root preparations. Both heavy and light amylases hydrolyzed amylopectin, soluble starch, and amylose but did not hydrolyze pullulan or β-limit dextrin. The ratio of viscosity change to reducing power production during starch hydrolysis was identical for both alfalfa amylase fractions and sweet potato β-amylase, while that of bacterial α-amylase was considerably higher. The identification of maltose and β-limit dextrin as hydrolytic end-products confirmed that these alfalfa root amylases are all β-amylases. The pH optimum for both β-amylase fractions was 6.0. Both light and heavy β-amylases showed normal Michaelis-Menten kinetics, with soluble starch as substrate, and had respectively Km values of 5.9 and 6.8 milligrams starch per milliliter and Vmax of 640 and 130 international units per milligram protein. Arrhenius plots indicated that the energy of activation for the heavy β-amylase remained relatively unchanged (12.7 to 13.0 kilocalories per mole) from 0 to 30°C, whereas the energy of activation for the light amylase increased from 12.0 to about 28.0 kilocalories per mole at 8.7°C as temperature was lowered. The light amylase was shown to be inhibited by maltose. Images PMID:16662350

  14. Asparagine Biosynthesis in Alfalfa (Medicago sativa L.) Root Nodules 1

    Science.gov (United States)

    Snapp, Sieglinde S.; Vance, Carroll P.

    1986-01-01

    Rapid direct conversion of exogenously supplied [14C]aspartate to [14C] asparagine and to tricarboxylic cycle acids was observed in alfalfa (Medicago sativa L.) nodules. Aspartate aminotransferase activity readily converted carbon from exogenously applied [14C]aspartate into the tricarboxylic acid cycle with subsequent conversion to the organic acids malate, succinate, and fumarate. Aminooxyacetate, an inhibitor of aminotransferase activity, reduced the flow of carbon from [14C]aspartate into tricarboxylic cycle acids and decreased 14CO2 evolution by 99%. Concurrently, maximum conversion of aspartate to asparagine was observed in aminooxyacetate treated nodules (30 nanomoles asparagine per gram fresh weight per hour. Metabolism of [14C]aspartate and distribution of nodulefixed 14CO2 suggest that two pools of aspartate occur in alfalfa nodules: (a) one involved in asparagine biosynthesis, and (b) another supplying a malate/aspartate shuttle. Conversion of [14C]aspartate to [14C]asparagine was not inhibited by methionine sulfoximine, a glutamine synthetase inhibitor, or azaserine, a glutmate synthetase, inhibitor. The data did not indicate that asparagine biosynthesis in alfalfa nodules has an absolute requirement for glutamine. Radioactivity in the xylem sap, derived from nodule 14CO2 fixation, was markedly decreased by treating nodulated roots with aminooxyacetate, methionine sulfoximine, and azaserine. Inhibitors decreased the [14C]aspartate and [14]asparagine content of xylem sap by greater than 80% and reduced the total amino nitrogen content of xylem sap (including nonradiolabeled amino acids) by 50 to 80%. Asparagine biosynthesis in alfalfa nodules and transport in xylem sap are dependent upon continued aminotransferase activity and an uninterrupted assimilation of ammonia via the glutamine synthetase/glutamate synthase pathway. Continued assimilation of ammonia apparently appears crucial to continued root nodule CO2 fixation in alfalfa. PMID:16665039

  15. Bioaccumulation and degradation of pentachloronitrobenzene in Medicago sativa.

    Science.gov (United States)

    Li, Ying Ying; Yang, Hong

    2013-04-15

    Pentachloronitrobenzene (PCNB) is a fungicide belonging to the organochlorine family and used extensively in agriculture for crop production. Many studies have implied that PCNB has become an environmental concern due to its widespread contamination in eco-systems. However, whether PCNB is bioaccumulated, degraded and phytotoxic in plants is poorly understood. In this study, several alfalfa (Medicago sativa) cultivars were grown in soil with PCNB to investigate their absorption and catabolism, including PCNB residues in the soil and PCNB-induced toxic responses in plants. Alfalfa plants varied widely in their ability to accumulate and degrade PCNB. The degradation rate of PCNB was 66.26-77.68% after alfalfa growth in the soils for 20 d, while the rates in the control (soil without alfalfa) were only 48.42%. Moreover, concentrations of PCNB residues in the rhizosphere soil were significantly higher than those in the non-rhizosphere soils. Alfalfa exposed to 10 mg kg(-1) PCNB showed inhibited growth and oxidative damage, but the effects of PCNB on the cultivars differed significantly, indicating that the alfalfa cultivars have different tolerance to PCNB. Activities of invertase (INV), urease (URE), polyphenol oxidase (PPO), alkaline phosphatase (ALP) and acid phosphatase (ACP) were assayed in the treated soils and showed that the enzyme activities were altered after PCNB exposure. The URE, PPO, ALP and ACP activities were increased in soil following the planting of alfalfa. The objective of the study was to analyze the potential of different cultivars of alfalfa to accumulate and degrade PCNB from the contaminated soil. Copyright © 2013 Elsevier Ltd. All rights reserved.

  16. Biological nitrogen fixation by lucerne (Medicago sativa L.) in acid soils

    NARCIS (Netherlands)

    Pijnenborg, J.

    1990-01-01

    Growth of lucerne( Medicago sativa L.) is poor in soils with values of pH-H2O below 6. This is often due to nitrogen deficiency, resulting from a hampered performance of the symbiosis withRhizobium

  17. Integration and Expression of gfp in the plastid of Medicago sativa L.

    Science.gov (United States)

    Xing, Shaochen; Wei, Zhengyi; Wang, Yunpeng; Liu, Yanzhi; Lin, Chunjing

    2014-01-01

    Here we describe a protocol of alfalfa (Medicago sativa L.) plastid transformation by which gfp, a gene encoding the green fluorescent protein (GFP), is inserted into plastid genome via particle bombardment and homoplastomic plant is obtained. Plastid engineering is likely to make a significant contribution to the genetic improvement of this crop and the production of vaccines and therapeutic proteins.

  18. The effects of ascorbic acid on salt induced alfalfa (Medicago sativa L.)

    African Journals Online (AJOL)

    Dr. J. T. Ekanem

    Ascorbic acid as an antioxidant agent has already been used for increasing of stress tolerance. Callus was produced from stem segments of alfalfa (Medicago sativa L.) on MS medium supplemented with 2,4 dichlorophenoxy acetic acid, naphthalene acetic acid and kinetin (2 mg/l each).Calli were then transferred to the ...

  19. First report of race 2 of Colletotrichum trifolii causing anthracnose on alfalfa (Medicago sativa) in Wisconsin

    Science.gov (United States)

    Anthracnose of alfalfa (Medicago sativa), caused by Colletotrichum trifolii, is widespread in the United States. Three physiological races have been described. Race 1 is reported to be the dominant race that is present wherever alfalfa is grown, while race 2 was reported in a limited area in the Mid...

  20. The effects of ascorbic acid on salt induced alfalfa ( Medicago sativa ...

    African Journals Online (AJOL)

    Ascorbic acid as an antioxidant agent has already been used for increasing of stress tolerance. Callus was produced from stem segments of alfalfa (Medicago sativa L.) on MS medium supplemented with 2,4 dichlorophenoxy acetic acid, naphthalene acetic acid and kinetin (2 mg/l each).Calli were then transferred to the ...

  1. Lucerne (Medicago sativa) or grass-clover as cut-and-carry fertilizers in organic agriculture

    NARCIS (Netherlands)

    Burgt, van der G.J.H.M.; Eekeren, van N.J.M.; Scholberg, J.M.S.; Koopmans, C.J.

    2013-01-01

    Onfarm nitrogen fixation is a driving force in organic agriculture. The efficiency with which this nitrogen is used can be increased by using lucerne (Medicago sativa) or grassclover directly as sources of fertilizer on arable land: cutandcarry fertilizers. In two arable crops, the use of lucerne

  2. Identification of novel RNA viruses in alfalfa (Medicago sativa): an Alphapartitivirus, a Deltapartitivirus, and a Marafivirus.

    Science.gov (United States)

    Kim, Hyein; Park, Dongbin; Hahn, Yoonsoo

    2018-01-05

    Genomic RNA molecules of plant RNA viruses are often co-isolated with the host RNAs, and their sequences can be detected in plant transcriptome datasets. Here, an alfalfa (Medicago sativa) transcriptome dataset was analyzed and three new RNA viruses were identified, which were named Medicago sativa alphapartitivirus 1 (MsAPV1), Medicago sativa deltapartitivirus 1 (MsDPV1), and Medicago sativa marafivirus 1 (MsMV1). The RNA-dependent RNA polymerases of MsAPV1, MsDPV1, and MsMV1 showed about 68%, 58%, and 46% amino acid sequence identity, respectively, with their closest virus species. Sequence similarity and phylogenetic analyses indicated that MsAPV1, MsDPV1, and MsMV1 were novel RNA virus species that belong to the genus Alphapartitivirus of the family Partitiviridae, the genus Deltapartitivirus of the family Partitiviridae, and the genus Marafivirus of the family Tymoviridae, respectively. The bioinformatics procedure applied in this study may facilitate the identification of novel RNA viruses from plant transcriptome data. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. The nutritive value of South African Medicago sativa L. hay | Scholtz ...

    African Journals Online (AJOL)

    A study was conducted to evaluate the variation and expand the existing and limiting nutritive value database of. Medicago sativa L. hay (168 near-infrared reflectance spectroscopy spectrally selected samples) in South Africa. The highest moisture content recorded (140 g/kg) was safely below the critical moisture level of ...

  4. Comparison of the phytoremediation potentials of Medicago falcata L. And Medicago sativa L. in aged oil-sludge-contaminated soil.

    Science.gov (United States)

    Panchenko, Leonid; Muratova, Anna; Turkovskaya, Olga

    2017-01-01

    Thirteen-year monitoring of the vegetation growing in the industrial and adjacent areas of an oil refinery showed the prevalence of yellow medick (Medicago falcata L.) over other plant species, including alfalfa (Medicago sativa L.). A comparative field study of the two Medicago species established that yellow medick and alfalfa exhibited similar resistance to soil petroleum hydrocarbons and that the pollutant concentration in their rhizosphere was 30% lower than that in the surrounding bulk soil. In laboratory pot experiments, yellow medick reduced the contaminant content by 18% owing to the degradation of the major heavy oil fractions, such as paraffins, naphthenes, and alcohol and benzene tars; and it was more successful than alfalfa. Both species were equally effective in stimulating the total number of soil microorganisms, but the number of hydrocarbon-oxidizing microorganisms, including polycyclic aromatic hydrocarbon degraders, was larger in the root zone of alfalfa. In turn, yellow medick provided a favorable balance of available nitrogen. Both Medicago species equally stimulated the dehydrogenase and peroxidase activities of the soil, and yellow medick increased the activity of soil polyphenol oxidase but reduced the activity of catalase. The root tissue activity of catalase, ascorbate oxidase, and tyrosinase was grater in alfalfa than in yellow medick. The peroxidase activity of plant roots was similar in both species, but nondenaturing polyacrylamide gel electrophoresis showed some differences in the peroxidase profiles of the root extracts of alfalfa and yellow medick. Overall, this study suggests that the phytoremediation potentials of yellow medick and alfalfa are similar, with some differences.

  5. The efficiency of nitrogen fixation of the model legume Medicago truncatula (Jemalong A17) is low compared to Medicago sativa.

    Science.gov (United States)

    Sulieman, Saad; Schulze, Joachim

    2010-06-15

    Medicago truncatula (Gaertn.) (barrel medic) serves as a model legume in plant biology. Numerous studies have addressed molecular aspects of the biology of M. truncatula, while comparatively little is known about the efficiency of N(2) fixation at the whole plant level. The objective of the present study was to compare the efficiency of N(2) fixation of M. truncatula to the genetically closely related Medicago sativa (L.) (alfalfa). The relative growth of both species relying exclusively on N(2) fixation versus nitrate nutrition, H(2) evolution, nitrogen assimilation, the concentration of amino acids and organic acids in nodules, and (15)N(2) uptake and distribution were studied. M. truncatula showed much lower efficiency of N(2) fixation. Nodule-specific activity was several-fold lower when compared to M. sativa, partially as a result of a lower electron allocation to N(2) versus H(+). M. truncatula or M. sativa plants grown solely on N(2) fixation as a nitrogen source reached about 30% or 80% of growth, respectively, when compared to plants supplied with sufficient nitrate. Moreover, M. truncatula had low %N in shoots and a lower allocation of (15)N to shoots during 1h (15)N(2) labeling period. Amino acid concentration was about 20% higher in M. sativa nodules, largely as a result of more asparagine, while the organic acid concentration was about double in M. sativa, coinciding with a six-fold higher concentration of malate. Total soluble protein in nodules was about three times lower in M. truncatula and the pattern of enzyme activity in that fraction was strongly different. Sucrose cleaving enzymes displayed higher activity in M. truncatula nodules, while the activity of phosphoenolpyruvate carboxylase (PEPC) was much lower. It is concluded that the low efficiency of the M. truncatula symbiotic system is related to a low capacity of organic acid formation and limited nitrogen export from nodules. Copyright 2010 Elsevier GmbH. All rights reserved.

  6. Identification of molecular markers associated with Verticillium wilt resistance in alfalfa (Medicago sativa L. using high-resolution melting.

    Directory of Open Access Journals (Sweden)

    Tiejun Zhang

    Full Text Available Verticillium wilt, caused by the soilborne fungus, Verticillium alfalfae, is one of the most serious diseases of alfalfa (Medicago sativa L. worldwide. To identify loci associated with resistance to Verticillium wilt, a bulk segregant analysis was conducted in susceptible or resistant pools constructed from 13 synthetic alfalfa populations, followed by association mapping in two F1 populations consisted of 352 individuals. Simple sequence repeat (SSR and single nucleotide polymorphism (SNP markers were used for genotyping. Phenotyping was done by manual inoculation of the pathogen to replicated cloned plants of each individual and disease severity was scored using a standard scale. Marker-trait association was analyzed by TASSEL. Seventeen SNP markers significantly associated with Verticillium wilt resistance were identified and they were located on chromosomes 1, 2, 4, 7 and 8. SNP markers identified on chromosomes 2, 4 and 7 co-locate with regions of Verticillium wilt resistance loci reported in M. truncatula. Additional markers identified on chromosomes 1 and 8 located the regions where no Verticillium resistance locus has been reported. This study highlights the value of SNP genotyping by high resolution melting to identify the disease resistance loci in tetraploid alfalfa. With further validation, the markers identified in this study could be used for improving resistance to Verticillium wilt in alfalfa breeding programs.

  7. Identification of molecular markers associated with Verticillium wilt resistance in alfalfa (Medicago sativa L.) using high-resolution melting.

    Science.gov (United States)

    Zhang, Tiejun; Yu, Long-Xi; McCord, Per; Miller, David; Bhamidimarri, Suresh; Johnson, David; Monteros, Maria J; Ho, Julie; Reisen, Peter; Samac, Deborah A

    2014-01-01

    Verticillium wilt, caused by the soilborne fungus, Verticillium alfalfae, is one of the most serious diseases of alfalfa (Medicago sativa L.) worldwide. To identify loci associated with resistance to Verticillium wilt, a bulk segregant analysis was conducted in susceptible or resistant pools constructed from 13 synthetic alfalfa populations, followed by association mapping in two F1 populations consisted of 352 individuals. Simple sequence repeat (SSR) and single nucleotide polymorphism (SNP) markers were used for genotyping. Phenotyping was done by manual inoculation of the pathogen to replicated cloned plants of each individual and disease severity was scored using a standard scale. Marker-trait association was analyzed by TASSEL. Seventeen SNP markers significantly associated with Verticillium wilt resistance were identified and they were located on chromosomes 1, 2, 4, 7 and 8. SNP markers identified on chromosomes 2, 4 and 7 co-locate with regions of Verticillium wilt resistance loci reported in M. truncatula. Additional markers identified on chromosomes 1 and 8 located the regions where no Verticillium resistance locus has been reported. This study highlights the value of SNP genotyping by high resolution melting to identify the disease resistance loci in tetraploid alfalfa. With further validation, the markers identified in this study could be used for improving resistance to Verticillium wilt in alfalfa breeding programs.

  8. Detection of Norspermidine and Norspermine in Medicago sativa L. (Alfalfa) 1

    Science.gov (United States)

    Rodriguez-Garay, Benjamin; Phillips, Gregory C.; Kuehn, Glenn D.

    1989-01-01

    Shoot meristem tissues of alfalfa, Medicago sativa L., were found by high performance liquid chromatography analyses to contain the uncommon polyamines, norspermidine and norspermine. The chemical structures of norspermidine and norspermine, purified from alfalfa, were confirmed by comparison of mass spectra with those from authentic standards. The discovery of norspermidine and norspermine in alfalfa implicates the presence of at least two biosynthetic enzymes, a polyamine oxidase and a previously uncharacterized aminopropyltransferase. PMID:16666576

  9. Accumulation of heavy metals in Medicago sativa L. and Trifolium pratense L. at the contaminated fluvisol

    Directory of Open Access Journals (Sweden)

    Jakšić Snežana P.

    2013-01-01

    Full Text Available Recently, heavy metals concentrations increased in some agricultural areas due to the consequences of anthropogenic impacts. The aim of this study was to determine the level of heavy metals (As, Cr, Ni and Pb in Medicago sativa L. and Trifolium pratense L. grown on fluvisol, in order to obtain information on safety of these nutrients. The total content of Pb, As, Cr and Ni in the samples of fluvisol was above the maximum allowable amount. The content of heavy metals in Medicago sativa L. and Trifolium pratense L. was below the critical and toxic concentrations in all samples originating from contaminated soil. It was concluded that the accumulation of heavy metals in plants did not depend only on the total content in soil, but also the affinity of the plant, and individual and interactive effects of various soil properties. No statistically significant differences in the accumulation of heavy metals between Medicago sativa L. and Trifolium pratense L were observed. It is necessary to further control of heavy metals in the investigated area, in order to prevent their entry into the food chain and provide healthy food.

  10. Lead phytoextraction from printed circuit computer boards by Lolium perenne L. and Medicago sativa L.

    Science.gov (United States)

    Díaz Martínez, María Esther; Argumedo-Delira, Rosalba; Sánchez-Viveros, Gabriela; Alarcón, Alejandro; Trejo-Téllez, Libia Iris

    2017-10-20

    This work assessed the ability of Lolium perenne and Medicago sativa for extracting lead (Pb) from particulate printed circuit computer boards (PCB) mixed in sand with the following concentrations: 0.5, 1.0 and 1.5 g of PCB, and including a control treatment without PCB. The PCB were obtained from computers, and grinded in two particle sizes: 0.0594 mm (PCB1) and 0.0706 mm (PCB2). The PCB particle sizes at their corresponding concentrations were applied to L. perenne and M. sativa by using three experimental assays. In assay II, PCB2 affected the biomass production for both plants. For assay III, the PCB1 increased the biomass of M. sativa (236.5%) and L. perenne (142.2%) when applying either 0.5 or 1.0 g, respectively. In regards to phytoextraction, assay I showed the highest Pb-extraction by roots of L. perenne (4.7%) when exposed to 1.5 g of PCB1. At assay I, L. perenne showed a Pb-bioconcentration factor higher than 1.0 when growing at 0.5 g of PCB1, and when HNO3 was used as digestion solution; moreover, in assay III both plants showed a Pb-translocation factor higher than 1.0. Therefore, Lolium perenne and Medicago sativa are able to recover Pb from electronic wastes (PCB).

  11. The Effects of Medicago Sativa and Allium Porrum on Iron Overload in Rats.

    Science.gov (United States)

    Mirzaei, Ali; Delaviz, Hamdollah; Mirzaei, Mahsa; Tolooei, Mohsen

    2015-04-23

    Iron overload may occur due to regular blood transfusions and high intestinal iron absorption. Currently, there is no effective drug without side effects for the treatment of iron excess in thalassemia and other iron storage diseases, except chelation therapy, which is the only safe method for iron excretion. Thus, scientists are more focused on medicinal plants rich in phytochemical compounds for the removal of iron in thalassemia. Therefore this study was managed to discover the therapeutic potential of hydro-alcoholic extract of Allium porrum and Medicago sativa for iron chelating potential. Aerial parts of Allium porrum and Medicago sativa were collected in Yasuj Iran. Rats were divided into seven groups each containing six. Extracts were administrated in four groups (two groups for each extract) by single doses of each plant with 200 and 400 mg/kg body weight by (i.p.) route every other day for 28 days. Group 1 as negative control received saline (0.5 ml/kg) by (i.p.) route. Positive control received iron dextran 200 mg/kg body weight. Experimental groups 1 and 2 for each plant extract were fed with 200 and 400 mg/kg, hydro-alcoholic extract respectively via (i.p.) route, 1 h after the injection of iron dextran. Standard group was treated with deferoxamine (DF) 50 mg/kg by (i.p.) route 1 h after the injection of iron dextran. Serum iron (SI) and serum total iron binding capacity (TIBC) were determined .The serum ferritin was then measured using enzyme immunoassay ELISA kit for rat. For Analysis of data ANOVA test was used. Hydro-alcoholic extract of Medicago sativa and Allium porrum at 400 mg/kg showed significant (pMedicago sativapsativa and A. Porrum against iron overload in rat model was reported. Significant decrease in serum ferritin and iron concentration was reported in iron overload rats which induced by iron dextran.

  12. Isolation and functional characterization of salt-stress induced RCI2-like genes from Medicago sativa and Medicago truncatula.

    Science.gov (United States)

    Long, Ruicai; Zhang, Fan; Li, Zhenyi; Li, Mingna; Cong, Lili; Kang, Junmei; Zhang, Tiejun; Zhao, Zhongxiang; Sun, Yan; Yang, Qingchuan

    2015-07-01

    Salt stress is one of the most significant adverse abiotic factors, causing crop failure worldwide. So far, a number of salt stress-induced genes, and genes improving salt tolerance have been characterized in a range of plants. Here, we report the isolation and characterization of a salt stress-induced Medicago sativa (alfalfa) gene (MsRCI2A), which showed a high similarity to the yeast plasma membrane protein 3 gene (PMP3) and Arabidopsis RCI2A. The sequence comparisons revealed that five genes of MtRCI2(A-E) showed a high similarity to MsRCI2A in the Medicago truncatula genome. MsRCI2A and MtRCI2(A-E) encode small, highly hydrophobic proteins containing two putative transmembrane domains, predominantly localized in the plasma membrane. The transcript analysis results suggest that MsRCI2A and MtRCI2(A-D) genes are highly induced by salt stress. The expression of MsRCI2A and MtRCI2(A-C) in yeast mutants lacking the PMP3 gene can functionally complement the salt sensitivity phenotype resulting from PMP3 deletion. Overexpression of MsRCI2A in Arabidopsis plants showed improved salt tolerance suggesting the important role of MsRCI2A in salt stress tolerance in alfalfa.

  13. Natural occurrence of entomophthoroid fungi of aphid pests on Medicago sativa L. in Argentina

    Directory of Open Access Journals (Sweden)

    Romina G Manfrino

    Full Text Available Four species of entomophthoroid fungi, Pandora neoaphidis (Entomophthorales: Entomophthoraceae, Zoophthora radicans (Entomophthorales: Entomophthoraceae, Entomophthora planchoniana (Entomophthorales: Entomophthoraceae and Neozygites fresenii (Neozygitales: Neozygitaceae were found to infect Aphis craccivora, Therioaphis trifolii, and Acyrthosiphon pisum and unidentified species of Acyrthosiphon on lucerne in Argentina. Samples were collected from five sites (Ceres, Rafaela, Sarmiento, Monte Vera and Bernardo de Irigoyen in the province of Santa Fe. In this study, Zoophthora radicans was the most important pathogen and was recorded mainly on Acyrthosiphon sp. Zoophthora radicans was successfully isolated and maintained in pure cultures. This study is the first report of entomophthoroid fungi infecting lucerne (Medicago sativa L. aphids in Argentina.

  14. Biogenic Pt uptake and nanoparticle formation in Medicago sativa and Brassica juncea

    Science.gov (United States)

    Bali, Roza; Siegele, Rainer; Harris, Andrew T.

    2010-10-01

    The ability of the facultative metallophyte plants, Medicago sativa ( M. sativa) and Brassica juncea ( B. juncea) to accumulate and translocate platinum (Pt) from aqueous substrates is reported. The influence of Pt concentration in the substrate (5, 10, 20, 40 and 80 ppm), exposure time (24, 48 and 72 h) and substrate pH (2, 3, 5, 7 and 9) was determined. In both plants the concentration of Pt increased with substrate concentration and exposure time. Greater accumulation was detected in the roots of M. sativa than B. juncea, up to a maximum of 94.19 mg Pt g-1 (dry biomass) compared with 38.5 mg Pt g-1 (dry biomass) following exposure to 80 ppm Pt after 72 h exposure, respectively. However, at lower substrate concentrations (5 and 20 ppm) greater quantities of Pt were detected in the shoots of B. juncea, ranging between 0.02 and 0.32 mg Pt g-1 (dry biomass) at 5 ppm across the different time intervals studied, compared with 0.02-0.14 mg Pt g-1 (dry biomass) for M. sativa, suggesting B. juncea to be a better translocator of Pt under idealised conditions at low concentrations. Higher Pt uptake was also observed in acidic media, with a maximum at pH 2 for M. sativa and pH 3 for B. juncea, indicating the role of net surface charge on the bioaccumulation of Pt. Once sequestered Pt(II) was reduced to Pt(0) due to the action of local metabolites. TEM images of M. sativa root samples showed the in vivo formation of Pt nanoparticles between 3 and 100 nm in size and of varying morphologies in the epidermal root cells. In vivo Pt distribution profiles were assessed using proton induced X-ray emission (μ-PIXE) spectroscopy, which showed even distribution across all tissue systems (epidermal, cortical and vascular) within the roots of both M. sativa and B. juncea.

  15. Kinetics and Strain Specificity of Rhizosphere and Endophytic Colonization by Enteric Bacteria on Seedlings of Medicago sativa and Medicago truncatula

    Science.gov (United States)

    Dong, Yuemei; Iniguez, A. Leonardo; Ahmer, Brian M. M.; Triplett, Eric W.

    2003-01-01

    The presence of human-pathogenic, enteric bacteria on the surface and in the interior of raw produce is a significant health concern. Several aspects of the biology of the interaction between these bacteria and alfalfa (Medicago sativa) seedlings are addressed here. A collection of enteric bacteria associated with alfalfa sprout contaminations, along with Escherichia coli K-12, Salmonella enterica serotype Typhimurium strain ATCC 14028, and an endophyte of maize, Klebsiella pneumoniae 342, were labeled with green fluorescent protein, and their abilities to colonize the rhizosphere and the interior of the plant were compared. These strains differed widely in their endophytic colonization abilities, with K. pneumoniae 342 and E. coli K-12 being the best and worst colonizers, respectively. The abilities of the pathogens were between those of K. pneumoniae 342 and E. coli K-12. All Salmonella bacteria colonized the interiors of the seedlings in high numbers with an inoculum of 102 CFU, although infection characteristics were different for each strain. For most strains, a strong correlation between endophytic colonization and rhizosphere colonization was observed. These results show significant strain specificity for plant entry by these strains. Significant colonization of lateral root cracks was observed, suggesting that this may be the site of entry into the plant for these bacteria. At low inoculum levels, a symbiosis mutant of Medicago truncatula, dmi1, was colonized in higher numbers on the rhizosphere and in the interior by a Salmonella endophyte than was the wild-type host. Endophytic entry of M. truncatula appears to occur by a mechanism independent of the symbiotic infections by Sinorhizobium meliloti or mycorrhizal fungi. PMID:12620870

  16. Phenanthrene uptake by Medicago sativa L. under the influence of an arbuscular mycorrhizal fungus

    Energy Technology Data Exchange (ETDEWEB)

    Wu Naiying [State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Centre for Eco-Environmental Sciences, Chinese Academy of Sciences, PO Box 2871, Beijing 100085 (China); Department of Chemistry, Shangqiu Normal College, Shangqiu 476000 (China); Huang Honglin [State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Centre for Eco-Environmental Sciences, Chinese Academy of Sciences, PO Box 2871, Beijing 100085 (China); Zhang Shuzhen, E-mail: szzhang@rcees.ac.c [State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Centre for Eco-Environmental Sciences, Chinese Academy of Sciences, PO Box 2871, Beijing 100085 (China); Zhu Yongguan [State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Centre for Eco-Environmental Sciences, Chinese Academy of Sciences, PO Box 2871, Beijing 100085 (China); Christie, Peter [Agri-Environment Branch, Agriculture Food and Environmental Science Division, Agri-Food and Biosciences Institute, Newforge Lane, Belfast BT9 5PX (United Kingdom); Zhang Yong [State Key Laboratory of Marine Environmental Science, Environmental Science Research Centre, Xiamen University, Xiamen 361005 (China)

    2009-05-15

    Phenanthrene uptake by Medicago sativa L. was investigated under the influence of an arbuscular mycorrhizal fungus. Inoculation of lucerne with the arbuscular mycorrhizal fungus Glomus etunicatum L. resulted in higher phenanthrene accumulation in the roots and lower accumulation in the shoots compared to non-mycorrhizal controls. Studies on sorption and desorption of phenanthrene by roots and characterization of heterogeneity of mycorrhizal and non-mycorrhizal roots using solid-state {sup 13}C nuclear magnetic resonance spectroscopy ({sup 13}C NMR) demonstrated that increased aromatic components due to mycorrhizal inoculation resulted in enhanced phenanthrene uptake by the roots but lower translocation to the shoots. Direct visualization using two-photon excitation microscopy (TPEM) revealed higher phenanthrene accumulation in epidermal cells of roots and lower transport into the root interior and stem in mycorrhizal plants than in non-mycorrhizal controls. These results provide some insight into the mechanisms by which arbuscular mycorrhizal inoculation may influence the uptake of organic contaminants by plants. - Colonization by an arbuscular mycorrhizal fungus promoted root uptake and decreased shoot uptake of phenanthrene by Medicago sativa L.

  17. Resistant and susceptible responses in alfalfa (Medicago sativa) to bacterial stem blight caused by Pseudomonas syringae pv. syringae

    Science.gov (United States)

    Bacterial stem blight caused by Pseudomonas syringae pv. syringae is a common disease of alfalfa (Medicago sativa L.) in the central and western U.S. and has been reported in Australia and Europe. The disease is not always recognized because symptoms are often associated with frost damage. Two culti...

  18. Effect of wheel traffic and green manure treatments on forage yield and crown rot in alfalfa (Medicago sativa)

    Science.gov (United States)

    Harvesting alfalfa (Medicago sativa) results in mechanical wheel traffic on plants, which may damage crowns and increase the opportunity for entry of pathogens causing crown rot. Developing resistance to crown rot is problematic due to the large number of pathogens involved. Incorporation of plant b...

  19. Alfalfa (Medicago sativa L.) forage production, tissue and soil nutrient concentration under three N based broiler litter regimes

    Science.gov (United States)

    Alfalfa (Medicago sativa L.) is considered as most important forage legume grown in Kentucky. Alfalfa supports many livestock production systems including the beef, dairy, and horse industries in Kentucky. Being a legume, alfalfa typically meets its N requirement through symbiotic N2 fixation, but h...

  20. Mapping fall dormancy and winter injury in tetraploid alfalfa (Medicago sativa L.)

    Science.gov (United States)

    Alfalfa is a widely planted perennial forage crop. Dormancy in autumn (fall dormancy) is generally negatively correlated with winter injury in alfalfa. To understand the genetic basis of the two traits, we identified quantitative trait loci (QTL) controlling autumn growth and winter injury using a...

  1. Characterization of two novel cold-inducible K3 dehydrin genes from alfalfa (Medicago sativa spp. sativa L.).

    Science.gov (United States)

    Dubé, Marie-Pier; Castonguay, Yves; Cloutier, Jean; Michaud, Josée; Bertrand, Annick

    2013-03-01

    Dehydrin defines a complex family of intrinsically disordered proteins with potential adaptive value with regard to freeze-induced cell dehydration. Search within an expressed sequence tags library from cDNAs of cold-acclimated crowns of alfalfa (Medicago sativa spp. sativa L.) identified transcripts putatively encoding K(3)-type dehydrins. Analysis of full-length coding sequences unveiled two highly homologous sequence variants, K(3)-A and K(3)-B. An increase in the frequency of genotypes yielding positive genomic amplification of the K(3)-dehydrin variants in response to selection for superior tolerance to freezing and the induction of their expression at low temperature strongly support a link with cold adaptation. The presence of multiple allelic forms within single genotypes and independent segregation indicate that the two K(3) dehydrin variants are encoded by distinct genes located at unlinked loci. The co-inheritance of the K(3)-A dehydrin with a Y(2)K(4) dehydrin restriction fragment length polymorphism with a demonstrated impact on freezing tolerance suggests the presence of a genome domain where these functionally related genes are located. These results provide additional evidence that dehydrin play important roles with regard to tolerance to subfreezing temperatures. They also underscore the value of recurrent selection to help identify variants within a large multigene family in allopolyploid species like alfalfa.

  2. Toxic effects of copper-based nanoparticles or compounds to lettuce (Lactuca sativa) and alfalfa (Medicago sativa).

    Science.gov (United States)

    Hong, Jie; Rico, Cyren M; Zhao, Lijuan; Adeleye, Adeyemi S; Keller, Arturo A; Peralta-Videa, Jose R; Gardea-Torresdey, Jorge L

    2015-01-01

    The increased production and use of nanoparticles (NPs) has generated concerns about their impact on living organisms. In this study, nCu, bulk Cu, nCuO, bulk CuO, Cu(OH)2 (CuPRO 2005, Kocide 3000), and CuCl2 were exposed for 15 days to 10 days-old hydroponically grown lettuce (Lactuca sativa) and alfalfa (Medicago sativa). Each compound was applied at 0, 5, 10, and 20 mg L(-1). At harvest, we measured the size of the plants and determined the concentration of Cu, macro and microelements by using ICP-OES. Catalase and ascorbate peroxidase activity was also determined. Results showed that all Cu NPs/compounds reduced the root length by 49% in both plant species. All Cu NPs/compounds increased Cu, P, and S (>100%, >50%, and >20%, respectively) in alfalfa shoots and decreased P and Fe in lettuce shoot (>50% and >50%, respectively, excluding Fe in CuCl2 treatment). Biochemical assays showed reduced catalase activity in alfalfa (root and shoot) and increased ascorbate peroxidase activity in roots of both plant species. Results suggest that Cu NPs/compounds not only reduced the size of the plants but altered nutrient content and enzyme activity in both plant species.

  3. Mixed Nodule Infection in Sinorhizobium meliloti-Medicago sativa Symbiosis Suggest the Presence of Cheating Behavior.

    Science.gov (United States)

    Checcucci, Alice; Azzarello, Elisa; Bazzicalupo, Marco; Galardini, Marco; Lagomarsino, Alessandra; Mancuso, Stefano; Marti, Lucia; Marzano, Maria C; Mocali, Stefano; Squartini, Andrea; Zanardo, Marina; Mengoni, Alessio

    2016-01-01

    In the symbiosis between rhizobia and legumes, host plants can form symbiotic root nodules with multiple rhizobial strains, potentially showing different symbiotic performances in nitrogen fixation. Here, we investigated the presence of mixed nodules, containing rhizobia with different degrees of mutualisms, and evaluate their relative fitness in the Sinorhizobium meliloti-Medicago sativa model symbiosis. We used three S. meliloti strains, the mutualist strains Rm1021 and BL225C and the non-mutualist AK83. We performed competition experiments involving both in vitro and in vivo symbiotic assays with M. sativa host plants. We show the occurrence of a high number (from 27 to 100%) of mixed nodules with no negative effect on both nitrogen fixation and plant growth. The estimation of the relative fitness as non-mutualist/mutualist ratios in single nodules shows that in some nodules the non-mutualist strain efficiently colonized root nodules along with the mutualist ones. In conclusion, we can support the hypothesis that in S. meliloti-M. sativa symbiosis mixed nodules are formed and allow non-mutualist or less-mutualist bacterial partners to be less or not sanctioned by the host plant, hence allowing a potential form of cheating behavior to be present in the nitrogen fixing symbiosis.

  4. Transcriptomic and physiological analyses of Medicago sativa L. roots in response to lead stress.

    Science.gov (United States)

    Xu, Bo; Wang, Yingzhe; Zhang, Shichao; Guo, Qiang; Jin, Yan; Chen, Jingjing; Gao, Yunhang; Ma, Hongxia

    2017-01-01

    Lead (Pb) is one of the nonessential and toxic metals that threaten the environment and human health. Medicago sativa L. is a legume with high salt tolerance and high biomass production. It is not only a globally important forage crop but is also an ideal plant for phytoremediation. However, the biological and molecular mechanisms that respond to heavy metals are still not well defined in M. sativa. In this study, de novo and strand-specific RNA-sequencing was performed to identify genes involved in the Pb stress response in M. sativa roots. A total of 415,350 unigenes were obtained from the assembled cDNA libraries, among which 5,416 were identified as significantly differentially expressed genes (DEGs) (false discovery rate < 0.005) between cDNA libraries from control and Pb-treated plants. Gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of the DEGs showed they mainly clustered with terms associated with binding, transport, membranes, and the pathways related to signal and energy metabolism. Moreover, a number of candidate genes included antioxidant enzymes, metal transporters, and transcription factors involved in heavy metal response were upregulated under Pb stress. Quantitative real-time PCR(qRT-PCR) validation of the expression patterns of 10 randomly selected candidate DEGs were consistent with the transcriptome analysis results. Thus, this study offers new information towards the investigation of biological changes and molecular mechanisms related to Pb stress response in plants.

  5. Medicago sativa--Sinorhizobium meliloti Symbiosis Promotes the Bioaccumulation of Zinc in Nodulated Roots.

    Science.gov (United States)

    Zribi, Kais; Nouairi, Issam; Slama, Ines; Talbi-Zribi, Ons; Mhadhbi, Haythem

    2015-01-01

    In this study we investigated effects of Zn supply on germination, growth, inorganic solutes (Zn, Ca, Fe, and Mg) partitioning and nodulation of Medicago sativa This plant was cultivated with and without Zn (2 mM). Treatments were plants without (control) and with Zn tolerant strain (S532), Zn intolerant strain (S112) and 2 mM urea nitrogen fertilisation. Results showed that M. sativa germinates at rates of 50% at 2 mM Zn. For plants given nitrogen fertilisation, Zn increased plant biomass production. When grown with symbionts, Zn supply had no effect on nodulation. Moreover, plants with S112 showed a decrease of shoot and roots biomasses. However, in symbiosis with S532, an increase of roots biomass was observed. Plants in symbiosis with S. meliloti accumulated more Zn in their roots than nitrogen fertilised plants. Zn supply results in an increase of Ca concentration in roots of fertilised nitrogen plants. However, under Zn supply, Fe concentration decreased in roots and increased in nodules of plants with S112. Zn supply showed contrasting effects on Mg concentrations for plants with nitrogen fertilisation (increase) and plants with S112 (decrease). The capacity of M. sativa to accumulate Zn in their nodulated roots encouraged its use in phytostabilisation processes.

  6. Biogenic Pt uptake and nanoparticle formation in Medicago sativa and Brassica juncea

    Energy Technology Data Exchange (ETDEWEB)

    Bali, Roza [University of Sydney, Laboratory for Sustainable Technology, School of Chemical and Biomolecular Engineering (Australia); Siegele, Rainer [Australian Nuclear Science and Technology Organization, Institute for Environmental Research (Australia); Harris, Andrew T., E-mail: a.harris@usyd.edu.a [University of Sydney, Laboratory for Sustainable Technology, School of Chemical and Biomolecular Engineering (Australia)

    2010-10-15

    The ability of the facultative metallophyte plants, Medicago sativa (M. sativa) and Brassica juncea (B. juncea) to accumulate and translocate platinum (Pt) from aqueous substrates is reported. The influence of Pt concentration in the substrate (5, 10, 20, 40 and 80 ppm), exposure time (24, 48 and 72 h) and substrate pH (2, 3, 5, 7 and 9) was determined. In both plants the concentration of Pt increased with substrate concentration and exposure time. Greater accumulation was detected in the roots of M. sativa than B. juncea, up to a maximum of 94.19 mg Pt g{sup -1} (dry biomass) compared with 38.5 mg Pt g{sup -1} (dry biomass) following exposure to 80 ppm Pt after 72 h exposure, respectively. However, at lower substrate concentrations (5 and 20 ppm) greater quantities of Pt were detected in the shoots of B. juncea, ranging between 0.02 and 0.32 mg Pt g{sup -1} (dry biomass) at 5 ppm across the different time intervals studied, compared with 0.02-0.14 mg Pt g{sup -1} (dry biomass) for M. sativa, suggesting B. juncea to be a better translocator of Pt under idealised conditions at low concentrations. Higher Pt uptake was also observed in acidic media, with a maximum at pH 2 for M. sativa and pH 3 for B. juncea, indicating the role of net surface charge on the bioaccumulation of Pt. Once sequestered Pt(II) was reduced to Pt(0) due to the action of local metabolites. TEM images of M. sativa root samples showed the in vivo formation of Pt nanoparticles between 3 and 100 nm in size and of varying morphologies in the epidermal root cells. In vivo Pt distribution profiles were assessed using proton induced X-ray emission ({mu}-PIXE) spectroscopy, which showed even distribution across all tissue systems (epidermal, cortical and vascular) within the roots of both M. sativa and B. juncea.

  7. [Natural nucleotide polymorphism of the Srlk gene that determines salt stress tolerance in alfalfa (Medicago sativa L)].

    Science.gov (United States)

    Vishnevskaia, M S; Pavlov, A V; Dziubenko, E A; Dziubenko, N I; Potokina, E K

    2014-04-01

    Based on legume genome syntheny, the nucleotide sequence of Srlk gene, key role of which in response to salt stress was demonstrated for the model species Medicago truncatula, was identified in the major forage and siderate crop alfalfa (Medicago sativa). In twelve alfalfa samples originating from regions with contrasting growing conditions, 19 SNPs were revealed in the Srlk gene. For two nonsynonymous SNPs, molecular markers were designed that could be further used to analyze the association between Srlk gene nucleotide polymorphism and the variability in salt stress tolerance among alfalfa cultivars.

  8. Photoperiodic Effects on the Emanation of Volatiles from Alfalfa (Medicago sativa L.) Florets 1

    Science.gov (United States)

    Loper, G. M.; Lapioli, A. M.

    1972-01-01

    Alfalfa (Medicago sativa L.) plants acclimated to photoperiods of 18 hours light, 6 hour dark in plant growth chambers exhibited a daily cyclic pattern of floret volatile emanation with a maximum emanation of about 6.5 nanograms of hydrocarbons/floret·30 minutes. This maximum was reached about 6 to 8 hours into the light period. After 8 hours of light, emanation of volatiles decreased rapidly to less than 0.1 ng/floret·30 min even though light and temperature remained constant. Under continuous illumination, only a small increase of volatile emanation occurred during the following 24 hours. It appeared that a dark period was necessary to promote floret volatile emanation. Floret volatile emanation was drastically affected for at least 7 days following a photoperiod change. A photoperiod change caused 6-fold concentration oscillations every 2 hours. The results are interpreted on the basis of a very active floral metabolism controlled by photoperiodically induced rhythms. PMID:16658038

  9. [Control of the fermentation activity during ensilage of Lolium italicum A. Br. and Medicago sativa L].

    Science.gov (United States)

    Ozino Marletto, O I; Ambrosoli, R; Piccone, G; Biasiol, B

    1982-01-01

    Samples with different dry matter contents of Lolium italicum A. Br., Medicago sativa L., have been ensiled in hermetically sealed containers, in order to study the evolution of microflora and its activity. The principal microbial groups (coliforms, proteolytics , lactic acid bacteria, clostridia, yeasts) have been detected and enumerated, in anaerobic atmosphere, after ensiling periods of 3, 6, 10, 13, 20, 100 days. At the same time, the samples were chemically analyzed for the detection of: pH, moisture, ashes, total and ammoniacal nitrogen, total reducing sugars, lactic acid, volatile fatty acids, short chain alcohols. The relations found between chemical and microbiological data, show that a strong lactic fermentation is not enough for the inhibition of silage spoiling microorganisms, such as coliforms, yeasts, and (less) proteolytics . This phenomenon may be related to the "quality" of the lactic acid microflora, more than to the characteristics of the environment.

  10. A PROPOSITO DE UN ENSAYO CON ABONOS (cal y harina de huesos EN ALFALFAl (Medicago sativa

    Directory of Open Access Journals (Sweden)

    Isidoro Mogilner

    1960-01-01

    Full Text Available En este trabajo, los autores estudian el efecto que sobre el rendimiento de la alfalfa (Medicago sativa produce la incorporación de diferentes dosis de cal y harina de huesos, en un suelo pobre de P y Ca; de una textura pesada con alto contenido de arcilla en todos los horizontes, de un pH ácido (5.1 a 6.2 y que es característico de la zona donde fué hecha la experiencia.Se han obtenido resultados referentes a la influencia de la inoculación de la semilla con Rhizobium y el agregado de cal y harina de huesos sobre los rendimientos.

  11. Natural occurrence of entomophthoroid fungi of aphid pests on Medicago sativa L. in Argentina.

    Science.gov (United States)

    Manfrino, Romina G; Zumoffen, Leticia; Salto, César E; Lastra, Claudia C López

    2014-01-01

    Four species of entomophthoroid fungi, Pandora neoaphidis (Entomophthorales: Entomophthoraceae), Zoophthora radicans (Entomophthorales: Entomophthoraceae), Entomophthora planchoniana (Entomophthorales: Entomophthoraceae) and Neozygites fresenii (Neozygitales: Neozygitaceae) were found to infect Aphis craccivora, Therioaphis trifolii, and Acyrthosiphon pisum and unidentified species of Acyrthosiphon on lucerne in Argentina. Samples were collected from five sites (Ceres, Rafaela, Sarmiento, Monte Vera and Bernardo de Irigoyen) in the province of Santa Fe. In this study, Zoophthora radicans was the most important pathogen and was recorded mainly on Acyrthosiphon sp. Zoophthora radicans was successfully isolated and maintained in pure cultures. This study is the first report of entomophthoroid fungi infecting lucerne (Medicago sativa L.) aphids in Argentina. Copyright © 2014 Asociación Colombiana de Psiquiatría. Publicado por Elsevier España. All rights reserved.

  12. Nodulation of Medicago Sativa L. by Different Rhizobium Strains in the Sultanate of Oman

    Directory of Open Access Journals (Sweden)

    A.E. Elshafie

    1997-12-01

    Full Text Available Three native Rhizobium strains were isolated from Musandam. Buraimi and the Agricultural Experiment Station at Sultan Qaboos University in the Sultanate of Oman. An internationally known Rhizobium strain (TAI. 1373} was obtained from the Nitrogen Fixation for Tropical Agriculture I.egume Collection at the University of Hawaii. U.S.A-. for comparison. The different abilities of the strains to nodulate  Medicago sativa I. were investigated. Different plant parameters namely, the fresh and dry weights of the shoot (g/plant . the number of nodules per plant and the nitrogenase  activity (u mole C2H2/sample/3hours were measured. The results showed that the native Rhizobia were capable of nodulating M saliva. But inoculation with effective known strains such as TAL1373 enhanced nodulation.

  13. Modulation of endogenous indole-3-acetic acid biosynthesis in bacteroids within Medicago sativa nodules.

    Science.gov (United States)

    Bianco, C; Senatore, B; Arbucci, S; Pieraccini, G; Defez, R

    2014-07-01

    To evaluate the dose-response effects of endogenous indole-3-acetic acid (IAA) on Medicago plant growth and dry weight production, we increased the synthesis of IAA in both free-living and symbiosis-stage rhizobial bacteroids during Rhizobium-legume symbiosis. For this purpose, site-directed mutagenesis was applied to modify an 85-bp promoter sequence, driving the expression of iaaM and tms2 genes for IAA biosynthesis. A positive correlation was found between the higher expression of IAA biosynthetic genes in free-living bacteria and the increased production of IAA under both free-living and symbiotic conditions. Plants nodulated by RD65 and RD66 strains, synthetizing the highest IAA concentration, showed a significant (up to 73%) increase in the shoot fresh weight and upregulation of nitrogenase gene, nifH, compared to plants nodulated by the wild-type strain. When these plants were analyzed by confocal microscopy, using an anti-IAA antibody, the strongest signal was observed in bacteroids of Medicago sativa RD66 (Ms-RD66) plants, even when they were located in the senescent nodule zone. We show here a simple system to modulate endogenous IAA biosynthesis in bacteria nodulating legumes suitable to investigate which is the maximum level of IAA biosynthesis, resulting in the maximal increase of plant growth. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  14. SPL13 regulates shoot branching and flowering time in Medicago sativa.

    Science.gov (United States)

    Gao, Ruimin; Gruber, Margaret Y; Amyot, Lisa; Hannoufa, Abdelali

    2018-01-01

    Our results show SPL13 plays a crucial role in regulating vegetative and reproductive development in Medicago sativa L. (alfalfa), and that MYB112 is targeted and downregulated by SPL13 in alfalfa. We previously showed that transgenic Medicago sativa (alfalfa) plants overexpressing microRNA156 (miR156) show a bushy phenotype, reduced internodal length, delayed flowering time, and enhanced biomass yield. In alfalfa, transcripts of seven SQUAMOSA-PROMOTER BINDING PROTEIN-LIKE (SPL) transcription factors, including SPL13, are targeted for cleavage by miR156. Thus, association of each target SPL gene to a trait or set of traits is essential for developing molecular markers for alfalfa breeding. In this study, we investigated SPL13 function using SPL13 overexpression and silenced alfalfa plants. Severe growth retardation, distorted branches and up-curled leaves were observed in miR156-impervious 35S::SPL13m over-expression plants. In contrast, more lateral branches and delayed flowering time were observed in SPL13 silenced plants. SPL13 transcripts were predominantly present in the plant meristems, indicating that SPL13 is involved in regulating shoot branch development. Accordingly, the shoot branching-related CAROTENOID CLEAVAGE DIOXYGENASE 8 gene was found to be significantly downregulated in SPL13 RNAi silencing plants. A R2R3-MYB gene MYB112 was also identified as being directly silenced by SPL13 based on Next Generation Sequencing-mediated transcriptome analysis and chromatin immunoprecipitation assays, suggesting that MYB112 may be involved in regulating alfalfa vegetative growth.

  15. Transfer of anthracnose resistance and pod coiling traits from Medicago arborea to M. sativa by sexual reproduction.

    Science.gov (United States)

    Armour, D J; Mackie, J M; Musial, J M; Irwin, J A G

    2008-07-01

    Five asymmetric hybrid plants were obtained between Medicago sativa (2n = 4x = 32) and Medicago arborea (2n = 4x = 32) through sexual reproduction and the use of a cytoplasmically male sterile M. sativa genotype. Over 2,000 pollinations were made to obtain these hybrids. Amplified fragment length polymorphism (AFLP) analysis showed that in the most studied hybrid (WA2273), 4% of the bands unique to the M. arborea parent were present, versus 72% for the unique M. sativa bands. This suggests that only a single M. arborea chromosome or chromosome parts has been transferred. WA2273 had 7% of AFLP bands which were not present in either parent, which is suggestive of chromosome rearrangements as would be expected if only chromosome parts or a single part had been transferred from M. arborea. Phenotypic evidence for hybridity was obtained for pod coiling (1.4 coils in WA2273 versus three coils in the M. sativa parent and its self and testcross populations, and one coil in M. arborea), and Colletotrichum trifolii race 2 resistance (transferred from the resistant M. arborea parent, as the M. sativa parent and the self populations were highly susceptible). The hybrids were self sterile, but were female fertile to a high level when crossed with 4x, but not 2x, M. sativa, indicating they were at or near 4x. Both the pod coiling trait and anthracnose resistance segregated in the progeny of testcrosses between WA2273 and M. sativa. The work demonstrates that agronomically useful traits can be introgressed into M. sativa from M. arborea by use of male sterile M. sativa and sexual reproduction.

  16. Occurrence of Transgenic Feral Alfalfa (Medicago sativa subsp. sativa L. in Alfalfa Seed Production Areas in the United States.

    Directory of Open Access Journals (Sweden)

    Stephanie L Greene

    Full Text Available The potential environmental risks of transgene exposure are not clear for alfalfa (Medicago sativa subsp. sativa, a perennial crop that is cross-pollinated by insects. We gathered data on feral alfalfa in major alfalfa seed-production areas in the western United States to (1 evaluate evidence that feral transgenic plants spread transgenes and (2 determine environmental and agricultural production factors influencing the location of feral alfalfa, especially transgenic plants. Road verges in Fresno, California; Canyon, Idaho; and Walla Walla, Washington were surveyed in 2011 and 2012 for feral plants, and samples were tested for the CP4 EPSPS protein that conveys resistance to glyphosate. Of 4580 sites surveyed, feral plants were observed at 404 sites. Twenty-seven percent of these sites had transgenic plants. The frequency of sites having transgenic feral plants varied among our study areas. Transgenic plants were found in 32.7%, 21.4.7% and 8.3% of feral plant sites in Fresno, Canyon and Walla Walla, respectively. Spatial analysis suggested that feral populations started independently and tended to cluster in seed and hay production areas, places where seed tended to drop. Significant but low spatial auto correlation suggested that in some instances, plants colonized nearby locations. Neighboring feral plants were frequently within pollinator foraging range; however, further research is needed to confirm transgene flow. Locations of feral plant clusters were not well predicted by environmental and production variables. However, the likelihood of seed spillage during production and transport had predictive value in explaining the occurrence of transgenic feral populations. Our study confirms that genetically engineered alfalfa has dispersed into the environment, and suggests that minimizing seed spillage and eradicating feral alfalfa along road sides would be effective strategies to minimize transgene dispersal.

  17. Occurrence of Transgenic Feral Alfalfa (Medicago sativa subsp. sativa L.) in Alfalfa Seed Production Areas in the United States.

    Science.gov (United States)

    Greene, Stephanie L; Kesoju, Sandya R; Martin, Ruth C; Kramer, Matthew

    2015-01-01

    The potential environmental risks of transgene exposure are not clear for alfalfa (Medicago sativa subsp. sativa), a perennial crop that is cross-pollinated by insects. We gathered data on feral alfalfa in major alfalfa seed-production areas in the western United States to (1) evaluate evidence that feral transgenic plants spread transgenes and (2) determine environmental and agricultural production factors influencing the location of feral alfalfa, especially transgenic plants. Road verges in Fresno, California; Canyon, Idaho; and Walla Walla, Washington were surveyed in 2011 and 2012 for feral plants, and samples were tested for the CP4 EPSPS protein that conveys resistance to glyphosate. Of 4580 sites surveyed, feral plants were observed at 404 sites. Twenty-seven percent of these sites had transgenic plants. The frequency of sites having transgenic feral plants varied among our study areas. Transgenic plants were found in 32.7%, 21.4.7% and 8.3% of feral plant sites in Fresno, Canyon and Walla Walla, respectively. Spatial analysis suggested that feral populations started independently and tended to cluster in seed and hay production areas, places where seed tended to drop. Significant but low spatial auto correlation suggested that in some instances, plants colonized nearby locations. Neighboring feral plants were frequently within pollinator foraging range; however, further research is needed to confirm transgene flow. Locations of feral plant clusters were not well predicted by environmental and production variables. However, the likelihood of seed spillage during production and transport had predictive value in explaining the occurrence of transgenic feral populations. Our study confirms that genetically engineered alfalfa has dispersed into the environment, and suggests that minimizing seed spillage and eradicating feral alfalfa along road sides would be effective strategies to minimize transgene dispersal.

  18. Development of simple sequence repeat markers and diversity analysis in alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Wang, Zan; Yan, Hongwei; Fu, Xinnian; Li, Xuehui; Gao, Hongwen

    2013-04-01

    Efficient and robust molecular markers are essential for molecular breeding in plant. Compared to dominant and bi-allelic markers, multiple alleles of simple sequence repeat (SSR) markers are particularly informative and superior in genetic linkage map and QTL mapping in autotetraploid species like alfalfa. The objective of this study was to enrich SSR markers directly from alfalfa expressed sequence tags (ESTs). A total of 12,371 alfalfa ESTs were retrieved from the National Center for Biotechnology Information. Total 774 SSR-containing ESTs were identified from 716 ESTs. On average, one SSR was found per 7.7 kb of EST sequences. Tri-nucleotide repeats (48.8 %) was the most abundant motif type, followed by di-(26.1 %), tetra-(11.5 %), penta-(9.7 %), and hexanucleotide (3.9 %). One hundred EST-SSR primer pairs were successfully designed and 29 exhibited polymorphism among 28 alfalfa accessions. The allele number per marker ranged from two to 21 with an average of 6.8. The PIC values ranged from 0.195 to 0.896 with an average of 0.608, indicating a high level of polymorphism of the EST-SSR markers. Based on the 29 EST-SSR markers, assessment of genetic diversity was conducted and found that Medicago sativa ssp. sativa was clearly different from the other subspecies. The high transferability of those EST-SSR markers was also found for relative species.

  19. Accumulation and residue of napropamide in alfalfa (Medicago sativa) and soil involved in toxic response.

    Science.gov (United States)

    Cui, Li E; Yang, Hong

    2011-06-15

    Napropamide belongs to the amide herbicide family and widely used to control weeds in farmland. Intensive use of the herbicide has resulted in widespread contamination to ecosystems. The present study demonstrated an analysis on accumulation of the toxic pesticide napropamide in six genotypes of alfalfa (Medicago sativa), along with biological parameters and its residues in soils. Soil was treated with napropamide at 3 mg kg(-1) dry soil and alfalfa plants were cultured for 10 or 30 d, respectively. The maximum value for napropamide accumulation is 0.426 mg kg(-1) in shoots and 2.444 mg kg(-1) in roots. The napropamide-contaminated soil with alfalfa cultivation had much lower napropamide concentrations than the control (soil without alfalfa cultivation). Also, the content of napropamide residue in the rhizosphere was significantly lower than that in the non-rhizosphere soil. M. sativa exposed to 3 mg kg(-1) napropamide showed inhibited growth. Further analysis revealed that plants treated with napropamide accumulated more reactive oxygen species (O(2)(-) and H(2)O(2)) and less amounts of chlorophyll. However, not all cultivars showed oxidative injury, suggesting that the alfalfa cultivars display different tolerance to napropamide. Copyright © 2011 Elsevier B.V. All rights reserved.

  20. Allelopatic effects of cyanobacteria extracts containing microcystins on Medicago sativa-Rhizobia symbiosis.

    Science.gov (United States)

    El Khalloufi, Fatima; Oufdou, Khalid; Lahrouni, Majida; El Ghazali, Issam; Saqrane, Sanaa; Vasconcelos, Vitor; Oudra, Brahim

    2011-03-01

    The eutrophication of water leads to massive blooms of cyanobacteria potentially producers of highly toxic substances: cyanotoxins, especially microcystins (MC). The contamination of water used for irrigation by these toxins, can cause several adverse effects on plants and microorganisms. In this work, we report the phytotoxic effects of microcystins on the development of symbiosis between the leguminous plant Medicago sativa (Alfalfa) and rhizobia strains. The exposure of rhizobial strains to three different concentrations 0.01, 0.05 and 0.1 μg MC ml(-1) led to decrease on the bacteria growth. The strains of rhizobia Rh L1, Rh L2, Rh L3 and Rh L4 reduced their growth to, respectively, 20.85%, 20.80%, 33.19% and 25.65%. The chronic exposure of alfalfa seeds and seedlings to different MC concentrations affects the whole stages of plant development. The germination process has also been disrupted with an inhibition, which reaches 68.34% for a 22.24 μg MC ml(-1). Further, seedlings growth and photosynthetic process were also disrupted. The toxins reduced significantly the roots length and nodule formation and leads to an oxidative stress. Thus, the MCs contained in lake water and used for irrigation affect the development of symbiosis between M. sativa and Rhizobia. Copyright © 2010 Elsevier Inc. All rights reserved.

  1. Characterization of Chalcones from Medicago sativa L. and Their Hypolipidemic and Antiangiogenic Activities.

    Science.gov (United States)

    Ma, Qin-Ge; Li, Ting; Wei, Rong-Rui; Liu, Wen-Min; Sang, Zhi-Pei; Song, Zhong-Wen

    2016-11-02

    Medicago sativa L. is the most important cultivated herbage, known as "the king of forage" and "feed queen", in the world. A total of 8 new chalcones (1-8), and 12 known chalcones (9-20) were isolated from the aerial parts of M. sativa for the first time. Their structures were identified by extensive spectral data and references. The hypolipidemic and antiangiogenic activities of compounds (1-20) were evaluated for the first time. Compounds 3, 4, 11, 12, and 20 (10 μM) exhibited significant hypolipidemic activities by measuring the triglyceride content in HepG2 cells, with simvastatin as the positive control. Moreover, compounds 6, 8, 18, and 19 exhibited moderate antiangiogenic activities, which inhibited vascular-endothelial-growth-factor-induced human umbilical vein endothelial cell proliferation in vitro, with IC50 values of 13.86 ± 0.43, 15.53 ± 0.19, 39.52 ± 0.24, and 45.04 ± 0.51 μM, respectively. These research results may guide the search for new natural products with hypolipidemic and antiangiogenic attributes.

  2. Evaluation of Medicago sativa L. sprouts as antihyperlipidemic and antihyperglycemic agent.

    Science.gov (United States)

    Seida, Ahmed; El-Hefnawy, Hala; Abou-Hussein, Dina; Mokhtar, Fatma Alzahraa; Abdel-Naim, Ashraf

    2015-11-01

    Medicago sativa L. (Alfalfa) is traditionally used to treat diabetes. This study was designed to investigate the potential antihyperlipidemic and antihyperglycemic activity of M. sativa sprouts in streptozotocin (STZ) induced diabetes via i.p. injection of 55 mg/kg of STZ. Experimental animals were divided into the following groups: GP1 (normal), GP2 (STZ-hyperlipidemic), GP3 (rouvastatin), GP4 (metformin), GP 5-9 (diabetic treated with methanolic, petroleum ether, chloroform, ethyl acetate and butanol extracts). The administration of the total methanolic extract (500 mg/kg), the petroleum ether (32.5mg) and butanol fractions (60 mg) for 4 weeks significantly decreased (p<0.05) triglycerides (TG), total cholesterol (TC), low-density lipoproteins (LDL) and very low density lipoproteins (VLDL) in comparison to rouvastatin. Petroleum ether fraction proved to exhibit the best activity as antihyperlipidemic agent (12.23%). On the other hand, ethyl acetate fraction retained the best activity (vs. metformin) as antihyperglycemic agent. Histopathological evidences on liver, pancreas and spleen were in agreement with the above mentioned results. Purification, characterization, and identification of isolated compounds from the active fractions afforded 9 compounds: β-sitosterol and stigmasterol from the petroleum ether fraction; 10-hydroxy-coumestrol, apigenin, genistein, p-hydroxy-benzoic-acid, 7, 4'- dihydroxyflavone, quercetin-3-glucoside and sissotrin from the ethyl acetate fraction.

  3. Micromonospora from nitrogen fixing nodules of alfalfa (Medicago sativa L.). A new promising Plant Probiotic Bacteria.

    Science.gov (United States)

    Martínez-Hidalgo, Pilar; Galindo-Villardón, Purificación; Trujillo, Martha E; Igual, José M; Martínez-Molina, Eustoquio

    2014-09-17

    Biotic interactions can improve agricultural productivity without costly and environmentally challenging inputs. Micromonospora strains have recently been reported as natural endophytes of legume nodules but their significance for plant development and productivity has not yet been established. The aim of this study was to determine the diversity and function of Micromonospora isolated from Medicago sativa root nodules. Micromonospora-like strains from field alfalfa nodules were characterized by BOX-PCR fingerprinting and 16S rRNA gene sequencing. The ecological role of the interaction of the 15 selected representative Micromonospora strains was tested in M. sativa. Nodulation, plant growth and nutrition parameters were analyzed. Alfalfa nodules naturally contain abundant and highly diverse populations of Micromonospora, both at the intra- and at interspecific level. Selected Micromonospora isolates significantly increase the nodulation of alfalfa by Ensifer meliloti 1021 and also the efficiency of the plant for nitrogen nutrition. Moreover, they promote aerial growth, the shoot-to-root ratio, and raise the level of essential nutrients. Our results indicate that Micromonospora acts as a Rhizobia Helper Bacteria (RHB) agent and has probiotic effects, promoting plant growth and increasing nutrition efficiency. Its ecological role, biotechnological potential and advantages as a plant probiotic bacterium (PPB) are also discussed.

  4. Small RNA deep sequencing identifies novel and salt-stress-regulated microRNAs from roots of Medicago sativa and Medicago truncatula.

    Science.gov (United States)

    Long, Rui-Cai; Li, Ming-Na; Kang, Jun-Mei; Zhang, Tie-Jun; Sun, Yan; Yang, Qing-Chuan

    2015-05-01

    Small 21- to 24-nucleotide (nt) ribonucleic acids (RNAs), notably the microRNA (miRNA), are emerging as a posttranscriptional regulation mechanism. Salt stress is one of the primary abiotic stresses that cause the crop losses worldwide. In saline lands, root growth and function of plant are determined by the action of environmental salt stress through specific genes that adapt root development to the restrictive condition. To elucidate the role of miRNAs in salt stress regulation in Medicago, we used a high-throughput sequencing approach to analyze four small RNA libraries from roots of Zhongmu-1 (Medicago sativa) and Jemalong A17 (Medicago truncatula), which were treated with 300 mM NaCl for 0 and 8 h. Each library generated about 20 million short sequences and contained predominantly small RNAs of 24-nt length, followed by 21-nt and 22-nt small RNAs. Using sequence analysis, we identified 385 conserved miRNAs from 96 families, along with 68 novel candidate miRNAs. Of all the 68 predicted novel miRNAs, 15 miRNAs were identified to have miRNA*. Statistical analysis on abundance of sequencing read revealed specific miRNA showing contrasting expression patterns between M. sativa and M. truncatula roots, as well as between roots treated for 0 and 8 h. The expression of 10 conserved and novel miRNAs was also quantified by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). The miRNA precursor and target genes were predicted by bioinformatics analysis. We concluded that the salt stress related conserved and novel miRNAs may have a large variety of target mRNAs, some of which might play key roles in salt stress regulation of Medicago. © 2014 Scandinavian Plant Physiology Society.

  5. WATER DEFICIT EFFECT ON YIELD AND FORAGE QUALITY OF MEDICAGO SATIVA POPULATIONS UNDER FIELD CONDITIONS IN MARRAKESH AREA (MOROCCO)

    OpenAIRE

    Mohamed FARISSI; Cherki GHOULAM; Abdelaziz BOUIZGAREN

    2014-01-01

    The present study focused the effect of water deficit on agronomic potential and some traits related to forage quality in plants of Moroccan Alfalfa (Medicago sativa L.) populations (Taf 1, Taf 2, Dem and Tata) originated from Oasis and High Atlas of Morocco and an introduced variety from Australia (Siriver). The experiment was conducted under field conditions in experimental station of INRA-Marrakech and under two irrigation treatments. The first treatment was normal irrigation, providing an...

  6. The Hybridization Barrier between Herbaceous Medicago sativa and Woody M. arborea Is Weakened by Selection of Seed Parents

    Directory of Open Access Journals (Sweden)

    Edwin Bingham

    2013-05-01

    Full Text Available Medicago sativa, alfalfa or lucerne, and M. arborea were considered reproductively isolated until recently. Then, in 2003, an alfalfa genotype was identified that produced a few seeds and progeny with hybrid traits after a large number of pollinations by M. arborea. A derivative of this alfalfa genotype also produced a low frequency of progeny with hybrid traits. Thus, the hybridization barrier was weakened by selection of seed parents. Hybrids from both events expressed traits from M. arborea and M. arborea-specific DNA bands, although more of the M. sativa genome was retained, based on the DNA results. Thus, there was chromatin elimination during embryogenesis, resulting in partial hybrids (hereafter hybrids. However, more than 30 hybrids with an array of M. arborea traits have been obtained thus far, and research continues on the nature of the hybrids. Traits have been genetically transmitted in crosses, and selected traits are in use for alfalfa breeding. This paper reviews the first hybrids and then focuses on further weakening of the hybridization barrier with the discovery of a more efficient hybridizer derived from crossing Medicago sativa subspecies, sativa, coerulea and falcata. This genotype was found to have reproductive abnormalities associated with its complex subspecies origin that are best described as hybrid breakdown. In effect, this subspecies derivative is a bridge-cross parent that consistently produces hybrids. Reproductive abnormalities in the bridge-cross parent are reported and discussed.

  7. The Hybridization Barrier between Herbaceous Medicago sativa and Woody M. arborea Is Weakened by Selection of Seed Parents

    Science.gov (United States)

    Bingham, Edwin; Armour, David; Irwin, John

    2013-01-01

    Medicago sativa, alfalfa or lucerne, and M. arborea were considered reproductively isolated until recently. Then, in 2003, an alfalfa genotype was identified that produced a few seeds and progeny with hybrid traits after a large number of pollinations by M. arborea. A derivative of this alfalfa genotype also produced a low frequency of progeny with hybrid traits. Thus, the hybridization barrier was weakened by selection of seed parents. Hybrids from both events expressed traits from M. arborea and M. arborea-specific DNA bands, although more of the M. sativa genome was retained, based on the DNA results. Thus, there was chromatin elimination during embryogenesis, resulting in partial hybrids (hereafter hybrids). However, more than 30 hybrids with an array of M. arborea traits have been obtained thus far, and research continues on the nature of the hybrids. Traits have been genetically transmitted in crosses, and selected traits are in use for alfalfa breeding. This paper reviews the first hybrids and then focuses on further weakening of the hybridization barrier with the discovery of a more efficient hybridizer derived from crossing Medicago sativa subspecies, sativa, coerulea and falcata. This genotype was found to have reproductive abnormalities associated with its complex subspecies origin that are best described as hybrid breakdown. In effect, this subspecies derivative is a bridge-cross parent that consistently produces hybrids. Reproductive abnormalities in the bridge-cross parent are reported and discussed. PMID:27137379

  8. Genetic transformation and analysis of rice OsAPx2 gene in Medicago sativa.

    Directory of Open Access Journals (Sweden)

    Qingjie Guan

    Full Text Available The OsAPx2 gene from rice was cloned to produce PBI121::OsAPx2 dual-expression plants, of which expression level would be increasing under stressful conditions. The enzyme ascorbate peroxidase (APX in the leaves and roots of the plants increased with increasing exposure time to different sodium chloride (NaCl and hydrogen peroxide (H(2O(2concentrations, as indicated by protein gel blot analysis. The increased enzyme yield improved the ability of the plants to resist the stress treatments. The OsAPx2 gene was localized in the cytoplasm of epidermal onion cells as indicated by the instantaneous expression of green fluorescence. An 80% regeneration rate was observed in Medicago sativa L. plants transformed with the OsAPx2 gene using Agrobacterium tumefaciens, as indicated by specific primer PCR. The OsAPx2 gene was expressed at the mRNA level and the individual M. sativa (T#1,T#2,T#5 were obtained through assaying the generation of positive T2 using RNA gel blot analysis. When the seeds of the wild type (WT and the T2 (T#1,T#5 were incubated in culture containing MS with NaCl for 7 days, the results as shown of following: the root length of transgenic plant was longer than WT plants, the H(2O(2 content in roots of WT was more than of transgenic plants, the APX activity under stresses increased by 2.89 times compared with the WT, the malondialdehyde (MDA content of the WT was higher than the transgenic plants, the leaves of the WT turned yellow, but those of the transgenic plants remained green and remained healthy. The chlorophyll content in the WT leaves was less than in the transgenic plants, after soaking in solutions of H(2O(2, sodium sulfite (Na(2SO(3, and sodium bicarbonate (NaHCO(3. Therefore, the OsAPx2 gene overexpression in transgenic M. sativa improves the removal of H(2O(2 and the salt-resistance compared with WT plants. A novel strain of M. sativa carrying a salt-resistance gene was obtained.

  9. Genetic transformation and analysis of rice OsAPx2 gene in Medicago sativa.

    Science.gov (United States)

    Guan, Qingjie; Takano, Tetsuo; Liu, Shenkui

    2012-01-01

    The OsAPx2 gene from rice was cloned to produce PBI121::OsAPx2 dual-expression plants, of which expression level would be increasing under stressful conditions. The enzyme ascorbate peroxidase (APX) in the leaves and roots of the plants increased with increasing exposure time to different sodium chloride (NaCl) and hydrogen peroxide (H(2)O(2))concentrations, as indicated by protein gel blot analysis. The increased enzyme yield improved the ability of the plants to resist the stress treatments. The OsAPx2 gene was localized in the cytoplasm of epidermal onion cells as indicated by the instantaneous expression of green fluorescence. An 80% regeneration rate was observed in Medicago sativa L. plants transformed with the OsAPx2 gene using Agrobacterium tumefaciens, as indicated by specific primer PCR. The OsAPx2 gene was expressed at the mRNA level and the individual M. sativa (T#1,T#2,T#5) were obtained through assaying the generation of positive T2 using RNA gel blot analysis. When the seeds of the wild type (WT) and the T2 (T#1,T#5) were incubated in culture containing MS with NaCl for 7 days, the results as shown of following: the root length of transgenic plant was longer than WT plants, the H(2)O(2) content in roots of WT was more than of transgenic plants, the APX activity under stresses increased by 2.89 times compared with the WT, the malondialdehyde (MDA) content of the WT was higher than the transgenic plants, the leaves of the WT turned yellow, but those of the transgenic plants remained green and remained healthy. The chlorophyll content in the WT leaves was less than in the transgenic plants, after soaking in solutions of H(2)O(2), sodium sulfite (Na(2)SO(3)), and sodium bicarbonate (NaHCO(3)). Therefore, the OsAPx2 gene overexpression in transgenic M. sativa improves the removal of H(2)O(2) and the salt-resistance compared with WT plants. A novel strain of M. sativa carrying a salt-resistance gene was obtained.

  10. Cloning and characterization of chromosomal markers in alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Yu, Feng; Lei, Yunting; Li, Yuan; Dou, Quanwen; Wang, Haiqing; Chen, Zhiguo

    2013-07-01

    Eleven tandemly repetitive sequences were identified from a Cot-1 library by FISH and sequence analysis of alfalfa (Medicago sativa). Five repetitive sequences (MsCR-1, MsCR-2, MsCR-3, MsCR-4, and MsCR-5) were centromeric or pericentromeric, of which three were satellite DNAs and two were minisatellite DNAs. Monomers of 144, 148, and 168 bp were identified in MsCR-1, MsCR-2, and MsCR-3, respectively, while 15 and 39 bp monomers were identified in MsCR-4 and MsCR-5, respectively. Three repetitive sequences were characterized as subtelomeric; one repetitive sequence, MsTR-1, had a 184 bp monomer, and two repetitive sequences had fragments of 204 and 327 bp. Sequence analysis revealed homology (70-80 %) between MsTR-1 and a highly repeated sequence (C300) isolated from M. ssp. caerulea. Three identified repetitive sequences produced hybridization signals at multiple sites in a few of the chromosomes; one repetitive sequence was identified as the E180 satellite DNA previously isolated from M. sativa, while the other 163 and 227 bp fragments had distinct sequences. Physical mapping of the repetitive sequences with double-target FISH revealed different patterns. Thus, nine novel tandemly repetitive sequences that can be adopted as distinct chromosome markers in alfalfa were identified in this study. Furthermore, the chromosome distribution of each sequence was well described. Though significant chromosome variations were detected within and between cultivars, a molecular karyotype of alfalfa was suggested with the chromosome markers we identified. Therefore, these novel chromosome markers will still be a powerful tool for genome composition analysis, phylogenetic studies, and breeding applications.

  11. Physiological and antioxidant responses of Medicago sativa-rhizobia symbiosis to cyanobacterial toxins (Microcystins) exposure.

    Science.gov (United States)

    El Khalloufi, Fatima; Oufdou, Khalid; Lahrouni, Majida; Faghire, Mustapha; Peix, Alvaro; Ramírez-Bahena, Martha Helena; Vasconcelos, Vitor; Oudra, Brahim

    2013-12-15

    Toxic cyanobacteria in freshwaters can induce potent harmful effects on growth and development of plants irrigated with contaminated water. In this study, the effect of cyanobacteria extract containing Microcystins (MC) on Medicago sativa-rhizobia symbiosis was investigated in order to explore plants response through biomass production, photosynthetic pigment and antioxidant enzymes analysis: Peroxidase (POD), Polyphenoloxidase (PPO) and Catalase (CAT). Alfalfa plants were inoculated with two endosymbiotic rhizobial strains: RhOL1 (MC less sensitive strain) and RhOL3 (MC more sensitive strain), to evaluate the rhizobial contribution on the plant response cultured under cyanobacterial toxins stress. The two rhizobia strains were identified as Ensifer meliloti by sequence analysis of their rrs and atpD genes. The chronic exposure to MC extract showed shoot, root and nodules dry weight decrease, in both symbiosis cultures. The rate of decline in plants inoculated with RhOL3 was higher than that in symbiosis with RhOL1 mainly at 20 μg L(-1) of MC. Cyanotoxins also reduced photosynthetic pigment content and generated an oxidative stress observed at cellular level. POD, PPO and CAT activities were significantly increased in leaves, roots and nodules of alfalfa plants exposed to MC. These enzyme activities were higher in plants inoculated with RhOL3 especially when alfalfa plants were exposed to 20 μg L(-1) of MC. The present paper reports new scientific finding related to the behavior of rhizobia-M. sativa associations to MC (Microcystins) for later recommendation concerning the possible use of these symbiosis face to crops exposure to MC contaminated water irrigation. Copyright © 2013 Elsevier Ltd. All rights reserved.

  12. Asymmetric somatic hybrid plants between Medicago sativa L. (alfalfa, lucerne) and Onobrychis viciifolia Scop. (sainfoin).

    Science.gov (United States)

    Li, Y G; Tanner, G J; Delves, A C; Larkin, P J

    1993-12-01

    This paper reports on the production of intergeneric somatic hybrid plants between two sexually incompatible legume species. Medicago sativa (alfalfa, lucerne) leaf protoplasts were inactivated by lethal doses of iodoacetamide. Onobrychis viciifolia (sainfoin) suspension-cell protoplasts were gamma-irradiated at lethal doses. Following electrofusion under optimized conditions about 50,000 viable heterokaryons were produced in each test. The fusion products were cultured with the help of alfalfa nurse protoplasts. Functional complementation permitted only the heterokaryons to survive. A total of 706 putative heterokaryon-derived plantlets were regenerated and 570 survived transplantation to soil. Experimentation was aimed at the introduction of proanthocyanidins (condensed tannins) from sainfoin, a bloat-safe plant, to alfalfa, a bloat-causing forage crop; however, no tannin-positive regenerant plants were detected. Most regenerant plants have shown morphological differences from the fusion parents, although, as expected, all resembled the "recipient" parent, alfalfa. Southern analysis using an improved total-genomic probing technique has shown low levels of sainfoin-specific DNA in 43 out of 158 tested regenerants. Cytogenetic analysis of these asymmetric hybrids has confirmed the existence of euploid (2n=32; 17%) as well as aneuploid (2n=30, 33-78; 83%) plants. Pollen germination tests have indicated that the majority of the hybrids were fertile, while 35% had either reduced fertility or were completely sterile.

  13. Nodule and Leaf Nitrate Reductases and Nitrogen Fixation in Medicago sativa L. under Water Stress

    Science.gov (United States)

    Aparicio-Tejo, P.; Sánchez-Díaz, Manuel

    1982-01-01

    The effect of water stress on patterns of nitrate reductase activity in the leaves and nodules and on nitrogen fixation were investigated in Medicago sativa L. plants watered 1 week before drought with or without NO3−. Nitrogen fixation was decreased by water stress and also inhibited strongly by the presence of NO3−. During drought, leaf nitrate reductase activity (NRA) decreased significantly particularly in plants watered with NO3−, while with rewatering, leaf NRA recovery was quite important especially in the NO3−-watered plants. As water stress progressed, the nodular NRA increased both in plants watered with NO3− and in those without NO3− contrary to the behavior of the leaves. Beyond −15.105 pascal, nodular NRA began to decrease in plants watered with NO3−. This phenomenon was not observed in nodules of plants given water only. Upon rewatering, it was observed that in plants watered with NO3− the nodular NRA increased again, while in plants watered but not given NO3−, such activity began to decrease. Nitrogen fixation increased only in plants without NO3−. PMID:16662233

  14. Uptake of oxytetracycline and its phytotoxicity to alfalfa (Medicago sativa L.)

    Energy Technology Data Exchange (ETDEWEB)

    Kong, W.D. [Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085 (China); Zhu, Y.G. [Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085 (China)]. E-mail: ygzhu@rcees.ac.cn; Liang, Y.C. [Ministry of Agriculture Key Laboratory of Plant Nutrition and Nutrient Cycling, Institute of Soils and Fertilizers, Chinese Academy of Agricultural Sciences, Beijing 100081 (China); Zhang, J. [Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085 (China); Smith, F.A. [Soil and Land Systems, School of Earth and Environmental Sciences, University of Adelaide, DP 636, Adelaide, SA 5005 (Australia); Yang, M. [Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085 (China)

    2007-05-15

    A series of experiments were conducted in a hydroponic system to investigate the uptake of oxytetracycline (OTC) and its toxicity to alfalfa (Medicago sativa L.). OTC inhibited alfalfa shoot and root growth by up to 61% and 85%, respectively. The kinetics of OTC uptake could be well described by Michaelis-Menten equation with V {sub max} of 2.25 {mu}mol g{sup -1} fresh weight h{sup -1}, and K {sub m} of 0.036 mM. The uptake of OTC by alfalfa was strongly inhibited by the metabolic inhibitor, 2,4-DNP (2,4-dinitrophenol), at pH 3.5 and 6.0, but not by the aquaporin competitors, glycerol and Ag{sup +}. OTC uptake, however, was significantly inhibited by Hg{sup 2+}, suggesting that the inhibition of influx was due to general cellular stress rather than the specific action of Hg{sup 2+} on aquaporins. Results from the present study suggested that OTC uptake into alfalfa is an energy-dependent process. - Plant uptake of antibiotic oxytetracycline is energy-dependent.

  15. Early somatic embryo induction events in alfalfa callus cultures. [Medicago sativa

    Energy Technology Data Exchange (ETDEWEB)

    El-Bakry, A.A.; Hildebrand, D.F.

    1987-04-01

    High and low regenerating alfalfa Medicago sativa L. cv Regen S full sibs were isolated from a callus culture screen on modified Blaydes medium. The average number of embryos per ovary were thirty and zero for the high and low genotypes respectively after six weeks in culture. Proembryonic cell masses (4-8 celled) were observed after 4-5 days in culture and maximum meristematic activity was at 6-7 days in culture, for the high regenerating genotypes. Well formed globular embryos, both epidermal and subepidermal in origin, were observed after 2 weeks is culture. Samples in culture for 3, 6 and 14 days from the high and low regenerating genotypes were radiolabeled in vivo with /sup 35/S-methionine and run both on one and two dimension gels. The results will be discussed in relation to differences in proteins between the high and low regenerating genotypes at the stage of maximum meristematic activity (day 6) and differences occurring relative to the appearance of globular stage embryos (day 14) will be presented.

  16. Water transfer in an alfalfa/maize association. [Medicago sativa; Zea mays

    Energy Technology Data Exchange (ETDEWEB)

    Corak, S.J.; Blevins, D.G.; Pallardy, S.G.

    1987-07-01

    The authors investigated the possibility of interspecific water transfer in an alfalfa (Medicago sativa L.) and maize (Zea mays L.) association. An alfalfa plant was grown through two vertically stacked plastic tubes. A 5 centimeter air gap between tubes was bridged by alfalfa roots. Five-week old maize plants with roots confined to the top tube were not watered, while associated alfalfa roots had free access to water in the bottom tube (the -/+ treatment). Additional treatments included: top and bottom tubes watered (+/+), top and bottom tubes droughted (-/-), and top tube droughted after removal of alfalfa root bridges and routine removal of alfalfa tillers (-*). Predawn leaf water potential of maize in the -/+ treatment fell to -1.5 megapascals 13 days after the start of drought; thereafter, predawn and midday potentials were maintained near -1.9 megapascals. Leaf water potentials of maize in the -/- and -* treatments declined steadily; all plants in these treatments were completely desiccated before day 50. High levels of tritium activity were detected in water extracted from both alfalfa and maize leaves after /sup 3/H/sub 2/O was injected into the bottom -/+ tube at day 70 or later. Maize in the -/+ treatment was able to survive an otherwise lethal period of drought by utilizing water lost by alfalfa roots.

  17. Detection of polycyclic aromatic hydrocarbons (PAHs) in Medicago sativa L. by fluorescence microscopy.

    Science.gov (United States)

    Alves, Wilber S; Manoel, Evelin A; Santos, Noemi S; Nunes, Rosane O; Domiciano, Giselli C; Soares, Marcia R

    2017-04-01

    Green technologies, such as phytoremediation, are effective for removing organic pollutants derived from oil and oil products, including polycyclic aromatic hydrocarbons (PAHs). Given the increasing popularity of these sustainable remediation techniques, methods based on fluorescence microscopy and multiphoton microscopy for the environmental monitoring of such pollutants have emerged in recent decades as effective tools for phytoremediation studies aimed at understanding the fate of these contaminants in plants. However, little is known about the cellular and molecular mechanisms involved in PAH uptake, responses and degradation by plants. Thus, the present study aimed to detect the location of pyrene, anthracene and phenanthrene using fluorescence microscopy techniques in shoots and roots of Medicago sativa L. (alfalfa) plants grown in artificially contaminated soil (150ppm PAHs) for 40days. Leaflet and root samples were then collected and observed under a fluorescence microscope to detect the presence of PAHs in various tissues. One important finding of the present study was intense fluorescence in the glandular secreting trichomes (GSTs) of plants grown in contaminated soil. These trichomes, with a previously unknown function, may be sites of PAH conjugation and degradation. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. [Determination of Hard Rate of Alfalfa (Medicago sativa L.) Seeds with Near Infrared Spectroscopy].

    Science.gov (United States)

    Wang, Xin-xun; Chen, Ling-ling; Zhang, Yun-wei; Mao, Pei-sheng

    2016-03-01

    Alfalfa (Medicago sativa L.) is the most commonly grown forage crop due to its better quality characteristics and high adaptability in China. However, there was 20%-80% hard seeds in alfalfa which could not be identified easily from non hard seeds which would cause the loss of seed utilization value and plant production. This experiment was designed for 121 samples of alfalfa. Seeds were collected according to different regions, harvested year and varieties. 31 samples were artificial matched as hard rates ranging from 20% to 80% to establish a model for hard seed rate by near infrared spectroscopy (NIRS) with Partial Least Square (PLS). The objective of this study was to establish a model and to estimate the efficiency of NIRS for determining hard rate of alfalfa seeds. The results showed that the correlation coefficient (R2(cal)) of calibration model was 0.981 6, root mean square error of cross validation (RMSECV) was 5.32, and the ratio of prediction to deviation (RPD) was 3.58. The forecast model in this experiment presented the satisfied precision. The proposed method using NIRS technology is feasible for identification and classification of hard seed in alfalfa. A new method, as nondestructive testing of hard seed rate, was provided to theoretical basis for fast nondestructive detection of hard seed rates in alfalfa.

  19. Screening of Cd tolerant genotypes and isolation of metallothionein genes in alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Wang, Xiaojuan; Song, Yu; Ma, Yanhua; Zhuo, Renying; Jin, Liang

    2011-12-01

    In order to evaluate Cd tolerance in wide-ranging sources of alfalfa (Medicago sativa) and to identify Cd tolerant genotypes which may potentially be useful for restoring Cd-contaminated environments, thirty-six accessions of alfalfa were screened under hydroponic culture. Our results showed that the relative root growth rate varied from 0.48 to 1.0, which indicated that different alfalfa accessions had various responses to Cd stress. The candidate fragments derived from differentially expressed metallothionein (MT) genes were cloned from leaves of two Cd tolerant genotypes, YE and LZ. DNA sequence and the deduced protein sequence showed that MsMT2a and MsMT2b had high similarity to those in leguminous plants. DDRT-PCR analysis showed that MsMT2a expressed in both YE and LZ plants under control and Cd stress treatment, but MsMT2b only expressed under Cd stress treatment. This suggested that MsMT2a was universally expressed in leaves of alfalfa but expression of MsMT2b was Cadmium (Cd) inducible. Copyright © 2011 Elsevier Ltd. All rights reserved.

  20. Over-Expression of Arabidopsis EDT1 Gene Confers Drought Tolerance in Alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Zheng, Guangshun; Fan, Cunying; Di, Shaokang; Wang, Xuemin; Xiang, Chengbin; Pang, Yongzhen

    2017-01-01

    Alfalfa (Medicago sativa L.) is an important legume forage crop with great economic value. However, as the growth of alfalfa is seriously affected by an inadequate supply of water, drought is probably the major abiotic environmental factor that most severely affects alfalfa production worldwide. In an effort to enhance alfalfa drought tolerance, we transformed the Arabidopsis Enhanced Drought Tolerance 1 (AtEDT1) gene into alfalfa via Agrobacterium-mediated transformation. Compared with wild type plants, drought stress treatment resulted in higher survival rates and biomass, but reduced water loss rates in the transgenic plants. Furthermore, transgenic alfalfa plants had increased stomatal size, but reduced stomatal density, and these stomatal changes contributed greatly to reduced water loss from leaves. Importantly, transgenic alfalfa plants exhibited larger root systems with larger root lengths, root weight, and root diameters than wild type plants. The transgenic alfalfa plants had reduced membrane permeability and malondialdehyde content, but higher soluble sugar and proline content, higher superoxide dismutase activity, higher chlorophyll content, enhanced expression of drought-responsive genes, as compared with wild type plants. Notably, transgenic alfalfa plants grew better in a 2-year field trial and showed enhanced growth performance with increased biomass yield. All of our morphological, physiological, and molecular analyses demonstrated that the ectopic expression of AtEDT1 improved growth and enhanced drought tolerance in alfalfa. Our study provides alfalfa germplasm for use in forage improvement programs, and may help to increase alfalfa production in arid lands.

  1. Determination of sugars and cyclitols isolated from various morphological parts of Medicago sativa L.

    Science.gov (United States)

    Al-Suod, Hossam; Ratiu, Ileana-Andreea; Ligor, Magdalena; Ligor, Tomasz; Buszewski, Bogusław

    2017-12-18

    Plant research interest has increased all over the world, and a large body of evidence has been collected to show the huge potential of medicinal plants in various disease treatments. Medicago sativa L., known as alfalfa, is a rich source of biologically active components and secondary metabolites and was frequently used from the ancient times both as fodder crop and as a traditional medicine in the treatment of various diseases. Cyclitols, naturally occurring in this plant, have a particular interest for us due to their significant anti-diabetic, antioxidant, anti-inflammatory and anti-cancer properties. In the present study we revealed the isolation, the identification and the quantification of some cyclitols and sugars extracted from different morphological parts of alfalfa plant. Soxhlet extraction and solid phase extraction were used as extraction and purification methods, while for the analyses derivatization followed by gas chromatography with mass spectrometry was involved. The obtained results showed significant differences in the quantities of cyclitols and sugars found in the investigated morphological parts, ranging between 0.02 and 13.86 mg/g of plant in case of cyclitols, and in the range of 0.09 and 40.09 mg/g of plant for sugars. However, roots have the richest part of cyclitols and sugars in contrast to the leaves. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  2. The thiol compounds glutathione and homoglutathione differentially affect cell development in alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Pasternak, Taras; Asard, Han; Potters, Geert; Jansen, Marcel A K

    2014-01-01

    Glutathione (GSH) is an important scavenger of Reactive Oxygen Species (ROS), precursor of metal chelating phytochelatins, xenobiotic defence compound and regulator of cell proliferation. Homoglutathione (hGSH) is a GSH homologue that is present in several taxa in the family of Fabaceae. It is thought that hGSH performs many of the stress-defence roles typically ascribed to GSH, yet little is known about the potential involvement of hGSH in controlling cell proliferation. Here we show that hGSH/GSH ratios vary across organs and cells and that these changes in hGSH/GSH ratio occur during dedifferentiation and/or cell cycle activation events. The use of a GSH/hGSH biosynthesis inhibitor resulted in impaired cytokinesis in isolated protoplasts, showing the critical importance of these thiol-compounds for cell division. However, exposure of isolated protoplasts to exogenous GSH accelerated cytokinesis, while exogenous hGSH was found to inhibit the same process. We conclude that GSH and hGSH have distinct functional roles in cell cycle regulation in Medicago sativa L. GSH is associated with meristemic cells, and promotes cell cycle activation and induction of somatic embryogenesis, while hGSH is associated with differentiated cells and embryo proliferation. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  3. Metabolomic Analysis of Alfalfa (Medicago sativa L.) Root-Symbiotic Rhizobia Responses under Alkali Stress.

    Science.gov (United States)

    Song, Tingting; Xu, Huihui; Sun, Na; Jiang, Liu; Tian, Pu; Yong, Yueyuan; Yang, Weiwei; Cai, Hua; Cui, Guowen

    2017-01-01

    Alkaline salts (e.g., NaHCO3 and Na2CO3) causes more severe morphological and physiological damage to plants than neutral salts (e.g., NaCl and Na2SO4) due to differences in pH. The mechanism by which plants respond to alkali stress is not fully understood, especially in plants having symbotic relationships such as alfalfa (Medicago sativa L.). Therefore, a study was designed to evaluate the metabolic response of the root-nodule symbiosis in alfalfa under alkali stress using comparative metabolomics. Rhizobium-nodulized (RI group) and non-nodulized (NI group) alfalfa roots were treated with 200 mmol/L NaHCO3 and, roots samples were analyzed for malondialdehydyde (MDA), proline, glutathione (GSH), superoxide dismutase (SOD), and peroxidase (POD) content. Additionally, metabolite profiling was conducted using gas chromatography combined with time-of-flight mass spectrometry (GC/TOF-MS). Phenotypically, the RI alfalfa exhibited a greater resistance to alkali stress than the NI plants examined. Physiological analysis and metabolic profiling revealed that RI plants accumulated more antioxidants (SOD, POD, GSH), osmolytes (sugar, glycols, proline), organic acids (succinic acid, fumaric acid, and alpha-ketoglutaric acid), and metabolites that are involved in nitrogen fixation. Our pairwise metabolomics comparisons revealed that RI alfalfa plants exhibited a distinct metabolic profile associated with alkali putative tolerance relative to NI alfalfa plants. Data provide new information about the relationship between non-nodulized, rhizobium-nodulized alfalfa and alkali resistance.

  4. Effect of cumulative growing degree day on accumulation of alfalfa (Medicago sativa L. root reserve saccharides

    Directory of Open Access Journals (Sweden)

    Josef Hakl

    2008-01-01

    Full Text Available The aim of this study was to investigate the relation between growing degree day method and accumulation of root reserve saccharides before over wintering by alfalfa (Medicago sativa L. in the condition of Czech Republic. In 2002–2004, the field experiment was conducted at the Research station of the Czech University of Life Sciences in central Bohemia. The interval between summer and last autumn harvest was 40–50 days or 60–70 days, respectively. These intervals were expressed as cumulative growing degree-days (GDD for each year. The plants were sampled in each autumn with four replicates for each variant; the average depth of sampling was 150 mm. The weight of roots, amount of starch, and water soluble saccharides (WSC per m2 was determined. The total accumulation of root reserve saccharides was determined mainly by conditions during all vegetation period. The length of the interval or cumulative GDD influenced only variation of this basic amount. The GDD was corresponded better with starch content whilst WSC was more related to length of the interval. In central Bohemia condition, the GDD 600–700 °C was preliminary determined for maximal accumulation of root reserve saccharides. The GDD above this level did not significantly increase the root reserve accumulation. For exact verify of this model, other experiments in more sites are needed.

  5. Metabolomic Analysis of Alfalfa (Medicago sativa L. Root-Symbiotic Rhizobia Responses under Alkali Stress

    Directory of Open Access Journals (Sweden)

    Tingting Song

    2017-07-01

    Full Text Available Alkaline salts (e.g., NaHCO3 and Na2CO3 causes more severe morphological and physiological damage to plants than neutral salts (e.g., NaCl and Na2SO4 due to differences in pH. The mechanism by which plants respond to alkali stress is not fully understood, especially in plants having symbotic relationships such as alfalfa (Medicago sativa L.. Therefore, a study was designed to evaluate the metabolic response of the root-nodule symbiosis in alfalfa under alkali stress using comparative metabolomics. Rhizobium-nodulized (RI group and non-nodulized (NI group alfalfa roots were treated with 200 mmol/L NaHCO3 and, roots samples were analyzed for malondialdehydyde (MDA, proline, glutathione (GSH, superoxide dismutase (SOD, and peroxidase (POD content. Additionally, metabolite profiling was conducted using gas chromatography combined with time-of-flight mass spectrometry (GC/TOF-MS. Phenotypically, the RI alfalfa exhibited a greater resistance to alkali stress than the NI plants examined. Physiological analysis and metabolic profiling revealed that RI plants accumulated more antioxidants (SOD, POD, GSH, osmolytes (sugar, glycols, proline, organic acids (succinic acid, fumaric acid, and alpha-ketoglutaric acid, and metabolites that are involved in nitrogen fixation. Our pairwise metabolomics comparisons revealed that RI alfalfa plants exhibited a distinct metabolic profile associated with alkali putative tolerance relative to NI alfalfa plants. Data provide new information about the relationship between non-nodulized, rhizobium-nodulized alfalfa and alkali resistance.

  6. Nutraceutical Potential of New Alfalfa (Medicago sativa) Ingredients for Beverage Preparations.

    Science.gov (United States)

    Soto-Zarazúa, Maria Guadalupe; Bah, Moustapha; Costa, Anabela Silvia Gomes; Rodrigues, Francisca; Pimentel, Filipa Botelho; Rojas-Molina, Isela; Rojas, Alejandra; Oliveira, Maria Beatriz Prior Pinto

    2017-10-01

    Alfalfa (Medicago sativa) has been extensively used as animal feed, due to its fiber, protein, minerals, and vitamins, being also a useful source of phenolic compounds with potential therapeutic benefits. Nevertheless, its potential use as human ingredient is scarce. The aim of this work was to assess the nutritional composition, amino acid profile, and antioxidant capacity (AOC) of freeze-dried juice (FDJ) and fibrous residual material (RM), two new alfalfa-derived products (Adps) recently launched as ingredients for beverage preparations. Results demonstrated a high content of proteins (23-30 g/100 g FDJ and 13-17 g/100 g RM), crude fiber (29 g/100 g RM), and minerals (such as sodium, calcium, iron, and zinc). No significant difference was found in caloric content (4 kcal/g). Essential and nonessential amino acids were quantified in both Adps being leucine and lysine the most abundant. Total phenolic and total flavonoid contents (TPC and TFC, respectively) and their changes along the different harvesting periods of the year were also examined. FDJ presented the highest TPC in May (19 mg gallic acid equivalents/g dry weight [dw]), while in October TFC had the maximum value (4 mg catechin equivalents/g dw). Both products exhibited an interesting AOC by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ferric reducing antioxidant power assays. This study reports the nutraceutical potential of two new types of Adps.

  7. Concerted changes in N and C primary metabolism in alfalfa (Medicago sativa) under water restriction.

    Science.gov (United States)

    Aranjuelo, Iker; Tcherkez, Guillaume; Molero, Gemma; Gilard, Françoise; Avice, Jean-Christophe; Nogués, Salvador

    2013-02-01

    Although the mechanisms of nodule N(2) fixation in legumes are now well documented, some uncertainty remains on the metabolic consequences of water deficit. In most cases, little consideration is given to other organs and, therefore, the coordinated changes in metabolism in leaves, roots, and nodules are not well known. Here, the effect of water restriction on exclusively N(2)-fixing alfalfa (Medicago sativa L.) plants was investigated, and proteomic, metabolomic, and physiological analyses were carried out. It is shown that the inhibition of nitrogenase activity caused by water restriction was accompanied by concerted alterations in metabolic pathways in nodules, leaves, and roots. The data suggest that nodule metabolism and metabolic exchange between plant organs nearly reached homeostasis in asparagine synthesis and partitioning, as well as the N demand from leaves. Typically, there was (i) a stimulation of the anaplerotic pathway to sustain the provision of C skeletons for amino acid (e.g. glutamate and proline) synthesis; (ii) re-allocation of glycolytic products to alanine and serine/glycine; and (iii) subtle changes in redox metabolites suggesting the implication of a slight oxidative stress. Furthermore, water restriction caused little change in both photosynthetic efficiency and respiratory cost of N(2) fixation by nodules. In other words, the results suggest that under water stress, nodule metabolism follows a compromise between physiological imperatives (N demand, oxidative stress) and the lower input to sustain catabolism.

  8. Over-Expression of Arabidopsis EDT1 Gene Confers Drought Tolerance in Alfalfa (Medicago sativa L.

    Directory of Open Access Journals (Sweden)

    Guangshun Zheng

    2017-12-01

    Full Text Available Alfalfa (Medicago sativa L. is an important legume forage crop with great economic value. However, as the growth of alfalfa is seriously affected by an inadequate supply of water, drought is probably the major abiotic environmental factor that most severely affects alfalfa production worldwide. In an effort to enhance alfalfa drought tolerance, we transformed the Arabidopsis Enhanced Drought Tolerance 1 (AtEDT1 gene into alfalfa via Agrobacterium-mediated transformation. Compared with wild type plants, drought stress treatment resulted in higher survival rates and biomass, but reduced water loss rates in the transgenic plants. Furthermore, transgenic alfalfa plants had increased stomatal size, but reduced stomatal density, and these stomatal changes contributed greatly to reduced water loss from leaves. Importantly, transgenic alfalfa plants exhibited larger root systems with larger root lengths, root weight, and root diameters than wild type plants. The transgenic alfalfa plants had reduced membrane permeability and malondialdehyde content, but higher soluble sugar and proline content, higher superoxide dismutase activity, higher chlorophyll content, enhanced expression of drought-responsive genes, as compared with wild type plants. Notably, transgenic alfalfa plants grew better in a 2-year field trial and showed enhanced growth performance with increased biomass yield. All of our morphological, physiological, and molecular analyses demonstrated that the ectopic expression of AtEDT1 improved growth and enhanced drought tolerance in alfalfa. Our study provides alfalfa germplasm for use in forage improvement programs, and may help to increase alfalfa production in arid lands.

  9. Toxic effects of arsenic on Sinorhizobium-Medicago sativa symbiotic interaction

    Energy Technology Data Exchange (ETDEWEB)

    Pajuelo, Eloisa [Department of Microbiology and Parasitology, Faculty of Pharmacy, University of Seville, 41012 Seville (Spain); Rodriguez-Llorente, Ignacio D. [Department of Microbiology and Parasitology, Faculty of Pharmacy, University of Seville, 41012 Seville (Spain)], E-mail: irodri@us.es; Dary, Mohammed; Palomares, Antonio J. [Department of Microbiology and Parasitology, Faculty of Pharmacy, University of Seville, 41012 Seville (Spain)

    2008-07-15

    Recently, the Rhizobium-legume symbiotic interaction has been proposed as an interesting tool in bioremediation. However, little is known about the effect of most common contaminants on this process. The phytotoxic effects of arsenic on nodulation of Medicago sativa have been examined in vitro using the highly arsenic resistant and symbiotically effective Sinorhizobium sp. strain MA11. The bacteria were able to grow on plates containing As concentrations as high as 10 mM. Nevertheless, as little as 25-35 {mu}M arsenite produced a 75% decrease in the total number of nodules, due to a 90% reduction in the number of rhizobial infections, as could be determined using the strain MA11 carrying a lacZ reporter gene. This effect was associated to root hair damage and a shorter infective root zone. However, once nodulation was established nodule development seemed to continue normally, although earlier senescence could be observed in nodules of arsenic-grown plants. - First steps of nodulation of alfalfa, in particular infection thread formation, are more sensitive to As than nitrogen fixation due to plant effects.

  10. In silico identification of transcription factors in Medicago sativa using available transcriptomic resources.

    Science.gov (United States)

    Postnikova, Olga A; Shao, Jonathan; Nemchinov, Lev G

    2014-06-01

    Transcription factors (TFs) are proteins that govern organismal development and response to the environment by regulating gene expression. Information on the amount and diversity of TFs within individual plant species is critical for understanding of their biological roles and evolutionary history across the plant kingdom. Currently, only scattered information on separate TFs is available for alfalfa, the most extensively cultivated forage legume in the world. In the meantime, several large transcriptomic resources that can be used to identify and characterize alfalfa TF genes are freely accessible online. In this study, we have performed an in silico analysis of transcriptome data generated in our laboratory and publicly acquirable from other sources to reveal and systematize alfalfa transcription factors. Transcriptome-wide mining enabled prediction of 983 TFs along with their sequence features and putative phylogenies of the largest families. All data were assembled into a simple open-access database named AlfalfaTFDB ( http://plantpathology.ba.ars.usda.gov/alfalfatfdb.html ). Transcriptomic analysis used in this work represents an effective approach for the identification of TF genes in plants with incomplete genomes, such as alfalfa. Integrated TF repertoires of Medicago sativa will provide an important tool for studying regulation of gene expression in other complex non-model species of agricultural significance.

  11. The Current Status, Problems, and Prospects of Alfalfa (Medicago sativa L. Breeding in China

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    Shangli Shi

    2017-01-01

    Full Text Available This paper reviews the current status, methodology, achievements, and prospects of alfalfa (Medicago sativa L. breeding in China. There are 77 cultivars that have been registered in the country, these include 36 cultivars bred through breeding programs, 17 introduced from overseas, 5 domesticated from wild ecotypes, and 19 through regional collection/breeding programs. Cultivars have been selected for cold resistance, disease resistance, salt tolerance, grazing tolerance, high yield, and early maturity. Most of these cultivars have been developed through conventional breeding techniques, such as selective and cross breeding, and some are now being evaluated that have been developed through the application of transgenic technology. The major problems for alfalfa breeding in China include low success rate, slow progress to breed resistant varieties, lack of breeding materials and their systematic collection, storage and evaluation, lack of advanced breeding techniques, and low adoption rate of new cultivars. There are gaps in alfalfa breeding between China and the developed world. Improvement of alfalfa breeding techniques, development of cultivars with adaptations to different regions within China, and the protection and utilization of alfalfa germplasm resources have been identified as major strategies to improve the efficiency of alfalfa breeding in China.

  12. Enhanced salt tolerance of alfalfa (Medicago sativa) by rstB gene transformation.

    Science.gov (United States)

    Zhang, Wan-Jun; Wang, Tao

    2015-05-01

    Generating salt tolerance forage plant is essential for use of the land affected by high salinity. A salt tolerance gene rstB was used as a selectable marker gene in Agrobacterium-mediated transformation of tobacco under a selective regime of 170mM NaCl. The transgenic plants showed clear improvement in salt tolerance. To improve salt tolerance of alfalfa (Medicago sativa L.), rstB gene was introduced into alfalfa genome by Agrobacterium-mediated transformation. No abnormal phenotype was observed among the transgenic plants when compared with wild type (wt) plants. Significant enhancement of resistance to salt-shock treatment was noted on the rstB transgenic (T0) plants. Transgenic second-generation (T1) seeds showed improved germination rate and seedling growth under salt-stress condition. Hindered Na(+) accumulation, but enhanced Ca(2+) accumulation was observed on the rstB T1 plants when subjected to salt-stresses. Enhanced calcium accumulation in transgenic plants was also verified by cytohistochemical localization of calcium. Under salt-stress of 50mM NaCl, about 15% of the transgenic plants finished their life-cycle but the wt plants had no flower formation. The results demonstrated that the expression of rstB gene improved salt tolerance in transgenic alfalfa. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  13. Selective lignin downregulation leads to constitutive defense response expression in alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Gallego-Giraldo, Lina; Jikumaru, Yusuke; Kamiya, Yuji; Tang, Yuhong; Dixon, Richard A

    2011-05-01

    • Downregulation of hydroxycinnamoyl CoA: shikimate hydroxycinnamoyl transferase (HCT) in alfalfa (Medicago sativa) reduces lignin levels and improves forage quality and saccharification efficiency for bioethanol production. However, the plants have reduced stature. It was previously reported that HCT-down-regulated Arabidopsis have impaired auxin transport, but this has recently been disproved. • To address the basis for the phenotypes of lignin-modified alfalfa, we measured auxin transport, profiled a range of metabolites including flavonoids and hormones, and performed in depth transcriptome analyses. • Auxin transport is unaffected in HCT antisense alfalfa despite increased flavonoid biosynthesis. The plants show increased cytokinin and reduced auxin levels, and gibberellin levels and sensitivity are both reduced. Levels of salicylic, jasmonic and abscisic acids are elevated, associated with massive upregulation of pathogenesis and abiotic stress-related genes and enhanced tolerance to fungal infection and drought. • We suggest that HCT downregulated alfalfa plants exhibit constitutive activation of defense responses, triggered by release of bioactive cell wall fragments and production of hydrogen peroxide as a result of impaired secondary cell wall integrity. © 2011 The Authors. New Phytologist © 2011 New Phytologist Trust.

  14. Transcriptome analysis of Glomus mosseae/Medicago sativa mycorrhiza on atrazine stress.

    Science.gov (United States)

    Song, Fuqiang; Li, Jize; Fan, Xiaoxu; Zhang, Quan; Chang, Wei; Yang, Fengshan; Geng, Gui

    2016-02-02

    Arbuscular mycorrhizal fungi (AMF) protect host plants against diverse biotic and abiotic stresses, and promote biodegradation of various contaminants. In this study effect of Glomus mosseae/Medicago sativa mycorrhiza on atrazine degradation was investigated. It was observed that the atrazine degradation rates with any addition level in mycorrhizal treatments were all significantly higher than those in non-mycorrhizal treatments. When atrazine was applied at 20 mg kg(-1), the removal efficiency was up to 74.65%. Therefore, G. mosseae can be considered as ideal inhabitants of technical installations to facilitate phytoremediation. Furthermore, a total of 10.4 Gb was used for de novo transcriptome assembly, resulting in a comprehensive data set for the identification of genes corresponding to atrazine stress in the AM association. After comparative analysis with edgeR, a total of 2,060 differential expressed genes were identified, including 570 up-regulated genes and 1490 down-regulated genes. After excluding 'function unknown' and 'general function predictions only' genes, 172 up-regulated genes were obtained. The differentially expressed genes in AM association with and without atrazine stress were associated with molecular processes/other proteins, zinc finger protein, intracellular/extracellular enzymes, structural proteins, anti-stress/anti-disease protein, electron transport-related protein, and plant growth associated protein. Our results not only prove AMF has important ecological significance on atrazine degradation but also provide evidence for the molecular mechanisms of atrazine degradation by AMF.

  15. MicroRNA156 improves drought stress tolerance in alfalfa (Medicago sativa) by silencing SPL13.

    Science.gov (United States)

    Arshad, Muhammad; Feyissa, Biruk A; Amyot, Lisa; Aung, Banyar; Hannoufa, Abdelali

    2017-05-01

    Alfalfa (Medicago sativa) is an important forage crop that is often grown in areas that frequently experience drought and water shortage. MicroRNA156 (miR156) is an emerging tool for improving various traits in plants. We tested the role of miR156d in drought response of alfalfa, and observed a significant improvement in drought tolerance of miR156 overexpression (miR156OE) alfalfa genotypes compared to the wild type control (WT). In addition to higher survival and reduced water loss, miR156OE genotypes also maintained higher stomatal conductance compared to WT during drought stress. Furthermore, we observed an enhanced accumulation of compatible solute (proline) and increased levels of abscisic acid (ABA) and antioxidants in miR156OE genotypes. Similarly, alfalfa plants with reduced expression of miR156-targeted SPL13 showed reduced water loss and enhanced stomatal conductance, chlorophyll content and photosynthetic assimilation. Several genes known to be involved in drought tolerance were differentially expressed in leaf and root of miR156 overexpression plants. Taken together, our findings reveal that miR156 improves drought tolerance in alfalfa at least partially by silencing SPL13. Crown Copyright © 2017. Published by Elsevier B.V. All rights reserved.

  16. Ractopamine up take by alfalfa (Medicago sativa) and wheat (Triticum aestivum) from soil.

    Science.gov (United States)

    Shelver, Weilin L; DeSutter, Thomas M

    2015-08-01

    Ractopamine is a beta adrenergic agonist used as a growth promoter in swine, cattle and turkeys. To test whether ractopamine has the potential to accumulate in plants grown in contaminated soil, a greenhouse study was conducted with alfalfa (Medicago sativa) and wheat (Triticum aestivum) grown in two soils having different concentrations of organic matter (1.3% and 2.1%), amended with 0, 0.5, and 10 μg/g of ractopamine. Plant growth ranged from 2.7 to 8.8 g dry weight (dw) for alfalfa, and 8.7 to 40 g dw for wheat and was generally greater in the higher organic matter content soil. The uptake of ractopamine in plant tissues ranged from non-detectable to 897 ng/g and was strongly dependent on soil ractopamine concentration across soil and plant tissue. When adjusted to the total fortified quantities, the amount of ractopamine taken up by the plant tissue was low, <0.01% for either soil. Copyright © 2015. Published by Elsevier B.V.

  17. Chemical modification and degradation of atrazine in Medicago sativa through multiple pathways.

    Science.gov (United States)

    Zhang, Jing Jing; Lu, Yi Chen; Yang, Hong

    2014-10-08

    Atrazine is a member of the triazine herbicide family intensively used to control weeds for crop production. In this study, atrazine residues and its degraded products in alfalfa (Medicago sativa) were characterized using UPLC-TOF-MS/MS. Most of atrazine absorbed in plants was found as chemically modified derivatives like deisopropylated atrazine (DIA), dehydrogenated atrazine (DHA), or methylated atrazine (MEA), and some atrazine derivatives were conjugated through different functional groups such as sugar, glutathione, and amino acids. Interestingly, the specific conjugates DHA+hGSH (homoglutathione) and MEA-HCl+hGSH in alfalfa were detected. These results suggest that atrazine in alfalfa can be degraded through different pathways. The increased activities of glycosyltransferase and glutathione S-transferase were determined to support the atrazine degradation models. The outcome of the work uncovered the detailed mechanism for the residual atrazine accumulation and degradation in alfalfa and will help to evaluate whether the crop is suitable to be cultivated in the atrazine-polluted soil.

  18. Stem nematode counteracts plant resistance of aphids in alfalfa, Medicago sativa.

    Science.gov (United States)

    Ramirez, Ricardo A; Spears, Lori R

    2014-10-01

    Plants are exploited by a diverse community of insect herbivores and phytopathogens that interact indirectly through plant-mediated interactions. Generally, plants are thought to respond to insects and pathogens through different defensive signaling pathways. As plants are selected for resistance to one phytophagous organism type (insect vs. pathogen) in managed systems, it is not clear how this selection may affect community interactions. This study examined the effect of nematode-resistant varieties on aphid (Acyrthosiphon pisum) suppression, and then determined how infection by the stem nematode, Ditylenchus dipsaci, mediated ecological effects on aphids and on plant defense proteins. Four alfalfa (Medicago sativa) varieties were selected with resistance to nematodes only (+,-), aphids only (-,+), nematodes and aphids (+,+), and susceptibility to nematodes and aphids (-,-). Field and greenhouse experiments were conducted to isolate the effect of nematode infection and aphid abundance on each variety. We found that varieties resistant to nematode, regardless of aphid resistance, had the lowest aphid counts, suggesting possible cross-resistance. Aphid abundance, however, increased when plants were exposed to nematodes. Resistant varieties were associated with elevated saponins but these compounds were not affected by insect or pathogen feeding. Concentrations of peroxidases and trypsin inhibitors, however, were increased in nematode resistant varieties when exposed to nematodes and aphids, respectively. The patterns of plant defense were variable, and a combination of resistance traits and changes in nutrient availability may drive positive interactions between nematodes and aphids aboveground.

  19. Stoichiometric characteristics of carbon, nitrogen and phosphorus in leaves of differently aged lucerne (Medicago sativa stands

    Directory of Open Access Journals (Sweden)

    Zhennan eWang

    2015-12-01

    Full Text Available Element concentration within a plant which is vital to function maintenance and adaptation to environment, may change with plant growth. However, how carbon (C, nitrogen (N and phosphorus (P vary stoichiometrically with stand growth, i.e. ages or cuts, was still untouched in perennial species. This study tested the hypothesis that lucerne (Medicago sativa C:N, C:P and N:P should change with stand age and cut. Leaf C:N, C:P and N:P changed with stand age, showing various trends in different cuts of lucerne. Generally the greatest stoichiometric ratios were measured in 8 year stand and in the 2nd cut. They were affected significantly and negatively by total N and P concentrations of leaf, but not by organic C concentration. There were significantly positive correlations among leaf C:N, C:P and N:P. However, leaf C:N, C:P and N:P were hardly affected by soil features. Conclusively, lucerne C, N and P stoichiometry are age- and cut-specific, and regulated mainly by leaf N, P concentrations and stoichiometry. There are few correlations with soil fertility. To our knowledge, it is the first try to elucidate the stoichiometry in the viewpoint of age and cut with a perennial herbaceous legume.

  20. Allelopathic interference of alfalfa (Medicago sativa L.) genotypes to annual ryegrass (Lolium rigidum).

    Science.gov (United States)

    Zubair, Hasan Muhammad; Pratley, James E; Sandral, G A; Humphries, A

    2017-07-01

    Alfalfa (Medicago sativa L.) genotypes at varying densities were investigated for allelopathic impact using annual ryegrass (Lolium rigidum) as the target species in a laboratory bioassay. Three densities (15, 30, and 50 seedlings/beaker) and 40 alfalfa genotypes were evaluated by the equal compartment agar method (ECAM). Alfalfa genotypes displayed a range of allelopathic interference in ryegrass seedlings, reducing root length from 5 to 65%. The growth of ryegrass decreased in response to increasing density of alfalfa seedlings. At the lowest density, Q75 and Titan9 were the least allelopathic genotypes. An overall inhibition index was calculated to rank each alfalfa genotype. Reduction in seed germination of annual ryegrass occurred in the presence of several alfalfa genotypes including Force 10, Haymaster7 and SARDI Five. A comprehensive metabolomic analysis using Quadruple Time of Flight (Q-TOF), was conducted to compare six alfalfa genotypes. Variation in chemical compounds was found between alfalfa root extracts and exudates and also between genotypes. Further individual compound assessments and quantitative study at greater chemical concentrations are needed to clarify the allelopathic activity. Considerable genetic variation exists among alfalfa genotypes for allelopathic activity creating the opportunity for its use in weed suppression through selection.

  1. Exploring the plant-associated bacterial communities in Medicago sativa L.

    Science.gov (United States)

    Pini, Francesco; Frascella, Arcangela; Santopolo, Luisa; Bazzicalupo, Marco; Biondi, Emanuele G; Scotti, Carla; Mengoni, Alessio

    2012-05-20

    Plant-associated bacterial communities caught the attention of several investigators which study the relationships between plants and soil and the potential application of selected bacterial species in crop improvement and protection. Medicago sativa L. is a legume crop of high economic importance as forage in temperate areas and one of the most popular model plants for investigations on the symbiosis with nitrogen fixing rhizobia (mainly belonging to the alphaproteobacterial species Sinorhizobium meliloti). However, despite its importance, no studies have been carried out looking at the total bacterial community associated with the plant. In this work we explored for the first time the total bacterial community associated with M. sativa plants grown in mesocosms conditions, looking at a wide taxonomic spectrum, from the class to the single species (S. meliloti) level. Results, obtained by using Terminal-Restriction Fragment Length Polymorphism (T-RFLP) analysis, quantitative PCR and sequencing of 16 S rRNA gene libraries, showed a high taxonomic diversity as well as a dominance by members of the class Alphaproteobacteria in plant tissues. Within Alphaproteobacteria the families Sphingomonadaceae and Methylobacteriaceae were abundant inside plant tissues, while soil Alphaproteobacteria were represented by the families of Hyphomicrobiaceae, Methylocystaceae, Bradyirhizobiaceae and Caulobacteraceae. At the single species level, we were able to detect the presence of S. meliloti populations in aerial tissues, nodules and soil. An analysis of population diversity on nodules and soil showed a relatively low sharing of haplotypes (30-40%) between the two environments and between replicate mesocosms, suggesting drift as main force shaping S. meliloti population at least in this system. In this work we shed some light on the bacterial communities associated with M. sativa plants, showing that Alphaproteobacteria may constitute an important part of biodiversity in this

  2. NIVELES DE ABSORCIÓN DEL ZINC PROVENIENTE DE CALAMINAS DE VIVIENDAS POR MEDICAGO SATIVA Y SU METABOLISMO EN CAVIA PORCELLUS

    OpenAIRE

    MALCA ACUÑA, LEOCADIO

    2009-01-01

    The levels of absorption of zinc had been determined coming from leaf blades of the houses` roof by Medicago sativa; and its metabolism in Cavia porcellus, are presented. Leaf blades were made with an alloy of zinc. The content of zinc in the agricultural soil, and in Medicago sativa; irrigated with the rainfall collected from the houses` roofs, from Tumbes city –Perú. Two places of Tumbes were selected for the collection of the rainfall water. The pH, temperature, electrical conductivi...

  3. The succinoglycan endoglycanase encoded by exoK is required for efficient symbiosis of Sinorhizobium meliloti 1021 with the host plants Medicago truncatula and Medicago sativa (Alfalfa).

    Science.gov (United States)

    Mendis, Hajeewaka C; Queiroux, Clothilde; Brewer, Tess E; Davis, Olivia M; Washburn, Brian K; Jones, Kathryn M

    2013-09-01

    The acidic polysaccharide succinoglycan produced by the nitrogen-fixing rhizobial symbiont Sinorhizobium meliloti 1021 is required for this bacterium to invade the host plant Medicago truncatula and to efficiently invade the host plant M. sativa (alfalfa). The β-glucanase enzyme encoded by exoK has previously been demonstrated to cleave succinoglycan and participate in producing the low molecular weight form of this polysaccharide. Here, we show that exoK is required for efficient S. meliloti invasion of both M. truncatula and alfalfa. Deletion mutants of exoK have a substantial reduction in symbiotic productivity on both of these plant hosts. Insertion mutants of exoK have an even less productive symbiosis than the deletion mutants with the host M. truncatula that is caused by a secondary effect of the insertion itself, and may be due to a polar effect on the expression of the downstream exoLAMON genes.

  4. Complete nucleotide sequence of Alfalfa mosaic virus isolated from alfalfa (Medicago sativa L.) in Argentina.

    Science.gov (United States)

    Trucco, Verónica; de Breuil, Soledad; Bejerman, Nicolás; Lenardon, Sergio; Giolitti, Fabián

    2014-06-01

    The complete nucleotide sequence of an Alfalfa mosaic virus (AMV) isolate infecting alfalfa (Medicago sativa L.) in Argentina, AMV-Arg, was determined. The virus genome has the typical organization described for AMV, and comprises 3,643, 2,593, and 2,038 nucleotides for RNA1, 2 and 3, respectively. The whole genome sequence and each encoding region were compared with those of other four isolates that have been completely sequenced from China, Italy, Spain and USA. The nucleotide identity percentages ranged from 95.9 to 99.1 % for the three RNAs and from 93.7 to 99 % for the protein 1 (P1), protein 2 (P2), movement protein and coat protein (CP) encoding regions, whereas the amino acid identity percentages of these proteins ranged from 93.4 to 99.5 %, the lowest value corresponding to P2. CP sequences of AMV-Arg were compared with those of other 25 available isolates, and the phylogenetic analysis based on the CP gene was carried out. The highest percentage of nucleotide sequence identity of the CP gene was 98.3 % with a Chinese isolate and 98.6 % at the amino acid level with four isolates, two from Italy, one from Brazil and the remaining one from China. The phylogenetic analysis showed that AMV-Arg is closely related to subgroup I of AMV isolates. To our knowledge, this is the first report of a complete nucleotide sequence of AMV from South America and the first worldwide report of complete nucleotide sequence of AMV isolated from alfalfa as natural host.

  5. Nonphotosynthetic CO/sub 2/ fixation by alfalfa (Medicago sativa L. ) roots and nodules

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, M.P.; Heichel, G.H.; Vance, C.P.

    1987-09-01

    The dependence of alfalfa (Medicago sativa L.) root and nodule nonphotosynthetic CO/sub 2/ fixation on the supply of currently produced photosynthate and nodule nitrogenase activity was examined a various times after phloem-girdling and exposure of nodules to Ar:O/sub 2/. Phloem-girdling was effected 20 hours and exposure to Ar:O/sub 2/ was effected 2 to 3 hours before initiation of experiments. Nodule and root CO/sub 2/ fixation rates of phloem-girdled plants were reduced to 38 and 50%, respectively, of those of control plants. Exposure to Ar:O/sub 2/ decreased nodule CO/sub 2/ fixation rates to 45%, respiration rates to 55%, and nitrogenase activities to 51% of those of the controls. The products of nodule CO/sub 2/ fixation were exported through the xylem to the shoot mainly as amino acids within 30 to 60 minutes after exposure to /sup 14/CO/sub 2/. In contrast to nodules, roots exported very little radioactivity, and most of the /sup 14/C was exported as organic acids. The nonphotosynthetic CO/sub 2/ fixation rate of roots and nodules averaged 26% of the gross respiration rate, i.e. the sum of net respiration and nonphotosynthetic CO/sub 2/ assimilation. Nodules fixed CO/sub 2/ at a rate 5.6 times that of roots, but since nodules comprised a small portion of root system mass, roots accounted for 76% of the nodulated roots system CO/sub 2/ fixation. The results indicate that nodule CO/sub 2/ fixation in alfalfa is associated with N assimilation.

  6. Superoxide dismutase enhances tolerance of freezing stress in transgenic alfalfa (Medicago sativa L.).

    Science.gov (United States)

    McKersie, B D; Chen, Y; de Beus, M; Bowley, S R; Bowler, C; Inzé, D; D'Halluin, K; Botterman, J

    1993-01-01

    Activated oxygen or oxygen free radicals have been implicated in a number of physiological disorders in plants including freezing injury. Superoxide dismutase (SOD) catalyzes the dismutation of superoxide into O2 and H2O2 and thereby reduces the titer of activated oxygen molecules in the cell. To further examine the relationship between oxidative and freezing stresses, the expression of SOD was modified in transgenic alfalfa (Medicago sativa L.). The Mn-SOD cDNA from Nicotiana plumbaginifolia under the control of the cauliflower mosaic virus 35S promoter was introduced into alfalfa using Agrobacterium tumefaciens-mediated transformation. Two plasmid vectors, pMitSOD and pChlSOD, contained a chimeric Mn-SOD construct with a transit peptide for targeting to the mitochondria or one for targeting to the chloroplast, respectively. The putatively transgenic plants were selected for resistance to kanamycin and screened for neomycin phosphotransferase activity and the presence of an additional Mn-SOD isozyme. Detailed analysis of a set of four selected transformants indicated that some had enhanced SOD activity, increased tolerance to the diphenyl ether herbicide, acifluorfen, and increased regrowth after freezing stress. The F1 progeny of one line, RA3-ChlSOD-30, were analyzed by SOD isozyme activity, by polymerase chain reaction for the Mn-SOD gene, and by polymerase chain reaction for the neo gene. RA3-ChlSOD-30 had three sites of insertion of pChlSOD, but only one gave a functional Mn-SOD isozyme; the other two were apparently partial insertions. The progeny with a functional Mn-SOD transgene had more rapid regrowth following freezing stress than those progeny lacking the functional Mn-SOD transgene, suggesting that Mn-SOD serves a protective role by minimizing oxygen free radical production after freezing stress. PMID:8290627

  7. Root Nodule Enzymes of Ammonia Assimilation in Alfalfa (Medicago sativa L.) 1

    Science.gov (United States)

    Groat, R. Gene; Vance, Carroll P.

    1981-01-01

    Nitrogenase-dependent acetylene reduction activity of glasshouse-grown alfalfa (Medicago sativa L.) decreased rapidly in response both to harvesting (80% shoot removal) and applied NO3− at 40 and 80 kilograms N per hectare. Acetylene reduction activity of harvested plants grown on 0 kilogram N per hectare began to recover by day 15 as shoot regrowth became significant. In contrast, acetylene reduction activity of all plants treated with 80 kilograms NO3−-N per hectare and harvested plants treated with 40 kilograms NO3−-N per hectare remained low for the duration of the experiment. Acetylene reduction of unharvested alfalfa treated with 40 kilograms N per hectare declined to an intermediate level and appeared to recover slightly by day 15. Changes in N2-fixing capacity were accompanied by similar changes in levels of nodule soluble protein. After an initial lag of 24 hours, specific activities of alfalfa nodule glutamine synthetase, NADH-glutamate synthase, and NAD-glutamate dehydrogenase (oxidative amination) decreased similar to but less rapidly than acetylene reduction activity. Increased specific activities of these nodule enzymes occurred as acetylene reduction activity increased and shoot growth resumed. The observed rates of glutamine synthetase and glutamate synthase were sufficient to assimilate ammonia produced via symbiotic N2 fixation. Nodule NADH-dependent glutamate dehydrogenase (reductive amination) specific activity was not associated with changes in acetylene reduction activity. The data indicate that host plant glutamine synthetase and NADH-glutamate synthase function to assimilate symbiotically fixed N and that NADH-dependent glutamate dehydrogenase may function in ammonia assimilation during senescence in alfalfa nodules. PMID:16661836

  8. Proteomic Identification of Differentially Expressed Proteins during Alfalfa (Medicago sativa L.) Flower Development.

    Science.gov (United States)

    Chen, Lingling; Chen, Quanzhu; Zhu, Yanqiao; Hou, Longyu; Mao, Peisheng

    2016-01-01

    Flower development, pollination, and fertilization are important stages in the sexual reproduction process of plants; they are also critical steps in the control of seed formation and development. During alfalfa (Medicago sativa L.) seed production, some distinct phenomena such as a low seed setting ratio, serious flower falling, and seed abortion commonly occur. However, the causes of these phenomena are complicated and largely unknown. An understanding of the mechanisms that regulate alfalfa flowering is important in order to increase seed yield. Hence, proteomic technology was used to analyze changes in protein expression during the stages of alfalfa flower development. Flower samples were collected at pre-pollination (S1), pollination (S2), and the post-pollination senescence period (S3). Twenty-four differentially expressed proteins were successfully identified, including 17 down-regulated in pollinated flowers, one up-regulated in pollinated and senesced flowers, and six up-regulated in senesced flowers. The largest proportions of the identified proteins were involved in metabolism, signal transduction, defense response, oxidation reduction, cell death, and programmed cell death (PCD). Their expression profiles demonstrated that energy metabolism, carbohydrate metabolism, and amino acid metabolism provided the nutrient foundation for pollination in alfalfa. Furthermore, there were three proteins involved in multiple metabolic pathways: dual specificity kinase splA-like protein (kinase splALs), carbonic anhydrase, and NADPH: quinone oxidoreductase-like protein. Expression patterns of these proteins indicated that MAPK cascades regulated multiple processes, such as signal transduction, stress response, and cell death. PCD also played an important role in the alfalfa flower developmental process, and regulated both pollination and flower senescence. The current study sheds some light on protein expression profiles during alfalfa flower development and

  9. Screening of Cd tolerant genotypes and isolation of metallothionein genes in alfalfa (Medicago sativa L.)

    Energy Technology Data Exchange (ETDEWEB)

    Wang Xiaojuan, E-mail: xiaojuanwang@lzu.edu.cn [School of Pastoral Agriculture Science and Technology, Lanzhou University, P.O. Box 61, Lanzhou 730020 (China); Song, Yu [School of Pastoral Agriculture Science and Technology, Lanzhou University, P.O. Box 61, Lanzhou 730020 (China); Environment Management College of China, Qinhuangdao 066004 (China); Ma Yanhua [Hebei Normal University of Science and Technology, Qinhuangdao 066004 (China); Zhuo Renying [Key Lab of Tree Genomics, Research Institute of Subtropical of Forest, Chinese Academy of Forest, Fuyang 311400 (China); Jin Liang [School of Pastoral Agriculture Science and Technology, Lanzhou University, P.O. Box 61, Lanzhou 730020 (China)

    2011-12-15

    In order to evaluate Cd tolerance in wide-ranging sources of alfalfa (Medicago sativa) and to identify Cd tolerant genotypes which may potentially be useful for restoring Cd-contaminated environments, thirty-six accessions of alfalfa were screened under hydroponic culture. Our results showed that the relative root growth rate varied from 0.48 to 1.0, which indicated that different alfalfa accessions had various responses to Cd stress. The candidate fragments derived from differentially expressed metallothionein (MT) genes were cloned from leaves of two Cd tolerant genotypes, YE and LZ. DNA sequence and the deduced protein sequence showed that MsMT2a and MsMT2b had high similarity to those in leguminous plants. DDRT-PCR analysis showed that MsMT2a expressed in both YE and LZ plants under control and Cd stress treatment, but MsMT2b only expressed under Cd stress treatment. This suggested that MsMT2a was universally expressed in leaves of alfalfa but expression of MsMT2b was Cadmium (Cd) inducible. - Highlights: > Evaluate Cd tolerance in wide sources of alfalfa accessions. > Identify Cd-hyperaccumulators potentially useful for restoring Cd-contaminated environments. > Cloned differentially expressed metallothionein (MT) genes. > Characteristics and deduced protein sequence of MsMT2a and MsMT2b were analyzed. > MsMT2a might be a universally gene of alfalfa but MsMT2b might be an inductive gene. - Two Cd tolerant alfalfa genotypes were screened and their metallothionein genes were cloned which showed that MsMT2a was universally expressed but MsMT2b was Cd inducible expression.

  10. Carbohydrate Metabolism in Taproots of Medicago sativa L. during Winter Adaptation and Spring Regrowth 1

    Science.gov (United States)

    Volenec, Jeffrey J.; Boyce, Patricia J.; Hendershot, Kerry L.

    1991-01-01

    Our objective was to identify amylases that may participate in starch degradation in alfalfa (Medicago sativa L.) taproots during winter hardening and subsequent spring regrowth. Taproots from field-grown plants were sampled at intervals throughout fall, winter, and early spring. In experiment 1, taproots were separated into bark and wood tissues. Concentrations of soluble sugars, starch, and buffer-soluble proteins and activities of endo- and exoamylase were determined. Starch concentrations declined in late fall, whereas concentrations of sucrose increased. Total amylolytic activity (primarily exoamylase) was not consistently associated with starch degradation but followed trends in soluble protein concentration of taproots. This was especially evident in spring when both declined as starch degradation increased and shoot growth resumed. Activity of endoamylase increased during periods of starch degradation, especially in bark tissues. In experiment 2, a low starch line had higher specific activity of taproot amylases. This line depleted its taproot starch by late winter, after which taproot sugar concentrations declined. As in experiment 1, total amylolytic activity declined in spring in both lines, whereas that of endoamylase increased in both lines even though little starch remained in taproots of the low starch line. Several isoforms of both amylases were distinguished using native polyacrylamide electrophoresis, with isoforms being similar in bark and wood tissues. The slowest migrating isoform of endoamylase was most prominent at each sampling. Activity of all endoamylase isoforms increased during winter adaptation and in spring when shoot growth resumed. Endoamylase activity consistently increased at times of starch utilization in alfalfa taproots (hardening, spring regrowth, after defoliation), indicating that it may serve an important role in starch degradation. ImagesFigure 3Figure 6Figure 7 PMID:16668255

  11. Isolation and Immunochemical Characterization of Plant Glutamine Synthetase in Alfalfa (Medicago sativa L.) Nodules 1

    Science.gov (United States)

    Groat, R. Gene; Schrader, Larry E.

    1982-01-01

    Host plant glutamine synthetase (GS) has been purified 100-fold from N2-fixing alfalfa (Medicago sativa L.) nodules by a new procedure involving preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as a final step. An SDS-polypeptide fraction corresponding to plant GS was identified and consisted of two major polypeptides of 40,000 to 45,000 molecular weight. Antibodies to the SDS-polypeptide fraction were raised in mice by intraperitoneal injection, and antisera were collected as ascitic fluid. Crude extracts of soluble protein from the plant fraction of nodules were resolved by SDS-PAGE and then subjected to electrophoresis in the second dimension into antibody-containing agarose gel. A single immunochemically active protein species was observed using this crossed immunoelectrophoresis method, even though both major GS SDS-polypeptides were apparently resolved in the first (SDS-PAGE) dimension. Plant GS protein in crude nodule extracts was quantitated immunochemically by comparison with immunoprecipitin arcs of similarly treated amounts of pure antigen. Using this technique, it was determined that plant GS was present at 150 micrograms per gram fresh weight or 1.2% of total plant soluble protein in N2-fixing alfalfa nodules. Results suggest that alfalfa nodule plant GS consists of two major subunit polypeptides, but only a single immunochemically active native protein was observed. The crossed immunoelectrophoresis procedure described here should be generally applicable for immunochemical detection of lower abundance components of crude plant extracts. Images Fig. 2 Fig. 3 Fig. 4 PMID:16662757

  12. Composition and Structure of Starch from Taproots of Contrasting Genotypes of Medicago sativa L. 1

    Science.gov (United States)

    Fankhauser, Jerry J.; Volenec, Jeffrey J.; Brown, Geoffrey A.

    1989-01-01

    The objective of this study was to examine the composition and branch chain lengths of alfalfa (Medicago sativa L.) taproot starch during starch utilization and reaccumulation in response to defoliation. Genotypes were propagated vegetatively and well-established plants were sampled at defoliation and at weekly intervals thereafter. Starch granules from root tissues were dispersed in dimethyl sulfoxide and starch components separated using gel permeation chromatography. Root starches also were debranched enzymically, and branch chain lengths were examined. Results indicate that, irrespective of starch concentration, starch from taproots of the high starch genotype was composed of approximately 80% high molecular weight starch with I2-Kl absorbance characteristics similar to amylopectin. The remaining 20% of the starch was low molecular weight with I2-Kl absorbance characteristics similar to amylose. Starches of the low starch genotype contained approximately 85% high molecular weight polysaccharide at high root starch concentrations (>50 grams per kilogram). At low root starch concentrations (<10 grams per kilogram), starch from the low starch genotype had nearly equal proportions of low and high molecular weight polysaccharide. The I2-Kl absorbance properties of the low molecular weight starches from roots of the low starch genotype indicated that some branching may be present. The distribution of chain lengths from amylopectin did not change during starch degradation and reaccumulation for the high starch genotype. In the low starch genotype, the proportion of low molecular weight branches having a degree of polymerization between 1 and 30 was decreased at the very low starch concentrations observed on the 14th day of regrowth. Higher concentrations and/or quantities of starch in roots of the high starch genotype were not associated with greater rate of herbage regrowth, when compared to the low starch genotype. PMID:16666871

  13. Nitrogen Reserve Mobilization during Regrowth of Medicago sativa L. (Relationships between Availability and Regrowth Yield).

    Science.gov (United States)

    Ourry, A.; Kim, T. H.; Boucaud, J.

    1994-01-01

    An experiment was designed to study the role of N and C reserves on regrowth of the shoots following defoliation of forage species. Starch and N accumulation in root and crown tissue of nonnodulated Medicago sativa L. were modified during regrowth by applying different levels of N and different cutting heights. Plants were obtained with similar crown and root dry weights, but having either low starch and high tissue N or high starch and low tissue N. The plants were then submitted to a second defoliation and supplied with optimal N nutrition, and N flow from reserve was quantified using pulse-chase 15N labeling. Maximum yields following the second regrowth were obtained from those plants having a high tissue N, despite their low level of nonstructural carbohydrate. When N in the roots and crown exceeded 5 mg N plant-1 at the beginning of regrowth, about 68% was translocated to regrowing shoots. Highly significant correlations were also found between the amounts of N available in roots and crown at the beginning of regrowth and (a) the amount of N that was mobilized to new tissues, (b) the amount of N taken up during the regrowth period, and (c) the final shoot yield after 24 d of regrowth. No similar correlations were found for plants that varied in their initial starch content of roots and crown. It is suggested that N reserves were used mainly during the first 10 d after defoliation, and that the resulting aerial growth during this period should be sufficient to restore N2 fixation and/or N uptake to levels equal to those prior to defoliation. These data emphasize (a) the importance of root N reserves in initiating and sustaining new shoot growth, and (b) the need for a re-evaluation of the contribution of C reserves to shoot regrowth. PMID:12232246

  14. Cohnella formosensis sp. nov., a xylanolytic bacterium isolated from the rhizosphere of Medicago sativa L.

    Science.gov (United States)

    Hameed, Asif; Hung, Mei-Hua; Lin, Shih-Yao; Hsu, Yi-Han; Liu, You-Cheng; Shahina, Mariyam; Lai, Wei-An; Huang, Hsin-Chieh; Young, Li-Sen; Young, Chiu-Chung

    2013-08-01

    A Gram-positive, spore-forming, aerobic, rod-shaped, xylanolytic bacterium designated strain CC-Alfalfa-35(T) was isolated from the rhizosphere of Medicago sativa L. in Taiwan. Phylogenetic analysis based on 16S rRNA gene sequence showed that strain CC-Alfalfa-35(T) was affiliated to the genus Cohnella. Strain CC-Alfalfa-35(T) shared 95.3 % pairwise 16S rRNA gene sequence similarity to the type strain of the type species of the genus Cohnella (Cohnella thermotolerans DSM 17683(T)) besides showing a similarity of 97.4-93.6 % with other recognized species of the genus Cohnella. The DNA-DNA hybridization value between CC-Alfalfa-35(T) and Cohnella thailandensis KCTC 22296(T) was 37.7 % ± 1.7 % (reciprocal value, 55.7 % ± 3.0 %). Predominant cellular fatty acids were iso-C16 : 0 and anteiso-C15 : 0. The polar lipid profile constituted diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, lysyl-phosphatidylglycerol, three unidentified phospholipids and three unidentified aminophospholipids. The major respiratory quinone was MK-7 and the DNA G+C content was 58.3 mol%. Strain CC-Alfalfa-35(T) contained meso-diaminopimelic acid as the major diamino acid in the cell-wall peptidoglycan. Based on the polar lipid and fatty acid profiles, which were in line with those of C. thermotolerans DSM 17683(T), coupled with additional distinguishing genotypic, phenotypic and chemotaxonomic features, strain CC-Alfalfa-35(T) is proposed to represent a novel species within the genus Cohnella, for which the name Cohnella formosensis sp. nov. is proposed. The type strain is CC-Alfalfa-35(T) ( = JCM 18405(T) = BCRC 80428(T)).

  15. Speciation Matters: Bioavailability of Silver and Silver Sulfide Nanoparticles to Alfalfa (Medicago sativa).

    Science.gov (United States)

    Stegemeier, John P; Schwab, Fabienne; Colman, Benjamin P; Webb, Samuel M; Newville, Matthew; Lanzirotti, Antonio; Winkler, Christopher; Wiesner, Mark R; Lowry, Gregory V

    2015-07-21

    Terrestrial crops are directly exposed to silver nanoparticles (Ag-NPs) and their environmentally transformed analog silver sulfide nanoparticles (Ag2S-NPs) when wastewater treatment biosolids are applied as fertilizer to agricultural soils. This leads to a need to understand their bioavailability to plants. In the present study, the mechanisms of uptake and distribution of silver in alfalfa (Medicago sativa) were quantified and visualized upon hydroponic exposure to Ag-NPs, Ag2S-NPs, and AgNO3 at 3 mg total Ag/L. Total silver uptake was measured in dried roots and shoots, and the spatial distribution of elements was investigated using transmission electron microscopy (TEM) and synchrotron-based X-ray imaging techniques. Despite large differences in release of Ag(+) ions from the particles, Ag-NPs, Ag2S-NPs, and Ag(+) became associated with plant roots to a similar degree, and exhibited similarly limited (<1%) amounts of translocation of silver into the shoot system. X-ray fluorescence (XRF) mapping revealed differences in the distribution of Ag into roots for each treatment. Silver nanoparticles mainly accumulated in the (columella) border cells and elongation zone, whereas Ag(+) accumulated more uniformly throughout the root. In contrast, Ag2S-NPs remained largely adhered to the root exterior, and the presence of cytoplasmic nano-SixOy aggregates was observed. Exclusively in roots exposed to particulate silver, NPs smaller than the originally dosed NPs were identified by TEM in the cell walls. The apparent accumulation of Ag in the root apoplast determined by XRF, and the presence of small NPs in root cell walls suggests uptake of partially dissolved NPs and translocation along the apoplast.

  16. Proteomic identification of differentially expressed proteins during alfalfa (Medicago sativa L. flower development

    Directory of Open Access Journals (Sweden)

    Lingling Chen

    2016-10-01

    Full Text Available Flower development, pollination, and fertilization are important stages in the sexual reproduction process of plants; they are also critical steps in the control of seed formation and development. During alfalfa (Medicago sativa L. seed production, some distinct phenomena such as a low seed setting ratio, serious flower falling, and seed abortion commonly occur. However, the causes of these phenomena are complicated and largely unknown. An understanding of the mechanisms that regulate alfalfa flowering is important in order to increase seed yield. Hence, proteomic technology was used to analyze changes in protein expression during the stages of alfalfa flower development. Flower samples were collected at pre-pollination (S1, pollination (S2, and the post-pollination senescence period (S3. Twenty-four differentially expressed proteins were successfully identified, including 17 down-regulated in pollinated flowers, one up-regulated in pollinated and senesced flowers, and six up-regulated in senesced flowers. The largest proportions of the identified proteins were involved in metabolism, signal transduction, defense response, oxidation reduction, cell death, and programmed cell death (PCD. Their expression profiles demonstrated that energy metabolism, carbohydrate metabolism, and amino acid metabolism provided the nutrient foundation for pollination in alfalfa. Furthermore, there were three proteins involved in multiple metabolic pathways: dual specificity kinase splA-like protein (kinase splALs, carbonic anhydrase (CA, and NADPH: quinone oxidoreductase-like protein (NQOLs. Expression patterns of these proteins indicated that MAPK cascades regulated multiple processes, such as signal transduction, stress response, and cell death. PCD also played an important role in the alfalfa flower developmental process, and regulated both pollination and flower senescence. The current study sheds some light on protein expression profiles during alfalfa flower

  17. Weed control in dormant alfalfa (Medicago sativa L. with active ingredients’ metribuzin, imazetapyr and pronamide

    Directory of Open Access Journals (Sweden)

    Zvonko Pacanoski

    2017-03-01

    Full Text Available Field trials were conducted during 2008 – 2010 to evaluate weed control in dormant alfalfa (Medicago sativa L. with metribuzin, imazetapyr and pronamide. The weed population in all experimental years was consisted mainly of annual winter and spring grass and broadleaf weeds, and some perennial weeds. The number of weed species and weed density increased with the years of alfalfa growing, from second to the fourth year. Weed density in the untreated control plots was 201.0, 217.2 and 240.5 plants per m2 in 2008, 2009 and 2010, respectively. The most dominant weeds were Anthemis cotula, Capsell bursa-pastoris and Taraxacum officinale in 2008, Alopecurus myosuroides and Poa pratensis in 2009 and Millium vernale and Arabidopsis thaliana in 2010. Efficacy of herbicides in control of weeds was ranged of 91.8% (pronamide to 98.4% (metribuzin 1.0 kg*ha-1 in 2008, 93.1% (imazetapyr to 97.3% (metribuzin 1.0 kg*ha-1 in 2009 and 92.1% (imazetapyr to 97.3% (metribuzin 1.0 kg*ha-1 in 2010, respectively. Efficacy of herbicides in control of prevailing weeds during the 3 years field trial period was ranged of 48.5% to 100.0%. No visual alfalfa injured was determined by any rates during the experimental period, and consequently, none of the applied herbicides reduced first-harvest alfalfa yields. Alfalfa yield was markedly affected by herbicide efficacy in all experimental years, particularly in the second year, where yields of herbicide treatments were similar to that of the weed free control.

  18. The coupling of the plant and microbial catabolisms of phenanthrene in the rhizosphere of Medicago sativa.

    Science.gov (United States)

    Muratova, Anna; Dubrovskaya, Ekaterina; Golubev, Sergey; Grinev, Vyacheslav; Chernyshova, Marina; Turkovskaya, Olga

    2015-09-01

    We studied the catabolism of the polycyclic aromatic hydrocarbon phenanthrene by four rhizobacterial strains and the possibility of enzymatic oxidation of this compound and its microbial metabolites by the root exudates of alfalfa (Medicago sativa L.) in order to detect the possible coupling of the plant and microbial metabolisms under the rhizospheric degradation of the organic pollutant. A comparative study of phenanthrene degradation pathways in the PAH-degrading rhizobacteria Ensifer meliloti, Pseudomonas kunmingensis, Rhizobium petrolearium, and Stenotrophomonas sp. allowed us to identify the key metabolites from the microbial transformation of phenanthrene, including 9,10-phenanthrenequinone, 2-carboxybenzaldehyde, and 1-hydroxy-2-naphthoic, salicylic, and o-phthalic acids. Sterile alfalfa plants were grown in the presence and absence of phenanthrene (0.03 g kg(-1)) in quartz sand under controlled environmental conditions to obtain plant root exudates. The root exudates were collected, concentrated by ultrafiltration, and the activity of oxidoreductases was detected spectrophotometrically by the oxidation rate for various substrates. The most marked activity was that of peroxidase, whereas the presence of oxidase and tyrosinase was detected on the verge of the assay sensitivity. Using alfalfa root exudates as a crude enzyme preparation, we found that in the presence of the synthetic mediator, the plant peroxidase could oxidize phenanthrene and its microbial metabolites. The results indicate the possibility of active participation of plants in the rhizospheric degradation of polycyclic aromatic hydrocarbons and their microbial metabolites, which makes it possible to speak about the coupling of the plant and microbial catabolisms of these contaminants in the rhizosphere. Copyright © 2015 Elsevier GmbH. All rights reserved.

  19. Genome-Wide Association Mapping and Genomic Selection for Alfalfa (Medicago sativa) Forage Quality Traits.

    Science.gov (United States)

    Biazzi, Elisa; Nazzicari, Nelson; Pecetti, Luciano; Brummer, E Charles; Palmonari, Alberto; Tava, Aldo; Annicchiarico, Paolo

    2017-01-01

    Genetic progress for forage quality has been poor in alfalfa (Medicago sativa L.), the most-grown forage legume worldwide. This study aimed at exploring opportunities for marker-assisted selection (MAS) and genomic selection of forage quality traits based on breeding values of parent plants. Some 154 genotypes from a broadly-based reference population were genotyped by genotyping-by-sequencing (GBS), and phenotyped for leaf-to-stem ratio, leaf and stem contents of protein, neutral detergent fiber (NDF) and acid detergent lignin (ADL), and leaf and stem NDF digestibility after 24 hours (NDFD), of their dense-planted half-sib progenies in three growing conditions (summer harvest, full irrigation; summer harvest, suspended irrigation; autumn harvest). Trait-marker analyses were performed on progeny values averaged over conditions, owing to modest germplasm × condition interaction. Genomic selection exploited 11,450 polymorphic SNP markers, whereas a subset of 8,494 M. truncatula-aligned markers were used for a genome-wide association study (GWAS). GWAS confirmed the polygenic control of quality traits and, in agreement with phenotypic correlations, indicated substantially different genetic control of a given trait in stems and leaves. It detected several SNPs in different annotated genes that were highly linked to stem protein content. Also, it identified a small genomic region on chromosome 8 with high concentration of annotated genes associated with leaf ADL, including one gene probably involved in the lignin pathway. Three genomic selection models, i.e., Ridge-regression BLUP, Bayes B and Bayesian Lasso, displayed similar prediction accuracy, whereas SVR-lin was less accurate. Accuracy values were moderate (0.3-0.4) for stem NDFD and leaf protein content, modest for leaf ADL and NDFD, and low to very low for the other traits. Along with previous results for the same germplasm set, this study indicates that GBS data can be exploited to improve both quality traits

  20. Influence of pCO2 on carbon allocation in nodulated Medicago sativa L.

    Science.gov (United States)

    Pereyra, Gabriela; Hartmann, Henrik; Ziegler, Waldemar; Michalzik, Beate; Gonzalez-Meler, Miquel; Trumbore, Susan

    2016-04-01

    Atmospheric CO2 concentrations (pCO_2) have been related to changes in plant carbon (C) availability and photosynthetic capacity, yet there is no clear consensus as to the effect of pCO2 on the plant C balance and on nitrogen fixation in symbiotic systems. We investigated how different pCO2 (Pleistocene: 170 ppm, ambient: 400 ppm and projected future: 700 ppm) influence C allocation in nodulated Medicago sativa L. We labeled 17 week old plants with depleted 13C (-34.7±1.2‰) and traced the label over a 9-day period, to assess the redistribution of newly assimilated C across different sinks, including nodules. We analyzed N concentrations in plant tissues and found no significant differences in leaves and roots across treatments. However, growth and C fixation rates increased with pCO_2, and differences were greatest between 170 ppm and 700 ppm. Across pCO2 treatments we observed a 13C-enrichment in roots compared to leaves. We further observed the highest 13C depletion of non-structural carbohydrates (NSCs) and respired CO2 in tissues of plants grown at 700 ppm, especially in leaves and nodules. Our preliminary results suggest that sink organs like roots and nodules are fed with newly-assimilated NSCs from leaves to support respiration, and especially in 170 ppm plants represented a major respiratory loss of newly assimilated C (≈ 35{%} of the total plant respiration). Our results suggest that although plant metabolic processes like photosynthesis and respiration are affected by changes in pCO_2, nitrogen acquisition in such a symbiotic system is not.

  1. Salicylic acid improves the salinity tolerance of Medicago sativa in symbiosis with Sinorhizobium meliloti by preventing nitrogen fixation inhibition.

    Science.gov (United States)

    Palma, F; López-Gómez, M; Tejera, N A; Lluch, C

    2013-07-01

    In this work we have investigated the contribution of pretreatment with 0.1 and 0.5mM salicylic acid (SA) to the protection against salt stress in root nodules of Medicago sativa in symbiosis with Sinorhizobium meliloti. SA alleviated the inhibition induced by salinity in the plant growth and photosynthetic capacity of M. sativa-S. meliloti symbiosis. In addition, SA prevented the inhibition of the nitrogen fixation capacity under salt stress since nodule biomass was not affected by salinity in SA pretreated plants. Antioxidant enzymes peroxidase (POX), superoxide dismutase (SOD), ascorbate peroxidase (APX), dehidroascorbate reductase (DHAR) and glutathione reductase (GR), key in the main pathway that scavenges H2O2 in plants, were induced by SA pretreatments which suggest that SA may participate in the redox balance in root nodules under salt stress. Catalase activity (CAT) was inhibited around 40% by SA which could be behind the increase of H2O2 detected in nodules of plants pretreated with SA. The accumulation of polyamines (PAs) synthesized in response to salinity was prevented by SA which together with the induction of 1-aminocyclopropane-l-carboxylic acid (ACC) content suggest the prevalence of the ethylene signaling pathway induced by SA in detriment of the synthesis of PAs. In conclusion, SA alleviated the negative effect of salt stress in the M. sativa-S. meliloti symbiosis through the increased level of nodule biomass and the induction of the nodular antioxidant metabolism under salt stress. The H2O2 accumulation and the PAs inhibition induced by SA in nodules of M. sativa suggest that SA activates a hypersensitive response dependent on ethylene. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  2. The antibacterial effects of aqueous extract of Medicago sativa L. in comparsion with three antibiotics (Gentamicin, Ciprofloxacin and Penicillin

    Directory of Open Access Journals (Sweden)

    maryam Rezaei

    2013-08-01

    Full Text Available Background: In recent years, it has been recommended to use natural materials, instead of chemically synthesized drugs with side effects, in the control and treatment of infections. Increasing use of medicinal plants for medical treatment made for this branch of complementary medicine a unique position. The present research was done to study anti-microbial effect of Medicago sativa exctract in comparison with Ciprofloxacin, Gentamicin and Penicillin antibiotics on clinical strains and phatogenic bacteria such as P. aeruginosa , E. coli, Staphylococcus aureus and Listeria monocytogenes. Materials and Methods: Experiments were conducted using micro broth dilution and disc diffusion methods, and MIC and MBC of the plant have been evaluatead. Results: MIC Medicago sativa extract was effective on gram-positive bacteria Staphylococcus aureus and Listeria monocytogenes on the 206.25 and 6600 µg/ml and on the gram-negative bacteria tested in this study had no effect. Conclusion: Based on the research findings, mentioned herbal extract can be a good candidates for laboratory studies to separate active compounds in this plant to achieve effective antimicrobial drugs.

  3. Analysis of Cell Wall-Related Genes in Organs of Medicago sativa L. under Different Abiotic Stresses.

    Science.gov (United States)

    Behr, Marc; Legay, Sylvain; Hausman, Jean-Francois; Guerriero, Gea

    2015-07-16

    Abiotic constraints are a source of concern in agriculture, because they can have a strong impact on plant growth and development, thereby affecting crop yield. The response of plants to abiotic constraints varies depending on the type of stress, on the species and on the organs. Although many studies have addressed different aspects of the plant response to abiotic stresses, only a handful has focused on the role of the cell wall. A targeted approach has been used here to study the expression of cell wall-related genes in different organs of alfalfa plants subjected for four days to three different abiotic stress treatments, namely salt, cold and heat stress. Genes involved in different steps of cell wall formation (cellulose biosynthesis, monolignol biosynthesis and polymerization) have been analyzed in different organs of Medicago sativa L. Prior to this analysis, an in silico classification of dirigent/dirigent-like proteins and class III peroxidases has been performed in Medicago truncatula and M. sativa. The final goal of this study is to infer and compare the expression patterns of cell wall-related genes in response to different abiotic stressors in the organs of an important legume crop.

  4. In vivo characterization of the effects of abscisic acid and drying protocols associated with the acquisition of desiccation tolerance in alfalfa (Medicago sativa L.) somatic embryos

    NARCIS (Netherlands)

    Sreedhar, L.; Wolkers, W.F.; Hoekstra, F.A.; Bewley, J.D.

    2002-01-01

    Although somatic embryos of alfalfa (Medicago sativa L.) had acquired some tolerance to desiccation at the cotyledonary stage of development (22 d after plating), additional culturing in 20 ?M abscisic acid (ABA) for 8 d induced greater desiccation tolerance, as determined by increased germination.

  5. How planting configuration influences plant secondary metabolites and total N in tall fescue (Festuca arundinacea Schreb.), alfalfa (Medicago sativa L.) and birdsfoot trefoil (Lotus corniculatus L.)

    Science.gov (United States)

    Theories suggest that incorporating alfalfa (Medicago sativa L.; Alf) or birdsfoot trefoil (Lotus corniculatus L.; BFT) into endophyte-infected tall fescue (Festuca arundinaceas Schreb.; E+TF) pasturelands may improve livestock production. We investigated how planting configuration might influence p...

  6. Starch Grain Distribution in Taproots of Defoliated Medicago sativa L. 1

    Science.gov (United States)

    Habben, Jeffrey E.; Volenec, Jeffrey J.

    1990-01-01

    Defoliation of alfalfa (Medicago sativa L.) results in a cyclic pattern of starch degradation followed by reaccumulation in taproots. Characterization of changes in anatomical distribution of starch grains in taproots will aid our understanding of biochemical and physiological mechanisms involved in starch metabolism in taproots of this species. Our objectives were to determine the influence of defoliation on starch grain distribution and size variation in taproots of two alfalfa lines selected for contrasting concentrations of taproot starch. In addition, we used electron microscopy to examine the cellular environment of starch grains, and computer-based image optical analysis to determine how cross-sectional area of tissues influenced starch accumulation. Taproots of field-grown plants were sampled at defoliation and weekly thereafter over a 28-day period. Taproot segments were fixed in glutaraldehyde and prepared for either light or electron microscopy. Transverse sections were examined for number and size of starch grains and tissue areas were measured. Starch grains were located throughout bark tissues, but were confined primarily to ray parenchyma cells in wood tissues. During the first week of foliar regrowth after defoliation, starch grains in ray cells near the cambium disappeared first, while degradation of those near the center of the taproot was delayed. During the third and fourth weeks of regrowth, there was a uniform increase in number of starch grains per cell profile across the rays, but by 28 days after defoliation there were more starch grains in ray cells near the cambium than in cells near the center of the taproot (low starch line only). Bark tissues from both lines showed synchronous degradation and synthesis of starch grains that was not influenced greatly by cell location. Diameter of starch grains varied with cell location in medullary rays during rapid starch degradation, but was not influenced by cell position in bark tissues. Therefore

  7. Genome-Wide Association Mapping and Genomic Selection for Alfalfa (Medicago sativa) Forage Quality Traits

    Science.gov (United States)

    Pecetti, Luciano; Brummer, E. Charles; Palmonari, Alberto; Tava, Aldo

    2017-01-01

    Genetic progress for forage quality has been poor in alfalfa (Medicago sativa L.), the most-grown forage legume worldwide. This study aimed at exploring opportunities for marker-assisted selection (MAS) and genomic selection of forage quality traits based on breeding values of parent plants. Some 154 genotypes from a broadly-based reference population were genotyped by genotyping-by-sequencing (GBS), and phenotyped for leaf-to-stem ratio, leaf and stem contents of protein, neutral detergent fiber (NDF) and acid detergent lignin (ADL), and leaf and stem NDF digestibility after 24 hours (NDFD), of their dense-planted half-sib progenies in three growing conditions (summer harvest, full irrigation; summer harvest, suspended irrigation; autumn harvest). Trait-marker analyses were performed on progeny values averaged over conditions, owing to modest germplasm × condition interaction. Genomic selection exploited 11,450 polymorphic SNP markers, whereas a subset of 8,494 M. truncatula-aligned markers were used for a genome-wide association study (GWAS). GWAS confirmed the polygenic control of quality traits and, in agreement with phenotypic correlations, indicated substantially different genetic control of a given trait in stems and leaves. It detected several SNPs in different annotated genes that were highly linked to stem protein content. Also, it identified a small genomic region on chromosome 8 with high concentration of annotated genes associated with leaf ADL, including one gene probably involved in the lignin pathway. Three genomic selection models, i.e., Ridge-regression BLUP, Bayes B and Bayesian Lasso, displayed similar prediction accuracy, whereas SVR-lin was less accurate. Accuracy values were moderate (0.3–0.4) for stem NDFD and leaf protein content, modest for leaf ADL and NDFD, and low to very low for the other traits. Along with previous results for the same germplasm set, this study indicates that GBS data can be exploited to improve both quality traits

  8. New pharmacological properties of Medicago sativa and Saponaria officinalis saponin-rich fractions addressed to Candida albicans.

    Science.gov (United States)

    Sadowska, Beata; Budzyńska, Aleksandra; Więckowska-Szakiel, Marzena; Paszkiewicz, Małgorzata; Stochmal, Anna; Moniuszko-Szajwaj, Barbara; Kowalczyk, Mariusz; Różalska, Barbara

    2014-08-01

    The antifungal activity of the saponin-rich fractions (SFs) from Medicago sativa (aerial parts and roots) and Saponaria officinalis (used as a well-known source of plant saponins) against Candida albicans reference and clinical strains, their yeast-to-hyphal conversion, adhesion, and biofilm formation was investigated. Direct fungicidal/fungistatic properties of the tested phytochemicals used alone, as well as their synergy with azoles (probably resulting from yeast cell wall instability) were demonstrated. Here, to the best of our knowledge, we report for the first time the ability of saponin-rich extracts of M. sativa and S. officinalis to inhibit C. albicans germ tube formation, limit hyphal growth, reduce yeast adherence and biofilm formation, and eradicate mature (24 h) Candida biofilm. Moreover, M. sativa SFs (mainly obtained from aerial parts), in the range of concentrations which were active modulators of Candida virulence factors, exhibited low cytotoxicity against the mouse fibroblast line L929. These properties seem to be very promising in the context of using plant-derived SFs as potential novel antifungal therapeutics supporting classic drugs or as ingredients of disinfectants. © 2014 The Authors.

  9. [MEDICAGO SATIVA L: IMPROVEMENT AND NEW APPROACHES OF ITS NUTRITIONAL AND FUNCTIONAL VALUE BY BACTERIAL CO-INOCULATION].

    Science.gov (United States)

    Martínez, Rosario; Nebot, Elena; Porres, Jesús María; Kapravelou, Garyfallia; Del Moral, Ana; Talbi, Chouhra; Bedmar, Eulogio Jose; López-Jurado, María

    2015-12-01

    to study the effect of co-inoculation with Ensifer meliloti and Halomonas maura of the leguminous Medicago sativa L., on growth, nutritional and functional value, grown under salinity conditions. plants of M. sativa were grown in a solution with a mixture of salts (CaSO4, MgCl, NaCl and NaHCO 3) and were co-inoculated with its specific rhizobium and the halophilic moderated bacterium H. maura. Different physiologic parameters were determined, as well as, nitrogen and minerals content. Furthermore, an assay of in vitro digestibility was carried out. salinity had a negative effect on the plants; however, co-inoculation increased yield, nitrogen content, total minerals, Ca and Mg. Moreover, physiologic parameters as water potential and leghemoglobin content in fresh nodules were higher compared to those of plants inoculated only with E. meliloti. Both, salinity and bacterial treatment with E. meliloti and H. maura increased the antioxidant capacity of the legume, in dialyzates and retentates collected after an in vitro digestibility assay. co-inoculation of plants with E. meliloti and H. maura could improve the alfalfa yield under specific salinity conditions, increasing the nutritional and functional value of the plants. M. sativa could be considered in the formulations of nutritional supplements for the human diet. Copyright AULA MEDICA EDICIONES 2014. Published by AULA MEDICA. All rights reserved.

  10. Effects of Shepherd’s Purse (Capsella bursa pastoris (L. Medic. on the Chemical Composition of Lucerne (Medicago sativa L.

    Directory of Open Access Journals (Sweden)

    Tsvetanka Dimitrova

    2008-01-01

    Full Text Available A study was conducted in a pure stand of lucerne (variety Viktoria under natural weed infestation with shepherd’s purse (Capsella bursa pastoris (L. Medic. on a slightly leached chernozem soil under nonirrigated conditions in the experimental field of the Institute ofForage Crops – Pleven during the 2006-2007 period. The effect of shepherd’s purse Capsella bursa pastoris (L. Medic. on the chemical composition of lucerne Medicago sativa (L. was analyzed.Statistically significant (P<0.05 functional relations were found between the chemical characteristics and percentage of Capsella bursa pastoris (L. Medic. participation in the lucerne sward, and forage quality. These relations indicated a multiple practical relevance and a necessity to control Capsella bursa pastoris (L. Medic. in lucerne stands in order to decrease weed density and improve forage quality.

  11. Citrate, Malate, and Succinate Concentration in Exudates from P-Sufficient and P-Stressed Medicago sativa L. Seedlings 1

    Science.gov (United States)

    Lipton, Doug S.; Blanchar, Robert W.; Blevins, Dale G.

    1987-01-01

    Under certain stress conditions roots exude organic molecules, which may facilitate the uptake of nutrients. The objective of this research was to identify and measure the effect of low P upon the exudation of organic acids by roots of alfalfa (Medicago sativa L.) seedlings. Surface-sterilized alfalfa seeds were grown aseptically in sterile sand using an apparatus specially designed for the addition of ±P nutrient solutions and for the collection of root exudates. Citric, malic, and succinic acids were detected in the root exudates of 24-day-old alfalfa seedlings. Citrate exudation from the roots of P-stressed alfalfa was 182% that of plants receiving a complete nutrient solution. The increased release of citrate may provide a mechanism by which P-stressed plants enhance the availability of P in the rhizosphere. PMID:16665693

  12. Effects of salinity and drought stress on germination, biomass and growth in three varieties of Medicago sativa L.

    OpenAIRE

    A. Castroluna; Ruiz, O. M.; A. M. Quiroga; H. E. Pedranzani

    2014-01-01

    En plántulas de alfalfa (Medicago sativa L.) se evaluó la germinación y el crecimiento y su tolerancia al estrés osmótico inducido por Polietilenglicol 6000 (PEG6000) y salinidad (NaCl) en tres genotipos distintos: Dk166, Verdor y Salina. Esta investigación se llevó a cabo con un diseño experimental completamente aleatorizado, con cinco réplicas y los parámetros que se determinaron fueron: energía y poder germinativo, largo de hojas y peso fresco y peso seco de raíces y hojas. Los resultados ...

  13. MsPG3, a Medicago sativa polygalacturonase gene expressed during the alfalfa–Rhizobium meliloti interaction

    Science.gov (United States)

    Muñoz, Jose A.; Coronado, Carmen; Pérez-Hormaeche, Javier; Kondorosi, Adam; Ratet, Pascal; Palomares, Antonio J.

    1998-01-01

    Polygalacturonase (PG) is one of the most important enzymes associated with plant cell wall degradation. It has been proposed to participate in the early steps of the Rhizobium–legume interaction. We have identified two classes of cDNA fragments corresponding to two classes of PG genes in the Medicago genome. One of this class, represented by E2 in M. truncatula and Pl1 in M. sativa, seems to be related to previously characterized plant PG genes expressed in pollen. We have isolated the genomic clone containing the entire gene corresponding to the second class (E3). We showed that MsPG3 is a single gene in the Medicago genome coding for PG. By reverse transcription-PCR, MsPG3 expression was detected in roots 1 day after Rhizobium inoculation. The early induction of the MsPG3, as also seen by in situ hybridization experiments, supports its involvement in the early stages of the Rhizobium-legume infection process. In addition, by analyzing the expression of a MsPG3 promoter-gus construct in Vicia hirsuta-transgenic root nodules, we showed that MsPG3 was expressed in all cells of nodule primordia and in the cells of the invasion zone. By Northern blot, MsPG3 transcripts are not detected in various Medicago tissues, indicating that the function of this gene is related closely to symbiosis. Thus, our results strongly suggest the involvement of MsPG3 gene during meristem formation and/or in the infection process, probably by facilitating cell wall rearrangement, penetration of the bacteria through the root hair wall, or infection thread formation and release of bacteria in plant cells. MsPG3 represents a class of PG genes, distinct from the pollen-specific genes, and it is the first pectic encoded enzyme demonstrated to be involved in Rhizobium-legume symbiosis. PMID:9689142

  14. Molecular cloning and functional analysis of the drought tolerance gene MsHSP70 from alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Li, Zhenyi; Long, Ruicai; Zhang, Tiejun; Wang, Zhen; Zhang, Fan; Yang, Qingchuan; Kang, Junmei; Sun, Yan

    2017-03-01

    Heat shock proteins (HSPs) are a ubiquitously expressed class of protective proteins that play a key role in plant response to stressful conditions. This study aimed to characterize and investigate the function of an HSP gene in alfalfa (Medicago sativa). MsHSP70, which contains a 2028-bp open reading frame, was identified through homology cloning. MsHSP70 shares high sequence identity (94.47%) with HSP70 from Medicago truncatula. Expression analysis of MsHSP70 in alfalfa organs revealed a relatively higher expression level in aerial organs such as flowers, stems and leaves than in roots. MsHSP70 was induced by heat shock, abscisic acid (ABA) and hydrogen peroxide. Transgenic Arabidopsis seedlings overexpressing MsHSP70 were hyposensitive to polyethylene glycol (PEG) and ABA treatments, suggesting that exogenous expression of MsHSP70 enhanced Arabidopsis tolerance to these stresses. Examination of physiological indexes related to drought and ABA stress demonstrated that in comparison with non-transgenic plants, T3 transgenic Arabidopsis plants had an increased proline content, higher superoxide dismutase (SOD) activity, and decreased malondialdehyde (MDA) content. Furthermore, higher relative water content (RWC) was detected in transgenic plants compared with non-transgenic plants under drought stress. These findings clearly indicate that molecular manipulation of MsHSP70 in plants can have substantial effects on stress tolerance.

  15. Effects of salinity and drought stress on germination, biomass and growth in three varieties of Medicago sativa L.

    Directory of Open Access Journals (Sweden)

    A. Castroluna

    2014-01-01

    Full Text Available En plántulas de alfalfa (Medicago sativa L. se evaluó la germinación y el crecimiento y su tolerancia al estrés osmótico inducido por Polietilenglicol 6000 (PEG6000 y salinidad (NaCl en tres genotipos distintos: Dk166, Verdor y Salina. Esta investigación se llevó a cabo con un diseño experimental completamente aleatorizado, con cinco réplicas y los parámetros que se determinaron fueron: energía y poder germinativo, largo de hojas y peso fresco y peso seco de raíces y hojas. Los resultados mostraron que la germinación fue retrasada en todas las variedades con ambas soluciones, con diferencias entre genotipos. El crecimiento de las plántulas, bajo un ligero estrés mostraron un incremento de la biomasa aérea y en un estrés hídrico y salino severo, muerte de las plantas excepto en Var. Salina (tolerante. PEG causó muerte en la variedad Verdor que mostró más sensibilidad a PEG que a NaCl. En raíces, los genotipos más sensibles frente a leves estrés salinos detuvieron su crecimiento y bajo leves situaciones de estrés hídrico lo incrementaron. Nuestros estudios nos permiten definir las variedades comerciales de Medicago sativa como moderadamente tolerantes a salinidad y sequía la variedad DK166, sensible o intolerante a salinidad y sequía la variedad Verdor y con una alta tolerancia a salinidad e insensible a sequía la variedad Salina.

  16. Identification of genetic loci associated with crude protein and mineral concentrations in alfalfa (Medicago sativa) using association mapping.

    Science.gov (United States)

    Jia, Congjun; Wu, Xinming; Chen, Min; Wang, Yunqi; Liu, Xiqiang; Gong, Pan; Xu, Qingfang; Wang, Xuemin; Gao, Hongwen; Wang, Zan

    2017-06-06

    Alfalfa (Medicago sativa) is one of the most important legume forage species in China and many other countries of the world. It provides a quality source of proteins and minerals to animals. Genetic underpinnings for these important traits, however, are elusive. An alfalfa (M. sativa) association mapping study for six traits, namely crude protein (CP), rumen undegraded protein (RUP), and four mineral elements (Ca, K, Mg and P), was conducted in three consecutive years using a large collection encompassing 336 genotypes genotyped with 85 simple sequence repeat (SSR) markers. All the traits were significantly influenced by genotype, environment, and genotype × environment interaction. Eight-five significant associations (P < 0.005) were identified. Among these, five associations with Ca were repeatedly observed and six co-localized associations were identified. The identified marker alleles significantly associated with the traits provided important information for understanding genetic controls of alfalfa quality. The markers could be used in assisting selection for the individual traits in breeding populations for developing new alfalfa cultivars.

  17. Detection of Changes in the Medicago sativa Retinoblastoma-Related Protein (MsRBR1) Phosphorylation During Cell Cycle Progression in Synchronized Cell Suspension Culture.

    Science.gov (United States)

    Ayaydin, Ferhan; Kotogány, Edit; Ábrahám, Edit; Horváth, Gábor V

    2017-01-01

    Deepening our knowledge on the regulation of the plant cell division cycle depends on techniques that allow for the enrichment of cell populations in defined cell cycle phases. Synchronization of cell division can be achieved using different plant tissues; however, well-established cell suspension cultures provide large amount of biological sample for further analyses. Here, we describe the methodology of the establishment, propagation, and analysis of a Medicago sativa suspension culture that can be used for efficient synchronization of the cell division. A novel 5-ethynyl-2'-deoxyuridine (EdU)-based method is used for the estimation of cell fraction that enters DNA synthesis phase of the cell cycle and we also demonstrate the changes in the phosphorylation level of Medicago sativa retinoblastoma-related protein (MsRBR1) during cell cycle progression.

  18. Contributions of Sinorhizobium meliloti Transcriptional Regulator DksA to Bacterial Growth and Efficient Symbiosis with Medicago sativa.

    Science.gov (United States)

    Wippel, Kathrin; Long, Sharon R

    2016-05-01

    The stringent response, mediated by the (p)ppGpp synthetase RelA and the RNA polymerase-binding protein DksA, is triggered by limiting nutrient conditions. For some bacteria, it is involved in regulation of virulence. We investigated the role of two DksA-like proteins from the Gram-negative nitrogen-fixing symbiont Sinorhizobium meliloti in free-living culture and in interaction with its host plant Medicago sativa The two paralogs, encoded by the genes SMc00469 and SMc00049, differ in the constitution of two major domains required for function in canonical DksA: the DXXDXA motif at the tip of a coiled-coil domain and a zinc finger domain. Using mutant analyses of single, double, and triple deletions for SMc00469(designated dksA),SMc00049, and relA, we found that the ΔdksA mutant but not the ΔSMc00049 mutant showed impaired growth on minimal medium, reduced nodulation on the host plant, and lower nitrogen fixation activity in early nodules, while its nod gene expression was normal. The ΔrelA mutant showed severe pleiotropic phenotypes under all conditions tested. Only S. meliloti dksA complemented the metabolic defects of an Escherichia coli dksA mutant. Modifications of the DXXDXA motif in SMc00049 failed to establish DksA function. Our results imply a role for transcriptional regulator DksA in the S. meliloti-M. sativa symbiosis. The stringent response is a bacterial transcription regulation process triggered upon nutritional stress.Sinorhizobium meliloti, a soil bacterium establishing agriculturally important root nodule symbioses with legume plants, undergoes constant molecular adjustment during host interaction. Analyzing the components of the stringent response in this alphaproteobacterium helps understand molecular control regarding the development of plant interaction. Using mutant analyses, we describe how the lack of DksA influences symbiosis with Medicago sativa and show that a second paralogous S. meliloti protein cannot substitute for this missing

  19. PHYSIOLOGICAL, CYTOLOGICAL AND BIOCHEMICAL STABILITY OF Medicago sativa L. CELL CULTURE AFTER 27 YEARS OF CRYOGENIC STORAGE.

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    Volkova, L A; Urmantseva, V V; Popova, E V; Nosov, A M

    2015-01-01

    The efficiency of long-term cryogenic storage to prevent somaclonal variations in plant cell cultures and retain their major cytogenetic and biochemical traits remains under debate. In particular, it is not clear how stress conditions associated with cryopreservation, such as low temperature, dehydration and toxic action of some cryoprotectants (DMSO in particular), affect post-storage regrowth and genetic integrity of cell samples. We assessed growth, cytogenetic and biochemical characteristics of the peroxidase-producing strain of Medicago sativa L. cell culture recovered after 27 years of cryogenic storage as compared to the same culture before cryopreservation. In 1984, M. sativa L. cell culture was cryopreserved using programmed freezing and 7% DMSO as a cryoprotectant. In 2011, after rewarming in a water bath at 40 degree C for 90 s, cell culture was recovered and proliferated. Viability, growth profile, mitotic index, ploidy level, peroxidase activity and cell response to hypothermia and osmotic stress were compared between the recovered and the initial cell cultures using the records available from 1984. Viability of alfalfa cell culture after rewarming was below 20% but it increased to 80% by the 27th subculture cycle. Recovered culture showed higher mitotic activity and increased number of haploid and diploid cells compared to the initial cell line. Both peroxidase activity and response to abiotic stress in the recovered cell culture were similar to that of the initial culture. Cryopreservation by programmed freezing was effective at retaining the main characteristics of M. sativa undifferentiated cell culture after 27 years of storage. According to available data, this is longest period of successful cryopreservation of plant cell cultures reported so far. After storage, there was no evidence that DMSO had any detrimental effect on cell viability, growth or cytogenetics.

  20. A saturated genetic linkage map of autotetraploid alfalfa (Medicago sativa L.) developed using genotyping-by-sequencing is highly syntenous with the Medicago truncatula genome.

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    Li, Xuehui; Wei, Yanling; Acharya, Ananta; Jiang, Qingzhen; Kang, Junmei; Brummer, E Charles

    2014-08-21

    A genetic linkage map is a valuable tool for quantitative trait locus mapping, map-based gene cloning, comparative mapping, and whole-genome assembly. Alfalfa, one of the most important forage crops in the world, is autotetraploid, allogamous, and highly heterozygous, characteristics that have impeded the construction of a high-density linkage map using traditional genetic marker systems. Using genotyping-by-sequencing (GBS), we constructed low-cost, reasonably high-density linkage maps for both maternal and paternal parental genomes of an autotetraploid alfalfa F1 population. The resulting maps contain 3591 single-nucleotide polymorphism markers on 64 linkage groups across both parents, with an average density of one marker per 1.5 and 1.0 cM for the maternal and paternal haplotype maps, respectively. Chromosome assignments were made based on homology of markers to the M. truncatula genome. Four linkage groups representing the four haplotypes of each alfalfa chromosome were assigned to each of the eight Medicago chromosomes in both the maternal and paternal parents. The alfalfa linkage groups were highly syntenous with M. truncatula, and clearly identified the known translocation between Chromosomes 4 and 8. In addition, a small inversion on Chromosome 1 was identified between M. truncatula and M. sativa. GBS enabled us to develop a saturated linkage map for alfalfa that greatly improved genome coverage relative to previous maps and that will facilitate investigation of genome structure. GBS could be used in breeding populations to accelerate molecular breeding in alfalfa. Copyright © 2014 Li et al.

  1. Effects of Plant Density and Water Stress on Competitive Ability and Yield of Medicago Sativa L. and Bromus tomentellus Boiss.in Mono and Mixed Cropping

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    S. Barati

    2016-03-01

    Full Text Available The effects of plant density and water stress on yield of Medicago sativa and Bromus tomentellus was studied. A greenhouse experiment was conducted at Isfahan University of Technology in 2013. The experiment included 18 treatments, three crop compositions (M.sativa, B. tomentellus or mixture of the two, two plant density levels, three watering regimes and four replicates, arranged in a completely randomized block design. Results showed that total yield of M. sativa mono-cropping was higher than mixed cropping and it was higher than B. tomentellus mono-cropping. Land Equivalent Ratio (LER values were less than 1 for all mixed cropping treatments, indicated the interspecific competition in mixed cropping. The biomass production per plant decreased with increasing density, competition for resource utilization and water stress. Compare of above-ground and below-ground dry matter showed that M. sativa appeared to be more constrained by intraspecific than by interspecific competition, Conversely, B. tomentellus was more suppressed by interspecific competition exerted by M. sativa than by intraspecific competition. Relative competition intensity (RCI values were positive for B. tomentellus and negative for M.sativa, implying that competitive ability of M.sativa was higher than B. tomentellus in mixed cropping .

  2. A cryptic species produced by autopolyploidy and subsequent introgression involving Medicago prostrata (Fabaceae).

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    Eriksson, J S; Blanco-Pastor, J L; Sousa, F; Bertrand, Y J K; Pfeil, B E

    2017-02-01

    Although hybridisation through genome duplication is well known, hybridisation without genome duplication (homoploid hybrid speciation, HHS) is not. Few well-documented cases have been reported. A possible instance of HHS in Medicago prostrata Jacq. was suggested previously, based on only two genes and one individual. We tested whether this species was formed through HHS by sampling eight nuclear loci and 22 individuals, with additional individuals from related species, using gene capture and Illumina sequencing. Phylogenetic inference and coalescent simulations were performed to infer the causes of gene tree incongruence. We found no evidence that phylogenetic differences among M. prostrata individuals were the result of HHS. Instead, an autopolyploid origin of tetraploids with introgression from tetraploids of the M. sativa complex is likely. We argue that tetraploid M. prostrata individuals constitute a new species, characterised by a partially non-overlapping distribution and distinctive alleles (from the M. sativa complex). No gene flow from tetraploid to diploid M. prostrata is apparent, suggesting partial reproductive isolation. Thus, speciation via autopolyploidy appears to have been reinforced by introgression. This raises the intriguing possibility that introgressed alleles may be responsible for the increased range exploited by tetraploid M. prostrata with respect to that of the diploids. Copyright © 2016 Elsevier Inc. All rights reserved.

  3. Genome-Wide Identification of Different Dormant Medicago sativa L. MicroRNAs in Response to Fall Dormancy

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    Du, Hongqi; Sun, Xiaoge; Shi, Yinghua; Wang, Chengzhang

    2014-01-01

    Background MicroRNAs (miRNAs) are a class of regulatory small RNAs (sRNAs) that regulate gene post-transcriptional expression in plants and animals. High-throughput sequencing technology is capable of identifying small RNAs in plant species. Alfalfa (Medicago sativa L.) is one of the most widely cultivated perennial forage legumes worldwide, and fall dormancy is an adaptive characteristic related to the biomass production and winter survival in alfalfa. Here, we applied high-throughput sRNA sequencing to identify some miRNAs that were responsive to fall dormancy in standard variety (Maverick and CUF101) of alfalfa. Results Four sRNA libraries were generated and sequenced from alfalfa leaves in two typical varieties at distinct seasons. Through integrative analysis, we identified 51 novel miRNA candidates of 206 families. Additionally, we identified 28 miRNAs associated with fall dormancy in standard variety (Maverick and CUF101), including 20 known miRNAs and eight novel miRNAs. Both high-throughput sequencing and RT-qPCR confirmed that eight known miRNA members were up-regulated and six known miRNA members were down-regulated in response to fall dormancy in standard variety (Maverick and CUF101). Among the 51 novel miRNA candidates, five miRNAs were up-regulated and three miRNAs were down-regulated in response to fall dormancy in standard variety (Maverick and CUF101), and five of them were confirmed by Northern blot analysis. Conclusion We identified 20 known miRNAs and eight new miRNA candidates that were responsive to fall dormancy in standard variety (Maverick and CUF101) by high-throughput sequencing of small RNAs from Medicago sativa. Our data provide a useful resource for investigating miRNA-mediated regulatory mechanisms of fall dormancy in alfalfa, and these findings are important for our understanding of the roles played by miRNAs in the response of plants to abiotic stress in general and fall dormancy in alfalfa. PMID:25473944

  4. Genome-wide identification of different dormant Medicago sativa L. MicroRNAs in response to fall dormancy.

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    Fan, Wenna; Zhang, Senhao; Du, Hongqi; Sun, Xiaoge; Shi, Yinghua; Wang, Chengzhang

    2014-01-01

    MicroRNAs (miRNAs) are a class of regulatory small RNAs (sRNAs) that regulate gene post-transcriptional expression in plants and animals. High-throughput sequencing technology is capable of identifying small RNAs in plant species. Alfalfa (Medicago sativa L.) is one of the most widely cultivated perennial forage legumes worldwide, and fall dormancy is an adaptive characteristic related to the biomass production and winter survival in alfalfa. Here, we applied high-throughput sRNA sequencing to identify some miRNAs that were responsive to fall dormancy in standard variety (Maverick and CUF101) of alfalfa. Four sRNA libraries were generated and sequenced from alfalfa leaves in two typical varieties at distinct seasons. Through integrative analysis, we identified 51 novel miRNA candidates of 206 families. Additionally, we identified 28 miRNAs associated with fall dormancy in standard variety (Maverick and CUF101), including 20 known miRNAs and eight novel miRNAs. Both high-throughput sequencing and RT-qPCR confirmed that eight known miRNA members were up-regulated and six known miRNA members were down-regulated in response to fall dormancy in standard variety (Maverick and CUF101). Among the 51 novel miRNA candidates, five miRNAs were up-regulated and three miRNAs were down-regulated in response to fall dormancy in standard variety (Maverick and CUF101), and five of them were confirmed by Northern blot analysis. We identified 20 known miRNAs and eight new miRNA candidates that were responsive to fall dormancy in standard variety (Maverick and CUF101) by high-throughput sequencing of small RNAs from Medicago sativa. Our data provide a useful resource for investigating miRNA-mediated regulatory mechanisms of fall dormancy in alfalfa, and these findings are important for our understanding of the roles played by miRNAs in the response of plants to abiotic stress in general and fall dormancy in alfalfa.

  5. The effect of Medicago arabica, M. hybrida and M. sativa saponins on the growth and development of Fusarium oxysporum Schlecht f. sp. tulipae apt.

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    Anna Jarecka

    2012-12-01

    Full Text Available In the present work it was shown that total saponins originated from M. hybrida and M. sativa substantially limited mycelium growth of F. oxysporum f. sp. tulipae and symptoms of fusariosis on tulip bulbs. Out of 15 individual tested saponins originated from M. arabica, M. hybrida and M. sativa, four compounds: 3-O-[β-D-glucopyranosyl (1→2α-L-arabinopyranosyl] hederagenin, hederagenin 3-O-β-D-glucopyranoside, medicagenic acid, medicagenic acid 3-O-β-D-glucopyranoside had the strongest inhibitory effect on mycelium growth of Fusarium oxysporum f. sp. tulipae on PDA medium. The total saponins from M. arabica, M. hybrida and M. sativa inhibited the number of colony forming units of Fusarium oxysporum f. sp. tulipae in artificially infested substrate. The use of saponins originated from Medicago as a fungicide is suggested.

  6. Therapeutic Potential of Hydroxypropyl-β-Cyclodextrin-Based Extract of Medicago sativa in the Treatment of Mucopolysaccharidoses.

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    Fumić, Barbara; Končić, Marijana Zovko; Jug, Mario

    2017-01-01

    Mucopolysaccharidoses are inherited metabolic disorders resulting in the dysfunction of enzymes involved in the degradation of glycosaminoglycans, leading to severe clinical symptoms and a significantly shortened life span of patients. Flavonoids are recognized as glycosaminoglycan metabolism modulators, able to correct glycosaminoglycan cell storage. Therefore, the aim of this work was the development of an efficient and eco-friendly extraction process of phytochemicals from Medicago sativa by simultaneous use of ultrasound extraction and hydroxypropyl-β-cyclodextrin complexation, and investigation of the potential of such an extract as a glycosaminoglycan metabolism modulator. The Box-Behnken design and response surface methodology were used in order to optimize the extraction process, considering hydroxypropyl-β-cyclodextrin concentration, ultrasonic power, and extraction time as the key parameters. The dependent variables included total phenolicand total flavonoid content, DPPH radical scavenging activity, and Fe2+ chelating activity, due to the importance of oxidative stress in the pathology of mucopolysaccharidoses. The developed technology using hydroxypropyl-β-cyclodextrin led to more selective flavonoid extraction from M. sativa than obtained either by the use of water or ethanol. The lyophilization of extracts resulted in products with high radical scavenging activity, suitable for further use. The application of 20 mM hydroxypropyl-β-cyclodextrin solution, 432 W ultrasonic power, and an extraction time of 45 min resulted in an extract with both the highest total flavonoid content and the lowest radical scavenging activity IC50. This extract reduced the levels of glycosaminoglycans in skin fibroblasts of mucopolysaccharidose III patient in a dose-dependent manner. At concentrations of 3 and 6 µg/mL, the observed levels of glycosaminoglycans were reduced by 41.2 and 51.1 %, respectively, clearly demonstrating the validity of the selected

  7. Effect of Row Spacing and Seeding Rate on Yield Component and Seed Yield of Alfalfa (Medicago sativa L.

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    Gholamreza ABADOUZ

    2010-03-01

    Full Text Available In order to investigation the effect of row spacing and seeding rate on alfalfa (Medicago sativa L. cv. Mesasirsa an experiment was carried out at the research farm of Khouzestan Agricultural Research Center from 2000 to 2004. This research was conducted in the form of a split plot design in a randomized complete block, with four replications. Main plot was three row spaces (50, 60 and 70 cm and sub plots were five seeding rates (5, 10, 15, 20, and 25 kg seed ha-1. Results showed that the highest seed yield obtained in the second year. Seed yield and raceme per m2 increased with row spacing decreased. Seeding rate did not have any significant effect on pod per raceme, seed per pod and seed weight. The interaction of year and treatments on whole of traits were not significant. It was observed that correlation among seed yield and yield components was positive and significant. It was concluded that with using 15 kg seed ha-1 obtained highest seed yield due to increase in raceme per m2 and seed weight.

  8. Paenibacillus medicaginis sp. nov. a chitinolytic endophyte isolated from the root nodule of alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Lai, Wei-An; Hameed, Asif; Lin, Shih-Yao; Hung, Mei-Hua; Hsu, Yi-Han; Liu, You-Cheng; Shahina, Mariyan; Shen, Fo-Ting; Young, Chiu-Chung

    2015-08-04

    A Gram-stain-variable, short-rod-shaped, endospore-forming, strictly aerobic, non-motile, chitinolytic and endophytic bacterium, designated strain CC-Alfalfa-19T, exhibiting unusual bipolar appendages was isolated from the root nodule of alfalfa (Medicago sativa L.) in Taiwan and subjected to a polyphasic taxonomy. Based on 16S rRNA gene sequence analysis, strain CC-Alfalfa-19T was found to be most closely related to Paenibacillus puldeungensis CAU 9324T (95.2 %), whereas other Paenibacillus species shared ≤95.0 % sequence similarity. The phylogenetic analysis revealed a distinct phyletic lineage established by strain CC-Alfalfa-19T with respect to other Paenibacillus species. Fatty acids comprised predominantly anteiso-C15:0, C16:0, anteiso-C17:0 and iso-C16. Menaquinone 7 (MK-7) was identified as the sole respiratory quinone and the genomic DNA G+C content was 42.7 mol%. Polar lipids included diphosphatidylglycerol, phosphatidylglycerol, phosphatidylmonomethylethanolamine, phosphatidylethanolamine, an unidentified glycolipid and an unidentified lipid. The diagnostic diamino acid found in the cell-wall peptidoglycan was meso-diaminopimelic acid. Based on the polyphasic taxonomic evidences that were in line with the genus Paenibacillus and additional distinguishing characteristics, strain CC-Alfalfa-19T is considered to represent a novel species, for which the name Paenibacillus medicaginis sp. nov. (type strain CC-Alfalfa-19T = BCRC 80441T = JCM 18446T) is proposed.

  9. Concurrent synthesis and release of nod-gene-inducing flavonoids from alfalfa roots. [Medicago sativa L. ; Rhizobium meliloti

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    Maxwell, C.A.; Phillips, D.A. (Univ. of California, Davis (USA))

    1990-08-01

    Flavonoid signals from alfalfa (Medicago sativa L.) induce transcription of nodulation (nod) genes in Rhizobium meliloti. Alfalfa roots release three major nod-gene inducers: 4{prime},7-dihydroxyflavanone, 4{prime},7-dihydroxyflavone, and 4,4{prime}-dihydroxy-2{prime}-methoxychalcone. The objective of the present study was to define temporal relationships between synthesis and exudation for those flavonoids. Requirements for concurrent flavonoid biosynthesis were assessed by treating roots of intact alfalfa seedlings with (U-{sup 14}C)-L-phenylalanine in the presence or absence of the phenylalanine ammonia-lyase inhibitor L-2-aminoxy-3-phenylpropionic acid (AOPP). In the absence of AOPP, each of the three flavonoids in exudates contained {sup 14}C. In the presence of AOPP, {sup 14}C labeling and release of all the exuded nod-gene inducers were reduced significantly. AOPP inhibited labeling and release of the strongest nod-gene inducer, methoxychalcone, by more than 90%. The release process responsible for exudation of nod-gene inducers appears to be specific rather than a general phenomenon such as a sloughing off of cells during root growth.

  10. Allelopathic potential of switchgrass (Panicum virgatum L.) on perennial ryegrass (Lolium perenne L.) and alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Shui, Junfeng; An, Yu; Ma, Yongqing; Ichizen, Nobumasa

    2010-10-01

    This study investigated allelopathy and its chemical basis in nine switchgrass (Panicum virgatum L.) accessions. Perennial ryegrass (Lolium perenne L.) and alfalfa (Medicago sativa L.) were used as test species. Undiluted aqueous extracts (5 g plant tissue in 50 ml water) from the shoots and roots of most of the switchgrass accessions inhibited the germination and growth of the test species. However, the allelopathic effect of switchgrass declined when extracts were diluted 5- or 50-fold. Seedling growth was more sensitive than seed germination as an indicator of allelopathic effect. Allelopathic effect was related to switchgrass ecotype but not related to ploidy level. Upland accessions displayed stronger allelopathic potential than lowland accessions. The aqueous extract from one switchgrass accession was separated into phenols, organic acids, neutral chemicals, and alkaloids, and then these fractions were bioassayed to test for allelopathic potential. Alkaloids had the strongest allelopathic effect among the four chemical fractions. In summary, the results indicated that switchgrass has allelopathic potential; however, there is not enough evidence to conclude that allelopathic advantage is the main factor that has contributed to the successful establishment of switchgrass on China's Loess Plateau.

  11. In vitro and in vivo antioxidant activity of alfalfa (Medicago sativa L.) on carbon tetrachloride intoxicated rats.

    Science.gov (United States)

    Al-Dosari, Mohammed S

    2012-01-01

    The present study was conducted to determine whether lyophilized aqueous extract of alfalfa, or Medicago sativa L. could exert antioxidant activity against carbon tetrachloride-induced oxidative stress and liver injury in rats. The hepatoprotective activity of alfalfa extract was determined by assessing the levels of serum transaminases, ALP, bilirubin and lipid profile. Further, the effect of the test substance on malondialdehyde (MDA), an end product of lipid peroxidation; antioxidant liver enzyme non-protein sulfhydryl (NP-SH); and total protein (TP) were also studied. Serum transaminase, ALP, bilirubin level, lipid profile and liver MDA were significantly elevated and the antioxidant status in liver NP-SH and TP contents were declined in animals treated with CCl (4) alone. Pretreatment with alfalfa and silymarin for three weeks prior to the administration of CCl (4) significantly prevented the increase in the serum levels of hepatic marker, LDL, VLDL levels enzymes and reduced oxidative stress indicated by elevated NP-SH and TP concentration. The histopathological examination of the livers also showed that the alfalfa extract reduced the incidence of liver lesions induced by CCl (4). The in vitro antioxidant assessment of alfalfa extract on DPPH and carotene-linoleic assays demonstrated a moderate antioxidant potential. Results suggest that the alfalfa extract possesses hepatoprotective and antioxidative stress properties possibly through its antioxidant phytochemical constituents and substantiates its use in various liver disorders as a hepatoprotector.

  12. Alfalfa (Medicago sativa L.) shoot saponins: identification and bio-activity by the assessment of aphid feeding.

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    Mazahery-Laghab, H; Yazdi-Samadi, B; Bagheri, M; Bagheri, A R

    2011-01-01

    Biochemical components in alfalfa (Medicago sativa L.), such as saponins, can act as protecting factors against bio-stresses. Saponins are also antifeedants and show oral toxicity towards higher and lower animals. Changes in saponins, such as variation in the carbon skeleton, or hydrolysis of saponin glycosides and other conjugates, may change their biological effects. The aims of this research were to study saponin variation in different growth stages of alfalfa and to investigate the biological role of saponins in the spotted alfalfa aphid, Therioaphis maculata. Saponins from alfalfa shoots in different growth stages were extracted, chemically purified and analysed by TLC. Specific saponins such as soyasaponin1 from root and shoot and two bisdesmosides of medicagenic acid, one from shoot and another from root tissues, were identified using reference compounds allowing changes in saponin composition during plant development in different shoot tissues of alfalfa to be assessed. The response of the alfalfa aphid to feeding on alfalfa in different growth stages was studied. No significant difference in the survival of aphids, from neonate to adult, was observed, but due to the antibiotic effects of saponins, two differences were found in the onset of nymph production and cumulative nymph production. The results show that the saponin composition in alfalfa changes with plant development and this, in turn, can often negatively affect the development of specific insect pests such as the spotted alfalfa aphid, suggesting a possible biological role of alfalfa saponins.

  13. Cytotoxicity and apoptosis induced by alfalfa (Medicago sativa) leaf extracts in sensitive and multidrug-resistant tumor cells.

    Science.gov (United States)

    Gatouillat, Grégory; Magid, Abdulmagid Alabdul; Bertin, Eric; Okiemy-Akeli, Marie-Genevieve; Morjani, Hamid; Lavaud, Catherine; Madoulet, Claudie

    2014-01-01

    Alfalfa (Medicago sativa) has been used to cure a wide variety of ailments. However, only a few studies have reported its anticancer effects. In this study, extracts were obtained from alfalfa leaves and their cytotoxic effects were assessed on several sensitive and multidrug-resistant tumor cells lines. Using the mouse leukaemia P388 cell line and its doxorubicin-resistant counterpart (P388/DOX), we showed that the inhibition of cell growth induced by alfalfa leaf extracts was mediated through the induction of apoptosis, as evidenced by DNA fragmentation analysis. The execution of programmed cell death was achieved via the activation of caspase-3, leading to PARP cleavage. Fractionation of toluene extract (To-1), the most active extract obtained from crude extract, led to the identification of 3 terpene derivatives and 5 flavonoids. Among them, (-)-medicarpin, (-)-melilotocarpan E, millepurpan, tricin, and chrysoeriol showed cytotoxic effects in P388 as well as P388/DOX cells. These results demonstrate that alfalfa leaf extract may have interesting potential in cancer chemoprevention and therapy.

  14. Effects of compost organic amendments on chemical composition and in vitro digestibility of alfalfa (Medicago sativa L.

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    Francesco Montemurro

    2010-01-01

    Full Text Available The following fertiliser treatments were compared during the years 2002 and 2003 on alfalfa forage (Medicago sativa L.: compost obtained from the organic fraction of the Municipal Solid Waste (MSW; olive pomace compost (OPC; mineral fertiliser (Min. All the treatments allowed a distribution of 75kg ha-1 of P2O5. Three cuttings occurred: at 168, 206 and 351 days after compost application (DAA in 2002; 119, 152 and 320 DAA in 2003. Cumulative biomass and dry matter yields were measured during each experimental year. Furthermore, chemical composition and in vitro digestibility of dry matter (DMd, organic matter (OMd, crude protein (CPd and NDF (NDFd were determined. MSW treatment showed a significantly (P<0.01 higher content of ADL than OPC and Min (77.0, 66.0 and 65.0g kg-1 DM, respectively. Fertiliser treatments also affected (P<0.01 digestibility parameters. In fact, DMd and OMd values showed the same trend with lower percentages in MSW treatment than in the OPC and Min ones. The NDFd differed in all treatments having the highest value in OPC (40.1%. The results indicated that the soil distribution of organic materials offer the possibility to reduce the application of mineral fertilisers and production costs without decreasing alfalfa yield, forage chemical composition and in vitro digestibility.

  15. Soil bulk electrical resistivity and forage ground cover: nonlinear models in an alfalfa (Medicago sativa L. case study

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    Roberta Rossi

    2015-12-01

    Full Text Available Alfalfa is a highly productive and fertility-building forage crop; its performance, can be highly variable as influenced by within-field soil spatial variability. Characterising the relations between soil and forage- variation is important for optimal management. The aim of this work was to model the relationship between soil electrical resistivity (ER and plant productivity in an alfalfa (Medicago sativa L. field in Southern Italy. ER mapping was accomplished by a multi-depth automatic resistivity profiler. Plant productivity was assessed through normalised difference vegetation index (NDVI at 2 dates. A non-linear relationship between NDVI and deep soil ER was modelled within the framework of generalised additive models. The best model explained 70% of the total variability. Soil profiles at six locations selected along a gradient of ER showed differences related to texture (ranging from clay to sandy-clay loam, gravel content (0 to 55% and to the presence of a petrocalcic horizon. Our results prove that multi-depth ER can be used to localise permanent soil features that drive plant productivity.

  16. The arbuscular mycorrhizal fungus Glomus mosseae gives contradictory effects on phosphorus and arsenic acquisition by Medicago sativa Linn

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    Chen Baodong [Department of Soil Environmental Sciences/State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085 (China); Xiao Xueyi [Department of Soil Environmental Sciences/State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085 (China); Zhu Yongguan [Department of Soil Environmental Sciences/State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085 (China)]. E-mail: ygzhu@rcees.ac.en; Smith, F. Andrew [Soil and Land Systems, School of Earth and Environmental Sciences, Waite Campus, University of Adelaide, South Australia 5005 (Australia); Miao Xie, Z. [Department of Environmental Science and Engineering, Hangzhou Dianzi University, Hangzhou 310018 (China); Smith, Sally E. [Soil and Land Systems, School of Earth and Environmental Sciences, Waite Campus, University of Adelaide, South Australia 5005 (Australia)

    2007-07-01

    Mycorrhizal fungi may play an important role in protecting plants against arsenic (As) contamination. However, little is known about the direct and indirect involvement of arbuscular mycorrhizal fungi (AMF) in detoxification mechanisms. A compartmented pot cultivation system ('cross-pots') is used here to investigate the roles of AMF Glomus mosseae in plant phosphorus (P) and As acquisition by Medicago sativa, and P-As interactions. The results indicate that fungal colonization dramatically increased plant dry weight by a factor of around 6, and also substantially increased both plant P and As contents (i.e. total uptake). Irrespective of P and As addition levels, AM plants had shoot and root P concentrations 2 fold higher, but As concentrations significantly lower, than corresponding uninoculated controls. The decreased shoot As concentrations were largely due to 'dilution effects' that resulted from stimulated growth of AM plants and reduced As partitioning to shoots. The study provides further evidence for the protective effects of AMF on host plants against As contamination, and have uncovered key aspects of underlying mechanisms. The possible application of AMF in remediation practices is discussed.

  17. WATER DEFICIT EFFECT ON YIELD AND FORAGE QUALITY OF MEDICAGO SATIVA POPULATIONS UNDER FIELD CONDITIONS IN MARRAKESH AREA (MOROCCO

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    Mohamed FARISSI

    2014-06-01

    Full Text Available The present study focused the effect of water deficit on agronomic potential and some traits related to forage quality in plants of Moroccan Alfalfa (Medicago sativa L. populations (Taf 1, Taf 2, Dem and Tata originated from Oasis and High Atlas of Morocco and an introduced variety from Australia (Siriver. The experiment was conducted under field conditions in experimental station of INRA-Marrakech and under two irrigation treatments. The first treatment was normal irrigation, providing an amount of water corresponding to the potential evapo-transpiration of the crop, and the second treatment was water deficit stress (one irrigation per cut. For each treatment, the experiment was conducted as a split plot based on a randomized complete block design with four replications. The plants were measured and analyzed over three cuts. Some agronomic traits as, plant height, fresh and dry forage yields were measured. The forage quality was evaluated by leaf:stem ratio and the contents of plants in proteins and nitrogen. The results indicated that the water deficit has negatively affected the plant height and forage yield. The decrease in leaf:stem ratio was observed under water deficit conditions. However, the proteins and nitrogen contents were unaffected. The behavior of tested alfalfa genotypes was significantly different. The Moroccan alfalfa populations were more adapted to water deficit conditions comparatively to Siriver variety and the Tata population was the most adapted one.

  18. Proteomic analysis provides insights into the molecular bases of hydrogen gas-induced cadmium resistance in Medicago sativa.

    Science.gov (United States)

    Dai, Chen; Cui, Weiti; Pan, Jincheng; Xie, Yanjie; Wang, Jin; Shen, Wenbiao

    2017-01-30

    Recently, molecular hydrogen (H2) has emerged as a bio-regulator both in animals and plants. Normally, functions of endogenous generated H2 could be mimicked by exogenously applied hydrogen-rich water (HRW) or hydrogen-rich saline (particularly in animals). Although alfalfa seedlings showed more cadmium (Cd) resistance after the administration with HRW, corresponding molecular mechanism is still elusive. To address this gap, iTRAQ-based quantitative proteomics was used. The results showed that a total of 2377 proteins were identified with Medicago sativa is a widely used forage around the world. Recently, hydrogen gas (H2) was suggested as a candidate of signal molecule, and found to effectively attenuate Cd-induced damage in alfalfa seedlings. However, the underlying molecular mechanism still needs to be further elucidated. In the present work, an iTRAQ-based quantitative proteomics was firstly carried out, and the results revealed the main molecular targets and metabolic processes associated with Cd resistance conferred by H2. This study may expand our understanding of hydrogen gas-medicated heavy metal tolerance in plants. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Allelopathic Potential of Switchgrass ( Panicum virgatum L.) on Perennial Ryegrass ( Lolium perenne L.) and Alfalfa ( Medicago sativa L.)

    Science.gov (United States)

    Shui, Junfeng; An, Yu; Ma, Yongqing; Ichizen, Nobumasa

    2010-10-01

    This study investigated allelopathy and its chemical basis in nine switchgrass ( Panicum virgatum L.) accessions. Perennial ryegrass ( Lolium perenne L.) and alfalfa ( Medicago sativa L.) were used as test species. Undiluted aqueous extracts (5 g plant tissue in 50 ml water) from the shoots and roots of most of the switchgrass accessions inhibited the germination and growth of the test species. However, the allelopathic effect of switchgrass declined when extracts were diluted 5- or 50-fold. Seedling growth was more sensitive than seed germination as an indicator of allelopathic effect. Allelopathic effect was related to switchgrass ecotype but not related to ploidy level. Upland accessions displayed stronger allelopathic potential than lowland accessions. The aqueous extract from one switchgrass accession was separated into phenols, organic acids, neutral chemicals, and alkaloids, and then these fractions were bioassayed to test for allelopathic potential. Alkaloids had the strongest allelopathic effect among the four chemical fractions. In summary, the results indicated that switchgrass has allelopathic potential; however, there is not enough evidence to conclude that allelopathic advantage is the main factor that has contributed to the successful establishment of switchgrass on China’s Loess Plateau.

  20. P-HYDROXYPHENYLPYRUVATE DIOXYGENASE from Medicago sativa is involved in vitamin E biosynthesis and abscisic acid-mediated seed germination

    Science.gov (United States)

    Jiang, Jishan; Chen, Zhihong; Ban, Liping; Wu, Yudi; Huang, Jianping; Chu, Jinfang; Fang, Shuang; Wang, Zan; Gao, Hongwen; Wang, Xuemin

    2017-01-01

    P-HYDROXYPHENYLPYRUVATE DIOXYGENASE (HPPD) is the first committed enzyme involved in the biosynthesis of vitamin E, and is characterized by catalyzing the conversion of p-hydroxyphenyl pyruvate (HPP) to homogentisic acid (HGA). Here, an HPPD gene was cloned from Medicago sativa L. and designated MsHPPD, which was expressed at high levels in alfalfa leaves. PEG 6000 (polyethylene glycol), NaCl, abscisic acid and salicylic acid were shown to significantly induce MsHPPD expression, especially in the cotyledons and root tissues. Overexpression of MsHPPD was found to significantly increase the level of β-tocotrienol and the total vitamin E content in Arabidopsis seeds. Furthermore, these transgenic Arabidopsis seeds exhibited an accelerated germination time, compared with wild-type seeds under normal conditions, as well as under NaCl and ABA treatments. Meanwhile, the expression level of several genes associated with ABA biosynthesis (NCED3, NCED5 and NCED9) and the ABA signaling pathway (RAB18, ABI3 and ABI5) were significantly down-regulated in MsHPPD-overexpressing transgenic lines, as well as the total free ABA content. Taken together, these results demonstrate that MsHPPD functions not only in the vitamin E biosynthetic pathway, but also plays a critical role in seed germination via affecting ABA biosynthesis and signaling. PMID:28084442

  1. Salicornia europaea L. Na⁺/H⁺ antiporter gene improves salt tolerance in transgenic alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Zhang, L Q; Niu, Y D; Huridu, H; Hao, J F; Qi, Z; Hasi, A

    2014-07-24

    In order to obtain a salt-tolerant perennial alfalfa (Medicago sativa L.), we transferred the halophyte Salicornia europaea L. Na(+)/H(+) antiporter gene, SeNHX1, to alfalfa by using the Agrobacterium-mediated transformation method. The transformants were confirmed by both PCR and RT-PCR analyses. Of 197 plants that were obtained after transformation, 36 were positive by PCR analysis using 2 primer pairs for the CaMV35S-SeNHX1 and SeNHX1-Nos fragments; 6 plants survived in a greenhouse. RT-PCR analysis revealed that SeNHX1 was expressed in 5 plants. The resultant transgenic alfalfa had better salt tolerance. After stress treatment for 21 days with 0.6% NaCl, the chlorophyll and MDA contents in transgenic plants were lower, but proline content and SOD, POD, and CAT activities were higher than those in wild-type plants. These results suggest that the salt tolerance of transgenic alfalfa was improved by the overexpression of the SeNHX1 gene.

  2. Deep-sequencing transcriptome analysis of field-grown Medicago sativa L. crown buds acclimated to freezing stress.

    Science.gov (United States)

    Song, Lili; Jiang, Lin; Chen, Yue; Shu, Yongjun; Bai, Yan; Guo, Changhong

    2016-09-01

    Medicago sativa L. (alfalfa) 'Zhaodong' is an important forage legume that can safely survive in northern China where winter temperatures reach as low as -30 °C. Survival of alfalfa following freezing stress depends on the amount and revival ability of crown buds. In order to investigate the molecular mechanisms of frost tolerance in alfalfa, we used transcriptome sequencing technology and bioinformatics strategies to analyze crown buds of field-grown alfalfa during winter. We statistically identified a total of 5605 differentially expressed genes (DEGs) involved in freezing stress including 1900 upregulated and 3705 downregulated DEGs. We validated 36 candidate DEGs using qPCR to confirm the accuracy of the RNA-seq data. Unlike other recent studies, this study employed alfalfa plants grown in the natural environment. Our results indicate that not only the CBF orthologs but also membrane proteins, hormone signal transduction pathways, and ubiquitin-mediated proteolysis pathways indicate the presence of a special freezing adaptation mechanism in alfalfa. The antioxidant defense system may rapidly confer freezing tolerance to alfalfa. Importantly, biosynthesis of secondary metabolites and phenylalanine metabolism, which is of potential importance in coordinating freezing tolerance with growth and development, were downregulated in subzero temperatures. The adaptive mechanism for frost tolerance is a complex multigenic process that is not well understood. This systematic analysis provided an in-depth view of stress tolerance mechanisms in alfalfa.

  3. Involvement of abscisic acid in the response of Medicago sativa plants in symbiosis with Sinorhizobium meliloti to salinity.

    Science.gov (United States)

    Palma, F; López-Gómez, M; Tejera, N A; Lluch, C

    2014-06-01

    Legumes are classified as salt-sensitive crops with their productivity particularly affected by salinity. Abcisic acid (ABA) plays an important role in the response to environmental stresses as signal molecule which led us to study its role in the response of nitrogen fixation and antioxidant metabolism in root nodules of Medicago sativa under salt stress conditions. Adult plants inoculated with Sinorhizobium meliloti were treated with 1 μM and 10 μM ABA two days before 200 mM salt addition. Exogenous ABA together with the salt treatment provoked a strong induction of the ABA content in the nodular tissue which alleviated the inhibition induced by salinity in the plant growth and nitrogen fixation. Antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR) were induced by ABA pre-treatments under salt stress conditions which together with the reduction of the lipid peroxidation, suggest a role for ABA as signal molecule in the activation of the nodular antioxidant metabolism. Interaction between ABA and polyamines (PAs), described as anti-stress molecules, was studied being detected an induction of the common polyamines spermidine (Spd) and spermine (Spm) levels by ABA under salt stress conditions. In conclusion, ABA pre-treatment improved the nitrogen fixation capacity under salt stress conditions by the induction of the nodular antioxidant defenses which may be mediated by the common PAs Spd and Spm that seems to be involved in the anti-stress response induced by ABA. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  4. Influence of papermill sludge on growth of Medicago sativa, Festuca rubra and Agropyron trachycaulum in gold mine tailings: A greenhouse study

    Energy Technology Data Exchange (ETDEWEB)

    Green, Scott [Department of Botany, University of Manitoba, 505 Buller building, Winnipeg, Manitoba R3T 2N2 (Canada); Renault, Sylvie [Department of Botany, University of Manitoba, 505 Buller building, Winnipeg, Manitoba R3T 2N2 (Canada)], E-mail: renaults@cc.umanitoba.ca

    2008-02-15

    A greenhouse study was undertaken to determine the suitability of adding papermill sludge to neutral/alkaline gold mine tailings to improve the establishment of Festuca rubra, Agropyron trachycaulum and Medicago sativa. Festuca rubra root and shoot biomass and A. Trachycaulum shoot biomass were increased with papermill sludge amendment. The addition of papermill sludge and fertilizer drastically increased the shoot and root biomass of M. sativa (20-30 times) while A. trachycaulum and F. rubra showed a more moderate increase in growth. Photosynthetic pigment content of the leaves was higher in papermill sludge treatments than in the treatments without papermill sludge. The organic carbon content, macro-aggregate content and field capacity of the gold mine tailings were increased while the bulk density was decreased by the addition of papermill sludge. This study suggests that addition of papermill sludge and adequate fertilization can alleviate some of the adverse conditions of neutral/alkaline gold mine tailings. - Addition of papermill sludge and adequate fertilization of neutral gold mine tailings increased growth of Medicago sativa, Festuca rubra and Agropyron trachycaulum.

  5. Evaluation of microbial biomass C and N content of the soils cultivated with vetch (Vicia sativa L. and alfalfa (Medicago sativa L.

    Directory of Open Access Journals (Sweden)

    İlyas Bolat

    2016-01-01

    Full Text Available Legume forage crops have the ability of retaining free nitrogen in the air through symbiotic Rhizobium bacteria found in their roots. Additionally, microbial biomass (MB–an essential living component of soil and a significant factor influencing plant nutrient dynamics–is considered to be accurate indicator of soil’s biological condition. Given the aforementioned aspects, soil MB C (Cmic and MB N (Nmic of different legume forage crops were investigated in this study. Soil samples were taken in order to identify certain physical and chemical characteristics of the soil using volume cylinders (0 – 6.5 cm depth from Vicia sativa L. (VSP and Medicago sativa L. planted (MSP areas. To determine the Cmic and Nmic contents, topsoil samples were also taken from 0 – 6.5 cm depth. Cmic and Nmic contents were identified using chloroform – fumigation – extraction method. There was no statistical significance for particle density, bulk density, electrical conductivity, CaCO3 %, and decomposition ratio (Corg/Ntotal of the VSP and MSP soil (P > 0.05. However, some other soil characteristics such as temperature, porosity, sand, silt and clay contents, pH, organic C and total N differed significantly (P < 0.05. Compared to VSP soil, the Cmic contents were determined to be 27 % higher (P < 0.05 in MSP soil. In VSP soil, the soil Nmic content ranged from 83.38 µg g-1 to 124.67 µg g-1, while it ranged from 91.62 µg g-1 to 187.07 µg g-1 in MSP soil. The Nmic content of the MSP soil was observed to be approximately 35 % higher than VSP soil, and a statistically significant difference (P < 0.05 was noticed between the two. Moreover, a significant positive correlation was found not only between the Cmic and organic C contents (r = 0.667; P < 0.05 but also between the Nmic and total N contents of MSP and VSP soil (r = 0.881; P < 0.01. The results of the study revealed that soil Cmic and Nmic values differ as the types of planted legume forage crops

  6. Comparative transcriptome investigation of global gene expression changes caused by miR156 overexpression in Medicago sativa.

    Science.gov (United States)

    Gao, Ruimin; Austin, Ryan S; Amyot, Lisa; Hannoufa, Abdelali

    2016-08-19

    Medicago sativa (alfalfa) is a low-input forage and potential bioenergy crop, and improving its yield and quality has always been a focus of the alfalfa breeding industry. Transgenic alfalfa plants overexpressing a precursor of alfalfa microRNA156 (MsmiR156) were recently generated by our group. These plants (miR156OE) showed enhanced biomass yield, reduced internodal length, increased shoot branching and trichome density, and a delay in flowering time. Transcripts of three SQUAMOSA-PROMOTER BINDING PROTEIN-LIKE (SPL) genes (MsSPL6, MsSPL12, and MsSPL13) were found to be targeted for cleavage by MsmiR156 in alfalfa. To further illustrate the molecular mechanisms underlying the effects of miR156 in alfalfa, two miR156OE genotypes (A11a and A17) were subjected to Next Generation RNA Sequencing with Illumina HiSeq. More than 1.11 billion clean reads were obtained from our available sequenced samples. A total of 160,472 transcripts were generated using Trinity de novo assembly and 4,985 significantly differentially expressed genes were detected in miR156OE plants A11a and A17 using the Medicago truncatula genome as reference. A total of 17 genes (including upregulated, downregulated, and unchanged) were selected for quantitative real-time PCR (qRT-PCR) validation, which showed that gene expression levels were largely consistent between qRT-PCR and RNA-Seq data. In addition to the established SPL genes MsSPL6, MsSPL12 and MsSPL13, four new SPLs; MsSPL2, MsSPL3, MsSPL4 and MsSPL9 were also down-regulated significantly in both miR156OE plants. These seven SPL genes belong to genes phylogeny clades VI, IV, VIII, V and VII, which have been reported to be targeted by miR156 in Arabidopsis thaliana. The gene ontology terms characterized electron transporter, starch synthase activity, sucrose transport, sucrose-phosphate synthase activity, chitin binding, sexual reproduction, flavonoid biosynthesis and lignin catabolism correlate well to the phenotypes of miR156OE alfalfa

  7. Fotossíntese em alfafa (Medicago sativa L. sob supressão e ressuprimento de fosfato Photosynthesis in alfalfa (Medicago sativa L. under phosphate suppression and ressuply

    Directory of Open Access Journals (Sweden)

    Fernando Teixeira Gomes

    2003-10-01

    Full Text Available Neste estudo, foram avaliados os efeitos da supressão e do ressuprimento de fosfato (Pi sobre a fotossíntese e eficiência fotoquímica de plantas de Medicago sativa cv. Flórida 77, em diferentes estádios do desenvolvimento vegetativo (V3, V4 e reprodutivo (R6, R8. O ensaio foi conduzido em casa de vegetação e as plantas cultivadas na solução nutritiva de HOAGLAND & ARNON (1950, contendo 0,14mmol L-1 de Pi. A supressão de Pi por dez dias reduziu os teores de fósforo nas folhas amostradas, em todos os estádios do desenvolvimento. Entretanto, com o ressuprimento, somente nos estádios vegetativos, os valores foram semelhantes ao tratamento controle. A fotossíntese por área foliar, em todos os estádios do desenvolvimento diminuiu com a supressão de Pi. De modo geral, o ressuprimento de Pi à solução nutritiva resultou em recuperação na fotossíntese, excetuando-se as plantas no estádio V3, uma indicação de que o período de supressão não causou danos permanentes no aparato fotossintético. Os teores dos pigmentos fotossintéticos e a eficiência fotoquímica do fotossistema II (FS II, avaliada pela relação Fv/Fm, não foram alterados quando as plantas foram submetidas à supressão de Pi. Esse resultado demonstra que o transporte de elétrons através do FS II não limitou a fotossíntese nas folhas amostradas, sob supressão de Pi, sugerindo que a supressão causou efeito mais pronunciado na etapa bioquímica da fotossíntese.This work evaluated the phosphate (Pi suppression and ressupply on photosynthesis and photochemical efficiency from Medicago sativa plants cv. Florida 77, in different growth stages (V3 and V4 and reproductive (R6 and R8. The experiment was performed in a greenhouse, the plants being cultivated in HOAGLAND & ARMOND (1950 nutritive solution containing 0,14mmol L-1 of Pi. Pi suppression for ten days reduced Pi levels in sampled leaves, in all growth stages. However, with the re-supply only in the

  8. Avaliação de fontes de amônia para conservação do feno de alfafa (Medicago sativa L. armazenado com alta umidade Ammonia sources to preserve high moisture alfalfa (Medicago sativa L. hay

    Directory of Open Access Journals (Sweden)

    Djalma de Freitas

    2002-04-01

    Full Text Available O estudo foi desenvolvido com o objetivo de avaliar duas fontes de amônia (amônia anidra-NH3 ou uréia para conservação do feno de alfafa (Medicago sativa L. armazenado com alta umidade. Foram estudados os seguintes tratamentos: A - feno com 12 a 15% de umidade e não-tratado; B - feno com 24 a 27% de umidade e tratado com 1,0% de NH3 na MS; C - feno com 24 a 27% de umidade e tratado com 0,9% de uréia na MS; D - feno com 24 a 27% de umidade e tratado com 1,8% de uréia na MS; E - feno com 34 a 37% de umidade e tratado com 0,9% de uréia na MS; e F - feno com 34 a 37% de umidade e tratado com 1,8% de uréia na MS. Os fenos permaneceram sob lona plástica, hermeticamente fechada, por 60 dias. Foram realizadas amostragens para identificação de fungos nos fenos, aos 0 e 60 dias pós-tratamento, e determinação da composição química, avaliando-se os teores de proteína bruta (PB e dos constituintes da parede celular. Nas quantidades testadas, somente a NH3 foi eficiente no controle dos fungos. Nos tratamentos com uréia, apesar de haver controle dos gêneros Aspergillus e Penicillium, os demais gêneros presentes foram suficientes para deterioração dos fenos. Merece destaque o gênero Paecilomyces, que apresentou alta incidência em todos os fenos tratados. A quantidade utilizada de NH3 foi insuficiente para promover mudanças significativas na composição química dos fenos, exceto nos teores de PB, que aumentaram com o uso de 1,0% de NH3, quando comparados com o não-tratado.The research was conducted to evaluate two sources of ammonia (anhydrous ammonia-NH3 or urea to preserve alfalfa (Medicago sativa L. hay stored with high moisture. The following treatments were studied: A - Control: untreated hay (12 to 15% moisture; B - hay (24 to 27% moisture treated with NH3 (1.0% of DM; C - hay (24 to 27% moisture treated with urea (0.9% DM; D - hay (24 to 27% moisture treated with urea (1.8% DM; E - hay (34 to 37% moisture treated with urea

  9. EFECTO ANTIBACTERIANO in vitro DEL EXTRACTO ALCOHÓLICO DE Psidium guajava (guayaba) y Medicago sativa (alfalfa) SOBRE Streptococcus mutans ATCC 25175

    OpenAIRE

    Chero Nepo, Diego Armando; Universidad Señor de Sipán; Ruiz Barrueto, Miguel Ángel; Universidad Señor de Sipán

    2016-01-01

    Objetivo: Determinar el efecto de antibacteriano in vitro del extracto alcohólico de Psidium guajava y Medicago sativa sobre Streptococcus mutans ATCC 25175. Material y métodos: Investigación cuantitativa con diseño experimental de estímulo creciente. Se emplearon 20 concentraciones volumétricas de cada extracto de 1 - 20 mg/ml, un control positivo que fue clorhexidina al 0,12% y el control negativo que fue etanol absoluto. Para evaluar el efecto antibacteriano de ambos extractos alcohólicos ...

  10. Molecular cloning and characterization of a gene regulating flowering time from Alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Zhang, Tiejun; Chao, Yuehui; Kang, Junmei; Ding, Wang; Yang, Qingchuan

    2013-07-01

    Genes that regulate flowering time play crucial roles in plant development and biomass formation. Based on the cDNA sequence of Medicago truncatula (accession no. AY690425), the LFY gene of alfalfa was cloned. Sequence similarity analysis revealed high homology with FLO/LFY family genes of other plants. When fused to the green fluorescent protein, MsLFY protein was localized in the nucleus of onion (Allium cepa L.) epidermal cells. The RT-qPCR analysis of MsLFY expression patterns showed that the expression of MsLFY gene was at a low level in roots, stems, leaves and pods, and the expression level in floral buds was the highest. The expression of MsLFY was induced by GA3 and long photoperiod. Plant expression vector was constructed and transformed into Arabidopsis by the agrobacterium-mediated methods. PCR amplification with the transgenic Arabidopsis genome DNA indicated that MsLFY gene had integrated in Arabidopsis genome. Overexpression of MsLFY specifically caused early flowering under long day conditions compared with non-transgenic plants. These results indicated MsLFY played roles in promoting flowering time.

  11. [Symbiosis between the nodule bacterium Sinorhizobium meliloti and alfalfa (Medicago sativa) under salinization conditions].

    Science.gov (United States)

    Ibragimova, M V; Rumiantseva, M L; Onishchuk, O P; Belova, V S; Kurchak, O N; Andronov, E E; Dziubenko, N I; Simarov, B V

    2006-01-01

    Two hundred forty-three isolates of alfalfa nodule bacteria (Sinorhizobium meliloti) were obtained from legume nodules and soils sampled in the northern Aral region, experiencing secondary salinization. Isolates obtained from nodules (N isolates) were significantly more salt-tolerant than those from soils (S isolates) when grown in a liquid medium with 3.5% NaCl. It was found that wild species of alfalfa, melilot, and trigonella preferably formed symbioses with salt-tolerant nodule bacteria in both salinized and nonsalinized soils. Only two alfalfa species, Medicago falcata and M. trautvetteri, formed efficient symbioses in soils contrasting in salinity. The formation of efficient symbiosis with alfalfa in the presence of 0.6% NaCl was studied in 36 isolates (N and S) differing in salt tolerance and symbiotic efficiency. Fifteen isolates formed efficient symbioses in the presence of salt. The increase in the dry weight of the plants was 25-68% higher than in the control group. The efficiency of symbiotic interaction under salinization conditions depended on the efficiency of the isolates under standard conditions but did not correlate with the source of nodule bacteria (soil or nodule) or their salt tolerance. The results indicate that nodule bacterium strains forming efficient symbioses under salinization conditions can be found.

  12. Isolation and characterization of a gene from Medicago sativa L., encoding a bZIP transcription factor.

    Science.gov (United States)

    Li, Yan; Sun, Yan; Yang, Qingchuan; Fang, Feng; Kang, Junmei; Zhang, Tiejun

    2013-02-01

    A full-length cDNA of 1,537 nucleotides was cloned from Medicago sativa L. cv. "Zhongmu No. 1" by rapid amplification of cDNA ends. It was designated as MsZIP, encoding a protein of 340 amino acids. The protein molecular weight was 36.43 kDa, and the theoretical isoelectric point was 5.72. The MsZIP preferentially localized in nucleus and have signal peptide. Blast analysis revealed that MsZIP shared the highest homology with some bZIP proteins of M. truncatula. The transcript of MsZIP was strongly enriched in leaf compared with root and stem of mature alfalfa plants. MsZIP was strongly induced by 15 % PEG6000 (polyethylene glycol), 50 μM abscisic acid, 200 mM NaCl, 70 μM gibberellic acid, 5 mM salicylic acid and 200 μM methyl jasmonate. Physiological resistance parameters were measured in the transgenic tobacco. Malondialdehyde content, relative water content, soluble sugar content, soluble protein content and proline content in transgenic tobacco increased compared with non-transgenic tobacco under salt stress or drought stress. The results showed that accumulation of the MsZIP protein in the vegetative tissues of transgenic plants enhanced their tolerance to osmotic pressure stress. These results demonstrate a role for the MsZIP protein in stress protection and suggest the potential of the MsZIP gene for genetic engineering of salt tolerance and drought tolerance.

  13. Paenibacillus medicaginis sp. nov. a chitinolytic endophyte isolated from a root nodule of alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Lai, Wei-An; Hameed, Asif; Lin, Shih-Yao; Hung, Mei-Hua; Hsu, Yi-Han; Liu, You-Cheng; Shahina, Mariyan; Shen, Fo-Ting; Young, Chiu-Chung

    2015-11-01

    A Gram-stain-variable, short-rod-shaped, endospore-forming, strictly aerobic, non-motile, chitinolytic and endophytic bacterium, designated strain CC-Alfalfa-19T, exhibiting unusual bipolar appendages was isolated from a root nodule of alfalfa (Medicago sativa L.) in Taiwan and subjected to a polyphasic taxonomic study. Based on 16S rRNA gene sequence analysis, strain CC-Alfalfa-19T was found to be most closely related to Paenibacillus puldeungensis CAU 9324T (95.2 %), whereas other species of the genus Paenibacillus shared ≤ 95.0 % sequence similarity. The phylogenetic analysis revealed a distinct phyletic lineage established by strain CC-Alfalfa-19T with respect to other species of the genus Paenibacillus. Fatty acids comprised predominantly anteiso-C15 : 0, C16 : 0, anteiso-C17 : 0 and iso-C16 : 0. Menaquinone 7 (MK-7) was identified as the sole respiratory quinone and the genomic DNA G+C content was 42.7 mol%. Polar lipids included diphosphatidylglycerol, phosphatidylglycerol, phosphatidylmonomethylethanolamine, phosphatidylethanolamine, an unidentified glycolipid and an unidentified lipid. The diagnostic diamino acid found in the cell-wall peptidoglycan was meso-diaminopimelic acid. Based on the polyphasic taxonomic evidence that was in line with the genus Paenibacillus and additional distinguishing characteristics, strain CC-Alfalfa-19T is considered to represent a novel species, for which the name Paenibacillus medicaginis sp. nov. (type strain CC-Alfalfa-19T = BCRC 80441T = JCM 18446T) is proposed.

  14. Low-fiber alfalfa (Medicago sativa L.) meal in the laying hen diet: effects on productive traits and egg quality.

    Science.gov (United States)

    Laudadio, V; Ceci, E; Lastella, N M B; Introna, M; Tufarelli, V

    2014-07-01

    This study was designed to determine the effects on laying performance and egg quality resulting from partial substitution of soybean meal (SBM) with low-fiber alfalfa (LFA; Medicago sativa L.) meal in the diet of early-phase laying hens. ISA Brown layers, 18 wk of age, were randomly allocated to 2 dietary treatments and fed for 10 wk. The hens were fed 2 wheat middling-based diets: a control diet, which contained SBM (15% of diet), and a test diet containing LFA (15% of diet) as the main protein source. Low-fiber alfalfa meal was obtained by a combination of sieving and air-classification processes. Feed intake was recorded daily, and egg production was calculated on a hen-day basis; eggs from each group were weekly collected to evaluate egg components and quality. The partial substitution of SBM with LFA had no adverse effect on growth performance of early-phase laying hens. Egg production and none of the egg-quality traits examined were influenced by dietary treatment, except for yolk color (P < 0.001) and yolk percentage (P < 0.05) as well as yolk cholesterol and β-carotene contents (P < 0.001), which were improved in hens fed the LFA diet. Including LFA increased serum β-carotene and reduced serum cholesterol concentrations (P < 0.001). Our results suggest that partially replacing conventional SBM as protein source with low-fiber alfalfa meal in the laying-hen diet can positively influence yolk quality without adversely affecting productive traits. © 2014 Poultry Science Association Inc.

  15. Arbuscular mycorrhizal mediation of biomass-density relationship of Medicago sativa L. under two water conditions in a field experiment.

    Science.gov (United States)

    Zhang, Qian; Xu, Liming; Tang, Jianjun; Bai, Minge; Chen, Xin

    2011-05-01

    The biomass-density relationship (whereby the biomass of individual plants decreases as plant density increases) has generally been explained by competition for resources. Arbuscular mycorrhizal fungi (AMF) are able to affect plant interactions by mediating resource utilization, but whether this AMF-mediated interaction will change the biomass-density relationship is unclear. We conducted an experiment to test the hypothesis that AMF will shift the biomass-density relationship by affecting intraspecific competition. Four population densities (10, 100, 1,000, or 10,000 seedlings per square meter) of Medicago sativa L. were planted in field plots. Water application (1,435 or 327.7 mm/year) simulated precipitation in wet areas (sufficient water) and arid areas (insufficient water). The fungicide benomyl was applied to suppress AMF in some plots ("low-AMF" treatment) and not in others ("high-AMF" treatment). The effect of the AMF treatment on the biomass-density relationship depended on water conditions. High AMF enhanced the decrease of individual biomass with increasing density (the biomass-density line had a steeper slope) when water was sufficient but not when water was insufficient. AMF treatment did not affect plant survival rate or population size but did affect absolute competition intensity (ACI). When water was sufficient, ACI was significantly higher in the high-AMF treatment than in the low-AMF treatment, but ACI was unaffected by AMF treatment when water was insufficient. Our results suggest that AMF status did not impact survival rate and population size but did shift the biomass-density relationship via effects on intraspecific competition. This effect of AMF on the biomass-density relationship depended on the availability of water.

  16. Distribution of hydrogen-metabolizing bacteria in alfalfa field soil. [Medicago sativa L. ; Convolvulus arvensis L. ; Rhizobium meliloti

    Energy Technology Data Exchange (ETDEWEB)

    Cunningham, S.D.; Kapulnik, Y.; Phillips, D.A.

    1986-11-01

    H/sub 2/ evolved by alfalfa root nodules during the process of N/sub 2/ fixation may be an important factor influencing the distribution of soil bacteria. To test this hypothesis under field conditions, over 700 bacterial isolates were obtained from fallow soil or from the 3-mm layer of soil surrounding alfalfa (Medicago sativa L.) root nodules, alfalfa roots, or bindweed (Convolvulus arvensis L.) roots. Bacteria were isolated under either aerobic or microaerophilic conditions and were tested for their capacity to metabolize H/sub 2/. Isolates showing net H/sub 2/ uptake and /sup 3/H/sub 2/ incorporation activity under laboratory conditions were assigned a Hup/sup +/ phenotype, whereas organisms with significant H/sub 2/ output capacity were designated as a Hout/sup +/ phenotype. Under aerobic isolation conditions two Hup/sup +/ isolates were obtained, whereas under microaerophilic conditions five Hup/sup +/ and two Hout/sup +/ isolates were found. The nine isolates differed on the basis of 24 standard bacteriological characteristics or fatty acid composition. Five of the nine organisms were isolated from soil around root nodules, whereas the other four were found distributed among the other three soil environments. On the basis of the microaerophilic isolations, 4.8% of the total procaryotic isolates from soil around root nodules were capable of oxidizing H/sub 2/, and 1.2% could produce H/sub 2/. Two of the Hup/sup +/ isolates were identified as Rhizobium meliloti by root nodulation tests, but the fact that none of the isolates reduced C/sub 2/H/sub 2/ under the assay conditions suggested that the H/sub 2/ metabolism traits were associated with various hydrogenase systems rather than with nitrogenase activity.

  17. Antisense inhibition of NADH glutamate synthase impairs carbon/nitrogen assimilation in nodules of alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Cordoba, Elizabeth; Shishkova, Svetlana; Vance, Carroll P; Hernández, Georgina

    2003-03-01

    Legumes acquire significant amounts of nitrogen for growth from symbiotic nitrogen fixation. The glutamine synthetase (GS)/NADH-dependent glutamate synthase (NADH-GOGAT) cycle catalyzes initial nitrogen assimilation. This report describes the impact of specifically reducing nodule NADH-GOGAT activity on symbiotic performance of alfalfa (Medicago sativa L.). Four independent transgenic alfalfa lines, designated GA89, GA87, GA88, and GA82 (for GOGATantisense), containing an antisense NADH-GOGAT cDNA fragment under the control of the soybean leghemoglobin (lbc3) promoter were evaluated. The GA plants were fertile and showed normal growth in non-symbiotic conditions. The NADH-GOGAT antisense transgene was heritable and the T1 plants showed phenotypic alterations - similar to primary transformants. Clonally propagated plants were inoculated with Sinorhizobium meliloti after rooting and the symbiotic phenotype was analyzed 21 days post-inoculation. Nodules of each GA line had reduced NADH-GOGAT activity, ranging from 33 to 87% of control plants, that was accompanied by comparable decreases in RNA and protein. Plants from the GA89 line, with the lowest NADH-GOGAT activity (c. 30%), presented a strikingly altered symbiotic phenotype: concomitantly activities of key enzyme for carbon and nitrogen assimilation decreased; nodule amino acids and amides were reduced while sucrose accumulated. Antisense GOGAT plants were chlorotic, reduced in fresh weight, and had a lower N content than control plants. Photosynthesis was also impaired in antisense plants. Specifically, reducing NADH-GOGAT in nodules resulted in plants having impaired nitrogen assimilation and altered carbon/nitrogen metabolic flux.

  18. Contrasting impacts of defoliation on root colonization by arbuscular mycorrhizal and dark septate endophytic fungi of Medicago sativa.

    Science.gov (United States)

    Saravesi, K; Ruotsalainen, A L; Cahill, J F

    2014-05-01

    Individual plants typically interact with multiple mutualists and enemies simultaneously. Plant roots encounter both arbuscular mycorrhizal (AM) and dark septate endophytic (DSE) fungi, while the leaves are exposed to herbivores. AMF are usually beneficial symbionts, while the functional role of DSE is largely unknown. Leaf herbivory may have a negative effect on root symbiotic fungi due to decreased carbon availability. However, evidence for this is ambiguous and no inoculation-based experiment on joint effects of herbivory on AM and DSE has been done to date. We investigated how artificial defoliation impacts root colonization by AM (Glomus intraradices) and DSE (Phialocephala fortinii) fungi and growth of Medicago sativa host in a factorial laboratory experiment. Defoliation affected fungi differentially, causing a decrease in arbuscular colonization and a slight increase in DSE-type colonization. However, the presence of one fungal species had no effect on colonization by the other or on plant growth. Defoliation reduced plant biomass, with this effect independent of the fungal treatments. Inoculation by either fungal species reduced root/shoot ratios, with this effect independent of the defoliation treatments. These results suggest AM colonization is limited by host carbon availability, while DSE may benefit from root dieback or exudation associated with defoliation. Reductions in root allocation associated with fungal inoculation combined with a lack of effect of fungi on plant biomass suggest DSE and AMF may be functional equivalent to the plant within this study. Combined, our results indicate different controls of colonization, but no apparent functional consequences between AM and DSE association in plant roots in this experimental setup.

  19. Resistant and susceptible responses in alfalfa (Medicago sativa) to bacterial stem blight caused by Pseudomonas syringae pv. syringae.

    Science.gov (United States)

    Nemchinov, Lev G; Shao, Jonathan; Lee, Maya N; Postnikova, Olga A; Samac, Deborah A

    2017-01-01

    Bacterial stem blight caused by Pseudomonas syringae pv. syringae is a common disease of alfalfa (Medicago sativa L). Little is known about host-pathogen interactions and host defense mechanisms. Here, individual resistant and susceptible plants were selected from cultivars Maverick and ZG9830 and used for transcript profiling at 24 and 72 hours after inoculation (hai) with the isolate PssALF3. Bioinformatic analysis revealed a number of differentially expressed genes (DEGs) in resistant and susceptible genotypes. Although resistant plants from each cultivar produced a hypersensitive response, transcriptome analyses indicated that they respond differently at the molecular level. The number of DEGs was higher in resistant plants of ZG9830 at 24 hai than in Maverick, suggesting that ZG9830 plants had a more rapid effector triggered immune response. Unique up-regulated genes in resistant ZG9830 plants included genes encoding putative nematode resistance HSPRO2-like proteins, orthologs for the rice Xa21 and soybean Rpg1-b resistance genes, and TIR-containing R genes lacking both NBS and LRR domains. The suite of R genes up-regulated in resistant Maverick plants had an over-representation of R genes in the CC-NBS-LRR family including two genes for atypical CCR domains and a putative ortholog of the Arabidopsis RPM1 gene. Resistance in both cultivars appears to be mediated primarily by WRKY family transcription factors and expression of genes involved in protein phosphorylation, regulation of transcription, defense response including synthesis of isoflavonoids, and oxidation-reduction processes. These results will further the identification of mechanisms involved in resistance to facilitate selection of parent populations and development of commercial varieties.

  20. Resistant and susceptible responses in alfalfa (Medicago sativa to bacterial stem blight caused by Pseudomonas syringae pv. syringae.

    Directory of Open Access Journals (Sweden)

    Lev G Nemchinov

    Full Text Available Bacterial stem blight caused by Pseudomonas syringae pv. syringae is a common disease of alfalfa (Medicago sativa L. Little is known about host-pathogen interactions and host defense mechanisms. Here, individual resistant and susceptible plants were selected from cultivars Maverick and ZG9830 and used for transcript profiling at 24 and 72 hours after inoculation (hai with the isolate PssALF3. Bioinformatic analysis revealed a number of differentially expressed genes (DEGs in resistant and susceptible genotypes. Although resistant plants from each cultivar produced a hypersensitive response, transcriptome analyses indicated that they respond differently at the molecular level. The number of DEGs was higher in resistant plants of ZG9830 at 24 hai than in Maverick, suggesting that ZG9830 plants had a more rapid effector triggered immune response. Unique up-regulated genes in resistant ZG9830 plants included genes encoding putative nematode resistance HSPRO2-like proteins, orthologs for the rice Xa21 and soybean Rpg1-b resistance genes, and TIR-containing R genes lacking both NBS and LRR domains. The suite of R genes up-regulated in resistant Maverick plants had an over-representation of R genes in the CC-NBS-LRR family including two genes for atypical CCR domains and a putative ortholog of the Arabidopsis RPM1 gene. Resistance in both cultivars appears to be mediated primarily by WRKY family transcription factors and expression of genes involved in protein phosphorylation, regulation of transcription, defense response including synthesis of isoflavonoids, and oxidation-reduction processes. These results will further the identification of mechanisms involved in resistance to facilitate selection of parent populations and development of commercial varieties.

  1. Identification of transcriptome involved in atrazine detoxification and degradation in alfalfa (Medicago sativa) exposed to realistic environmental contamination.

    Science.gov (United States)

    Zhang, Jing Jing; Lu, Yi Chen; Zhang, Shu Hao; Lu, Feng Fan; Yang, Hong

    2016-08-01

    Plants are constantly exposed to a variety of toxic compounds (or xenobiotics) such as pesticides (or herbicides). Atrazine (ATZ) as herbicide has become one of the environmental contaminants due to its intensive use during crop production. Plants have evolved strategies to cope with the adverse impact of ATZ. However, the mechanism for ATZ degradation and detoxification in plants is largely unknown. Here we employed a global RNA-sequencing (RNA-Seq) strategy to dissect transcriptome variation in alfalfa (Medicago sativa) exposed to ATZ. Four libraries were constructed including Root-ATZ (root control, ATZ-free), Shoot-ATZ, Root+ATZ (root treated with ATZ) and Shoot+ATZ. Hierarchical clustering was performed to display the expression patterns for all differentially expressed genes (DEGs) under ATZ exposure. Transcripts involved in ATZ detoxification, stress responses (e.g. oxidation and reduction, conjugation and hydrolytic reactions), and regulations of cysteine biosynthesis were identified. Several genes encoding glycosyltransferases, glutathione S-transferases or ABC transporters were up-regulated notably. Also, many other genes involved in oxidation-reduction, conjugation, and hydrolysis for herbicide degradation were differentially expressed. These results suggest that ATZ in alfalfa can be detoxified or degraded through different pathways. The expression patterns of some DEGs by high-throughput sequencing were well confirmed by qRT-PCR. Our results not only highlight the transcriptional complexity in alfalfa exposed to ATZ but represent a major improvement for analyzing transcriptional changes on a large scale as well. Copyright © 2016 Elsevier Inc. All rights reserved.

  2. Up-regulation of heme oxygenase-1 contributes to the amelioration of aluminum-induced oxidative stress in Medicago sativa.

    Science.gov (United States)

    Cui, Weiti; Zhang, Jing; Xuan, Wei; Xie, Yanjie

    2013-10-15

    In this report, pharmacological, histochemical and molecular approaches were used to investigate the effect of heme oxygenase-1 (HO-1) up-regulation on the alleviation of aluminum (Al)-induced oxidative stress in Medicago sativa. Exposure of alfalfa to AlCl3 (0-100 μM) resulted in a dose-dependent inhibition of root elongation as well as the enhancement of thiobarbituric acid reactive substances (TBARS) content. 1 and 10 μM (in particular) Al(3+) increased alfalfa HO-1 transcript or its protein level, and HO activity in comparison with the decreased changes in 100 μM Al-treated samples. After recuperation, however, TBARS levels in 1 and 10 μM Al-treated alfalfa roots returned to control values, which were accompanied with the higher levels of HO activity. Subsequently, exogenous CO, a byproduct of HO-1, could substitute for the cytoprotective effects of the up-regulation of HO-1 in alfalfa plants upon Al stress, which was confirmed by the alleviation of TBARS and Al accumulation, as well as the histochemical analysis of lipid peroxidation and loss of plasma membrane integrity. Theses results indicated that endogenous CO generated via heme degradation by HO-1 could contribute in a critical manner to its protective effects. Additionally, the pretreatments of butylated hydroxytoluene (BHT) and hemin, an inducer of HO-1, exhibited the similar cytoprotective roles in the alleviation of oxidative stress, both of which were impaired by the potent inhibitor of HO-1, zinc protoporphyrin IX (ZnPP). However, the Al-induced inhibition of root elongation was not influenced by CO, BHT and hemin, respectively. Together, the present results showed up-regulation of HO-1 expression could act as a mechanism of cell protection against oxidative stress induced by Al treatment. Copyright © 2013 Elsevier GmbH. All rights reserved.

  3. Ectopic Expression of GsPPCK3 and SCMRP in Medicago sativa Enhances Plant Alkaline Stress Tolerance and Methionine Content

    Science.gov (United States)

    Zhao, Yang; Zhao, Chaoyue; DuanMu, Huizi; Yu, Yang; Ji, Wei; Zhu, Yanming

    2014-01-01

    So far, it has been suggested that phosphoenolpyruvate carboxylases (PEPCs) and PEPC kinases (PPCKs) fulfill several important non-photosynthetic functions. However, the biological functions of soybean PPCKs, especially in alkali stress response, are not yet well known. In previous studies, we constructed a Glycine soja transcriptional profile, and identified three PPCK genes (GsPPCK1, GsPPCK2 and GsPPCK3) as potential alkali stress responsive genes. In this study, we confirmed the induced expression of GsPPCK3 under alkali stress and investigated its tissue expression specificity by using quantitative real-time PCR analysis. Then we ectopically expressed GsPPCK3 in Medicago sativa and found that GsPPCK3 overexpression improved plant alkali tolerance, as evidenced by lower levels of relative ion leakage and MDA content and higher levels of chlorophyll content and root activity. In this respect, we further co-transformed the GsPPCK3 and SCMRP genes into alfalfa, and demonstrated the increased alkali tolerance of GsPPCK3-SCMRP transgenic lines. Further investigation revealed that GsPPCK3-SCMRP co-overexpression promoted the PEPC activity, net photosynthetic rate and citric acid content of transgenic alfalfa under alkali stress. Moreover, we also observed the up-regulated expression of PEPC, CS (citrate synthase), H+-ATPase and NADP-ME genes in GsPPCK3-SCMRP transgenic alfalfa under alkali stress. As expected, we demonstrated that GsPPCK3-SCMRP transgenic lines displayed higher methionine content than wild type alfalfa. Taken together, results presented in this study supported the positive role of GsPPCK3 in plant response to alkali stress, and provided an effective way to simultaneously improve plant alkaline tolerance and methionine content, at least in legume crops. PMID:24586886

  4. Transgenic alfalfa (Medicago sativa) with increased sucrose phosphate synthase activity shows enhanced growth when grown under N2-fixing conditions.

    Science.gov (United States)

    Gebril, Sayed; Seger, Mark; Villanueva, Fabiola Muro; Ortega, Jose Luis; Bagga, Suman; Sengupta-Gopalan, Champa

    2015-10-01

    Overexpression of SPS in alfalfa is accompanied by early flowering, increased plant growth and an increase in elemental N and protein content when grown under N2-fixing conditions. Sucrose phosphate synthase (SPS; EC 2.3.1.14) is the key enzyme in the synthesis of sucrose in plants. The outcome of overexpression of SPS in different plants using transgenic approaches has been quite varied, but the general consensus is that increased SPS activity is associated with the production of new sinks and increased sink strength. In legumes, the root nodule is a strong C sink and in this study our objective was to see how increasing SPS activity in a legume would affect nodule number and function. Here we have transformed alfalfa (Medicago sativa, cv. Regen SY), with a maize SPS gene driven by the constitutive CaMV35S promoter. Our results showed that overexpression of SPS in alfalfa, is accompanied by an increase in nodule number and mass and an overall increase in nitrogenase activity at the whole plant level. The nodules exhibited an increase in the level of key enzymes contributing to N assimilation including glutamine synthetase and asparagine synthetase. Moreover, the stems of the transformants showed higher level of the transport amino acids, Asx, indicating increased export of N from the nodules. The transformants exhibited a dramatic increase in growth both of the shoots and roots, and earlier flowering time, leading to increased yields. Moreover, the transformants showed an increase in elemental N and protein content. The overall conclusion is that increased SPS activity improves the N status and plant performance, suggesting that the availability of more C in the form of sucrose enhances N acquisition and assimilation in the nodules.

  5. Responses of lucerne (Medicago sativa L.) and rhizobia to copper-based fungicide application in two contrasting soils

    Science.gov (United States)

    Schneider, Martin; Dober, Melanie; Jöchlinger, Lisa; Keiblinger, Katharina; Soja, Gerhard; Mentler, Axel; Zechmeister-Boltenstern, Sophie; Bruckner, Alexander; Golestani Fard, Alireza; Wenzel, Walter; Zehetner, Franz

    2016-04-01

    For more than 120 years, salts of copper (Cu) have been used in viticulture to prevent damages by fungal diseases. Due to restrictions in the use of synthetic fungicides and mineral fertilizers, organic viticulture depends on Cu as well as on biological nitrogen fixation. Here, we conducted an eco-toxicological pot experiment with an acidic, sandy soil and a calcareous, loamy soil and incrementally increasing fungicide application rates from 0 to 5000 mg Cu kg-1 soil. Lucerne (Medicago sativa L. cultivar. Plato) was grown in the pots for 3 months under greenhouse conditions. Acetylene reduction assays performed with harvested nodules showed no response to elevated soil Cu concentrations indicating that the nitrogen fixing capacity of rhizobia was not compromised by Cu in our experiment. Nevertheless, the nodule biomass was very sensitive to Cu and strongly decreased due to reduced amounts of fine roots and less energy supply by the plant. Legumes are known to be Cu-sensitive, and our contribution also showed a decrease in harvest by 50 % (EC50) at 21 mg kg-1 plant Cu tissue concentration in the acidic soil and at 30 mg kg-1 in the calcareous soil. This corresponded to diffusional fluxes measured by diffusive gradients in thin films (DGT) of 202 and 368 fmol cm-2 s-1, respectively. DGT measurements showed that in the acidic soil, Cu was 2 to 10 times more available for plants, depending on the concentration applied, than in the calcareous soil. A modeling approach for estimating the effective concentration (EC) by including the DGT-estimated plant Cu content and the pH produced more accurate values (NRMSE of 21.9 to 20.1 %) than EC directly estimated from DGT.

  6. Arbuscular mycorrhizal colonization alters subcellular distribution and chemical forms of cadmium in Medicago sativa L. and resists cadmium toxicity.

    Science.gov (United States)

    Wang, Yuanpeng; Huang, Jing; Gao, Yanzheng

    2012-01-01

    Some plants can tolerate and even detoxify soils contaminated with heavy metals. This detoxification ability may depend on what chemical forms of metals are taken up by plants and how the plants distribute the toxins in their tissues. This, in turn, may have an important impact on phytoremediation. We investigated the impact of arbuscular mycorrhizal (AM) fungus, Glomus intraradices, on the subcellular distribution and chemical forms of cadmium (Cd) in alfalfa (Medicago sativa L.) that were grown in Cd-added soils. The fungus significantly colonized alfalfa roots by day 25 after planting. Colonization of alfalfa by G. intraradices in soils contaminated with Cd ranged from 17% to 69% after 25-60 days and then decreased to 43%. The biomass of plant shoots with AM fungi showed significant 1.7-fold increases compared to no AM fungi addition under the treatment of 20 mg kg(-1) Cd. Concentrations of Cd in the shoots of alfalfa under 0.5, 5, and 20 mgkg(-1) Cd without AM fungal inoculation are 1.87, 2.92, and 2.38 times higher, respectively, than those of fungi-inoculated plants. Fungal inoculation increased Cd (37.2-80.5%) in the cell walls of roots and shoots and decreased in membranes after 80 days of incubation compared to untreated plants. The proportion of the inactive forms of Cd in roots was higher in fungi-treated plants than in controls. Furthermore, although fungi-treated plants had less overall Cd in subcellular fragments in shoots, they had more inactive Cd in shoots than did control plants. These results provide a basis for further research on plant-microbe symbioses in soils contaminated with heavy metals, which may potentially help us develop management regimes for phytoremediation.

  7. MsZEP, a novel zeaxanthin epoxidase gene from alfalfa (Medicago sativa), confers drought and salt tolerance in transgenic tobacco.

    Science.gov (United States)

    Zhang, Zhiqiang; Wang, Yafang; Chang, Leqin; Zhang, Tong; An, Jie; Liu, Yushi; Cao, Yuman; Zhao, Xia; Sha, Xuyang; Hu, Tianming; Yang, Peizhi

    2016-02-01

    The zeaxanthin epoxidase gene ( MsZEP ) was cloned and characterized from alfalfa and validated for its function of tolerance toward drought and salt stresses by heterologous expression in Nicotiana tabacum. Zeaxanthin epoxidase (ZEP) plays important roles in plant response to various environment stresses due to its functions in ABA biosynthetic and the xanthophyll cycle. To understand the expression characteristics and the biological functions of ZEP in alfalfa (Medicago sativa), a novel gene, designated as MsZEP (KM044311), was cloned, characterized and overexpressed in Nicotiana tabacum. The open reading frame of MsZEP contains 1992 bp nucleotides and encodes a 663-amino acid polypeptide. Amino acid sequence alignment indicated that deduced MsZEP protein was highly homologous to other plant ZEP sequences. Phylogenetic analysis showed that MsZEP was grouped into a branch with other legume plants. Real-time quantitative PCR revealed that MsZEP gene expression was clearly tissue-specific, and the expression levels were higher in green tissues (leaves and stems) than in roots. MsZEP expression decreased in shoots under drought, cold, heat and ABA treatment, while the expression levels in roots showed different trends. Besides, the results showed that nodules could up-regulate the MsZEP expression under non-stressful conditions and in the earlier stage of different abiotic stress. Heterologous expression of the MsZEP gene in N. tabacum could confer tolerance to drought and salt stress by affecting various physiological pathways, ABA levels and stress-responsive genes expression. Taken together, these results suggested that the MsZEP gene may be involved in alfalfa responses to different abiotic stresses and nodules, and could enhance drought and salt tolerance of transgenic tobacco by heterologous expression.

  8. Altered callose deposition during embryo sac formation of multi-pistil mutant (mp1) in Medicago sativa.

    Science.gov (United States)

    Zhou, H C; Jin, L; Li, J; Wang, X J

    2016-06-03

    Whether callose deposition is the cause or result of ovule sterility in Medicago sativa remains controversial, because it is unclear when and where changes in callose deposition and dissolution occur during fertile and sterile embryo sac formation. Here, alfalfa spontaneous multi-pistil mutant (mp1) and wild-type plants were used to compare the dynamics of callose deposition during embryo sac formation using microscopy. The results showed that both mutant and wild-type plants experienced megasporogenesis and megagametogenesis, and there was no significant difference during megasporogenesis. In contrast to the wild-type plants, in which the mature embryo sac was observed after three continuous cycles of mitosis, functional megaspores of mutant plants developed abnormally after the second round of mitosis, leading to degeneration of synergid, central, and antipodal cells. Callose deposition in both mutant and wild-type plants was first observed in the walls of megasporocytes, and then in the megaspore tetrad walls. After meiosis, the callose wall began to degrade as the functional megaspore underwent mitosis, and almost no callose was observed in the mature embryo sac in wild-type plants. However, callose deposition was observed in mp1 plants around the synergid, and increased with the development of the embryo sac, and was mainly deposited at the micropylar end. Our results indicate that synergid, central, and antipodal cells, which are surrounded by callose, may degrade owing to lack of nutrition. Callose accumulation around the synergid and at the micropylar end may hinder signals required for the pollen tube to enter the embryo sac, leading to abortion.

  9. Effect of arbuscular mycorrhizal fungi (AMF) on heavy metal tolerance of alfalfa (Medicago sativa L.) and oat (Avena sativa L.) on a sewage-sludge treated soil; Bedeutung der arbuskulaeren Mykorrhiza (AM) fuer die Schwermetalltoleranz von Luzerne (Medicago sativa L.) und Hafer (Avena sativa L.) auf einem klaerschlammgeduengten Boden

    Energy Technology Data Exchange (ETDEWEB)

    Ricken, B. [Institut fuer Pflanzenernaehrung, Giessen Univ. (Germany); Hoefner, W. [Institut fuer Pflanzenernaehrung, Giessen Univ. (Germany)

    1996-10-01

    In pot experiments with a sewage sludge treated soil, the influence of two arbuscular mycorrhizal fungi (AMF) isolates of Glomus sp. (T6 and D13) on plant growth and on the uptake of heavy metals by alfalfa (Medicago sativa L.) and oat (Avena sativa L.) was investigated. Alfalfa showed an increase of biomass with mycorrhizal infection only to a small extent. In oat AMF inoculation increased the growth of both root and shoot by up to 70% and 55% respectively. Mycorrhization raised the P-content and -uptake in alfalfa, but not in oat, in both roots and shoots. Mycorrhizal alfalfa showed lower Zn-, Cd- and Ni-contents and uptake in roots and shoots. The root length was significantly decreased in mycorrhizal alfalfa plants (up to 38%). The translocation of heavy metals into the shoot of mycorrhizal alfalfa was slightly increased. Mycorrhizal infection of oat led to higher concentrations of Zn, Cd and Ni in the root but to less Zn in the shoot. The translocation of heavy metals to the oat shoot was clearely decreased by mycorrhizal colonisation. This may be based on the ability of fungal tissues to complex heavy metals at the cell walls, thus excluding metals from the shoot. This conclusion is supported by the enhanced root length (up to 78%) of mycorrhizal oat plants in this experiment. The mycorrhizal infection seemed to protect plants against heavy metal pollution in soils. It was obvious that different host plants reacted in different ways. (orig.) [Deutsch] In Gefaessversuchen mit einem klaerschlammgeduengten Boden wurde der Einfluss der arbuskulaeren Mykorrhiza (AM)-Pilzisolate von Glomus sp. (T6 und D13) auf Pflanzenwachstum und Schwermetall (SM)-aufnahme von Luzerne (Medicago sativa L.) und Hafer (Avena sativa L.) untersucht. Das Wachstum von Luzerne wurde durch eine Mykorrhizierung nicht signifikant beeinflusst. Bei Hafer foerderte eine AM-Inokulation mit T6 das Wachstum von Wurzel und Spross bis zu 70% bzw. 55%, zur Reife aber ergab sich gleicher Sprossertrag

  10. Effect of Medicago sativa L. and compost on organic and inorganic pollutant removal from a mixed contaminated soil and risk assessment using ecotoxicological tests.

    Science.gov (United States)

    Marchand, Charlotte; Hogland, William; Kaczala, Fabio; Jani, Yahya; Marchand, Lilian; Augustsson, Anna; Hijri, Mohamed

    2016-11-01

    Several Gentle Remediation Options (GRO), e.g., plant-based options (phytoremediation), singly and combined with soil amendments, can be simultaneously efficient for degrading organic pollutants and either stabilizing or extracting trace elements (TEs). Here, a 5-month greenhouse trial was performed to test the efficiency of Medicago sativa L., singly and combined with a compost addition (30% w/w), to treat soils contaminated by petroleum hydrocarbons (PHC), Co and Pb collected at an auto scrap yard. After 5 months, total soil Pb significantly decreased in the compost-amended soil planted with M. sativa, but not total soil Co. Compost incorporation into the soil promoted PHC degradation, M. sativa growth and survival, and shoot Pb concentrations [3.8 mg kg(-1) dry weight (DW)]. Residual risk assessment after the phytoremediation trial showed a positive effect of compost amendment on plant growth and earthworm development. The O2 uptake by soil microorganisms was lower in the compost-amended soil, suggesting a decrease in microbial activity. This study underlined the benefits of the phytoremediation option based on M. sativa cultivation and compost amendment for remediating PHC- and Pb-contaminated soils.

  11. Cloning and characterization of a heme oxygenase-2 gene from alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Fu, Guang-Qing; Jin, Qi-Jiang; Lin, Yu-Ting; Feng, Jian-Fei; Nie, Li; Shen, Wen-Biao; Zheng, Tian-Qing

    2011-11-01

    Heme oxygenase (HO, EC 1.14.99.3) catalyzes the oxidation of heme and performs vital roles in plant development and stress responses. Two HO isozymes exist in plants. Between these, HO-1 is an oxidative stress-response protein, and HO-2 usually exhibited constitutive expression. Although alfalfa HO-1 gene (MsHO1) has been investigated previously, HO2 is still poorly understood. In this study, we report the cloning and characterization of HO2 gene, MsHO2, from alfalfa (Medica sativa L.). The full-length cDNA of MsHO2 contains an ORF of 870 bp and encodes for 290 amino acid residues with a predicted molecular mass of 33.3 kDa. Similar to MsHO1, MsHO2 also appears to have an N-terminal transit peptide sequence for chloroplast import. Many conserved residues in plant HO were also conserved in MsHO2. However, unlike HO-1, the conserved histidine (His) required for heme-iron binding and HO activity was replaced by tyrosine (Tyr) in MsHO2. Further biochemical activity analysis of purified mature MsHO2 showed no HO activity, suggesting that MsHO2 may not be a true HO in nature. Semi-quantitative RT-PCR confirmed its maximum expression in the germinating seeds. Importantly, the expression levels of MsHO2 were up-regulated under sodium nitroprusside (SNP) and H(2)O(2) (especially) treatment, respectively.

  12. Influence of atmospheric [CO2] on growth, carbon allocation and cost of plant tissues on leaf nitrogen concentration maintenance in nodulated Medicago sativa

    Science.gov (United States)

    Pereyra, Gabriela; Hartmann, Henrik; Ziegler, Waldemar; Michalzik, Beate; Gonzalez-Meler, Miquel; Trumbore, Susan

    2015-04-01

    Plant carbon (C) allocation and plant metabolic processes (i.e. photosynthesis and respiration) can be affected by changes in C availability, for example from changing atmospheric [CO2]. In nodulated plants, C availability may also influence nitrogen (N) fixation by bacteriods. But C allocation and N fixation are often studied independently and hence do not allow elucidating interactive effects. We investigated how different atmospheric [CO2] (Pleistocene: 170 ppm, ambient: 400 ppm and projected future: 700 ppm) influence plant growth, allocation to nodules, and the ratio of photosynthesis-to-respiration (R:A) as an indicator of C cost in Medicago sativa inoculated with Ensifer meliloti. M. sativa grew c. 38% more nodules at 400 ppm and 700 ppm than at 170 ppm. However, ratios of above- and belowground plant biomass to nodule biomass were constant over time and independent of atmospheric [CO2]. Total non-structural carbohydrate concentrations were not significantly different between plants grown at 400 and 700 ppm, but were four to five-fold higher than in 170 ppm plants. Leaf level N concentration was similar across treatments, but N-based photosynthetic rates were 82% and 93% higher in leaves of plants grown at 400 and 700 ppm, respectively, than plants grown at 170 ppm. In addition, leaf R:A was greater (48% or 55%) in plants grown at 170 ppm than plants grown at 400 and 700 ppm. Similarly, the greatest proportion of assimilated CO2 released by root respiration occurred in rhizobial plants growing at 170 ppm. Our results suggest that C limitation in nodulated Medicago sativa plants did not influence C allocation to nodule biomass but caused a proportionally greater allocation of C to belowground respiration, most likely to bacteriods. This suggests that N tissue concentration was maintained at low [CO2] by revving up bacteriod metabolism and at the expense of non-structural carbohydrate reserves.

  13. Contrasting Storage Protein Synthesis and Messenger RNA Accumulation during Development of Zygotic and Somatic Embryos of Alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Krochko, J E; Pramanik, S K; Bewley, J D

    1992-05-01

    During development on hormone-free media, somatic embryos pass through distinct morphological stages that superficially resemble those of zygotic embryo development (globular, heart, torpedo, cotyledonary stages). Despite these similarities, they differ from zygotic embryos in the extent of cotyledonary development and the patterns of synthesis and quantitative expression of seed-specific storage proteins (7S, 11S, and 2S proteins). Alfin (7S) is the first storage protein synthesized in developing zygotic embryos (stage IV). The 11S (medicagin) and 2S (Low Molecular Weight, LMW) storage proteins are not detectable until the following stage of development (stage V), although all three are present before the completion of embryo enlargement. Likewise, the 7S storage protein is the first to be synthesized in developing somatic embryos (day 5). Medicagin is evident by day 7 and the LMW protein by day 10. In contrast to zygotic embryos, alfin remains the predominant storage protein in somatic embryos throughout development. Not only are the relative amounts of medicagin and the LMW protein reduced in somatic embryos but the LMW protein is accumulated much later than the other proteins. Quantification of the storage protein mRNAs (7S, 11S, and 2S) by northern blot analysis confirms that there are substantial differences in the patterns of message accumulation in zygotic and somatic embryos of alfalfa (Medicago sativa). In zygotic embryos, the 7S, 11S, and 2S storage protein mRNAs are abundant during maturation and, in particular, during the stages of maximum protein synthesis (alfin, stages VI and VII; medicagin, stage VII; LMW, stage VII). In somatic embryos, the predominance of the 7S storage protein is correlated with increased accumulation of its mRNA, whereas the limited synthesis of the 11S storage protein is associated with much lower steady-state levels of its message. The mRNA for the LMW protein is present already by 3 days after transfer to hormone-free media

  14. Nodulation by Sinorhizobium meliloti originated from a mining soil alleviates Cd toxicity and increases Cd-phytoextraction in Medicago sativa L.

    Science.gov (United States)

    Ghnaya, Tahar; Mnassri, Majda; Ghabriche, Rim; Wali, Mariem; Poschenrieder, Charlotte; Lutts, Stanley; Abdelly, Chedly

    2015-01-01

    Besides their role in nitrogen supply to the host plants as a result of symbiotic N fixation, the association between legumes and Rhizobium could be useful for the rehabilitation of metal-contaminated soils by phytoextraction. A major limitation presents the metal-sensitivity of the bacterial strains. The aim of this work was to explore the usefulness of Sinorhizobium meliloti originated from a mining site for Cd phytoextraction by Medicago sativa. Inoculated and non-inoculated plants were cultivated for 60 d on soils containing 50 and/or 100 mg Cd kg(-1) soil. The inoculation hindered the occurrence of Cd- induced toxicity symptoms that appeared in the shoots of non-inoculated plants. This positive effect of S. meliloti colonization was accompanied by an increase in biomass production and improved nutrient acquisition comparatively to non-inoculated plants. Nodulation enhanced Cd absorption by the roots and Cd translocation to the shoots. The increase of plant biomass concomitantly with the increase of Cd shoot concentration in inoculated plants led to higher potential of Cd-phytoextraction in these plants. In the presence of 50 mg Cd kg(-1) in the soil, the amounts of Cd extracted in the shoots were 58 and 178 μg plant(-1) in non-inoculated and inoculated plants, respectively. This study demonstrates that this association M. sativa-S. meliloti may be an efficient biological system to extract Cd from contaminated soils.

  15. Nodulation by Sinorhizobium meliloti originated from a mining soil alleviates Cd toxicity and increases Cd-phytoextraction in Medicago sativa L.

    Directory of Open Access Journals (Sweden)

    Tahar eGhnaya

    2015-10-01

    Full Text Available Besides their role in nitrogen supply to the host plants by the process of symbiotic N fixation, the association between legumes and Rhizobium could be interesting also for the rehabilitation of metal contaminated soils by phytoextraction. A major limitation is however the metal sensitivity of the bacterial strains. The aim of this work was to explore the usefulness of Sinorhizobium meliloti originated from a mining site in phytoextraction of Cd by Medicago sativa. Inoculated and non-inoculated plants were cultivated for 60 d on soils containing 50 or 100 mg Cd kg-1 soil. The inoculation hindered the occurrence of Cd- induced toxicity symptoms that appeared in the shoots of non-inoculated plants. This positive effect of S.meliloti colonization was accompanied by an increase in biomass production and improved nutrient acquisition as compared to non-inoculated plants. Moreover, nodulation enhanced both Cd absorption by the roots and Cd transloaction to the shoots. The amelioration of plant biomass concomitantly with the increase in Cd shoot concentration in inoculated plants led to higher potential of Cd-phytoextraction in these plants. Hence, in the presence of 50 mg Cd kg-1 in the soil, the quantities of Cd extracted in the shoots were 58 and 178 µg plant-1 in non-inoculated and inoculated plants, respectively. Thus we suggest that this association M. sativa-S. meliloti is an efficient biological system to extract Cd.

  16. Nodulation by Sinorhizobium meliloti originated from a mining soil alleviates Cd toxicity and increases Cd-phytoextraction in Medicago sativa L.

    Science.gov (United States)

    Ghnaya, Tahar; Mnassri, Majda; Ghabriche, Rim; Wali, Mariem; Poschenrieder, Charlotte; Lutts, Stanley; Abdelly, Chedly

    2015-01-01

    Besides their role in nitrogen supply to the host plants as a result of symbiotic N fixation, the association between legumes and Rhizobium could be useful for the rehabilitation of metal-contaminated soils by phytoextraction. A major limitation presents the metal-sensitivity of the bacterial strains. The aim of this work was to explore the usefulness of Sinorhizobium meliloti originated from a mining site for Cd phytoextraction by Medicago sativa. Inoculated and non-inoculated plants were cultivated for 60 d on soils containing 50 and/or 100 mg Cd kg−1 soil. The inoculation hindered the occurrence of Cd- induced toxicity symptoms that appeared in the shoots of non-inoculated plants. This positive effect of S. meliloti colonization was accompanied by an increase in biomass production and improved nutrient acquisition comparatively to non-inoculated plants. Nodulation enhanced Cd absorption by the roots and Cd translocation to the shoots. The increase of plant biomass concomitantly with the increase of Cd shoot concentration in inoculated plants led to higher potential of Cd-phytoextraction in these plants. In the presence of 50 mg Cd kg−1 in the soil, the amounts of Cd extracted in the shoots were 58 and 178 μg plant−1 in non-inoculated and inoculated plants, respectively. This study demonstrates that this association M. sativa-S. meliloti may be an efficient biological system to extract Cd from contaminated soils. PMID:26528320

  17. Electrophoretic and Immunological Comparisons of Soluble Root Proteins of Medicago sativa L. Genotypes in the Cold Hardened and Non-Hardened Condition 1

    Science.gov (United States)

    Coleman, E. A.; Bula, R. J.; Davis, R. L.

    1966-01-01

    Electrophoretic and immunological properties of the soluble root protein complement of 6 Medicago sativa L. genotypes in the cold hardened and non-hardened physiological condition were compared. These 6 genotypes were chosen to represent a range of abilities to survive exposure to subfreezing temperatures when in the cold hardened condition. A zone of highly charged and/or low molecular weight protein components were found to be more prevalent in the protein complements of the cold-hardened material than the non-hardened material. Immunodiffusion plate tests were not so definitive as the electrophoretic patterns for identifying the genotypes or physiological conditions, but did corroborate the electrophoretic interpretations. Images Fig. 2 PMID:16656458

  18. Effects of Cold Hardening on the Regulation of Polyamine Levels in Wheat (Triticum aestivum L.) and Alfalfa (Medicago sativa L.) 1

    Science.gov (United States)

    Nadeau, Paul; Delaney, Sandra; Chouinard, Lucette

    1987-01-01

    When leaves of wheat (Triticum aestivum L.) are exposed to a cold hardening temperature, a major accumulation of putrescine (6-9 times) takes place. Spermidine accumulates to a lesser extent and, conversely, spermine decreases slightly. These variations are completely reversible when plants are returned to initial growing conditions. A similar response is obtained with crowns. During cold hardening, arginine decarboxylase activity remains near its initial level while a considerable loss of activity is observed in control plants. Ornithine decarboxylase and diamine oxidase activity levels are not substantially modified by the treatment. Alfalfa (Medicago sativa L.) also accumulates putrescine under low temperature stress, indicating that this phenomenon is not typical of cereals. The physiological significance of this accumulation of putrescine is still unexplained but the results obtained suggest the involvement of polyamines in the biochemical processes of cold hardening. PMID:16665409

  19. A PROPOSITO DE UN ENSAYO CON ABONOS (cal y harina de huesos EN ALFALFAl (Medicago sativai

    Directory of Open Access Journals (Sweden)

    Isidoro Mogilner

    2010-11-01

    Full Text Available

    En este trabajo, los autores estudian el efecto que sobre el rendimiento de la alfalfa (Medicago sativa produce la incorporación de diferentes dosis de cal y harina de huesos, en un suelo pobre de P y Ca; de una textura pesada con alto contenido de arcilla en todos los horizontes, de un pH ácido (5.1 a 6.2 y que es característico de la zona donde fué hecha la experiencia.
    Se han obtenido resultados referentes a la influencia de la inoculación de la semilla con Rhizobium y el agregado de cal y harina de huesos sobre los rendimientos.

  20. Comunidad de arañas (Arachnida, Araneae del cultivo de alfalfa (Medicago sativa en Buenos Aires, Argentina

    Directory of Open Access Journals (Sweden)

    Andrea Armendano

    2010-06-01

    Full Text Available En las últimas décadas se ha dado un interés creciente en el uso de enemigos naturales para controlar plagas de insectos, como arañas. Se estudió una comunidad de arañas en Argentina mediante un muestreo cada dos semanas durante el periodo 2004-2006 en lotes de una hectárea. En el estrato del suelo las arañas fueron colectadas con redes de arrastre y trampas de caída. Se recolecto un total de 6 229 ejemplares (15 familias y 50 especies. Siete familias se encuentran en el estrato herbáceo, las más abundantes fueron: Thomisidae (n=2 012, 32.30%, Araneidae (n=1 516, 24.33% y Oxyopidae (n=604, 9.70%. El suelo habían 14 familias, principalmente: Lycosidae (n=629, 10.10% y Linyphiidae (n=427, 6.85%. Predominaron las arañas cazadoras: por emboscadas (32.99%, al acecho (11.77%, corredoras de suelo (10.84% y tejedoras orbiculares (27.56%. Los índices de diversidad fueron: H´=2.97, Dsp=0.11 y J=0.79, evidenciando una comunidad de arañas moderadamente diversa, con predominio de Misumenops pallidus, Oxyopes salticus, Lycosa poliostoma and L. erythrognatha. Las arañas estuvieron presentes durante el desarrollo fenológico del cultivo con picos de abundancia en primavera y verano.Spider community (Arachnida, Araneae of alfalfa crops (Medicago sativa in Buenos Aires, Argentina. Over the last decades there has been an increasing interest in the use of natural enemies to control pest insects, including spiders. We studied a spider community in Argentina by sampling every two weeks during 2004-2006 in one-hectare lots. Soil stratum spiders were collected using nets and pitfall traps. A total of 6229 specimens were collected (15 families and 50 species. Seven families were found in the herbal stratum, the most abundant were Thomisidae (n=2012, 32.30%, Araneidae (n=1516, 24.33% and Oxyopidae (n=604, 9.70%. The soil had 14 families, mainly: Lycosidae (n=629, 10.10% and Linyphiidae (n=427, 6.85%. Hunting spiders predominated: ambushers (32

  1. Decomposition of olive mill waste compost, goat manure and Medicago sativa in Lebanese soils using the litterbag technique

    Science.gov (United States)

    Atallah, Therese

    2014-05-01

    Organic amendments, green manure and plant residues incorporation are the main sources of nutrients in organic farming, their decomposition rate is crucial for the accumulation and long-term storage of organic matter in soils. In this study the decomposition of compost from olive mill waste (N: 29.3 g kg-1; total dissolved nitrogen or TDN: 3.82 g kg-1), goat manure (N: 31.5 g kg-1; TDN: 0.94 g kg-1), the shoots (N: 33.6 g kg-1; TDN: 17.57 g kg-1) and roots (N: 22.12 g kg-1; TDN: 8.87 g kg-1) of Medicago sativa was followed in three Lebanese soils. The nitrogen, phosphorus and potassium released were followed over one year, starting in early winter (December-January). The mild sub-humid Mediterranean conditions allowed a rapid mass loss in alfalfa shoots 30 days after incorporation. Manure and compost were more persistent. Between 80 and 90% of TDN were released, after 30 days of in-situ incubation for compost, the release was over 90% for alfalfa shoots. The movement of P was slower, as the compost (6.99 g kg-1 of P) and manure (9.81 g kg-1 of P) lost 33% and 22%, respectively, during 30 days of incubation. After one year, 15 to 35% of P remained in the soils. The manure was the richest in potassium (19.66 g kg-1) followed by the alfalfa shoots (15.56 g kg-1), the compost (8.19 g kg-1) and the roots (5.96 g kg-1). The loss of potassium was important, as over 88% had disappeared over the year. All decomposition curves followed an exponential model. The calculated coefficients of decomposition for total nitrogen (lnfinal - lninitial/days) were significantly higher for alfalfa shoots (0.00547 day-1) and similar for the compost (0.00184 day-1) and the manure (0.00175 day-1). The ANOVA test showed a difference between two of the sites (Site A: 521 g kg-1 of clay and 42 g kg-1 of calcium carbonate; Site S: 260 g kg-1 of clay and 269 g kg-1 of CaCO3) and the third one (Site L: 315 g kg-1 of clay and 591 g kg-1 of CaCO3). The relationships between the soil calcium

  2. Validation of Genotyping-By-Sequencing Analysis in Populations of Tetraploid Alfalfa by 454 Sequencing

    Science.gov (United States)

    Rocher, Solen; Jean, Martine; Castonguay, Yves; Belzile, François

    2015-01-01

    Genotyping-by-sequencing (GBS) is a relatively low-cost high throughput genotyping technology based on next generation sequencing and is applicable to orphan species with no reference genome. A combination of genome complexity reduction and multiplexing with DNA barcoding provides a simple and affordable way to resolve allelic variation between plant samples or populations. GBS was performed on ApeKI libraries using DNA from 48 genotypes each of two heterogeneous populations of tetraploid alfalfa (Medicago sativa spp. sativa): the synthetic cultivar Apica (ATF0) and a derived population (ATF5) obtained after five cycles of recurrent selection for superior tolerance to freezing (TF). Nearly 400 million reads were obtained from two lanes of an Illumina HiSeq 2000 sequencer and analyzed with the Universal Network-Enabled Analysis Kit (UNEAK) pipeline designed for species with no reference genome. Following the application of whole dataset-level filters, 11,694 single nucleotide polymorphism (SNP) loci were obtained. About 60% had a significant match on the Medicago truncatula syntenic genome. The accuracy of allelic ratios and genotype calls based on GBS data was directly assessed using 454 sequencing on a subset of SNP loci scored in eight plant samples. Sequencing depth in this study was not sufficient for accurate tetraploid allelic dosage, but reliable genotype calls based on diploid allelic dosage were obtained when using additional quality filtering. Principal Component Analysis of SNP loci in plant samples revealed that a small proportion (<5%) of the genetic variability assessed by GBS is able to differentiate ATF0 and ATF5. Our results confirm that analysis of GBS data using UNEAK is a reliable approach for genome-wide discovery of SNP loci in outcrossed polyploids. PMID:26115486

  3. Phytotoxicity of three plant-based biodiesels, unmodified castor oil, and Diesel fuel to alfalfa (Medicago sativa L.), lettuce (Lactuca sativa L.), radish (Raphanus sativus), and wheatgrass (Triticum aestivum).

    Science.gov (United States)

    Bamgbose, Ifeoluwa; Anderson, Todd A

    2015-12-01

    The wide use of plant-based oils and their derivatives, in particular biodiesel, have increased extensively over the past decade to help alleviate demand for petroleum products and improve the greenhouse gas emissions profile of the transportation sector. Biodiesel is regarded as a clean burning alternative fuel produced from livestock feeds and various vegetable oils. Although in theory these animal and/or plant derived fuels should have less environmental impact in soil based on their simplified composition relative to Diesel, they pose an environmental risk like Diesel at high concentrations when disposed. The aim of the present study was to ascertain the phytotoxicity of three different plant-derived biodiesels relative to conventional Diesel. For phytotoxicological analysis, we used seeds of four crop plants, Medicago sativa, Lactuca sativa, Raphanus sativus, and Triticum aestivum to analyze the germination of seeds in contaminated soil samples. The toxicological experiment was conducted with two different soil textures: sandy loam soil and silt loam soil. The studied plant-based biodiesels were safflower methyl-ester, castor methyl ester, and castor ethyl-ester. Biodiesel toxicity was more evident at high concentrations, affecting the germination and survival of small-seeded plants to a greater extent. Tolerance of plants to the biodiesels varied between plant species and soil textures. With the exception of R. sativus, all plant species were affected and exhibited some sensitivity to the fuels, such as delayed seedling emergence and slow germination (average=10 days) at high soil concentrations (0.85% for Diesel and 1.76% for the biodiesels). Tolerance of plants to soil contamination had a species-specific nature, and on average, decreased in the following order: Raphanus sativus (0-20%)>Triticum aestivum (10-40%) ≥ Medicago sativa> Lactuca sativa (80-100%). Thus, we conclude that there is some phytotoxicity associated with plant-based biodiesels. Further

  4. medicago sativa l.

    African Journals Online (AJOL)

    user

    Awareness of lucerne's nutritional potential for human consumption should be raised since it could contribute to food security for sustainable development in Africa. — Dr A Mielmann*. Department of Consumer Sciences to School of. Physiology, Nutrition and Consumer Sciences. Private Bag X6001. North-West University.

  5. Medicago sativa spp. falcata

    African Journals Online (AJOL)

    STORAGESEVER

    2009-05-18

    May 18, 2009 ... For optimal utilization of the species, it is necessary to research and understand the genetic and .... extension step at 72°C for 10 min. PCR products (5 µl/lane) were denatured at 105°C for ..... satellite markers and their utilization in genetic diversity analysis of cultivated and wild populations of the mud carp.

  6. Medicago sativa spp. falcata

    African Journals Online (AJOL)

    STORAGESEVER

    2009-05-18

    Hanson, 1972b; Lesins and .... k jk ik. M. M. 2, where Mik and Mjk are means for the k character of populations i and j (k ranged from 1 to 7, representing the seven morphological characters detected in the experiment). Population ...

  7. Producción de biomasa aérea y uso equivalente de la tierra en intercultivos de alfalfa (Medicago sativa L.

    Directory of Open Access Journals (Sweden)

    T. W Pereyra

    Full Text Available Tradicionalmente, el incremento de la productividad se ha asociado al aumento del rendimiento a través del mejoramiento genético y las prácticas de manejo del cultivo. Sin embargo, si se considera la producción por unidad de área y de tiempo, el sistema de intercultivos puede ser otra forma de mejorar la rentabilidad. El objetivo del experimento fue determinar la biomasa producida y el uso equivalente de la tierra en monocultivo e intercultivos de alfalfa (Medicago sativa L. con sorgo sudán (Sorghum sudanense L. y avena (Avena sativa L.. Se determinó la biomasa aérea de todos los tratamientos (expresada por unidad de superficie y el uso equivalente de la tierra. El diseño fue completamente aleatorizado, dispuesto en bloques con dos repeticiones. Los resultados se sometieron a un ANOVA y las medias se compararon mediante la prueba de Duncan, a través del paquete estadístico INFOSTAT. El intercultivo alfalfa-sorgo triplicó la producción de alfalfa en relación con el monocultivo, mientras que alfalfa-avena no superó la producción de alfalfa pura en los meses de invierno. El intercultivo alfalfa-sorgo fue un 57 % más eficiente en el uso de la tierra que sus respectivos monocultivos, mientras que alfalfa-avena no logró superar la unidad.

  8. NaCl effect on the distribution of wall ingrowth polymers and arabinogalactan proteins in type A transfer cells of Medicago sativa Gabès leaves.

    Science.gov (United States)

    Boughanmi, Néziha; Thibault, Florence; Decou, Raphael; Fleurat-Lessard, Pierrette; Béré, Emile; Costa, Guy; Lhernould, Sabine

    2010-06-01

    We studied the distribution of wall ingrowth (WI) polymers by probing thin sections of companion cells specialized as transfer cells in minor veins of Medicago sativa cv Gabès blade with affinity probes and antibodies specific to polysaccharides and glycoproteins. The wall polymers in the controls were similar in WIs and in the primary wall but differently distributed. The extent of labeling in these papillate WIs differed for JIM5 and JIM7 homogalacturonans but was in the same range for LM5 and LM6 rhamnogalacturonans and xyloglucans. These data show that WI enhancement probably requires arabinogalactan proteins (JIM8) mainly localized on the outer part of the primary wall and WIs. By comparison, NaCl-treated plants exhibited cell wall polysaccharide modifications indicating (1) an increase in unesterified homogalacturonans (JIM5), probably implicated in Na(+) binding and/or polysaccharide network interaction for limiting turgor variations in mesophyll cells; (2) enhancement of the xyloglucan network with an accumulation of fucosylated xyloglucans (CCRC-M1) known to increase the capacity of cellulose binding; and (3) specific recognition of JIM8 arabinogalactan proteins that could participate in both wall enlargement and cohesion by increasing the number of molecular interactions with the other polymers. In conclusion, the cell wall polysaccharide distribution in enlarged WIs might (1) participate in wall resistance to sequestration of Na(+), allowing a better control of hydric homeostasis in mesophyll cells to maintain metabolic activity in source leaves, and (2) maintain tolerance of M. sativa to NaCl.

  9. Overexpression of Medicago sativa TMT elevates the α-tocopherol content in Arabidopsis seeds, alfalfa leaves, and delays dark-induced leaf senescence.

    Science.gov (United States)

    Jiang, Jishan; Jia, Huili; Feng, Guangyan; Wang, Zan; Li, Jun; Gao, Hongwen; Wang, Xuemin

    2016-08-01

    Alfalfa (Medicago sativa L.) is a major forage legume for livestock and a target for improving their dietary quality. Vitamin E is an essential vitamin that animals must obtain from their diet for proper growth and development. γ-tocopherol methyltransferase (γ-TMT), which catalyzes the conversion of δ- and γ-tocopherols (or tocotrienols) to β- and α-tocopherols (or tocotrienols), respectively, is the final enzyme involved in the vitamin E biosynthetic pathway. The overexpression of M. sativa L.'s γ-TMT (MsTMT) increased the α-tocopherol content 10-15 fold above that of wild type Arabidopsis seeds without altering the total content of vitamin E. Additionally, in response to osmotic stress, the biomass and the expression levels of several osmotic marker genes were significantly higher in the transgenic lines compared with wild type. Overexpression of MsTMT in alfalfa led to a modest, albeit significant, increase in α-tocopherol in leaves and was also responsible for a delayed leaf senescence phenotype. Additionally, the crude protein content was increased, while the acid and neutral detergent fiber contents were unchanged in these transgenic lines. Thus, increased α-tocopherol content occurred in transgenic alfalfa without compromising the nutritional qualities. The targeted metabolic engineering of vitamin E biosynthesis through MsTMT overexpression provides a promising approach to improve the α-tocopherol content of forage crops. Copyright © 2016 The Authors. Published by Elsevier Ireland Ltd.. All rights reserved.

  10. A sequence-based genetic map of Medicago truncatula and comparison of marker colinearity with M. sativa

    NARCIS (Netherlands)

    Choi, H.K.; Kim, D.; Uhm, T.; Limpens, E.H.M.; Lim, H.; Mun, J.H.; Kalo, P.; Penmetsa, R.V.; Seres, A.; Kulikova, O.; Roe, B.A.; Bisseling, T.; Kiss, G.B.; Cook, D.R.

    2004-01-01

    A core genetic map of the legume Medicago truncatula has been established by analyzing the segregation of 288 sequence-characterized genetic markers in an E, population composed of 93 individuals. These molecular markers correspond to 141 ESTs, 80 BAC end sequence tags, and 67 resistance gene

  11. Genome-Wide Association Study Identifies Loci for Salt Tolerance during Germination in Autotetraploid Alfalfa (Medicago sativa L.) Using Genotyping-by-Sequencing.

    Science.gov (United States)

    Yu, Long-Xi; Liu, Xinchun; Boge, William; Liu, Xiang-Ping

    2016-01-01

    Salinity is one of major abiotic stresses limiting alfalfa (Medicago sativa L.) production in the arid and semi-arid regions in US and other counties. In this study, we used a diverse panel of alfalfa accessions previously described by Zhang et al. (2015) to identify molecular markers associated with salt tolerance during germination using genome-wide association study (GWAS) and genotyping-by-sequencing (GBS). Phenotyping was done by germinating alfalfa seeds under different levels of salt stress. Phenotypic data of adjusted germination rates and SNP markers generated by GBS were used for marker-trait association. Thirty six markers were significantly associated with salt tolerance in at least one level of salt treatments. Alignment of sequence tags to the Medicago truncatula genome revealed genetic locations of the markers on all chromosomes except chromosome 3. Most significant markers were found on chromosomes 1, 2, and 4. BLAST search using the flanking sequences of significant markers identified 14 putative candidate genes linked to 23 significant markers. Most of them were repeatedly identified in two or three salt treatments. Several loci identified in the present study had similar genetic locations to the reported QTL associated with salt tolerance in M. truncatula. A locus identified on chromosome 6 by this study overlapped with that by drought in our previous study. To our knowledge, this is the first report on mapping loci associated with salt tolerance during germination in autotetraploid alfalfa. Further investigation on these loci and their linked genes would provide insight into understanding molecular mechanisms by which salt and drought stresses affect alfalfa growth. Functional markers closely linked to the resistance loci would be useful for MAS to improve alfalfa cultivars with enhanced resistance to drought and salt stresses.

  12. Genome-Wide Association Study Identifies Loci for Salt Tolerance during Germination in Autotetraploid Alfalfa (Medicago sativa L.) Using Genotyping-by-Sequencing

    Science.gov (United States)

    Yu, Long-Xi; Liu, Xinchun; Boge, William; Liu, Xiang-Ping

    2016-01-01

    Salinity is one of major abiotic stresses limiting alfalfa (Medicago sativa L.) production in the arid and semi-arid regions in US and other counties. In this study, we used a diverse panel of alfalfa accessions previously described by Zhang et al. (2015) to identify molecular markers associated with salt tolerance during germination using genome-wide association study (GWAS) and genotyping-by-sequencing (GBS). Phenotyping was done by germinating alfalfa seeds under different levels of salt stress. Phenotypic data of adjusted germination rates and SNP markers generated by GBS were used for marker-trait association. Thirty six markers were significantly associated with salt tolerance in at least one level of salt treatments. Alignment of sequence tags to the Medicago truncatula genome revealed genetic locations of the markers on all chromosomes except chromosome 3. Most significant markers were found on chromosomes 1, 2, and 4. BLAST search using the flanking sequences of significant markers identified 14 putative candidate genes linked to 23 significant markers. Most of them were repeatedly identified in two or three salt treatments. Several loci identified in the present study had similar genetic locations to the reported QTL associated with salt tolerance in M. truncatula. A locus identified on chromosome 6 by this study overlapped with that by drought in our previous study. To our knowledge, this is the first report on mapping loci associated with salt tolerance during germination in autotetraploid alfalfa. Further investigation on these loci and their linked genes would provide insight into understanding molecular mechanisms by which salt and drought stresses affect alfalfa growth. Functional markers closely linked to the resistance loci would be useful for MAS to improve alfalfa cultivars with enhanced resistance to drought and salt stresses. PMID:27446182

  13. Tolerancia experimental de las especies vegetales Nicotiana glauca, Jacaranda mimosifolia, Tecoma stans, Medicago sativa y Spinacea oleracea al boro, en Argentina

    Directory of Open Access Journals (Sweden)

    Marta L. de Viana

    2008-09-01

    Full Text Available La actividad de las industrias borateras constituye una fuente puntual y difusa de contaminación del aire, suelo y aguas superficiales y profundas. Por lo tanto, el estudio y experimentación acerca de las posibles formas de contrarrestar este impacto constituye una prioridad. Una técnica relativamente nueva para descontaminar suelos es la fitorremediación, que emplea plantas y microorganismos asociados. El primer paso es detectar las especies vegetales tolerantes, lo que constituye el objetivo de este trabajo. Se realizó un experimento en laboratorio para evaluar la germinación, la supervivencia y el crecimiento de distintas especies en diferentes concentraciones de boro. Al comienzo y al final del experimento se determinó la concentración de boro en el sustrato para cada tratamiento y para sustratos con y sin vegetación. Se encontraron diferencias significativas debidas al tratamiento, la especie y la interacción especie *tratamiento. M. sativa, N. glauca y J. mimosifolia fueron las especies de mayor tolerancia al boro. Las otras especies presentaron una disminución en todas las variables-respuesta en función de la concentración del contaminante. Todas presentaron una baja supervivencia en la máxima concentración. La disminución de boro fue máxima en el tratamiento de 30 ppm de boro con M. sativa y la menor se registró en los tratamiento de 20 ppm de boro con J. mimosifolia y de 30 ppm de boro con T. stans y S. oleraceae. Se concluye que N. glauca, M. sativa y J. mimosifolia podrían considerarse como prometedoras en remediación.Experimental tolerance to boron of the plant species Nicotiana glauca, Jacaranda mimosifolia, Tecoma stans, Medicago sativa y Spinacea oleracea in Argentina. The activity of boron industries is a punctual and diffuse source of air, soil and water pollution. Therefore, it is a priority to study possible ways of reducing this impact. A relatively new technology for reducing soil pollution is

  14. New Diversity Arrays Technology (DArT) markers for tetraploid oat (Avena magna Murphy et Terrell) provide the first complete oat linkage map and markers linked to domestication genes from hexaploid A. sativa L.

    Science.gov (United States)

    Oliver, R E; Jellen, E N; Ladizinsky, G; Korol, A B; Kilian, A; Beard, J L; Dumlupinar, Z; Wisniewski-Morehead, N H; Svedin, E; Coon, M; Redman, R R; Maughan, P J; Obert, D E; Jackson, E W

    2011-11-01

    Nutritional benefits of cultivated oat (Avena sativa L., 2n = 6x = 42, AACCDD) are well recognized; however, seed protein levels are modest and resources for genetic improvement are scarce. The wild tetraploid, A. magna Murphy et Terrell (syn A. maroccana Gdgr., 2n = 4x = 28, CCDD), which contains approximately 31% seed protein, was hybridized with cultivated oat to produce a domesticated A. magna. Wild and cultivated accessions were crossed to generate a recombinant inbred line (RIL) population. Although these materials could be used to develop domesticated, high-protein oat, mapping and quantitative trait loci introgression is hindered by a near absence of genetic markers. Objectives of this study were to develop high-throughput, A. magna-specific markers; generate a genetic linkage map based on the A. magna RIL population; and map genes controlling oat domestication. A Diversity Arrays Technology (DArT) array derived from 10 A. magna genotypes was used to generate 2,688 genome-specific probes. These, with 12,672 additional oat clones, produced 2,349 polymorphic markers, including 498 (21.2%) from A. magna arrays and 1,851 (78.8%) from other Avena libraries. Linkage analysis included 974 DArT markers, 26 microsatellites, 13 SNPs, and 4 phenotypic markers, and resulted in a 14-linkage-group map. Marker-to-marker correlation coefficient analysis allowed classification of shared markers as unique or redundant, and putative linkage-group-to-genome anchoring. Results of this study provide for the first time a collection of high-throughput tetraploid oat markers and a comprehensive map of the genome, providing insights to the genome ancestry of oat and affording a resource for study of oat domestication, gene transfer, and comparative genomics.

  15. Identification of the Large Subunit of Ribulose 1,5-Bisphosphate Carboxylase/Oxygenase as a Substrate for Transglutaminase in Medicago sativa L. (Alfalfa) 1

    Science.gov (United States)

    Margosiak, Stephen A.; Dharma, Abdi; Bruce-Carver, Megan R.; Gonzales, Andrea P.; Louie, Donna; Kuehn, Glenn D.

    1990-01-01

    Extracts prepared from floral meristematic tissue of alfalfa (Medicago sativa L.) were investigated for expression of the enzyme transglutaminase in order to identify the major protein substrate for transglutaminase-directed modifications among plant proteins. The large polymorphic subunits of ribulose 1,5-bisphosphate carboxylase/oxygenase in alfalfa, with molecular weights of 52,700 and 57,600, are major substrates for transglutaminase in these extracts. This was established by: (a) covalent conjugation of monodansylcadaverine to the large subunit followed by fluorescent detection in SDS-polyacrylamide gels; (b) covalent conjugation of [14C]putrescine to the large subunit with detection by autoradiography; (c) covalent conjugation of monodansylcadaverine to the large subunit and demonstration of immunocross-reactivity on nitrocellulose transblot of the modified large subunit with antibody prepared in rabbits against dansylated-ovalbumin; (d) demonstration of a direct dependence of the rate of transglutaminase-mediated, [14C]putrescine incorporation upon the concentration of ribulose, 1,5-bisphosphate carboxylase/oxygenase from alfalfa or spinach; and (e) presumptive evidence from size exclusion chromatography that transglutaminase may cofractionate with native molecules of ribulose 1,5-bisphosphate carboxylase/oxygenase in crude extracts. Analysis of the primary structure of plant large subunit has revealed numerous potential glutaminyl and lysyl sites for transglutaminase-directed modifications of ribulose 1,5-bisphosphate carboxylase/oxygenase. Images Figure 1 Figure 2 PMID:16667270

  16. Improvement of in vitro embryo maturation, plantlet regeneration and transformation efficiency from alfalfa (Medicago sativa L.) somatic embryos using Cuscuta campestris extract.

    Science.gov (United States)

    Amini, Massoume; Deljou, Ali; Nabiabad, Haidar Saify

    2016-07-01

    Developmental deficiency of somatic embryos and regeneration to plantlets, especially in the case of transformation, are major problems of somatic embryo regeneration in alfalfa. One of the ways to overcome these problems is the use of natural plant regulators and nutrients in the culture medium of somatic embryos. For investigating the influence of Cuscuta campestris extract on the efficiency of plant regeneration and transformation, chimeric tissue type plasminogen activator was transferred to explants using Agrobacterium tumefaciens, and transgenic plants were recovered using medium supplemented with different concentration of the extract. Transgenic plants were analyzed by PCR and RT-PCR. Somatic embryos of Medicago sativa L. developed into plantlets at high frequency level (52 %) in the maturation medium supplemented with 50 mg 1-1C. campestris extract as compared to the medium without extract (26 %). Transformation efficiency was 29.3 and 15.2 % for medium supplemented with dodder extract and without the extract, respectively. HPLC and GC/MS analysis of the extract indicated high level of ABA and some compounds such as Phytol, which can affect the somatic embryo maturation. The antibacterial assay showed that the extract was effective against some strains of A. tumefaciens. These results have provided a scientific basis for using of C. campestris extract as a good natural source of antimicrobial agents and plant growth regulator as well, that can be used in tissue culture of transgenic plants.

  17. [Interaction of biosurfactant-microorganism to enhance phytoremediation of aged polycyclic aromatic hydrocarbons (PAHS) contaminated soils with alfalfa (Medicago sativa L.)].

    Science.gov (United States)

    Liu, Wei-wei; Yin, Rui; Lin, Xian-gui; Zhang, Jing; Chen, Xiao-min; Li, Xuan-zhen; Yang, Ting

    2010-04-01

    A pot experiment in greenhouse was carried out to investigate the interactive effect of rhamnolipids (RH) addition and PAHs-specific degrading bacteria (DB) inoculation on the phytoremediation efficiency for removal polycyclic aromatic hydrocarbons (PAHs) from agricultural soils. Results indicated that RH addition and DB inoculation promoted alfalfa (Medicago sativa L. ) growth and PAHs degradation in the soil. After 90 days, residual PAHs concentration in soil reduced 30.0% and 49.6% for the treatment of RH and DB, respectively, but only 21.7% for control. For the treatment of RH + DB, residual PAHs concentration in soil reduced 53.9%, showing synergy effect of RH addition and DB inoculation. In addition, the average PAHs degradation gradually reduced with the increase of the PAHs rings, but DB inoculation promoted the degradation of four-ring PAHs and five-ring PAHs. The number of PAHs degrading bacteria and dehydrogenase activity increased with the removal ratio of PAHs. Therefore RH addition and PAHs-specific degrading bacteria inoculation were effective in enhancing the phytoremediation efficiency of the long-term PAHs contaminated soils.

  18. The photosynthetic and stomatal response of Medicago sativa cv. saranac to free-air CO{sub 2} enrichment (F.A.C.E.) and nitrogen

    Energy Technology Data Exchange (ETDEWEB)

    Bridson, N.P.

    1996-08-01

    Plots of Medicago sativa cv. saranac were grown in the field at ambient (355 {mu}mol CO{sub 2} mol{sup -1} air) or elevated (600{mu}mol CO{sub 2} mol{sup -1} air) CO{sub 2} concentrations. High (200kg yr{sup -1}) or low (20kg yr{sup -1}) nitrogen levels were applied to two isogeneic lines, one able and one unable to use nitrogen fixing bacteria. Plants were in the second year of field growth. Exposure to elevated CO{sub 2} was via a Free-Air CO{sub 2} Enrichment System (FACE). Elevated CO{sub 2} increased diurnal assimilation by between 12% and 92%. Analysis of A/C{sub i} responses showed that effective nitrogen fertilisation was more important to rubisCO and RuBP activity than elevated CO{sub 2}. No acclimation was consistently observed. Leaves lower down the canopy were found to have lower Vc{sub max} and J{sub max} values, though age may be the cause of the latter effect. FACE conditions have only a small effect on these responses. There was some evidence found for the down-regulation of photosynthesis in the late afternoon. The FACE conditions had no affect on stomatal density but did increase epidermal cell density.

  19. Fixed-Precision Sequential Sampling Plans for Estimating Alfalfa Caterpillar, Colias lesbia, Egg Density in Alfalfa, Medicago sativa, Fields in Córdoba, Argentina

    Science.gov (United States)

    Serra, Gerardo V.; Porta, Norma C. La; Avalos, Susana; Mazzuferi, Vilma

    2013-01-01

    The alfalfa caterpillar, Colias lesbia (Fabricius) (Lepidoptera: Pieridae), is a major pest of alfalfa, Medicago sativa L. (Fabales: Fabaceae), crops in Argentina. Its management is based mainly on chemical control of larvae whenever the larvae exceed the action threshold. To develop and validate fixed-precision sequential sampling plans, an intensive sampling programme for C. lesbia eggs was carried out in two alfalfa plots located in the Province of Córdoba, Argentina, from 1999 to 2002. Using Resampling for Validation of Sampling Plans software, 12 additional independent data sets were used to validate the sequential sampling plan with precision levels of 0.10 and 0.25 (SE/mean), respectively. For a range of mean densities of 0.10 to 8.35 eggs/sample, an average sample size of only 27 and 26 sample units was required to achieve a desired precision level of 0.25 for the sampling plans of Green and Kuno, respectively. As the precision level was increased to 0.10, average sample size increased to 161 and 157 sample units for the sampling plans of Green and Kuno, respectively. We recommend using Green's sequential sampling plan because it is less sensitive to changes in egg density. These sampling plans are a valuable tool for researchers to study population dynamics and to evaluate integrated pest management strategies. PMID:23909840

  20. The 3' untranslated region of the two cytosolic glutamine synthetase (GS(1)) genes in alfalfa (Medicago sativa) regulates transcript stability in response to glutamine.

    Science.gov (United States)

    Simon, Bindu; Sengupta-Gopalan, Champa

    2010-10-01

    Glutamine synthetase (GS) catalyzes the ATP-dependent condensation of ammonia with glutamate to produce glutamine. The GS enzyme is located either in the chloroplast (GS(2)) or in the cytoplasm (GS(1)). GS(1) is encoded by a small gene family and the members exhibit differential expression pattern mostly attributed to transcriptional regulation. Based on our recent finding that a soybean GS(1) gene, Gmglnβ ( 1 ) is subject to its 3'UTR-mediated post-transcriptional regulation as a transgene in alfalfa (Medicago sativa) we have raised the question of whether the 3'UTR-mediated transcript destabilization is a more universal phenomenon. Gene constructs consisting of the CaMV35S promoter driving the reporter gene, GUS, followed by the 3'UTRs of the two alfalfa GS(1) genes, MsGSa and MsGSb, were introduced into alfalfa and tobacco. The analysis of these transformants suggests that while both the 3'UTRs promote transcript turnover, the MsGSb 3'UTR is more effective than the MsGSa 3'UTR. However, both the 3'UTRs along with Gmglnβ ( 1 ) 3'UTR respond to nitrate as a trigger in transcript turnover. More detailed analysis points to glutamine rather than nitrate as the mediator of transcript turnover. Our data suggests that the 3'UTR-mediated regulation of GS(1) genes at the level of transcript turnover is probably universal and is used for fine-tuning the expression in keeping with the availability of the substrates.

  1. EFFECT OF SOIL pH ON P AND Ca UPTAKE TO THE ABOVE GROUND BIOMASS OF ALFALFA (Medicago sativa L.

    Directory of Open Access Journals (Sweden)

    Gordana Bukvić

    2002-06-01

    Full Text Available Alfalfa (Medicago sativa L. has been grown on two locations – Osijek (Eutric Cambisols with pH 1n KCl=5.4 and Petrijevci (Loessive pseudogley with pH 1n KCl=4.36 in eastern Croatia area during two years. Field experiments were set up as randomized block system in four replicates. Seeding was carried out by two new genotypes from gene collection of the Agricultural Institute in Osijek. Above ground biomass of alfalfa was removed in the budding-blooming stage. Three mowings were accomplished in the first investigation year on both locations. Five mowings were carried out on Osijek location in the second investigation year and three on Petrijevci. Total dry matter yield of 11.5 tha-1 was achieved on the average for genotypes and years in Osijek location whereas in Petrijevci it amounted to 6.95 tha-1. Concentration of P was 0.367%/DM and Ca 1.01%/DM on the average for genotypes, swaths and years on Osijek location while on Petrijevci the situation was as follows: P 0.321%/DM and Ca 0.94%/DM.

  2. Assessing quality of Medicago sativa silage by monitoring bacterial composition with single molecule, real-time sequencing technology and various physiological parameters.

    Science.gov (United States)

    Bao, Weichen; Mi, Zhihui; Xu, Haiyan; Zheng, Yi; Kwok, Lai Yu; Zhang, Heping; Zhang, Wenyi

    2016-06-24

    The present study applied the PacBio single molecule, real-time sequencing technology (SMRT) in evaluating the quality of silage production. Specifically, we produced four types of Medicago sativa silages by using four different lactic acid bacteria-based additives (AD-I, AD-II, AD-III and AD-IV). We monitored the changes in pH, organic acids (including butyric acid, the ratio of acetic acid/lactic acid, γ-aminobutyric acid, 4-hyroxy benzoic acid and phenyl lactic acid), mycotoxins, and bacterial microbiota during silage fermentation. Our results showed that the use of the additives was beneficial to the silage fermentation by enhancing a general pH and mycotoxin reduction, while increasing the organic acids content. By SMRT analysis of the microbial composition in eight silage samples, we found that the bacterial species number and relative abundances shifted apparently after fermentation. Such changes were specific to the LAB species in the additives. Particularly, Bacillus megaterium was the initial dominant species in the raw materials; and after the fermentation process, Pediococcus acidilactici and Lactobacillus plantarum became the most prevalent species, both of which were intrinsically present in the LAB additives. Our data have demonstrated that the SMRT sequencing platform is applicable in assessing the quality of silage.

  3. Co-expression of bacterial aspartate kinase and adenylylsulfate reductase genes substantially increases sulfur amino acid levels in transgenic alfalfa (Medicago sativa L..

    Directory of Open Access Journals (Sweden)

    Zongyong Tong

    Full Text Available Alfalfa (Medicago sativa L. is one of the most important forage crops used to feed livestock, such as cattle and sheep, and the sulfur amino acid (SAA content of alfalfa is used as an index of its nutritional value. Aspartate kinase (AK catalyzes the phosphorylation of aspartate to Asp-phosphate, the first step in the aspartate family biosynthesis pathway, and adenylylsulfate reductase (APR catalyzes the conversion of activated sulfate to sulfite, providing reduced sulfur for the synthesis of cysteine, methionine, and other essential metabolites and secondary compounds. To reduce the feedback inhibition of other metabolites, we cloned bacterial AK and APR genes, modified AK, and introduced them into alfalfa. Compared to the wild-type alfalfa, the content of cysteine increased by 30% and that of methionine increased substantially by 60%. In addition, a substantial increase in the abundance of essential amino acids (EAAs, such as aspartate and lysine, was found. The results also indicated a close connection between amino acid metabolism and the tricarboxylic acid (TCA cycle. The total amino acid content and the forage biomass tested showed no significant changes in the transgenic plants. This approach provides a new method for increasing SAAs and allows for the development of new genetically modified crops with enhanced nutritional value.

  4. Effects of cyanobacterial toxins and cyanobacterial cell-free crude extract on germination of alfalfa (Medicago sativa) and induction of oxidative stress.

    Science.gov (United States)

    Pflugmacher, Stephan; Jung, Katharina; Lundvall, Linn; Neumann, Stefanie; Peuthert, Anja

    2006-09-01

    Cyanobacterial toxins have adverse effects on both terrestrial and aquatic plants. Microcystins are cyclic heptapeptides and an important group of cyanotoxins. When lake water contaminated with cyanobacterial blooms is used for spray irrigation, these toxins can come in contact with agricultural plants. During the exposure to these toxins, reactive oxygen species can form. These reactive oxygen species have a strong reactivity and are able to interact with other cellular compounds (lipids, protein, and DNA). Plants have antioxidative systems that will limit the negative effects caused by reactive oxygen species. These systems consist of enzymes, such as superoxide dismutase, catalase, and ascorbate peroxidase, and nonenzymatic substances, such as reduced glutathione or vitamins. The aim of the present study was to investigate the effects of cyanobacterial toxins (microcystins and anatoxin-a) and cyanobacterial cell-free crude extract on alfalfa (Medicago sativa) seedlings. Inhibition of germination and root growth was observed with toxin concentrations of 5.0 microg/L. Also, oxidative damage, such as lipid peroxidation, was detected after the exposure of alfalfa seedlings to the toxin. Reactive oxygen detoxifying enzymes were elevated, showing a marked response in alfalfa to oxidative stress caused by the exposure to cyanobacterial metabolites that might influence the growth and development of these plants negatively.

  5. Co-transforming bar and CsLEA enhanced tolerance to drought and salt stress in transgenic alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Zhang, Jiyu; Duan, Zhen; Zhang, Daiyu; Zhang, Jianquan; Di, Hongyan; Wu, Fan; Wang, Yanrong

    2016-03-25

    Drought and high salinity are two major abiotic factors that restrict alfalfa productivity. A dehydrin protein, CsLEA, from the desert grass Cleistogenes songorica was transformed into alfalfa (Medicago sativa L.) via Agrobacterium-mediated transformation using the bar gene as a selectable marker, and the drought and salt stress tolerances of the transgenic plants were assessed. Thirty-nine of 119 transformants were positive, as screened by Basta, and further molecularly authenticated using PCR and RT-PCR. Phenotype observations revealed that the transgenic plants grew better than the wild-type (WT) plants after 15d of drought stress and 10d of salt stress: the leaves of WT alfalfa turned yellow, whereas the transgenic alfalfa leaves only wilted; after rewatering, the transgenic plants returned to a normal state, though the WT plants could not be restored. Evaluation of physiologic and biochemical indices during drought and salt stresses showed a relatively lower Na(+) content in the leaves of the transgenic plants, which would reduce toxic ion effects. In addition, the transgenic plants were able to maintain a higher relative water content (RWC), higher shoot biomass, fewer photosystem changes, decreased membrane injury, and a lower level of osmotic stress injury. These results demonstrate that overexpression of the CsLEA gene can enhance the drought and salt tolerance of transgenic alfalfa; in addition, carrying the bar gene in the genome may increase herbicide resistance. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. Effects of chlorpyrifos and chlorantraniliprole on fermentation quality of alfalfa (Medicago sativa L.) silage inoculated with or without Lactobacillus plantarum LP.

    Science.gov (United States)

    Zhang, Qing; Yu, Zhu; Wang, Xianguo; Na, Risu

    2017-03-01

    The effects of pesticides and Lactobacillus plantarum (LP) on fermentation quality of alfalfa (Medicago sativa L.) silage were investigated. Chlorpyrifos and chlorantraniliprole were sprayed on the surface of alfalfa plants at 658.6 and 45.0 g active ingredient/ha, respectively. Alfalfa plants were harvested on day 5 post-application and ensiled with or without LP. Chlorpyrifos and chlorantraniliprole decreased the yeast count of alfalfa material (P < 0.05). Both pesticides increased the butyric acid content of alfalfa silage (P < 0.001). Chlorpyrifos increased pH and decreased lactic acid, acetic acid and short-chain fatty acid contents (P < 0.05). LP decreased pH and butyric acid content, and increased lactic acid and short-chain fatty acid contents of alfalfa silage treated with pesticides (P < 0.05). LP increased the concentration of chlorpyrifos residue in alfalfa silage (P < 0.05). Chlorpyrifos and chlorantraniliprole affected the microbial communities of the material before ensiling, especially coliform bacteria and yeast; the two pesticide residues were reduced after the fermentation of alfalfa silage and affected the fermentation process, whereas LP improved the fermentation quality of pesticides-contaminated alfalfa silage and slowed down the dissipation of chlorpyrifos. © 2016 Japanese Society of Animal Science.

  7. Expression of an alfalfa (Medicago sativa L.) peroxidase gene in transgenic Arabidopsis thaliana enhances resistance to NaCl and H2O2.

    Science.gov (United States)

    Teng, K; Xiao, G Z; Guo, W E; Yuan, J B; Li, J; Chao, Y H; Han, L B

    2016-05-23

    Peroxidases (PODs) are enzymes that play important roles in catalyzing the reduction of H2O2 and the oxidation of various substrates. They function in many different and important biological processes, such as defense mechanisms, immune responses, and pathogeny. The POD genes have been cloned and identified in many plants, but their function in alfalfa (Medicago sativa L.) is not known, to date. Based on the POD gene sequence (GenBank accession No. L36157.1), we cloned the POD gene in alfalfa, which was named MsPOD. MsPOD expression increased with increasing H2O2. The gene was expressed in all of the tissues, including the roots, stems, leaves, and flowers, particularly in stems and leaves under light/dark conditions. A subcellular analysis showed that MsPOD was localized outside the cells. Transgenic Arabidopsis with MsPOD exhibited increased resistance to H2O2 and NaCl. Moreover, POD activity in the transgenic plants was significantly higher than that in wild-type Arabidopsis. These results show that MsPOD plays an important role in resistance to H2O2 and NaCl.

  8. Co-downregulation of the hydroxycinnamoyl-CoA:shikimate hydroxycinnamoyl transferase and coumarate 3-hydroxylase significantly increases cellulose content in transgenic alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Tong, Zongyong; Li, Heng; Zhang, Rongxue; Ma, Lei; Dong, Jiangli; Wang, Tao

    2015-10-01

    Lignin is a component of the cell wall that is essential for growth, development, structure and pathogen resistance in plants, but high lignin is an obstacle to the conversion of cellulose to ethanol for biofuel. Genetically modifying lignin and cellulose contents can be a good approach to overcoming that obstacle. Alfalfa (Medicago sativa L.) is rich in lignocellulose biomass and used as a model plant for the genetic modification of lignin in this study. Two key enzymes in the lignin biosynthesis pathway-hydroxycinnamoyl -CoA:shikimate hydroxycinnamoyl transferase (HCT) and coumarate 3-hydroxylase (C3H)-were co-downregulated. Compared to wild-type plants, the lignin content in the modified strain was reduced by 38%, cellulose was increased by 86.1%, enzyme saccharification efficiency was increased by 10.9%, and cell wall digestibility was increased by 13.0%. The modified alfalfa exhibited a dwarf phenotype, but normal above ground biomass. This approach provides a new strategy for reducing lignin and increasing cellulose contents and creates a new genetically modified crop with enhanced value for biofuel. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  9. A vapBC-type toxin-antitoxin module of Sinorhizobium meliloti influences symbiotic efficiency and nodule senescence of Medicago sativa.

    Science.gov (United States)

    Lipuma, Justine; Cinege, Gyöngyi; Bodogai, Monica; Oláh, Boglárka; Kiers, Aurélie; Endre, Gabriella; Dupont, Laurence; Dusha, Ilona

    2014-12-01

    The symbiotic nitrogen-fixing soil bacterium Sinorhizobium meliloti carries a large number of toxin-antitoxin (TA) modules both on the chromosome and megaplasmids. One of them, the vapBC-5 module that belongs to the type II systems was characterized here. It encodes an active toxin vapC-5, and was shown to be controlled negatively by the complex of its own proteins. Different mutants of the vapBC-5 genes exhibited diverse effects on symbiotic efficiency during interaction with the host plant Medicago sativa. The absence of the entire vapBC-5 region had no influence on nodule formation and nitrogen fixation properties. The strain carrying an insertion in the antitoxin gene showed a reduced nitrogen fixation capacity resulting in a lower plant yield. In contrast, when the toxin gene was mutated, the strain developed more efficient symbiosis with the host plant. The nitrogen fixing root nodules had a delayed senescent phenotype and contained elevated level of plant-derived molecules characteristic of later steps of nodule development. The longer bacteroid viability and abundance of active nitrogen fixing zone resulted in increased production of plant material. These data indicate that modification of the toxin/antitoxin production may influence bacteroid metabolism and may have an impact on the adaptation to changing environmental conditions. © 2014 Society for Applied Microbiology and John Wiley & Sons Ltd.

  10. Co-expression of bacterial aspartate kinase and adenylylsulfate reductase genes substantially increases sulfur amino acid levels in transgenic alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Tong, Zongyong; Xie, Can; Ma, Lei; Liu, Liping; Jin, Yongsheng; Dong, Jiangli; Wang, Tao

    2014-01-01

    Alfalfa (Medicago sativa L.) is one of the most important forage crops used to feed livestock, such as cattle and sheep, and the sulfur amino acid (SAA) content of alfalfa is used as an index of its nutritional value. Aspartate kinase (AK) catalyzes the phosphorylation of aspartate to Asp-phosphate, the first step in the aspartate family biosynthesis pathway, and adenylylsulfate reductase (APR) catalyzes the conversion of activated sulfate to sulfite, providing reduced sulfur for the synthesis of cysteine, methionine, and other essential metabolites and secondary compounds. To reduce the feedback inhibition of other metabolites, we cloned bacterial AK and APR genes, modified AK, and introduced them into alfalfa. Compared to the wild-type alfalfa, the content of cysteine increased by 30% and that of methionine increased substantially by 60%. In addition, a substantial increase in the abundance of essential amino acids (EAAs), such as aspartate and lysine, was found. The results also indicated a close connection between amino acid metabolism and the tricarboxylic acid (TCA) cycle. The total amino acid content and the forage biomass tested showed no significant changes in the transgenic plants. This approach provides a new method for increasing SAAs and allows for the development of new genetically modified crops with enhanced nutritional value.

  11. STUDY ON PHYTOEXTRACTION BALANCE OF ZN, CD, PB FROM MINE-WASTE POLLUTED SOILS BY USING MEDICAGO SATIVA AND TRIFOLIUM PRATENSE SPECIES

    Directory of Open Access Journals (Sweden)

    B. LIXANDRU

    2009-05-01

    Full Text Available For a term of two years was studied phytoextractive potential of Zn, Cd and Pb using successive culture of alfalfa (Medicago sativa and red clover (Trifolium pratense. In the experimental plot was incorporated a quantity of 20 kg mine waste per square meter, providing in soil 1209 mg Zn/kg d.s., 4.70 mg Cd/kg d.s. and 188.2 mg Pb/kg d.s. The metals content accumulated in plants was determined at the two moments of biomass harvesting, and through balance calculations we could establish the phytoextraction efficiency of the two forage-grasses species. The obtained results indicate that both perennial forage-legumes species have a good phytoextractive capacity and tolerance for Zn and Pb, especially Trifolium pratense specie. By using this species as phytoextractors on soil polluted with 3.76 times more Pb and 4.03 times more Zn, is provided the reduction of metallic ions concentration in soil to limits admitted by laws in a period of 3, respectively, 4 years.

  12. Effect of application rate of commercial lignite-derived amendments on early-stage growth of Medicago sativa and soil health, in acidic soil conditions

    Science.gov (United States)

    Patti, Antonio; Little, Karen; Rose, Michael; Jackson, Roy; Cavagnaro, Tim

    2013-04-01

    Commercially available lignite-derived amendments, sold mainly as humate preparations, have been promoted as plant growth stimulants leading to higher crop yields. These products are also claimed to improve soil properties such as pH. This study investigated the effect of application rate of three lignite-derived amendments on the early-stage growth of a pasture legume, lucerne (Medicago sativa), and soil health in a soil type common to south eastern Australia, in a glasshouse setting. An organic-mineral humate product and 'run of mine' lignite coal did not improve shoot or root growth despite application at a range of rates at, and in excess of, the manufacturers recommendation. Application of soluble K-humate product at 20 kg/ha (9.5 kg/ha C equivalent) produced an observable positive effect in shoot growth. At this application rate, a significant delay in the appearance of chlorotic symptoms was observed along with an increase in soil pH concurrent with decreased availability of soil Mn and Al. Higher root dry weight was associated with lower microbial biomass carbon which may indicate an effect on allocation of resources between the microbial community and the plant. An assessment of the effectiveness of lignite-derived amendments on plant growth, as well as their potential to improve the health of an acidic soil will assist farmers in making decisions regarding the use of these products.

  13. Effect of lead treatment on medicarpin accumulation and on the gene expression of key enzymes involved in medicarpin biosynthesis in Medicago sativa L.

    Science.gov (United States)

    Ghelich, Sima; Zarinkamar, Fatemeh; Soltani, Bahram Mohammad; Niknam, Vahid

    2014-12-01

    Lead (Pb) is the most common heavy metal contaminant in the environment. The present study was undertaken to determine the effect of Pb treatment on medicarpin production and accumulation in Medicago sativa L. To this aim, 7- and 30-day-old plants were treated with 0, 120, 240, 500, and 1,000 μM Pb during 10 days. The content of medicarpin was determined by HPLC, and the extent of medicarpin production was deduced from the result of semiquantitative RT-PCR performed on PAL, CHS, and VR genes. HPLC results indicated that medicarpin concentration has been reduced in the roots, while its exudation to the culture medium has been increased. RT-PCR results indicated that the transcript levels of PAL, CHS, and VR genes have not been affected following Pb stress in seedlings. At the vegetative stage, transcript levels of PAL and CHS genes have been reduced in the roots. However, the transcript level of VR gene increased at 120 and 240 μM Pb, while it decreased at higher concentrations. In the shoot, the transcript levels of PAL, CHS, and VR genes were increased following increased concentration of lead in the medium. Overall, q-PCR results suggest that medicarpin biosynthesis has been induced in the shoots and reduced in the roots of the plants treated with a toxic concentration of Pb.

  14. Is there sufficient Ensifer and Rhizobium species diversity in UK farmland soils to support red clover (Trifolium pratense), white clover (T. repens), lucerne (Medicago sativa) and black medic (M. lupulina)?

    Science.gov (United States)

    Roberts, Rachel; Jackson, Robert W; Mauchline, Tim H; Hirsch, Penny R; Shaw, Liz J; Döring, Thomas F; Jones, Hannah E

    2017-11-01

    Rhizobia play important roles in agriculture owing to their ability to fix nitrogen through a symbiosis with legumes. The specificity of rhizobia-legume associations means that underused legume species may depend on seed inoculation with their rhizobial partners. For black medic (Medicago lupulina) and lucerne (Medicago sativa) little is known about the natural prevalence of their rhizobial partner Ensifer meliloti in UK soils, so that the need for inoculating them is unclear. We analysed the site-dependence of rhizobial seed inoculation effects on the subsequent ability of rhizobial communities to form symbioses with four legume species (Medicago lupulina, M. sativa, Trifolium repens and T. pratense). At ten organic farms across the UK, a species-diverse legume based mixture (LBM) which included these four species was grown. The LBM seed was inoculated with a mix of commercial inocula specific for clover and lucerne. At each site, soil from the LBM treatment was compared to the soil sampled prior to the sowing of the LBM (the control). From each site and each of the two treatments, a suspension of soils was applied to seedlings of the four legume species and grown in axenic conditions for six weeks. Root nodules were counted and their rhizobia isolated. PCR and sequencing of a fragment of the gyrB gene from rhizobial isolates allowed identification of strains. The number of nodules on each of the four legume species was significantly increased when inoculated with soil from the LBM treatment compared to the control. Both the proportion of plants forming nodules and the number of nodules formed varied significantly by site, with sites significantly affecting the Medicago species but not the Trifolium species. These differences in nodulation were broadly reflected in plant biomass where site and treatment interacted; at some sites there was a significant advantage from inoculation with the commercial inoculum but not at others. In particular, this study has

  15. CdSe/ZnS Quantum Dots trigger DNA repair and antioxidant enzyme systems in Medicago sativa cells in suspension culture

    Science.gov (United States)

    2013-01-01

    Background Nanoparticles appear to be promising devices for application in the agriculture and food industries, but information regarding the response of plants to contact with nano-devices is scarce. Toxic effects may be imposed depending on the type and concentration of nanoparticle as well as time of exposure. A number of mechanisms may underlie the ability of nanoparticles to cause genotoxicity, besides the activation of ROS scavenging mechanisms. In a previous study, we showed that plant cells accumulate 3-Mercaptopropanoic acid-CdSe/ZnS quantum dots (MPA-CdSe/ZnS QD) in their cytosol and nucleus and increased production of ROS in a dose dependent manner when exposed to QD and that a concentration of 10 nM should be cyto-compatible. Results When Medicago sativa cells were exposed to 10, 50 and 100 nM MPA-CdSe/ZnS QD a correspondent increase in the activity of Superoxide dismutase, Catalase and Glutathione reductase was registered. Different versions of the COMET assay were used to assess the genotoxicity of MPA-CdSe/ZnS QD. The number of DNA single and double strand breaks increased with increasing concentrations of MPA-CdSe/ZnS QD. At the highest concentrations, tested purine bases were more oxidized than the pyrimidine ones. The transcription of the DNA repair enzymes Formamidopyrimidine DNA glycosylase, Tyrosyl-DNA phosphodiesterase I and DNA Topoisomerase I was up-regulated in the presence of increasing concentrations of MPA-CdSe/ZnS QD. Conclusions Concentrations as low as 10 nM MPA-CdSe/ZnS Quantum Dots are cytotoxic and genotoxic to plant cells, although not lethal. This sets a limit for the concentrations to be used when practical applications using nanodevices of this type on plants are being considered. This work describes for the first time the genotoxic effect of Quantum Dots in plant cells and demonstrates that both the DNA repair genes (Tdp1β, Top1β and Fpg) and the ROS scavenging mechanisms are activated when MPA-CdSe/ZnS QD contact M. sativa

  16. Co-transforming bar and CsALDH Genes Enhanced Resistance to Herbicide and Drought and Salt Stress in Transgenic Alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Duan, Zhen; Zhang, Daiyu; Zhang, Jianquan; Di, Hongyan; Wu, Fan; Hu, Xiaowen; Meng, Xuanchen; Luo, Kai; Zhang, Jiyu; Wang, Yanrong

    2015-01-01

    Drought and high salinity are two major abiotic factors that restrict the productivity of alfalfa. By application of the Agrobacterium-mediated transformation method, an oxidative responsive gene, CsALDH12A1, from the desert grass Cleistogenes songorica together with the bar gene associated with herbicide resistance, were co-transformed into alfalfa (Medicago sativa L.). From the all 90 transformants, 16 were positive as screened by spraying 1 mL L(-1) 10% Basta solution and molecularly diagnosis using PCR. Real-time PCR analysis indicated that drought and salt stress induced high CsALDH expression in the leaves of the transgenic plants. The CsALDH expression levels under drought (15 d) and salt stress (200 mM NaCl) were 6.11 and 6.87 times higher than in the control plants, respectively. In comparison to the WT plants, no abnormal phenotypes were observed among the transgenic plants, which showed significant enhancement of tolerance to 15 d of drought and 10 d of salinity treatment. Evaluation of the physiological and biochemical indices during drought and salt stress of the transgenic plants revealed relatively lower Na(+) content and higher K(+) content in the leaves relative to the WT plants, a reduction of toxic on effects and maintenance of osmotic adjustment. In addition, the transgenic plants could maintain a higher relative water content level, higher shoot biomass, fewer changes in the photosystem, decreased membrane injury, and a lower level of osmotic stress. These results indicate that the co-expression of the introduced bar and CsALDH genes enhanced the herbicide, drought and salt tolerance of alfalfa and therefore can potentially be used as a novel genetic resource for the future breeding programs to develop new cultivars.

  17. Root and Nodule Enzymes of Ammonia Assimilation in Two Plant-Conditioned Symbiotically Ineffective Genotypes of Alfalfa (Medicago sativa L.) 1

    Science.gov (United States)

    Groat, R. Gene; Vance, Carroll P.

    1982-01-01

    Biochemical and physiological parameters associated with nitrogen metabolism were measured in nodules and roots of glasshouse-grown clones of two symbiotically ineffective alfalfa (Medicago sativa L.) genotypes supplied with either NO3− or NH4+. Significant differences were observed between genotypes for nodule soluble protein concentrations and glutamine synthetase (GS) and glutamate synthase (GOGAT) specific activities, both in untreated controls and in response to applied N. Nodule soluble protein of both genotypes declined in response to applied N, while nodule GS, GOGAT, and glutamate dehydrogenase (GDH) specific activities either decreased or remained relatively constant. In contrast, no genotype differences were observed in roots for soluble protein concentrations and GS, GOGAT, and GDH specific activities, either in untreated controls or in response to applied N. Root soluble protein levels and GS and GOGAT specific activities of N-treated plants increased 2- to 4-fold within 4 days and then decreased between days 13 and 24. Root GDH specific activity of NH4+-treated plants increased steadily throughout the experiment and was 50 times greater than root GS or GOGAT specific activities by day 24. Enzymological data indicate that nodules of these ineffective alfalfa genotypes are uniquely differentiated plant organs. Decreasing or constant plant GS and GOGAT specific activities in ineffective nodules in response to applied N suggest that factors in addition to N supply are involved in the induction of high levels of plant ammonia-assimilating enzymes in nodules. Genotype differences observed for nodule enzyme specific activities support the concept that ineffectiveness may be expressed in different ways within the nodule. Senescence was evident in ineffective nodules of N-treated plants of both genotypes, indicating that nodule senescence induced by applied N may not be closely linked to symbiotic effectiveness in alfalfa. Data for ammonia

  18. De novo characterization of fall dormant and nondormant alfalfa (Medicago sativa L.) leaf transcriptome and identification of candidate genes related to fall dormancy.

    Science.gov (United States)

    Zhang, Senhao; Shi, Yinghua; Cheng, Ningning; Du, Hongqi; Fan, Wenna; Wang, Chengzhang

    2015-01-01

    Alfalfa (Medicago sativa L.) is one of the most widely cultivated perennial forage legumes worldwide. Fall dormancy is an adaptive character related to the biomass production and winter survival in alfalfa. The physiological, biochemical and molecular mechanisms causing fall dormancy and the related genes have not been well studied. In this study, we sequenced two standard varieties of alfalfa (dormant and non-dormant) at two time points and generated approximately 160 million high quality paired-end sequence reads using sequencing by synthesis (SBS) technology. The de novo transcriptome assembly generated a set of 192,875 transcripts with an average length of 856 bp representing about 165.1 Mb of the alfalfa leaf transcriptome. After assembly, 111,062 (57.6%) transcripts were annotated against the NCBI non-redundant database. A total of 30,165 (15.6%) transcripts were mapped to 323 Kyoto Encyclopedia of Genes and Genomes pathways. We also identified 41,973 simple sequence repeats, which can be used to generate markers for alfalfa, and 1,541 transcription factors were identified across 1,350 transcripts. Gene expression between dormant and non-dormant alfalfa at different time points were performed, and we identified several differentially expressed genes potentially related to fall dormancy. The Gene Ontology and pathways information were also identified. We sequenced and assembled the leaf transcriptome of alfalfa related to fall dormancy, and also identified some genes of interest involved in the fall dormancy mechanism. Thus, our research focused on studying fall dormancy in alfalfa through transcriptome sequencing. The sequencing and gene expression data generated in this study may be used further to elucidate the complete mechanisms governing fall dormancy in alfalfa.

  19. Chloroform extract of alfalfa (Medicago sativa) inhibits lipopolysaccharide-induced inflammation by downregulating ERK/NF-κB signaling and cytokine production.

    Science.gov (United States)

    Choi, Ki-Choon; Hwang, Jung-Min; Bang, Sung-Jun; Kim, Beom-Tae; Kim, Dong-Hern; Chae, Minseon; Lee, Seung-Ah; Choi, Gi Jun; Kim, Da Hye; Lee, Jeong-Chae

    2013-05-01

    Alfalfa (Medicago sativa L.) is commonly used as a traditional medicine and functional food. This study investigated the anti-inflammatory potential of alfalfa and the mechanisms involved. The chloroform extract of alfalfa aerial parts inhibited lipopolysaccharide (LPS)-stimulated immune responses more than ether, butanol, or water soluble extracts. Treatment with 1 μg/mL LPS increased nitrite concentrations to 44.3 μM in RAW267.4 macrophages, but it was reduced to 10.6 μM by adding 100 μg/mL chloroform extract. LPS treatment also increased the concentrations of tumor necrosis factor-α, interleukin (IL)-6, and IL-1β to 41.3, 11.6, and 0.78 ng/mL in culture supernatants of the cells, but these cytokine levels decreased to 12.5, 3.1, and 0.19 ng/mL, respectively, by pretreating with 100 μg/mL of the extract. ICR mice injected with LPS (30 mg/kg body weight) alone showed a 0% survival rate after 48 h of the injection, but 48-h survival of the mice increased to 60% after oral administration of the extract. Subfractions of the chloroform extract markedly suppressed LPS-mediated activation of the extracellular signal-regulated kinase and nuclear factor kappa-B. Cinnamic acid derivatives and fatty acids were found to be active constituents of the extract. This research demonstrated that alfalfa aerial parts exert anti-inflammatory activity and may be useful as a functional food for the prevention of inflammatory disorders.

  20. Co-transforming bar and CsALDH Genes Enhanced Resistance to Herbicide and Drought and Salt Stress in Transgenic Alfalfa (Medicago sativa L.)

    Science.gov (United States)

    Duan, Zhen; Zhang, Daiyu; Zhang, Jianquan; Di, Hongyan; Wu, Fan; Hu, Xiaowen; Meng, Xuanchen; Luo, Kai; Zhang, Jiyu; Wang, Yanrong

    2015-01-01

    Drought and high salinity are two major abiotic factors that restrict the productivity of alfalfa. By application of the Agrobacterium-mediated transformation method, an oxidative responsive gene, CsALDH12A1, from the desert grass Cleistogenes songorica together with the bar gene associated with herbicide resistance, were co-transformed into alfalfa (Medicago sativa L.). From the all 90 transformants, 16 were positive as screened by spraying 1 mL L-1 10% Basta solution and molecularly diagnosis using PCR. Real-time PCR analysis indicated that drought and salt stress induced high CsALDH expression in the leaves of the transgenic plants. The CsALDH expression levels under drought (15 d) and salt stress (200 mM NaCl) were 6.11 and 6.87 times higher than in the control plants, respectively. In comparison to the WT plants, no abnormal phenotypes were observed among the transgenic plants, which showed significant enhancement of tolerance to 15 d of drought and 10 d of salinity treatment. Evaluation of the physiological and biochemical indices during drought and salt stress of the transgenic plants revealed relatively lower Na+ content and higher K+ content in the leaves relative to the WT plants, a reduction of toxic on effects and maintenance of osmotic adjustment. In addition, the transgenic plants could maintain a higher relative water content level, higher shoot biomass, fewer changes in the photosystem, decreased membrane injury, and a lower level of osmotic stress. These results indicate that the co-expression of the introduced bar and CsALDH genes enhanced the herbicide, drought and salt tolerance of alfalfa and therefore can potentially be used as a novel genetic resource for the future breeding programs to develop new cultivars. PMID:26734025

  1. EFICIENCIA DEL AGUA DE RIEGO EN LA PRODUCCIÓN DE MAÍZ FORRAJERO (Zea mays L. Y ALFALFA (Medicago sativa: IMPACTO SOCIAL Y ECONÓMICO

    Directory of Open Access Journals (Sweden)

    Aurelio Pedroza Sandoval

    2014-07-01

    Full Text Available El maíz y la alfalfa, son los dos principales cultivos forrajeros en la Cuenca Lechera de la Laguna de los estados de Coahuila y Durango, México, donde el recurso hídrico es el mayor factor limitante. El objetivo de este estudio fue hacer una evaluación de la eficiencia biológica, económica y social del uso de agua de riego en los cultivos de maíz forrajero (Zea mays L. y alfalfa (Medicago sativa, mediante un análisis económico comparativo y de eficiencia de productividad. El maíz fue más eficiente que la alfalfa, puesto que un m3 de agua subterránea produjo 5.72 kg de biomasa, $0.67 de ganancia por m3 de agua usada y 100 000 m3 de agua produjeron 0.65 empleos durante el ciclo del cultivo; mientras que la alfalfa produjo 0.215 kg de biomasa, $0.90 de ganancia y se generaron 0.43 empleos con los mismos volúmenes de agua indicados en el maíz. El cultivo de maíz forrajero fue más eficiente en producción de biomasa por volumen de agua utilizado y empleos generados, pero menos eficiente en ganancias, donde la alfalfa fue mejor. Una combinación durante el año de cultivos forrajeros con calidad energética y proteica como el maíz y la alfalfa respectivamente, éstos producidos bajo un programa de rotación, habrán de redundar en un equilibrio entre calidad de forraje, productividad del mismo, un mayor beneficio social y un menor impacto ambiental.

  2. Transport and partitioning of CO/sub 2/ fixed by root nodules of ureide and amide producing legumes. [Vigna angularis; Glycine max; Medicago sativa

    Energy Technology Data Exchange (ETDEWEB)

    Vance, C.P.; Boylan, K.L.M.; Maxwell, C.A.; Heichel, G.H.; Hardman, L.L.

    1985-01-01

    Nodulated and denodulated roots of adzuki bean (Vigna angularis), soybean (Glycine max), and alfalfa (Medicago sativa) were exposed to /sup 14/CO/sub 2/ to investigate the contribution of nodule CO/sub 2/ fixation to assimilation and transport of fixed nitrogen. The distribution of radioactivity in xylem sap and partitioning of carbon fixed by nodules to the whole plant were measured. Radioactivity in the xylem sap of nodulated soybean and adzuki bean was located primarily (70 to 87%) in the acid fraction while the basic (amino acid) fraction contained 10 to 22%. In contrast radioactivity in the xylem sap of nodulated alfalfa was primarily in amino acids with about 20% in organic acids. Total ureide concentration was 8.1, 4.7, and 0.0 micromoles per milliliter xylem sap for soybean, adzuki bean, and alfalfa, respectively. While the major nitrogen transport products in soybeans and adzuki beans are ureides, this class of metabolites contained less than 20% of the the total radioactivity. When nodules of plants were removed, radioactivity in xylem sap decreased by 90% or more. Pulse-chase experiments indicated that CO/sub 2/ fixed by nodules was rapidly transported to shoots and incorporated into acid stable constituents. The data are consistent with a role for nodule CO/sub 2/ fixation providing carbon for the assimilation and transport of fixed nitrogen in amide-based legumes. In contrast, CO/sub 2/ fixation by nodules of ureide transporting legumes appears to contribute little to assimilation and transport of fixed nitrogen. 19 references, 2 figures, 5 tables.

  3. Citric acid- and Tween(®) 80-assisted phytoremediation of a co-contaminated soil: alfalfa (Medicago sativa L.) performance and remediation potential.

    Science.gov (United States)

    Agnello, A C; Huguenot, D; van Hullebusch, E D; Esposito, G

    2016-05-01

    A pot experiment was designed to assess the phytoremediation potential of alfalfa (Medicago sativa L.) in a co-contaminated (i.e., heavy metals and petroleum hydrocarbons) soil and the influence of citric acid and Tween(®) 80 (polyethylene glycol sorbitan monooleate), applied individually and combined together, for their possible use in chemically assisted phytoremediation. The results showed that alfalfa plants could tolerate and grow in a co-contaminated soil. Over a 90-day experimental time, shoot and root biomass increased and negligible plant mortality occurred. Heavy metals were uptaken by alfalfa to a limited extent, mostly by plant roots, and their concentration in plant tissues were in the following order: Zn > Cu > Pb. Microbial population (alkane-degrading microorganisms) and activity (lipase enzyme) were enhanced in the presence of alfalfa with rhizosphere effects of 9.1 and 1.5, respectively, after 90 days. Soil amendments did not significantly enhance plant metal concentration or total uptake. In contrast, the combination of citric acid and Tween(®) 80 significantly improved alkane-degrading microorganisms (2.4-fold increase) and lipase activity (5.3-fold increase) in the rhizosphere of amended plants, after 30 days of experiment. This evidence supports a favorable response of alfalfa in terms of tolerance to a co-contaminated soil and improvement of rhizosphere microbial number and activity, additionally enhanced by the joint application of citric acid and Tween(®) 80, which could be promising for future phytoremediation applications.

  4. Salt-tolerant and -sensitive alfalfa (Medicago sativa) cultivars have large variations in defense responses to the lepidopteran insect Spodoptera litura under normal and salt stress condition.

    Science.gov (United States)

    Lei, Yunting; Liu, Qing; Hettenhausen, Christian; Cao, Guoyan; Tan, Qing; Zhao, Weiye; Lin, Honghui; Wu, Jianqiang

    2017-01-01

    In nature, plants are often exposed to multiple stress factors at the same time. Yet, little is known about how plants modulate their physiology to counteract simultaneous abiotic and biotic stresses, such as soil salinity and insect herbivory. In this study, insect performance bioassays, phytohormone measurements, quantification of transcripts, and protein determination were employed to study the phenotypic variations of two alfalfa (Medicago sativa) cultivars in response to insect Spodoptera litura feeding under normal and salt stress condition. When being cultivated in normal soil, the salt-tolerant alfalfa cultivar Zhongmu-1 exhibited lower insect resistance than did the salt-sensitive cultivar Xinjiang Daye. Under salinity stress, the defense responses of Xinjiang Daye were repressed, whereas Zhongmu-1 did not show changes in resistance levels. It is likely that salinity influenced the resistance of Xinjiang Daye through suppressing the accumulation of jasmonic acid-isoleucine (JA-Ile), which is the bioactive hormone inducing herbivore defense responses, leading to attenuated trypsin proteinase inhibitor (TPI) activity. Furthermore, exogenous ABA supplementation suppressed the insect herbivory-induced JA/JA-Ile accumulation and levels of JAR1 (jasmonate resistant 1) and TPI, and further decreased the resistance of Xinjiang Daye, whereas Zhongmu-1 showed very little response to the increased ABA level. We propose a mechanism, in which high levels of abscisic acid induced by salt treatment may affect the expression levels of JAR1 and consequently decrease JA-Ile accumulation and thus partly suppress the defense of Xinjiang Daye against insects under salt stress. This study provides new insight into the mechanism by which alfalfa responds to concurrent abiotic and biotic stresses.

  5. Effects of four short-chain fatty acids or salts on fermentation characteristics and aerobic stability of alfalfa (Medicago sativa L.) silage.

    Science.gov (United States)

    Yuan, Xian J; Wen, Ai Y; Wang, Jian; Desta, Seare T; Dong, Zhi H; Shao, Tao

    2018-01-01

    The objective of the present study was to evaluate the effects of four chemicals on the fermentation quality and aerobic stability of alfalfa (Medicago sativa L.) silage. Wilted alfalfa was ensiled without additive (control), or with formic acid (FA), potassium diformate (KDF), sodium diacetate (SDA) or calcium propionate (CAP). After 60 days of ensiling, the pH values in FA, KDF and SDA silages were lower (P < 0.05) compared to that of control and CAP silages, and chemicals (P < 0.05) decreased butyric acid and ammonia N concentrations and populations of aerobic bacteria and yeasts compared to the control. The SDA and CAP silages had a higher (P < 0.05) lactic acid bacteria content compared to the FA and KDF silages. The SDA and CAP silages had higher (P < 0.05) acetic and propionic acid contents compared to the other silages, respectively. The ammonia N concentrations in the FA and KDF silages were lower compared to the other silages during the first 5 days of aerobic exposure, and then increased sharply to 105 and 100 g kg-1 total N, respectively, which was higher (P < 0.05) than that of the SDA and CAP silages on day 9 of aerobic exposure. Yeasts and aerobic bacteria counts in SDA silage slowly increased and remained at lower levels compared to the other silages after 7 days of aerobic exposure. Additives prolonged the aerobic stability duration compared to the control, and the SDA and CAP silages remained stable for more than 216 h, followed by the KDF and FA silages (202 and 196 h, respectively). © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  6. Risks and benefits of compost-like materials prepared by the thermal treatment of raw scallop hepatopancreas for supplying cadmium and the growth of alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Ushijima, Kensuke; Fukushima, Masami; Kanno, Shinya; Kanno, Itoko; Ohnishi, Mitsuhiro

    2016-01-01

    Scallop hepatopancreas, fishery waste, contains relatively high levels of Cd and organic nitrogen compounds, the latter of which represent a fertilizer. In this study, raw scallop hepatopancreas tissue was thermally treated with sawdust and red loam in the presence of an iron catalyst to produce compost-like materials (CLMs). Two CLM samples were prepared by varying the content of raw scallop hepatopancreas tissue: 46 wt.% for CLM-1 and 18 wt.% for CLM-2. Mixtures of control soil (CTL) and CLMs (CLM content: 10 and 25 wt.%) were examined for the growth of alfalfa (Medicago sativa L.) to evaluate the risks and benefits of using this material for fertilization. The Cd content in shoots and roots of alfalfa, that were grown in the presence of CLMs, was significantly higher than those for the plants grown in the CTL, indicating that Cd had accumulated in the plants from CLMs. The accumulation of Cd in the alfalfa roots was quite high in the case of the 25% CLM-1 sample. However, alfalfa growth was significantly promoted in the presence of 10% CLM-1. This can be attributed to the higher levels of nitrogen and humic substances, which serve as fertilizer components. Although the fertilization effect in case of CLM-1showed a potential benefit, the accumulation of Cd in alfalfa was clearly increased in the presence of both CLMs. In conclusion, the use of CLMs produced from raw scallop hepatopancreas tissue can be considered to have a desirable benefit from standpoint of its use as fertilizer, but is accompanied by a risk of the accumulation of Cd in alfalfa plants.

  7. De Novo Characterization of Fall Dormant and Nondormant Alfalfa (Medicago sativa L.) Leaf Transcriptome and Identification of Candidate Genes Related to Fall Dormancy

    Science.gov (United States)

    Cheng, Ningning; Du, Hongqi; Fan, Wenna; Wang, Chengzhang

    2015-01-01

    Alfalfa (Medicago sativa L.) is one of the most widely cultivated perennial forage legumes worldwide. Fall dormancy is an adaptive character related to the biomass production and winter survival in alfalfa. The physiological, biochemical and molecular mechanisms causing fall dormancy and the related genes have not been well studied. In this study, we sequenced two standard varieties of alfalfa (dormant and non-dormant) at two time points and generated approximately 160 million high quality paired-end sequence reads using sequencing by synthesis (SBS) technology. The de novo transcriptome assembly generated a set of 192,875 transcripts with an average length of 856 bp representing about 165.1 Mb of the alfalfa leaf transcriptome. After assembly, 111,062 (57.6%) transcripts were annotated against the NCBI non-redundant database. A total of 30,165 (15.6%) transcripts were mapped to 323 Kyoto Encyclopedia of Genes and Genomes pathways. We also identified 41,973 simple sequence repeats, which can be used to generate markers for alfalfa, and 1,541 transcription factors were identified across 1,350 transcripts. Gene expression between dormant and non-dormant alfalfa at different time points were performed, and we identified several differentially expressed genes potentially related to fall dormancy. The Gene Ontology and pathways information were also identified. We sequenced and assembled the leaf transcriptome of alfalfa related to fall dormancy, and also identified some genes of interest involved in the fall dormancy mechanism. Thus, our research focused on studying fall dormancy in alfalfa through transcriptome sequencing. The sequencing and gene expression data generated in this study may be used further to elucidate the complete mechanisms governing fall dormancy in alfalfa. PMID:25799491

  8. Melatonin systemically ameliorates drought stress-induced damage in Medicago sativa plants by modulating nitro-oxidative homeostasis and proline metabolism.

    Science.gov (United States)

    Antoniou, Chrystalla; Chatzimichail, Giannis; Xenofontos, Rafaella; Pavlou, Jan J; Panagiotou, Evangelia; Christou, Anastasis; Fotopoulos, Vasileios

    2017-05-01

    Recent reports have uncovered the multifunctional role of melatonin in plant physiological responses under optimal and suboptimal environmental conditions. In this study, we explored whether melatonin pretreatment could provoke priming effects in alfalfa (Medicago sativa L.) plants subsequently exposed to prolonged drought stress (7 days), by withholding watering. Results revealed that the rhizospheric application of melatonin (10 μmol L-1 ) remarkably enhanced the drought tolerance of alfalfa plants, as evidenced by the observed plant tolerant phenotype, as well as by the higher levels of chlorophyll fluorescence and stomatal conductance, compared with nontreated drought-stressed plants. In addition, lower levels of lipid peroxidation (MDA content) as well as of both H2 O2 and NO contents in primed compared with nonprimed stressed plants suggest that melatonin pretreatment resulted in the systemic mitigation of drought-induced nitro-oxidative stress. Nitro-oxidative homeostasis was achieved by melatonin through the regulation of reactive oxygen (SOD, GR, CAT, APX) and nitrogen species (NR, NADHde) metabolic enzymes at the enzymatic and/or transcript level. Moreover, melatonin pretreatment resulted in the limitation of cellular redox disruption through the regulation of the mRNA levels of antioxidant and redox-related components (ADH, AOX, GST7, GST17), as well via osmoprotection through the regulation of proline homeostasis, at both the enzymatic (P5CS) and gene expression level (P5CS, P5CR). Overall, novel results highlight the importance of melatonin as a promising priming agent for the enhancement of plant tolerance to drought conditions through the regulation of nitro-oxidative and osmoprotective homeostasis. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  9. Co-transforming bar and CsALDH genes enhanced resistance to herbicide and drought and salt stress in transgenic alfalfa (Medicago sativa L.

    Directory of Open Access Journals (Sweden)

    Zhen eDuan

    2015-12-01

    Full Text Available Drought and high salinity are two major abiotic factors that restrict the productivity of alfalfa. By application of the Agrobacterium-mediated transformation method, an oxidative responsive gene, CsALDH12A1, from the desert grass Cleistogenes songorica together with the bar gene associated with herbicide resistance, were co-transformed into alfalfa (Medicago sativa L.. From the all 90 transformants, 16 were positive as screened by spraying 1 mL L-1 10% Basta solution and molecularly diagnosis using PCR. Real-time PCR analysis indicated that drought and salt stress induced high CsALDH expression in the leaves of the transgenic plants. The CsALDH expression levels under drought (15 d and salt stress (200 mM NaCl were 6.11 and 6.87 times higher than in the control plants, respectively. In comparison to the WT plants, no abnormal phenotypes were observed among the transgenic plants, which showed significant enhancement of tolerance to 15 d of drought and 10 d of salinity treatment. Evaluation of the physiological and biochemical indices during drought and salt stress of the transgenic plants revealed relatively lower Na+ content and higher K+ content in the leaves relative to the WT plants, a reduction of toxic on effects and maintenance of osmotic adjustment. In addition, the transgenic plants could maintain a higher relative water content (RWC level, higher shoot biomass, fewer changes in the photosystem, decreased membrane injury, and a lower level of osmotic stress. These results indicate that the co-expression of the introduced bar and CsALDH genes enhanced the herbicide, drought and salt tolerance of alfalfa and therefore can potentially be used as a novel genetic resource for the future breeding programs to develop new cultivars.

  10. Impacts of coal fly ash on plant growth and accumulation of essential nutrients and trace elements by alfalfa (Medicago sativa) grown in a loessial soil.

    Science.gov (United States)

    He, Honghua; Dong, Zhigang; Peng, Qi; Wang, Xia; Fan, Chenbin; Zhang, Xingchang

    2017-07-15

    Coal fly ash (CFA) is a problematic solid waste all over the world. One distinct beneficial reuse of CFA is its utilization in land application as a soil amendment. A pot experiment was carried out to assess the feasibility of using CFA to improve plant growth and increase the supply of plant-essential elements and selenium (Se) of a loessial soil for agricultural purpose. Plants of alfalfa (Medicago sativa) were grown in a loessial soil amended with different rates (5%, 10%, 20% and 40%) of CFA for two years and subjected to four successive cuttings. Dry mass of shoots and roots, concentrations of plant-essential elements and Se in plants were measured. Shoot dry mass and root dry mass were always significantly increased by 5%, 10% and 20% CFA treatments, and by 40% CFA treatment in all harvests except the first one. The CFA had a higher supply of exchangeable phosphorus (P), magnesium (Mg), copper (Cu), zinc (Zn), molybdenum (Mo), and Se than the loessial soil. Shoot P, calcium (Ca), Mg, Mo, boron (B), and Se concentrations were generally markedly increased, but shoot potassium (K), Cu, and Zn concentrations were generally reduced. The CFA can be a promising source of some essential elements and Se for plants grown in the loessial soil, and an application rate of not higher than 5% should be safe for agricultural purpose without causing plant toxicity symptoms in the studied loessial soil and similar soils. Field trials will be carried out to confirm the results of the pot experiment. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Haem oxygenase-1 is involved in salicylic acid-induced alleviation of oxidative stress due to cadmium stress in Medicago sativa.

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    Cui, Weiti; Li, Le; Gao, Zhaozhou; Wu, Honghong; Xie, Yanjie; Shen, Wenbiao

    2012-09-01

    This work examines the involvement of haem oxygenase-1 (HO-1) in salicylic acid (SA)-induced alleviation of oxidative stress as a result of cadmium (Cd) stress in alfalfa (Medicago sativa L.) seedling roots. CdCl(2) exposure caused severe growth inhibition and Cd accumulation, which were potentiated by pre-treatment with zinc protoporphyrin (ZnPPIX), a potent HO-1 inhibitor. Pre-treatment of plants with the HO-1 inducer haemin or SA, both of which could induce MsHO1 gene expression, significantly reduced the inhibition of growth and Cd accumulation. The alleviation effects were also evidenced by a decreased content of thiobarbituric acid-reactive substances (TBARS). The antioxidant behaviour was confirmed by histochemical staining for the detection of lipid peroxidation and the loss of plasma membrane integrity. Furthermore, haemin and SA pre-treatment modulated the activities of ascorbate peroxidase (APX), superoxide dismutase (SOD), and guaiacol peroxidase (POD), or their corresponding transcripts. Significant enhancement of the ratios of reduced/oxidized homoglutathione (hGSH), ascorbic acid (ASA)/dehydroascorbate (DHA), and NAD(P)H/NAD(P)(+), and expression of their metabolism genes was observed, consistent with a decreased reactive oxygen species (ROS) distribution in the root tips. These effects are specific for HO-1, since ZnPPIX blocked the above actions, and the aggravated effects triggered by SA plus ZnPPIX were differentially reversed when carbon monoxide (CO) or bilirubin (BR), two catalytic by-products of HO-1, was added. Together, the results suggest that HO-1 is involved in the SA-induced alleviation of Cd-triggered oxidative stress by re-establishing redox homeostasis.

  12. Genome-wide SNP discovery in tetraploid alfalfa using 454 sequencing and high resolution melting analysis

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    Zhao Patrick X

    2011-07-01

    Full Text Available Abstract Background Single nucleotide polymorphisms (SNPs are the most common type of sequence variation among plants and are often functionally important. We describe the use of 454 technology and high resolution melting analysis (HRM for high throughput SNP discovery in tetraploid alfalfa (Medicago sativa L., a species with high economic value but limited genomic resources. Results The alfalfa genotypes selected from M. sativa subsp. sativa var. 'Chilean' and M. sativa subsp. falcata var. 'Wisfal', which differ in water stress sensitivity, were used to prepare cDNA from tissue of clonally-propagated plants grown under either well-watered or water-stressed conditions, and then pooled for 454 sequencing. Based on 125.2 Mb of raw sequence, a total of 54,216 unique sequences were obtained including 24,144 tentative consensus (TCs sequences and 30,072 singletons, ranging from 100 bp to 6,662 bp in length, with an average length of 541 bp. We identified 40,661 candidate SNPs distributed throughout the genome. A sample of candidate SNPs were evaluated and validated using high resolution melting (HRM analysis. A total of 3,491 TCs harboring 20,270 candidate SNPs were located on the M. truncatula (MT 3.5.1 chromosomes. Gene Ontology assignments indicate that sequences obtained cover a broad range of GO categories. Conclusions We describe an efficient method to identify thousands of SNPs distributed throughout the alfalfa genome covering a broad range of GO categories. Validated SNPs represent valuable molecular marker resources that can be used to enhance marker density in linkage maps, identify potential factors involved in heterosis and genetic variation, and as tools for association mapping and genomic selection in alfalfa.

  13. Design of optimal solvent for extraction of bio–active ingredients from six varieties of Medicago sativa

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    Caunii Angela

    2012-10-01

    Full Text Available Abstract Background Extensive research has been performed worldwide and important evidences were collected to show the immense potential of plants used in various traditional therapeutic systems. The aim of this work is to investigate the different extracting solvents in terms of the influence of their polarity on the extracting ability of bioactive molecules (phenolic compounds from the M. sativa flowers. Results The total phenolic content of samples was determined using the Folin Ciocalteu (FC procedure and their antioxidant activity was assayed through in vitro radical decomposing activity using the radical DPPH° assay (IUPAC name for DPPH is (phenyl–(2,4,6–trinitrophenyl iminoazanium. The results showed that water was better than methanol and acetic acid for extracting bioactive compounds, in particular for total phenolic compounds from the flowers of alfalfa. The average content of bioactive molecules in methanol extract was 263.5±1.02 mg GAE/100g of dry weight lyophilized extract. The total phenolic content of the tested plant extracts was highly correlated with the radical decomposing activity. However, all extracts were free–radical inhibitors, but the water extract was more potent than the acetic and the methanol ones. The order of inhibitor effectiveness (expressed by IC50 proved to be: water extract (0.924mg/mL > acetic acid extract (0.154mg/mL > methanol (0.079mg/mL. The profiles of each extract (fingerprint were characterized by FT–MIR spectroscopy. Conclusions The present study compares the fingerprint of different extracts of the M. sativa flowers, collected from the wild flora of Romania. The total phenolic content of the tested plant extracts was highly correlated with the radical decomposing activity. The dependence of the extract composition on the solvent polarity (acetic acid vs. methanol vs. water was revealed by UV–VIS spectrometry and Infrared fingerprint.

  14. Contrasting Storage Protein Synthesis and Messenger RNA Accumulation during Development of Zygotic and Somatic Embryos of Alfalfa (Medicago sativa L.) 1

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    Krochko, Joan E.; Pramanik, Saroj K.; Bewley, J. Derek

    1992-01-01

    During development on hormone-free media, somatic embryos pass through distinct morphological stages that superficially resemble those of zygotic embryo development (globular, heart, torpedo, cotyledonary stages). Despite these similarities, they differ from zygotic embryos in the extent of cotyledonary development and the patterns of synthesis and quantitative expression of seed-specific storage proteins (7S, 11S, and 2S proteins). Alfin (7S) is the first storage protein synthesized in developing zygotic embryos (stage IV). The 11S (medicagin) and 2S (Low Molecular Weight, LMW) storage proteins are not detectable until the following stage of development (stage V), although all three are present before the completion of embryo enlargement. Likewise, the 7S storage protein is the first to be synthesized in developing somatic embryos (day 5). Medicagin is evident by day 7 and the LMW protein by day 10. In contrast to zygotic embryos, alfin remains the predominant storage protein in somatic embryos throughout development. Not only are the relative amounts of medicagin and the LMW protein reduced in somatic embryos but the LMW protein is accumulated much later than the other proteins. Quantification of the storage protein mRNAs (7S, 11S, and 2S) by northern blot analysis confirms that there are substantial differences in the patterns of message accumulation in zygotic and somatic embryos of alfalfa (Medicago sativa). In zygotic embryos, the 7S, 11S, and 2S storage protein mRNAs are abundant during maturation and, in particular, during the stages of maximum protein synthesis (alfin, stages VI and VII; medicagin, stage VII; LMW, stage VII). In somatic embryos, the predominance of the 7S storage protein is correlated with increased accumulation of its mRNA, whereas the limited synthesis of the 11S storage protein is associated with much lower steady-state levels of its message. The mRNA for the LMW protein is present already by 3 days after transfer to hormone-free media

  15. Alfalfa (Medicago sativa L.) is tolerant to higher levels of salinity than previous guidelines indicated: Implications of field and greenhouse studies

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    Putnam, Daniel H.; Benes, Sharon; Galdi, Giuliano; Hutmacher, Bob; Grattan, Steve

    2017-04-01

    Alfalfa (Medicago sativa L.) is the most widely grown leguminous forage crop in North America and is valued for high productivity, quality, economic value, and for dairy productivity. Alfalfa has historically been classified as moderately sensitive to saline conditions, with yield declines predicted at >2 dS/m in the saturated soil paste extract. However, greenhouse, sand tank, and field studies over the past five years have confirmed that alfalfa can be grown with limited negative effects at much higher salinity levels. A broad collection of alfalfa varieties has exhibited a range of resistance at irrigation water salinities >5 dS/m ECw in greenhouse trials, with significant variation due to variety. USDA-ARS sand tank studies indicated similar or greater tolerances closer to 8 dS/m in the soil water, in addition to confirmation of significant varietal differences. A three-year field study on clay loam soil with applications of 5-7 dS/m ECw irrigation water indicated normal yields and excellent stand survivability. A second field study in the same soil type with levels from 8-10 dS/m ECw showed yield reductions of 10-15% but economic yields were still achieved at those levels. Field and greenhouse studies were conducted with mixed salt saline sodic waters typical of the San Joaquin Valley of California. Field evaluation of variety performance was subject to greater variation due to secondary salinity-soil interactions including water infiltration and crusting problems, not only salinity per-se. Thus, adequate irrigation water availability to the crop may be as important as salinity in impacting yields under field conditions. Once established, the deep-rooted characteristics of alfalfa enable utilization of deeper subsurface moisture, even at moderate to high salinity levels, as documented by USDA lysimeter studies. Significant advantages to salinity-tolerant varieties have been observed. It will be important to consider specific management factors which may enable

  16. Transgene silencing of sucrose synthase in alfalfa (Medicago sativa L.) stem vascular tissue suggests a role for invertase in cell wall cellulose synthesis.

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    Samac, Deborah A; Bucciarelli, Bruna; Miller, Susan S; Yang, S Samuel; O'Rourke, Jamie A; Shin, Sanghyun; Vance, Carroll P

    2015-12-01

    Alfalfa (Medicago sativa L.) is a widely adapted perennial forage crop that has high biomass production potential. Enhanced cellulose content in alfalfa stems would increase the value of the crop as a bioenergy feedstock. We examined if increased expression of sucrose synthase (SUS; EC 2.4.1.13) would increase cellulose in stem cell walls. Alfalfa plants were transformed with a truncated alfalfa phosphoenolpyruvate carboxylase gene promoter (PEPC7-P4) fused to an alfalfa nodule-enhanced SUS cDNA (MsSUS1) or the β-glucuronidase (GUS) gene. Strong GUS expression was detected in xylem and phloem indicating that the PEPC7-P4 promoter was active in stem vascular tissue. In contrast to expectations, MsSUS1 transcript accumulation was reduced 75-90 % in alfalfa plants containing the PEPC7-P4::MsSUS1 transgene compared to controls. Enzyme assays indicated that SUS activity in stems of selected down-regulated transformants was reduced by greater than 95 % compared to the controls. Although SUS activity was detected in xylem and phloem of control plants by in situ enzyme assays, plants with the PEPC7-P4::MsSUS1 transgene lacked detectable SUS activity in post-elongation stem (PES) internodes and had very low SUS activity in elongating stem (ES) internodes. Loss of SUS protein in PES internodes of down-regulated lines was confirmed by immunoblots. Down-regulation of SUS expression and activity in stem tissue resulted in no obvious phenotype or significant change in cell wall sugar composition. However, alkaline/neutral (A/N) invertase activity increased in SUS down-regulated lines and high levels of acid invertase activity were observed. In situ enzyme assays of stem tissue showed localization of neutral invertase in vascular tissues of ES and PES internodes. These results suggest that invertases play a primary role in providing glucose for cellulose biosynthesis or compensate for the loss of SUS1 activity in stem vascular tissue.

  17. Cloning and expression of 1-aminocyclopropane-1-carboxylate oxidase cDNA induced by thidiazuron during somatic embryogenesis of alfalfa (Medicago sativa).

    Science.gov (United States)

    Feng, Bi-Hong; Wu, Bei; Zhang, Chun-Rong; Huang, Xia; Chen, Yun-Feng; Huang, Xue-Lin

    2012-01-15

    Embryogenic callus (EC) induced from petioles of alfalfa (Medicago sativa L. cv. Jinnan) on B5h medium turned green, compact and non-embryogenic when the kinetin (KN) in the medium was replaced partially or completely by thidiazuron (TDZ). The application of CoCl₂, which is an inhibitor of 1-aminocyclopropane-1-carboxylate oxidase (ACO), counteracted the effect of TDZ. Ethylene has been shown to be involved in the modulation of TDZ-induced morphogenesis responses. However, very little is known about the genes involved in ethylene formation during somatic embryogenesis (SE). To investigate whether ethylene mediated by ACO is involved in the effect of TDZ on inhibition of embryogenic competence of the alfalfa callus. In this study we cloned full-length ACO cDNA from the alfalfa callus, named MsACO, and observed changes in this gene expression during callus formation and induction of SE under treatment with TDZ or TDZ plus CoCl₂. RNA blot analysis showed that during the EC subcultural period, the expression level of MsACO in EC was significantly increased on the 2nd day, rose to the highest level on the 8th day and remained at this high level until the 21st day. However, the ACO expression in the TDZ (0.93 μM)-treated callus was higher than in the EC especially on the 8th day. Moreover the ACO expression level increased with increasing TDZ concentration during the subcultural/maintenance period of the callus. It is worth noting that comparing the treatment with TDZ alone, the treatment with 0.93 μM TDZ plus 50 μM CoCl₂ reduced both of the ACO gene expressions and ACO activity in the treated callus. These results indicate that the effect of TDZ could be counteracted by CoCl₂ either on the ACO gene expression level or ACO activity. Thus, a TDZ inhibitory effect on embryogenic competence of alfalfa callus could be mediated by ACO gene expression. Crown Copyright © 2011. Published by Elsevier GmbH. All rights reserved.

  18. Expression of the R2R3-MYB Transcription Factor TaMYB14 from Trifolium arvense Activates Proanthocyanidin Biosynthesis in the Legumes Trifolium repens and Medicago sativa1[W][OA

    Science.gov (United States)

    Hancock, Kerry R.; Collette, Vern; Fraser, Karl; Greig, Margaret; Xue, Hong; Richardson, Kim; Jones, Chris; Rasmussen, Susanne

    2012-01-01

    Proanthocyanidins (PAs) are oligomeric flavonoids and one group of end products of the phenylpropanoid pathway. PAs have been reported to be beneficial for human and animal health and are particularly important in pastoral agricultural systems for improved animal production and reduced greenhouse gas emissions. However, the main forage legumes grown in these systems, such as Trifolium repens and Medicago sativa, do not contain any substantial amounts of PAs in leaves. We have identified from the foliar PA-accumulating legume Trifolium arvense an R2R3-MYB transcription factor, TaMYB14, and provide evidence that this transcription factor is involved in the regulation of PA biosynthesis in legumes. TaMYB14 expression is necessary and sufficient to up-regulate late steps of the phenylpropanoid pathway and to induce PA biosynthesis. RNA interference silencing of TaMYB14 resulted in almost complete cessation of PA biosynthesis in T. arvense, whereas Nicotiana tabacum, M. sativa, and T. repens plants constitutively expressing TaMYB14 synthesized and accumulated PAs in leaves up to 1.8% dry matter. Targeted liquid chromatography-multistage tandem mass spectrometry analysis identified foliar PAs up to degree of polymerization 6 in leaf extracts. Hence, genetically modified M. sativa and T. repens plants expressing TaMYB14 provide a viable option for improving animal health and mitigating the negative environmental impacts of pastoral animal production systems. PMID:22566493

  19. Efecto de las arañas (Arachnida: Araneae como depredadoras de insectos plaga en cultivos de alfalfa (Medicago sativa (Fabaceae en Argentina

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    Andrea Armendano

    2011-12-01

    Full Text Available Effect of spiders (Arachnida: Araneae as predators of insect pest in alfalfa crops (Medicago sativa (Fabaceae in Argentina. Spiders are predators that reduce insect pest populations in agroecosystems. Trials were conducted to measure the selectivity against different insect preys, the daily consumption, effect of predators alone and together with a known number of preys, and the indirect effect of predators on vegetation. For this, experimental units (1x1m were used covered with a fine plastic mesh. Misumenops pallidus, Oxyopes salticus and Araneus sp. were used as generalist predators, and aphids, weevils, locusts, chrysomelids and Lepidoptera larvae as their potential preys. Among the preys offered, the spiders preferred Lepidoptera larvae compared to the other two pests groups (weevils and aphids. The maximum consumption rate was of 93.33% for Lepidoptera larvae, 25.33% for aphids and 11.67% for weevils. The Q Index values for the three species of spiders showed a positive selectivity only for defoliating larvae. O. salticus showed the highest values of consumption rates while Rachiplusia nu was the most consumed. The maximum value of consumption in 24 hours was showed by O. salticus on R. nu (C=2.8. The association of several species of predatory spiders increased the total number of insects captured, and also showed that the addition of spiders caused a decrease in the number of leaves damaged by the effect of lepidopterous larvae. Rev. Biol. Trop. 59 (4: 1651-1662. Epub 2011 December 01Las arañas son depredadoras capaces de reducir las poblaciones de insectos plaga en agroecosistemas. Para medir la selectividad frente a distintas presas, se realizaron ensayos de consumo diario, efecto de los depredadores aisladamente y en conjunto sobre el número de presas y efecto indirecto de los depredadores sobre la vegetación; se utilizaron jaulas experimentales de 1x1m cubiertas con una fina malla plástica. Misumenops pallidus, Oxyopes salticus y

  20. Effects of untreated and treated wastewater at the morphological, physiological and biochemical levels on seed germination and development of sorghum (Sorghum bicolor (L.) Moench), alfalfa (Medicago sativa L.) and fescue (Festuca arundinacea Schreb.).

    Science.gov (United States)

    Rekik, Imen; Chaabane, Zayneb; Missaoui, Amara; Bouket, Ali Chenari; Luptakova, Lenka; Elleuch, Amine; Belbahri, Lassaad

    2017-03-15

    Wastewater reuse in agriculture may help mitigate water scarcity. This may be reached if high quality treatments removing harmful pollutants are applied. The aim of the present study was to compare the effect of untreated (UTW) and treated wastewater (TW) on germination and seedlings development of alfalfa (Medicago sativa L.), fescue (Festuca arundinacea Schreb.) and sorghum (Sorghum bicolor (L.) Moench). UTW presented high turbidity (130 NTU), chemical and biological oxygen demand (COD, 719mgL-1, BOD5, 291mgL-1) and metal concentrations. These levels caused mortality (18% for fescue), decreased germination speed in seeds (37.5% for alfalfa) and reductions of root and stem length in seedlings (80% and 22% respectively for alfalfa). Adverse effects on seeds germination were reflected at the biochemical level by increased H2O2 levels (6 times for sorghum after 5days) and by increased Malondialdehyde (MDA) levels (more than 600 times for sorghum roots) during seedlings development. When TW was used, these parameters were close to control seeds ones. They were also dependent on plant species and developmental stage. Therefore, for efficient reclaimed wastewater reuse in irrigation, suitable crops, displaying wide tolerance to toxic contents during germination and later seedling development stages have to be selected. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Nodule-enhanced expression of a sucrose phosphate synthase gene member (MsSPSA) has a role in carbon and nitrogen metabolism in the nodules of alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Aleman, Lorenzo; Ortega, Jose Luis; Martinez-Grimes, Martha; Seger, Mark; Holguin, Francisco Omar; Uribe, Diana J; Garcia-Ibilcieta, David; Sengupta-Gopalan, Champa

    2010-01-01

    Sucrose phosphate synthase (SPS) catalyzes the first step in the synthesis of sucrose in photosynthetic tissues. We characterized the expression of three different isoforms of SPS belonging to two different SPS gene families in alfalfa (Medicago sativa L.), a previously identified SPS (MsSPSA) and two novel isoforms belonging to class B (MsSPSB and MsSPSB3). While MsSPSA showed nodule-enhanced expression, both MsSPSB genes exhibited leaf-enhanced expression. Alfalfa leaf and nodule SPS enzymes showed differences in chromatographic and electrophoretic migration and differences in V (max) and allosteric regulation. The root nodules in legume plants are a strong sink for photosynthates with its need for ATP, reducing power and carbon skeletons for dinitrogen fixation and ammonia assimilation. The expression of genes encoding SPS and other key enzymes in sucrose metabolism, sucrose phosphate phosphatase and sucrose synthase, was analyzed in the leaves and nodules of plants inoculated with Sinorhizobium meliloti. Based on the expression pattern of these genes, the properties of the SPS isoforms and the concentration of starch and soluble sugars in nodules induced by a wild type and a nitrogen fixation deficient strain, we propose that SPS has an important role in the control of carbon flux into different metabolic pathways in the symbiotic nodules.

  2. Aspectos fisiológicos do crescimento da alfafa (Medicago sativa L. cv. Crioula após pulverização com ácido giberélico durante o período de seca invernal - DOI: 10.4025/actascianimsci.v25i1.2148 Physiological aspects of lucerne (Medicago sativa L. cv. Crioula growth after gibberellic acid application during the winter drought season - DOI: 10.4025/actascianimsci.v25i1.2148

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    Neli Mariza Azevedo Silva

    2003-04-01

    Full Text Available Os objetivos deste trabalho foram verificar alterações no crescimento e na produção da alfafa (Medicago sativa L. cv. Crioula, pulverizada com ácido giberélico (GA3 em época de crescimento reduzido (abril a setembro, em condições de campo. O experimento consistiu de três tratamentos (0, 5 e 10mg/kg GA3, com sete repetições, sendo que foram realizados seis cortes com intervalos de 28 dias. Os resultados obtidos indicaram que não houve diferença estatística (p 3 nas condições experimentais.The aim of this research was to evaluate, in field conditions, the growth and production of lucerne cv. Crioula sprayed with gibberellic acid (GA3 during slowly growth period (April to September. The experiment consisted of three treatments (0, 5 and 10mg/kg GA3 with seven repetitions each one and six cuts with 28 days intervals. The investigated parameters did not differ (p 3 application in experimental conditions

  3. Gene-Silencing-Induced Changes in Carbohydrate Conformation in Relation to Bioenergy Value and Carbohydrate Subfractions in Modeled Plant (Medicago sativa) with Down-Regulation of HB12 and TT8 Transcription Factors

    Science.gov (United States)

    Li, Xinxin; Hannoufa, Abdelali; Zhang, Yonggen; Yu, Peiqiang

    2016-01-01

    Gene silencing with RNA interference (RNAi) technology may be capable of modifying internal structure at a molecular level. This structural modification could affect biofunctions in terms of biodegradation, biochemical metabolism, and bioactive compound availability. The objectives of this study were to (1) Detect gene silencing-induced changes in carbohydrate molecular structure in an alfalfa forage (Medicago sativa spp. sativa: alfalfa) with down-regulation of genes that encode transcription factors TT8 and HB12; (2) Determine gene silencing-induced changes in nutrient bioutilization and bioavailability in the alfalfa forage (Medicago sativa); and (3) Quantify the correlation between gene silencing-induced molecular structure changes and the nutrient bioutilization and bioavailability in animals of ruminants. The experimental treatments included: T1 = Non-transgenic and no-gene silenced alfalfa forage (code “NT”); T2 = HB12-RNAi forage with HB12 gene down regulation (code “HB12”); T3 = TT8-RNAi forage with TT8 gene down regulation (code “TT8”). The HB12 and TT8 gene silencing-induced molecular structure changes were determined by non-invasive and non-destructive advanced molecular spectroscopy in a middle infrared radiation region that focused on structural, non-structural and total carbohydrate compounds. The nutrient bioutilization and bioavailability of the modified forage were determined using NRC-2001 system in terms of total digestive nutrient (TDN), truly digestible fiber (tdNDF), non-fiber carbohydrate (tdNDF), fatty acid (tdFA), crude protein (tdCP) and bioenergy profiles (digestible energy, metabolizable energy, net energy) for ruminants. The carbohydrate subfractions were evaluated using the updated CNCPS 6.0 system. The results showed that gene silencing significantly affected tdNFC (42.3 (NT) vs. 38.7 (HB12) vs. 37.4% Dry Matter (TT8); p = 0.016) and tdCP (20.8 (NT) vs. 19.4 (HB12) vs. 22.3% DM (TT8); p = 0.009). The gene-silencing also

  4. Gene-Silencing-Induced Changes in Carbohydrate Conformation in Relation to Bioenergy Value and Carbohydrate Subfractions in Modeled Plant (Medicago sativa with Down-Regulation of HB12 and TT8 Transcription Factors

    Directory of Open Access Journals (Sweden)

    Xinxin Li

    2016-05-01

    Full Text Available Gene silencing with RNA interference (RNAi technology may be capable of modifying internal structure at a molecular level. This structural modification could affect biofunctions in terms of biodegradation, biochemical metabolism, and bioactive compound availability. The objectives of this study were to (1 Detect gene silencing-induced changes in carbohydrate molecular structure in an alfalfa forage (Medicago sativa spp. sativa: alfalfa with down-regulation of genes that encode transcription factors TT8 and HB12; (2 Determine gene silencing-induced changes in nutrient bioutilization and bioavailability in the alfalfa forage (Medicago sativa; and (3 Quantify the correlation between gene silencing-induced molecular structure changes and the nutrient bioutilization and bioavailability in animals of ruminants. The experimental treatments included: T1 = Non-transgenic and no-gene silenced alfalfa forage (code “NT”; T2 = HB12-RNAi forage with HB12 gene down regulation (code “HB12”; T3 = TT8-RNAi forage with TT8 gene down regulation (code “TT8”. The HB12 and TT8 gene silencing-induced molecular structure changes were determined by non-invasive and non-destructive advanced molecular spectroscopy in a middle infrared radiation region that focused on structural, non-structural and total carbohydrate compounds. The nutrient bioutilization and bioavailability of the modified forage were determined using NRC-2001 system in terms of total digestive nutrient (TDN, truly digestible fiber (tdNDF, non-fiber carbohydrate (tdNDF, fatty acid (tdFA, crude protein (tdCP and bioenergy profiles (digestible energy, metabolizable energy, net energy for ruminants. The carbohydrate subfractions were evaluated using the updated CNCPS 6.0 system. The results showed that gene silencing significantly affected tdNFC (42.3 (NT vs. 38.7 (HB12 vs. 37.4% Dry Matter (TT8; p = 0.016 and tdCP (20.8 (NT vs. 19.4 (HB12 vs. 22.3% DM (TT8; p = 0.009. The gene-silencing also

  5. Gene-Silencing-Induced Changes in Carbohydrate Conformation in Relation to Bioenergy Value and Carbohydrate Subfractions in Modeled Plant (Medicago sativa) with Down-Regulation of HB12 and TT8 Transcription Factors.

    Science.gov (United States)

    Li, Xinxin; Hannoufa, Abdelali; Zhang, Yonggen; Yu, Peiqiang

    2016-05-13

    Gene silencing with RNA interference (RNAi) technology may be capable of modifying internal structure at a molecular level. This structural modification could affect biofunctions in terms of biodegradation, biochemical metabolism, and bioactive compound availability. The objectives of this study were to (1) Detect gene silencing-induced changes in carbohydrate molecular structure in an alfalfa forage (Medicago sativa spp. sativa: alfalfa) with down-regulation of genes that encode transcription factors TT8 and HB12; (2) Determine gene silencing-induced changes in nutrient bioutilization and bioavailability in the alfalfa forage (Medicago sativa); and (3) Quantify the correlation between gene silencing-induced molecular structure changes and the nutrient bioutilization and bioavailability in animals of ruminants. The experimental treatments included: T1 = Non-transgenic and no-gene silenced alfalfa forage (code "NT"); T2 = HB12-RNAi forage with HB12 gene down regulation (code "HB12"); T3 = TT8-RNAi forage with TT8 gene down regulation (code "TT8"). The HB12 and TT8 gene silencing-induced molecular structure changes were determined by non-invasive and non-destructive advanced molecular spectroscopy in a middle infrared radiation region that focused on structural, non-structural and total carbohydrate compounds. The nutrient bioutilization and bioavailability of the modified forage were determined using NRC-2001 system in terms of total digestive nutrient (TDN), truly digestible fiber (tdNDF), non-fiber carbohydrate (tdNDF), fatty acid (tdFA), crude protein (tdCP) and bioenergy profiles (digestible energy, metabolizable energy, net energy) for ruminants. The carbohydrate subfractions were evaluated using the updated CNCPS 6.0 system. The results showed that gene silencing significantly affected tdNFC (42.3 (NT) vs. 38.7 (HB12) vs. 37.4% Dry Matter (TT8); p = 0.016) and tdCP (20.8 (NT) vs. 19.4 (HB12) vs. 22.3% DM (TT8); p = 0.009). The gene-silencing also affected

  6. [Experimental tolerance to boron of the plant species Nicotiana glauca, Jacaranda mimosifolia, Tecoma stans, Medicago sativa y Spinacea oleracea in Argentina].

    Science.gov (United States)

    de Viana, Marta L; Albarracín Franco, Silvia

    2008-09-01

    The activity of boron industries is a punctual and diffuse source of air, soil and water pollution. Therefore, it is a priority to study possible ways of reducing this impact. A relatively new technology for reducing soil pollution is phytoremediation, which uses plants and associate microorganisms. The first step in phytoremediation is to detect tolerant plant species, which is the objective of this work. A laboratory experiment to assess the germination, survival and growth of different species at different boron concentrations was carried out following a factorial design with two factors: plant species and boron concentration. Boron concentrations were determined at the beginning and the end of the experiment, taking into account substrates with and without vegetation. We found significant differences for treatment, species and the interaction species*treatment. N. glauca, M. sativa and J. mimosifolia were the most tolerant species. The other species had a decrease in the response variables, with the concentration of the pollutant. All the species had a low survival at the highest boron concentration. The reduction in boron concentration at the end of the experiment was higher in the 30 ppm treatment with M. sativa and the lower was registered in the 20 ppm treatment with J. mimosifolia and in 30 ppm with T. stans and S. oleraceae. We conclude that N. glauca, M. sativa and J. mimosifolia can be considered in remediation plans.

  7. Genome-Wide Association Mapping of Loci Associated with Plant Growth and Forage Production under Salt Stress in Alfalfa (Medicago sativa L.

    Directory of Open Access Journals (Sweden)

    Xiang-Ping Liu

    2017-05-01

    Full Text Available Salinity tolerance is highly desirable to sustain alfalfa production in marginal lands that have been rendered saline. In this study, we used a diverse panel of 198 alfalfa accessions for mapping loci associated with plant growth and forage production under salt stress using genome-wide association studies (GWAS. The plants were genotyped using genotyping-by-sequencing (GBS. A greenhouse procedure was used for phenotyping four agronomic and physiological traits affected by salt stress, including dry weight (DW, plant height (PH, leaf chlorophyll content (LCC, and stomatal conductance (SC. For each trait, a stress susceptibility index (SSI was used to evaluate plant performance under stressed and non-stressed conditions. Marker-trait association identified a total of 42 markers significantly associated with salt tolerance. They were located on all chromosomes except chromosome 2 based on the alignment of their flanking sequences to the reference genome (Medicago truncatula. Of those identified, 13 were associated with multiple traits. Several loci identified in the present study were also identified in previous reports. BLAST search revealed that 19 putative candidate genes linked to 24 significant markers. Among them, B3 DNA-binding protein, Thiaminepyrophosphokinase and IQ calmodulin-binding motif protein were identified among multiple traits in the present and previous studies. With further investigation, these markers and candidates would be useful for developing markers for marker-assisted selection in breeding programs to improve alfalfa cultivars with enhanced tolerance to salt stress.

  8. Genome-Wide Association Mapping of Loci Associated with Plant Growth and Forage Production under Salt Stress in Alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Liu, Xiang-Ping; Yu, Long-Xi

    2017-01-01

    Salinity tolerance is highly desirable to sustain alfalfa production in marginal lands that have been rendered saline. In this study, we used a diverse panel of 198 alfalfa accessions for mapping loci associated with plant growth and forage production under salt stress using genome-wide association studies (GWAS). The plants were genotyped using genotyping-by-sequencing (GBS). A greenhouse procedure was used for phenotyping four agronomic and physiological traits affected by salt stress, including dry weight (DW), plant height (PH), leaf chlorophyll content (LCC), and stomatal conductance (SC). For each trait, a stress susceptibility index (SSI) was used to evaluate plant performance under stressed and non-stressed conditions. Marker-trait association identified a total of 42 markers significantly associated with salt tolerance. They were located on all chromosomes except chromosome 2 based on the alignment of their flanking sequences to the reference genome (Medicago truncatula). Of those identified, 13 were associated with multiple traits. Several loci identified in the present study were also identified in previous reports. BLAST search revealed that 19 putative candidate genes linked to 24 significant markers. Among them, B3 DNA-binding protein, Thiaminepyrophosphokinase and IQ calmodulin-binding motif protein were identified among multiple traits in the present and previous studies. With further investigation, these markers and candidates would be useful for developing markers for marker-assisted selection in breeding programs to improve alfalfa cultivars with enhanced tolerance to salt stress.

  9. Domestication via hybridization of the wild tetraploid oats Avena magna and A. murphyi.

    Science.gov (United States)

    Ladizinsky, G

    1995-09-01

    The wild tetraploid (2n=28) oat species Avena magna and A. murphyi have been domesticated by having been transferred from the common oat, A sativa (2n=42), the characteristics of non-shedding spikelets glabrous and yellow lemma, and reduced awn formation. Domestication has been achieved by crossing the common oat with either of the tetraploid species and then backcrossing the pentaploid hybrids with pollen of the tetraploid wild parent. Among the BC plants obtained only a few produced some seeds. Fertile tetraploids exhibiting the domesticated syndrome have been selected for in the F2 generation. Although morphologically they were almost indistinguishable from the common oat, they were tetraploids. Wild x domesticated A. magna hybrids were vigorous and fertile. They retained their spikelets at maturity, lemma color and pubescence were intermediate between the parental lines, and awns were formed only on the lower floret of the spikelet. Each of these characteristics segregated in a 3∶1 fashion, indicating single gene control, as in the common oat. These four characteristics formed a linkage group in one F2 family and two linkage groups in the other two families. The usefulness of the domesticated tetraploids for oat research and production has been discussed. Taxonomically, the domesticated tetraploids were ranked as subspecies: A. magna ssp. domestica, and A. murphyi ssp. rigida.

  10. Abscisic acid, H2O2 and nitric oxide interactions mediated cold-induced S-adenosylmethionine synthetase in Medicago sativa subsp. falcata that confers cold tolerance through up-regulating polyamine oxidation.

    Science.gov (United States)

    Guo, Zhenfei; Tan, Jiali; Zhuo, Chunliu; Wang, Congying; Xiang, Bin; Wang, Zengyu

    2014-06-01

    S-adenosylmethionine synthetase (SAMS) is the key enzyme catalysing the formation of S-adenosylmethionine (SAM), a precursor of polyamines and ethylene. To investigate the potential role of SAMS in cold tolerance, we isolated MfSAMS1 from the cold-tolerant germplasm Medicago sativa subsp. falcata and analysed the association of SAM-derived polyamines with cold tolerance. The expression of MfSAMS1 in leaves was greatly induced by cold, abscisic acid (ABA), H2O2 and nitric oxide (NO). Our data revealed that ABA, H2O2 and NO interactions mediated the cold-induced MfSAMS1 expression and cold acclimation in falcata. SAM, putrescine, spermidine and spermine levels, ethylene production and polyamine oxidation were sequentially altered in response to cold, indicating that SAMS-derived SAM is preferentially used in polyamine synthesis and homeostasis during cold acclimation. Antioxidant enzyme activities were also induced in response to cold and showed correlation with polyamine oxidation. Overexpression of MfSAMS1 in tobacco resulted in elevated SAM levels, but polyamine levels and ethylene production in the transgenic plants were not significantly changed. Compared to the wild type, transgenic plants had increased levels of apoplastic H2O2, higher transcript levels of genes involved in polyamine synthesis and oxidation, and higher activities of polyamine oxidation and antioxidant enzymes. The results showed that overexpression of MfSAMS1 promoted polyamine synthesis and oxidation, which in turn improved H2 O2 -induced antioxidant protection, as a result enhanced tolerance to freezing and chilling stress in transgenic plants. This is the first report demonstrating that SAMS plays an important role in plant tolerance to cold via up-regulating polyamine oxidation. © 2014 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.

  11. The Impact of Genotyping-by-Sequencing Pipelines on SNP Discovery and Identification of Markers Associated with Verticillium Wilt Resistance in Autotetraploid Alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Yu, Long-Xi; Zheng, Ping; Bhamidimarri, Suresh; Liu, Xiang-Ping; Main, Dorie

    2017-01-01

    Verticillium wilt (VW) of alfalfa is a soilborne disease causing severe yield loss in alfalfa. To identify molecular markers associated with VW resistance, we used an integrated framework of genome-wide association study (GWAS) with high-throughput genotyping by sequencing (GBS) to identify loci associated with VW resistance in an F1 full-sib alfalfa population. Phenotyping was performed using manual inoculation of the pathogen to cloned plants of each individual and disease severity was scored using a standard scale. Genotyping was done by GBS, followed by genotype calling using three bioinformatics pipelines including the TASSEL-GBS pipeline (TASSEL), the Universal Network Enabled Analysis Kit (UNEAK), and the haplotype-based FreeBayes pipeline (FreeBayes). The resulting numbers of SNPs, marker density, minor allele frequency (MAF) and heterozygosity were compared among the pipelines. The TASSEL pipeline generated more markers with the highest density and MAF, whereas the highest heterozygosity was obtained by the UNEAK pipeline. The FreeBayes pipeline generated tetraploid genotypes, with the least number of markers. SNP markers generated from each pipeline were used independently for marker-trait association. Markers significantly associated with VW resistance identified by each pipeline were compared. Similar marker loci were found on chromosomes 5, 6, and 7, whereas different loci on chromosome 1, 2, 3, and 4 were identified by different pipelines. Most significant markers were located on chromosome 6 and they were identified by all three pipelines. Of those identified, several loci were linked to known genes whose functions are involved in the plants' resistance to pathogens. Further investigation on these loci and their linked genes would provide insight into understanding molecular mechanisms of VW resistance in alfalfa. Functional markers closely linked to the resistance loci would be useful for MAS to improve alfalfa cultivars with enhanced resistance to

  12. Inoculação microbiana da silagem de alfafa (Medicago sativa e seu efeito sobre o consumo de matéria seca e sobre a fermentação ruminal em bovinos - DOI: 10.4025/actascianimsci.v25i2.2074 Microbial inoculation of alfalfa silage (Medicago sativa and its effect on dry matter intake and ruminal fermentation in bovines - DOI: 10.4025/actascianimsci.v25i2.2074

    Directory of Open Access Journals (Sweden)

    Paulo Henrique Mazza Rodrigues

    2003-04-01

    Full Text Available O objetivo do presente estudo foi avaliar os efeitos da inoculação microbiana da alfafa (Medicago sativa para ensilagem sobre o consumo de matéria seca, fermentação ruminal e taxa de passagem de líquidos em bovinos. Doze vacas não-gestantes e não-lactantes foram distribuídas em um delineamento em blocos, e os tratamentos corresponderam à silagem pré-secada de alfafa (60% de MS e 19,5% de PB controle ou inoculada com o produto Silobac® (Lactobacillus plantarum e Pediococcus pentosaceus. A dieta experimental continha 50% de silagem de alfafa e 50% de concentrado. O experimento teve duração total de 21 dias, sendo o 21º dia utilizado para colheitas de líquido ruminal realizadas às 0h, 2h, 4h, 6h, 8h, 10h e 12h, após a 1a refeição. A inoculação microbiana da silagem de alfafa não alterou o consumo de MS (inoculada = 2,56 vs. controle = 2,39% PV, o pH do líquido ruminal (6,15 vs. 6,27, a concentração ruminal de N-NH3 (19,0 vs. 18,2mg/dl, a concentração total de AGVs (122,5 vs. 113,8mM ou a proporção molar de ácido acético (66,1 vs. 66,8% molar, propiônico (21,1 vs. 19,6% molar e butírico (12,8 vs. 13,6% molar. Parâmetros relativos à dinâmica líquida ruminal, como o volume líquido (59,5 vs. 63,4 litros e a taxa de passagem de líquidos (8,6 vs. 8,0%/h, também não foram alterados com a inoculação.The objective of this study was to evaluate the effects of microbial inoculation of alfalfa (Medicago sativa for ensiling on dry matter intake, ruminal fermentation and liquid passage rate in twelve non pregnant dry cows. A randomized block design was used and the treatments were alfalfa haylage control (60% DM and 19.5% CP or inoculated with Silobac® product (Lactobacillus plantarum and Pediococcus pentosaceus. Experimental diet contained 50% of alfalfa silage and 50% of concentrate. Experimental period lasted for twenty-one days; the 21st day was used for ruminal liquid sampling at 0, 2, 4, 6, 8, 10 e 12 hours

  13. Effect of elevated CO₂ and temperature on the oxidative stress response to drought in Lolium perenne L. and Medicago sativa L.

    Science.gov (United States)

    Farfan-Vignolo, Evelyn Roxana; Asard, Han

    2012-10-01

    Studies addressing the combined impact of multiple climate factors on plant abiotic stress responses are still scarce. We investigated physiological and molecular (antioxidant), responses to water deficit, in grassland-model species, Lolium perenne L. and Medicago lupulina L., under future climate conditions, i.e. elevated CO₂ (+CO₂, +375 ppm) and elevated temperature (+T, +3 °C). Elevated CO₂, but not warming, significantly increased biomass (gDW) in L. perenne, but not in M. lupulina. Photosynthesis (A(sat)) and stomatal conductance (g(s)), were differently affected by climate in each species, L. perenne generally being more sensitive. Elevated CO₂ increased lipid peroxidation levels in M. lupulina, but not in L. perenne, and had no effect on protein oxidation and little effect on antioxidant levels. Drought stress caused severe inhibition in biomass and photosynthesis, most severely in L. perenne, and strongly increased oxidative damage. Elevated CO₂ protected against the drought-induced damage. Decreased activities of APX and POX may indicate lower levels of oxidative challenge (relaxation) at the level of H₂O₂ production. Polyphenols, tocopherols and antioxidant capacity, increased under drought stress, in all climate conditions. Elevated CO₂, increased reduced ascorbate (ASC) and reduced glutathione (GSH), and their redox status, in both species, although to different levels. Changes in activities of key ASC/GSH cycle enzymes, under stress and climate treatments, showed weak correlations with ASC and GSH levels, indicating the complexity of this network. Together this work supports the idea that redox changes are involved in responses to climate changes, in the absence and presence of water-deficit stress. Copyright © 2012 Elsevier Masson SAS. All rights reserved.

  14. Batı Akdeniz Sahil Kuşağından Toplanan Yonca (Medicago sativa L. Populasyonlarının Moleküler Karakterizasyonu

    Directory of Open Access Journals (Sweden)

    Mehmet ÖTEN

    2016-06-01

    Full Text Available Medicago tür ve populasyonlarında moleküler markırlar kullanılarak genotipik varyasyon seviyesini belirlemek yaygın olarak kullanılmaktadır. Bu çalışmada; Batı Akdeniz sahil kuşağında yer alan, Antalya iline ait 13 ilçede, 26 farklı lokasyonda, doğal vejetasyondan toplanan üstün yonca genotipleri klon yöntemi ile çoğaltılmıştır. Elde edilen 26 genotip ve 2 standart çeşidin aralarındaki benzerlikler, Basit Tekrarlı Sekanslar (SSR moleküler analiz tekniğiyle, 15 adet primer kullanılarak belirlenmiştir. SSR primerlerinin % 99,7’si polimorfik bant vermiştir. Araştırma sonucunda; kümeleme analizi sonucu oluşan dendograma göre yonca genotipleri 2 ana ve 4 alt gruba ayrılmıştır. Birinci ana grup % 60, ikinci ana grup ise % 65 seviyesinde kendi içerisinde iki alt gruba ayrılmıştır. Benzerlik katsayıları 0.58-0.88 değerleri arasında değişim göstermiştir. En yakın genetik benzerlik Kaş-2 ve Çeşit-1 arasında bulunurken, en uzak ise Gazipaşa-1-Kumluca-2 genotipleri arasında bulunmuştur. Elde edilen bu sonuca göre Batı Akdeniz sahil kuşağından toplanan yonca genotipleri arasında genetik olarak önemli bir varyasyonun olduğu tespit edilmiştir.

  15. Natural product biosynthesis in Medicago species.

    Science.gov (United States)

    Gholami, Azra; De Geyter, Nathan; Pollier, Jacob; Goormachtig, Sofie; Goossens, Alain

    2014-03-01

    The genus Medicago, a member of the legume (Fabaceae) family, comprises 87 species of flowering plants, including the forage crop M. sativa (alfalfa) and the model legume M. truncatula (barrel medic). Medicago species synthesize a variety of bioactive natural products that are used to engage into symbiotic interactions but also serve to deter pathogens and herbivores. For humans, these bioactive natural products often possess promising pharmaceutical properties. In this review, we focus on the two most interesting and well characterized secondary metabolite classes found in Medicago species, the triterpene saponins and the flavonoids, with a detailed overview of their biosynthesis, regulation, and profiling methods. Furthermore, their biological role within the plant as well as their potential utility for human health or other applications is discussed. Finally, we give an overview of the advances made in metabolic engineering in Medicago species and how the development of novel molecular and omics toolkits can influence a better understanding of this genus in terms of specialized metabolism and chemistry. Throughout, we critically analyze the current bottlenecks and speculate on future directions and opportunities for research and exploitation of Medicago metabolism.

  16. Differential response to water deficit stress in alfalfa ( Medicago ...

    African Journals Online (AJOL)

    The present study was fixed as objective to compare the response to water deficit (33% of field capacity, FC) stress of eight cultivars of Medicago sativa, originating from the Mediterranean basin. Comparison was performed on some key parameters such as growth, relative water content, leaf water potential, MDA tissue ...

  17. Получение трансгенных растений люцерны посевной (Medicago sativa L. ) для повышения эффективности фиторемедиации нефтезагрязненных почв

    OpenAIRE

    СТЕПАНОВА АННА ЮРЬЕВНА; ОРЛОВА ЕКАТЕРИНА ВЛАДИМИРОВНА; ТЕРЕШОНОК ДМИТРИЙ ВИКТОРОВИЧ; ДОЛГИХ ЮЛИЯ ИВАНОВНА

    2015-01-01

    The studies carried out in 2008-2015 revealed that such Medicago varia varieties as Vega 87, Nakhodka, Pastbishhnaya 88, Selena, grown on sod-podzolic soils for 5 years of use, had been forming stable yields of dry matter 6,24-9,25 t/ha. On the 6-7th year a strong thinning of Medicago varia grass stands was recorded because of suffering from diseases affection. A five-year period of economic use for such varieties was proved to found to be optimal. Medicago sativa varieties showed better stab...

  18. Impacto de una intervención alimentaria con un concentrado proteico de Medicago sativa L (alfalfa, en niños pre escolares con desnutrición crónica

    Directory of Open Access Journals (Sweden)

    María Soberón

    2009-09-01

    Full Text Available Introducción: Estudios realizados a nivel preclínico han demostrado el valor biológico del concentrado proteico obtenido de Medicago sativa L, alfalfa (CPA, por lo que este producto puede ser una alternativa para reducir la desnutrición. Objetivo: Evaluar el efecto del consumo de CPA por niños preescolares con desnutrición crónica, mediante indicadores bioquímicos y hematológicos. Diseño: Estudio analítico de intervención, de tipo longitudinal y prospectivo. Institución: Centro de Investigación de Bioquímica y Nutrición, Facultad de Medicina, Universidad Nacional Mayor de San Marcos. Participantes: Niños preescolares con desnutrición crónica, provenientes de una comunidad de Pampas, Huancavelica, Perú. Intervenciones: Previo consentimiento informado de los padres, se evaluó el estado alimentario-nutricional, bioquímico y hematológico de 23 niños menores de seis años, para establecer la condición de desnutrición. Luego, se desarrolló una intervención alimentario nutricional con el CPA como complemento alimenticio, por 25 días, en niños seleccionados. Finalmente, se repitió la evaluación una vez concluido el período de intervención. Las determinaciones bioquímicas fueron hemoglobina, proteínas totales, albúmina y globulina, creatinina, transaminasas, y las hematológicas, hematocrito, recuento de reticulocitos. Se utilizó el programa Excel y t-student, a fin de obtener la inferencia en los resultados. Resultados: Para el indicador talla/edad, 84,5% de los niños padecía desnutrición crónica; respecto a calorías y nutrientes consumidos por los niños, los valores de energía estuvieron entre 563 kcal y 2 597 kcal; de proteína, 9,7 g a 78 g; de grasas, 5,1 g a 53,8 g; carbohidratos, 79,8 g a 495,1 g; calcio, entre 180,9 mg y 986 mg; y hierro, 1,9 mg y 21,5 mg. Los valores de hemoglobina, proteínas totales, globulinas, creatinina, transaminasas y hematocrito obtenidos antes y después de la ingesta de

  19. Survival and growth of Alfalfa (Medicago sativa l.) inoculated with an am fungus (Glomus intraradices) in contaminated soils treated with two different remediation technologies (bio-pile and thermal desorption)

    Energy Technology Data Exchange (ETDEWEB)

    Norini, M.P.; Beguiristain, Th.; Leyval, C. [LIMOS UMR 7137 CNRS-UHP Nancy - Faculty of Sciences, 54 - Vandoeuvre-les-Nancy (France)

    2005-07-01

    (Medicago sativa L.), inoculated or not with an AM fungus (Glomus intraradices) and fertilized with Hewitt solution. Control non-inoculated and non-fertilized plants were included. There was four replicates per treatment. After 2 months, plant survival and growth was measured, as well as mycorrhizal colonization of roots. PAH in rhizosphere soils was analyzed using ASE and GC-FID. Major and trace elements in plants were analyzed after acid digestion using ICP. AM fungal and bacterial communities present in roots and rhizosphere were also analysed after DNA extraction with a PCR-TGGE technique. Plant survival rate was significantly lower in the soil treated with bio-pile than in untreated soil. Non-inoculated and non fertilized alfalfa biomass was lower with soil treated with bio-pile and thermal desorption than with untreated soil. Mycorrhizal inoculation as well as fertilization significantly improved alfalfa growth in the soil treated with bio-pile. In the soil treated with thermal desorption, shoot and root biomass were significantly higher with mycorrhizal inoculation and fertilization. Estimation of root colonization showed that the roots of un-inoculated plants growing on the untreated contaminated soil contained indigenous mycorrhizal fungi. With the soil treated with bio-pile, mycorrhizal colonization of un-inoculated plants was significantly lower, while no colonization was observed in the roots in the soil treated with thermal desorption. Inoculation with a mycorrhizal fungus increased root colonization rate in both treated soils. It also significantly increased the number of rhizobia nodules. Phosphorus concentration in plant roots and shoots was significantly improved by mycorrhizal inoculation in soil treated with thermal desorption. Data on PAH concentration in soils and fungal and bacterial communities will be also presented and discussed.

  20. Genetic Engineering of Alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Wang, Dan; Khurshid, Muhammad; Sun, Zhan Min; Tang, Yi Xiong; Zhou, Mei Liang; Wu, Yan Min

    2016-01-01

    Alfalfa is excellent perennial legume forage for its extensive ecological adaptability, high nutrition value, palatability and biological nitrogen fixation. It plays a very important role in the agriculture, animal husbandry and ecological construction. It is cultivated in all continents. With the development of modern plant breeding and genetic engineering techniques, a large amount of work has been carried out on alfalfa. Here we summarize the recent research advances in genetic engineering of alfalfa breeding, including transformation, quality improvement, stress resistance and as a bioreactor. The review article can enables us to understand the research method, direction and achievements of genetic engineering technology of Alfalfa.

  1. Effects of the weed density on grass yield of Alfalfa ( Medicago ...

    African Journals Online (AJOL)

    This study, in which the effects of different row spacing applications on weed density and on grass yield of Medicago sativa L. were investigated, was carried out in Van-Turkey from 2006 - 2008. Randomized blocks design was adopted with three replications. Row spacing applications of 20, 30, 40, 50, 60 and 70 cm were ...

  2. Meiotic behaviour of tetraploid wheats (Triticum turgidum L.) and ...

    Indian Academy of Sciences (India)

    Meiotic aberrations such as laggards, chromosome bridges, micronuclei, abnormal cytokines, chromatin pulling and meiotic restitution were observed and the studied genotypes were accordingly ranked as follows: triticale > synthetic hexaploid wheats > tetraploid wheats possessing meiotic restitution > tetraploid wheats ...

  3. Apomictic maternal diploids in tetraploid Job's tears.

    Science.gov (United States)

    Venkateswarlu, J; Rao, P N

    1975-06-01

    Two cases of reversion to diploidy were observed in autotetraploid Job's tears (Coix lacryma-jobi L. 4n = 40) out of a total of 1,112 plants examined over a period of 7 years. One of these was a trisomie (2n = 21) and the other a disomic (2n = 20), derived from apomictic development ofn+1 andn maternal gametes of the tetraploid, respectively. In some respects both these derivatives differed from the original diploid that gave rise to the tetraploid through colchicine treatment. The potentialities of such reversions in the evolution of new diploid races are discussed.

  4. Effect of the inherent variation in the mineral concentration of alfalfa cultivars on aphid populations Efeito da variação inata da concentração de minerais em cultivares de alfafa (Medicago sativa em população de afídeos (Hemiptera: Aphididae

    Directory of Open Access Journals (Sweden)

    Alexandre de Almeida e Silva

    2005-01-01

    Full Text Available Plants have inherent variability of mineral content which affects their physiology and consequently the herbivorous insects feeding on them. Besides, insects need considerable amounts of potassium, phosphorus and magnesium in their diets, whereas little calcium, sodium and chloride are required. In this study, the inherent variation on mineral (Ca, S, Mg, N, P, K and also C:N ratio concentrations and aphid (Acyrthosiphon spp., Therioaphis maculata, Aphis craccivora populations on three alfalfa (Medicago sativa cultivars (P3; Crioula, the most widely cultivated in Brazil, and CUF 101, an aphid-resistant were studied between September/1997 and August/1998. A significant variation on mineral concentrations and aphid populations was observed among different sampling times and cultivars. The correlations between C:N ratio, Mg, N, P and S concentrations and aphid density variation suggest that the mineral status affects aphid population dynamics under field conditions.As plantas têm variação inata do conteúdo de minerais e seu estado nutricional afeta sua fisiologia cloretos. A variação inata na concentração de minerais (Ca, S, Mg, N, P, K e também a razão C:N e na população de afídeos (Acyrthosiphon spp., Therioaphis maculata, Aphis craccivora em três cultivares de alfafa (M. sativa - P3; Crioula, as mais cultivadas no Brasil, e CUF 101, resistente a afídeos - foi estudada entre setembro/1997 a agosto/1998 neste trabalho. A concentração de minerais e as populações de pulgões variaram significativamente entre os diferentes períodos de coleta e cultivares. As correlações encontradas entre as concentrações de Mg, N, P, S e a razão C:N e a variação no número de pulgões sugerem que os minerais da planta afetam a dinâmica populacional dos pulgões em campo.

  5. Classifying Ethiopian Tetraploid Wheat ( Triticum turgidum L ...

    African Journals Online (AJOL)

    Thirty five accessions of Ethiopian tetraploid wheat (T. turgidum L.) landraces were grown in the greenhouse at IFA Tulln, Austria during spring 2009 for DNA extraction. The same accessions were already grown in spring 2008 at BOKU Vienna, Austraia for their phenotypical characterisation. DNA was extracted from each ...

  6. Comet assay on tetraploid yeast cells

    DEFF Research Database (Denmark)

    Rank, Jette; Syberg, Kristian; Jensen, Klara

    2009-01-01

    . Analytical problems that arose due to the small amount of DNA in the yeast nuclei in haploid and diploid cells, which contain 13 Mbp and 26 Mbp DNA per cell, respectively, were solved by using tetraploid yeast cells (52 Mbp) instead. DNA damage was shown after exposure to H2O2 and acrylamide. The lowest dose...

  7. The effect of tetraploidy induction on morphology and anatomy characteristics of Cannabis sativa L.

    Directory of Open Access Journals (Sweden)

    Hakimeh Mansouri

    2014-12-01

    Full Text Available The production of tetraploid plant was studied in Cannabis sativa L. with colchicine at three different concentrations (i.e., 0.0, 0.1 and 0.2% for about 24 and 48 h through dropping method. Flow cytometry analyses were used to confirm the ploidy level. Morphologic and anatomic characteristics between tetraploid and diploid control plants were compared. The results showed that 0.2% colchicine for 24 h had the best effect. The percentage of tetraploid plants and the survival rate were lowered by increasing the treatment time. In addition, the leaf index and height of tetraploid plants exhibited a significant decrease compared to the diploid plants. The size of leaves' epidermis stomata were larger in tetraploid plant compared to the diploid ones, in spite of their less density of stomata. However, the amount of total chlorophyll and carotenoids were almost the same in both of tetraploid and diploid plants. In addition, some differences were also observed in the cross section of stem of these plants from the descriptive structure point of view. On the whole, the results introduced usage of the stomata parameters as an effective and convenient method for detecting the tetraploid plants however, the flow cytometry analysis was more effective in assessing the ploidy percentage.

  8. INCREMENTO EN INVERNADERO DE LA CALIDAD Y CANTIDAD DEL FOLLAJE DE LA ALFALFA (Medicago Sativa L. VARIEDAD FLORIDA 77 CAUSADO POR LA COMBINACIÓN DE FERTILIZACIÓN BIOLÓGICA Y QUÍMICA EN UN SUELO DE LA SERIE BERMEO DE LA SABANA DE BOGOTÁ

    Directory of Open Access Journals (Sweden)

    J. Tovar-Franco

    2006-06-01

    Full Text Available En la Sabana de Bogotá existe la necesidad de incrementar la producción de forrajes que den buen rendimiento, con alta calidad nutricional y que podrían ser un aporte al desarrollo de la industria de concentrados de animales.El objetivo de este trabajo fue establecer el efecto de la inoculación rizobio-hongo micorriza arbuscular (MA sobre la fijación simbiótica de nitrógeno, la absorción de fósforo y el porcentaje de micorrización en alfalfa(Medicago sativa L. en un suelo andept de la serie Bermeo de la Sabana de Bogotá. Se seleccionó la variedad dealfalfa Florida 77 por su adaptabilidad a las condiciones del suelo, la cepa de Rhizobium (Sinorhizobium melilotiGR-4 por su habilidad para mejorar el rendimiento y estimular la fijación simbiótica de nitrógeno en esta variedad y el hongo MA Archaeospora leptoticha por su habilidad para mejorar el rendimiento y estimular la fijación defósforo en esta leguminosa.La inoculación dual (rizobio-hongo MA demostró eficacia de la fertilización biológica, al incrementar significativamente (p<0.05 los efectos del encalado y la fertilización química, aumentando el rendimiento en 26% y al mejorar en el follaje el contenido de nitrógeno (32% y fósforo (28%. Estos resultados destacan el estímulo de los dos microsimbiontes en el aprovechamiento de la fertilización química.La inoculación dual también estimuló la fijación biológica de nitrógeno incrementándose los efectos de la inoculación rizobial al aumentar el rendimiento en 25% y el contenido de nitrógeno del follaje en 22% (p<0.05. También superó el efecto de la inoculación fúngica aumentando el rendimiento en 31% (p<0.05 y la cantidad defósforo absorbido en 34% (p<0.05, demostrándose así los beneficios de la simbiosis tripartita alfalfa-rizobio-hongo MA en este Andisol.

  9. Viability of pollen grains of tetraploid banana

    Directory of Open Access Journals (Sweden)

    Taliane Leila Soares

    2016-01-01

    Full Text Available ABSTRACT Obtaining banana tetraploid cultivars from triploid strains results in total or partial reestablishment of fertility, allowing the occurrence of some fruits with seeds, a feature that is undesirable from a marketing perspective. The objective of this study was to assess the viability of pollen of 12 banana tetraploid hybrids (AAAB by means of in vitro germination and two histochemical tests (acetocarmine and 2,3,5-triphenyltetrazolium chloride. The pollen tube growth was evaluated by germinating grains in three culture media — M1: 0.03% Ca(NO3∙4H2O, 0.02% Mg(SO4∙7H2O, 0.01% KNO3, 0.01% H3BO3 and 15% sucrose; M2: 0.03% Ca(NO3∙4H2O, 0.01% KNO3, 0.01% H3BO3 and 10% sucrose; and M3: 0.015% H3BO3, 0.045% Ca3(PO42 and 25% sucrose. The acetocarmine staining indicated high viability (above 80%, except for the genotypes YB42-17 and Caprichosa, which were 76 and 70%, respectively. However, the in vitro germination rate was lower than 50% for all the genotypes, except for the hybrids YB42-17 (M1 and YB42-47 (M1. The medium M1 provided the greatest germination percentage and pollen tube growth. Among the genotypes assessed, YB42-47 presented the highest germination rate (61.5% and tube length (5.0 mm. On the other hand, the Vitória cultivar had the lowest germination percentage (8.2% in medium M1. Studies of meiosis can shed more light on the differences observed in the evaluated tetraploids, since meiotic irregularities can affect pollen viability.

  10. Embryo rescue of crosses between diploid and tetraploid grape ...

    African Journals Online (AJOL)

    ajl yemi

    2011-12-19

    Dec 19, 2011 ... tetraploid grapes can be a good way to obtain new triploid germplasm. However, there exists a severe mating obstacle in crosses between diploid and tetraploid grape. The embryo rescue technique may prevent the early stage abortion of triploid young embryo, so triploid plants can be produced (Pan et al., ...

  11. Genome sequence of the Medicago-nodulating Ensifer meliloti commercial inoculant strain RRI128

    Science.gov (United States)

    Reeve, Wayne; Ballard, Ross; Drew, Elizabeth; Tian, Rui; Bräu, Lambert; Goodwin, Lynne; Huntemann, Marcel; Han, James; Tatiparthi, Reddy; Chen, Amy; Mavrommatis, Konstantinos; Markowitz, Victor; Palaniappan, Krishna; Ivanova, Natalia; Pati, Amrita; Woyke, Tanja; Kyrpides, Nikos

    2014-01-01

    Ensifer meliloti strain RRI128 is an aerobic, motile, Gram-negative, non-spore-forming rod. RRI128 was isolated from a nodule recovered from the roots of barrel medic (Medicago truncatula) grown in the greenhouse and inoculated with soil collected from Victoria, Australia. The strain is used in commercial inoculants in Australia. RRI128 nodulates and forms an effective symbiosis with a diverse range of lucerne cultivars (Medicago sativa) and several species of annual medic (M. truncatula, Medicago littoralis and Medicago tornata), but forms an ineffective symbiosis with Medicago polymorpha. Here we describe the features of E. meliloti strain RRI128, together with genome sequence information and annotation. The 6,900,273 bp draft genome is arranged into 156 scaffolds of 157 contigs, contains 6,683 protein-coding genes and 87 RNA-only encoding genes, and is one of 100 rhizobial genomes sequenced as part of the DOE Joint Genome Institute 2010 Genomic Encyclopedia for Bacteria and Archaea-Root Nodule Bacteria (GEBA-RNB) project. PMID:25197447

  12. Probabilistic Multilocus Haplotype Reconstruction in Outcrossing Tetraploids.

    Science.gov (United States)

    Zheng, Chaozhi; Voorrips, Roeland E; Jansen, Johannes; Hackett, Christine A; Ho, Julie; Bink, Marco C A M

    2016-05-01

    For both plant (e.g., potato) and animal (e.g., salmon) species, unveiling the genetic architecture of complex traits is key to the genetic improvement of polyploids in agriculture. F1 progenies of a biparental cross are often used for quantitative trait loci (QTL) mapping in outcrossing polyploids, where haplotype reconstruction by identifying the parental origins of marker alleles is necessary. In this paper, we build a novel and integrated statistical framework for multilocus haplotype reconstruction in a full-sib tetraploid family from biallelic marker dosage data collected from single-nucleotide polymorphism (SNP) arrays or next-generation sequencing technology given a genetic linkage map. Compared to diploids, in tetraploids, additional complexity needs to be addressed, including double reduction and possible preferential pairing of chromosomes. We divide haplotype reconstruction into two stages: parental linkage phasing for reconstructing the most probable parental haplotypes and ancestral inference for probabilistically reconstructing the offspring haplotypes conditional on the reconstructed parental haplotypes. The simulation studies and the application to real data from potato show that the parental linkage phasing is robust to, and that the subsequent ancestral inference is accurate for, complex chromosome pairing behaviors during meiosis, various marker segregation types, erroneous genetic maps except for long-range disturbances of marker ordering, various amounts of offspring dosage errors (up to ∼20%), and various fractions of missing data in parents and offspring dosages. Copyright © 2016 by the Genetics Society of America.

  13. Initiating genomic selection in tetraploid potato

    DEFF Research Database (Denmark)

    Sverrisdóttir, Elsa; Janss, Luc; Byrne, Stephen

    Breeding for more space and resource efficient crops is important to feed the world’s increasing population. Potatoes produce approximately twice the amount of calories per hectare compared to cereals. The traditional “mate and phenotype” breeding approach is costly and time-consuming; however......, the completion of the genome sequence of potato has enabled the application of genomics-assisted breeding technologies. Genomic selection using genome-wide molecular markers is becoming increasingly applicable to crops as the genotyping costs continue to reduce and it is thus an attractive breeding alternative....... We have used genotyping-by-sequencing to genotype 762 individuals. The individuals were randomly selected from a population of 5,000 individuals derived from a poly-parental cross generated from 18 tetraploid cultivars and breeding clones (MASPot population). Phenotypic data have been established...

  14. The effect of lucerne (Medicago sativa) hay quality on milk ...

    African Journals Online (AJOL)

    Van Zyl, Marike

    2014-08-23

    Aug 23, 2014 ... Abstract. The influence of lucerne hay quality on the milk production potential and milk composition of Jersey cows was investigated. Three different grades of lucerne hay (Prime, Grade 1, Grade 2; selected according to the New Lucerne Quality Index) were included in a total mixed ration (TMR) and fed to ...

  15. Evaluation of Selection Indices for Alfalfa (Medicago sativa L.

    Directory of Open Access Journals (Sweden)

    Hassan MONIRIFAR

    2010-03-01

    Full Text Available In order to evaluate the efficiency of selection indices in alfalfa improvement, an experiment was conducted from 2000 to 2007 at East Azarbaijan Agricultural and Natural Resources Research Center, Iran. A set of 29 native ecotypes, which were collected in 2000 and 2001 from Azerbaijan (Iran, together with one improved variety were used to conduct a polycross nursery. For the purpose of random mating, a randomized complete block design was used with 12 replications arranged in this nursery. The resulting 30 half-sib families were planted individually in pots and the 30 days old seedlings were transplanted to the field in May, 2004. Each 3-row plot consisted of 45 HS plants. Several traits, such as individual plants fresh weight and individual plants dry weight (IPFW, IPDW, number of shoots per plant (NS and plant height (PH in each harvest and also, days until 10% flowering, the ratio of fresh and dry weight of leaves/plant and size of trifoliate leaves were measured for three cropping seasons. The results of analysis of variance showed large variation among polycross progenies. Six selection indices (Ii with different number of traits at adult plant stage were evaluated. Based on the result of this investigation, if number of shoots and height of adult plant, excluding yield, are recorded, I2 is suggested. If, in addition to fresh yield, height of adult plant is measured, I6 is recommended. I4 is useful when number of shoots and plant height with dry yield are included in the index. In conclusion, the importance of mature plant traits in selection indices was in the order of yield > plant height > number of shoots. The results provided more evidence that selection indices incorporating the component of dry yield are more advantageous. The most efficient selection index consisted of NS, IPDW and PH, having a relative efficiency of 280%.

  16. Studi Tentang Pola Produksi Alfalfa Tropis (Medicago Sativa L.)

    OpenAIRE

    Wahyuni, Rini Dwi; Kamaliyah, Siti Nurul

    2009-01-01

    The research aims to determine the growth pattern of alfalfa in the tropical region, which later can be used as a source of information for the people needed. Research methods used are experimental; the data acquired during the 90 days was showed in a graph and then analyzed using regression analysis with two variables, namely independent variables (cutting age) and dependent variables (the nutritive value, height of plants, and production of dry matter (DM), organic matter (OM), and crude pr...

  17. Toxicological responses in alfalfa (Medicago sativa) under joint ...

    African Journals Online (AJOL)

    AJL-021

    2012-11-13

    Nov 13, 2012 ... interaction of Cd2+ and napropamide would aggravate the toxic effects on chlorophyll synthesis in ... The study on biochemical responses of plants to joint ..... plants. Environ. Exp. Bot. 41:105–130. Sengupta-Gopalan C, Bagga S, Potenza C, Ortega JL (2007). Alfalfa transgenic crops, in: E.C. Pua, M.R. ...

  18. Biochemical responses of Alfalfa ( Medicago sativa L.) cultivars ...

    African Journals Online (AJOL)

    This investigation was conducted to determine NaCl salinity effects on antioxidant enzyme s activities, reducing sugar contents and lipid peroxidation in two alfalfa cultivars. Plants grown in solution cultures were subjected to 0, 100, 150 and 200 mM solutions of sodium chloride. Yazdi and Diabolourde alfalfa were used as ...

  19. Toxicological responses in alfalfa ( Medicago sativa ) under joint ...

    African Journals Online (AJOL)

    Joint effects of Cd2+ and napropamide in seeds, roots or leaves of alfalfa were investigated under different treatments. It was shown that single stress of Cd2+ or napropamide decreased chlorophyll content after 30 days of treatment in different concentrations. The decrease in chlorophyll content became insignificant under ...

  20. Metabolic responses of alfalfa ( Medicago Sativa L.) leaves to low ...

    African Journals Online (AJOL)

    To explore adaptation mechanism of alfalfa to cold and heat stresses, status of sucrose metabolism and relative water content (RWC) in leaves under low and high temperature treatments were studied. Seedlings (35 day old) were transferred to chambers for treatments. First group was subjected to 5°C as low temperature ...

  1. Metabolic responses of alfalfa (Medicago Sativa L.) leaves to low ...

    African Journals Online (AJOL)

    user

    2011-02-14

    Feb 14, 2011 ... Aranjuelo I, Irigoyen JJ, Perez P, Martinez-Carrasco R, Sanchez-Diaz. M (2006). Response of nodulated alfalfa to water supply, temperature and elevated CO2: productivity and water relations. Environ. Exp. Bot. 55: 130-141. Aranjuelo I, Irigoyen JJ, Sanchez-Diaz M (2007). Effect of elevated temperature ...

  2. Modeling of desorption of Alfalfa (Medicago sativa) stems and leaves.

    NARCIS (Netherlands)

    ArabHosseini, A.; Huisman, W.; Müller, J.

    2011-01-01

    The equilibrium moisture content of agricultural products is necessary to optimize drying process and helps to keep the quality of the product during the period of storage. The main aim of this research was to find the best model which could define well, the exchange of moisture between alfalfa

  3. The effect of lucerne ( Medicago sativa ) hay quality on milk ...

    African Journals Online (AJOL)

    The influence of lucerne hay quality on the milk production potential and milk composition of Jersey cows was investigated. Three different grades of lucerne hay (Prime, Grade 1, Grade 2; selected according to the New Lucerne Quality Index) were included in a total mixed ration (TMR) and fed to lactating cows. The three ...

  4. Performance of Medicago sativa under dryland conditions on the ...

    African Journals Online (AJOL)

    Reports the results of research conducted into pasture establishment using legumes. Examines possible strategies for integrating dryland lucerne into livestock production systems on the eastern Highveld. Focuses on livestock production from pure and mixed lucerne pastures; Increased livestock numbers and improved ...

  5. Evaluation of Selection Indices for Alfalfa (Medicago sativa L.

    Directory of Open Access Journals (Sweden)

    Hassan MONIRIFAR

    2010-03-01

    Full Text Available In order to evaluate the efficiency of selection indices in alfalfa improvement, an experiment was conducted from 2000 to 2007 at East Azarbaijan Agricultural and Natural Resources Research Center, Iran. A set of 29 native ecotypes, which were collected in 2000 and 2001 from Azerbaijan (Iran, together with one improved variety were used to conduct a polycross nursery. For the purpose of random mating, a randomized complete block design was used with 12 replications arranged in this nursery. The resulting 30 half-sib families were planted individually in pots and the 30 days old seedlings were transplanted to the field in May, 2004. Each 3-row plot consisted of 45 HS plants. Several traits, such as individual plants� fresh weight and individual plants� dry weight (IPFW, IPDW, number of shoots per plant (NS and plant height (PH in each harvest and also, days until 10% flowering, the ratio of fresh and dry weight of leaves/plant and size of trifoliate leaves were measured for three cropping seasons. The results of analysis of variance showed large variation among polycross progenies. Six selection indices (Ii with different number of traits at adult plant stage were evaluated. Based on the result of this investigation, if number of shoots and height of adult plant, excluding yield, are recorded, I2 is suggested. If, in addition to fresh yield, height of adult plant is measured, I6 is recommended. I4 is useful when number of shoots and plant height with dry yield are included in the index. In conclusion, the importance of mature plant traits in selection indices was in the order of yield > plant height > number of shoots. The results provided more evidence that selection indices incorporating the component of dry yield are more advantageous. The most efficient selection index consisted of NS, IPDW and PH, having a relative efficiency of 280%.

  6. PHENOTYPIC DIVERSITY OF ALFALFA (MEDICAGO SATIVA L. GERMPLASM

    Directory of Open Access Journals (Sweden)

    Marijana Tucak

    2011-06-01

    Full Text Available The objective of this study was to evaluate phenotypic diversity in the alfalfa germplasm collections using multivariate analysis to examine the extent of genetic diversity and contribution of selected characters to the total diversity and finally to select the most promising clusters/populations for further breeding work. Forty alfalfa populations/cultivars of different geographical origin were evaluated for 12 agro-morphological characters during two consecutive years. The populations/ cultivars were grouped into six clusters. In most cases populations/cultivars within clusters were not associated with their geographical origin. Intercluster distances were larger than the intracluster ones. This research revealed a broad phenotypic diversity within and between the alfalfa germplasm collections. The following characters contributed most to the total phenotypic diversity: dry matter yield in the first production year, plant height and length of central leaflet. Based on the mean value of the evaluated characters and determined distances between clusters, the most promising populations/cultivars belong to the clusters IV and V. Selected populations/cultivars could be considered as a valuable genetic material for the yield and quality improvement of alfalfa in our breeding programme.

  7. Amplification, contraction and genomic spread of a satellite DNA family (E180) in Medicago (Fabaceae) and allied genera.

    Science.gov (United States)

    Rosato, Marcela; Galián, José A; Rosselló, Josep A

    2012-03-01

    Satellite DNA is a genomic component present in virtually all eukaryotic organisms. The turnover of highly repetitive satellite DNA is an important element in genome organization and evolution in plants. Here we assess the presence and physical distribution of the repetitive DNA E180 family in Medicago and allied genera. Our goals were to gain insight into the karyotype evolution of Medicago using satellite DNA markers, and to evaluate the taxonomic and phylogenetic signal of a satellite DNA family in a genus hypothesized to have a complex evolutionary history. Seventy accessions from Medicago, Trigonella, Melilotus and Trifolium were analysed by PCR to assess the presence of the repetitive E180 family, and fluorescence in situ hybridization (FISH) was used for physical mapping in somatic chromosomes. The E180 repeat unit was PCR-amplified in 37 of 40 taxa in Medicago, eight of 12 species of Trigonella, six of seven species of Melilotus and in two of 11 Trifolium species. Examination of the mitotic chromosomes revealed that only 13 Medicago and two Trigonella species showed FISH signals using the E180 probe. Stronger hybridization signals were observed in subtelomeric and interstitial loci than in the pericentromeric loci, suggesting this satellite family has a preferential genomic location. Not all 13 Medicago species that showed FISH localization of the E180 repeat were phylogenetically related. However, nine of these species belong to the phylogenetically derived clade including the M. sativa and M. arborea complexes. The use of the E180 family as a phylogenetic marker in Medicago should be viewed with caution. Its amplification appears to have been produced through recurrent and independent evolutionary episodes in both annual and perennial Medicago species as well as in basal and derived clades.

  8. Manejo de água para estabelecimento de alfafa (Medicago sativa L. Water management for alfalfa (Medicago sativa L. establishment

    Directory of Open Access Journals (Sweden)

    Joaquim Bartolomeu Rassini

    2001-04-01

    Full Text Available O manejo de água para o desenvolvimento da alfafa cv. Crioula foi avaliado a campo, com a finalidade de verificar seus efeitos no estabelecimento da planta, até o 1º corte, em 1997. Para isso, verificou-se o comportamento em três estádios de desenvolvimento (E1 = início do estádio vegetativo, E2 = vegetativo pleno e E3 = reprodutivo até colheita em quatro níveis de água (A = 100%, B = 80%, C = 45-50% e D = 20-25% de uma lâmina de irrigação por aspersão. Foram avaliados oito experimentos: 111, 110, 100, 101, 001, 011, 010, 000, em que o primeiro dígito se refere a E1; o segundo, a E2; e o terceiro, a E3. O valor 0 representa a época em que o estádio não recebeu água, coberto por uma estrututura móvel, que deslizava sobre trilhos, e também protegia contra precipitações pluviais casuais. O valor 1 refere-se à época em que houve aplicação dos níveis de água de irrigação. Os estádios de desenvolvimento da planta, em termos de rendimento de matéria seca, responderam de maneira diferente às condições hídricas, sendo E2 e E3 (planta estabelecida os mais sensíveis e E1 (início do estádio vegetativo, o menos sensível, no qual a água foi prejudicial ao estabelecimento e rendimento da planta.The water management for growth of Crioula alfalfa cultivar was evaluated in a field study, in order to assess the effects in the plant establishment up to the first cut, at 1977. The study was carried out in 1997. So that, the behaviour at three growth stages (E1 = early vegetative stage, E2 = full vegetative stage, and E3 = reproductive stage until harvest in the presence of four water levels (A = 100%, B = 80%, C = 45-50%, and D = 20-25% from a sprinkler irrigation was studied. Eight combinations were evaluated: 111, 110, 100, 101, 001, 011, 010, 000, where the first digit is E1, the second is E2 and the third is E3. The number 0 in the figures represents the period in which growth stage did not receive water, cover by a mobile structure, which slide on trail, and also protected against occasional rainfall. The number 1 represents the time in which the irrigation water levels were applied. The growth stages of the plants as for dry matter yield, differently respond to the water conditions. The E2 and E3 were the most sensible to water supply and E1 the least sensible; in which the water was detrimental to the plant establishment and yield.

  9. Stress tolerance and symbiotic and phylogenic features of root nodule bacteria associated with Medicago species in different bioclimatic regions of Tunisia.

    Science.gov (United States)

    Djedidi, Salem; Yokoyama, Tadashi; Ohkama-Ohtsu, Naoko; Risal, Chandra Prasad; Abdelly, Chedly; Sekimoto, Hitoshi

    2011-01-01

    Thirty two rhizobial isolates were obtained from different bioclimatic regions of Tunisia using as trap plants, Medicago sativa, Medicago ciliaris, Medicago polymorpha and Medicago minima. To study their diversity and characterize them in relation to Mediterranean conditions, abiotic stress resistance, symbiotic properties and genetic diversity in terms of 16S rRNA and nodA sequences were assessed. Five isolates from M. sativa, three from M. ciliaris and three from M. minima could grow at 45°C. Only two isolates from M. sativa grew at 4% NaCl. The most stress tolerant isolates were obtained from arid soils. A phylogenetic analysis of 16S rRNA genes revealed 29 isolates to be closely related to Ensifer including one (Pl.3-9) that showed a 16S rRNA sequence similar to that of Ensifer meliloti and nodA sequence similar to that of Ensifer medicae. However, three isolates were categorized into Agrobacterium containing the nodA of Ensifer. Furthermore, these isolates developed nodules on original hosts. The results for the four isolates suggest horizontal gene transfer between the species.

  10. Production of tetraploid plants of some citrus genotypes

    Directory of Open Access Journals (Sweden)

    Berken ÇİMEN

    2016-12-01

    Full Text Available Improving new seedless citrus varieties is currently the main aspect of citrus breeding programs for both our country and the world. Triploid plants can be recovered directly from artificial and spontaneous mutations as well as crosses between two diploid genotypes resulting from the fertilization of 2n megagametophyte or by hybridization between diploid and tetraploid parents. Thus, production of tetraploid plants which are not naturally found in citrus germplasm has a great importance in terms of seedlessness breeding studies. This study covers production of tetraploid plants for parental usage in order to improve new varieties by ploidy manipulation. Buds of Clementine 22D, W. Murcott and Moro blood orange were used as plant material and effects of colchicine treatments on production of tetraploid forms of these genotypes were investigated. In this purpose, scion buds were treated with colchicine at concentration levels of 0.0%, 0.2%, 0.4%, 0.6% and 0.8% for 4, 6 and 8 hours. After treatments, survival rate (% of plants was recorded and ploidy levels of the plants were determined by flow cytometry analysis. In addition, stomatal observations were recorded such as stomata density, length, width, size and index on the leaves of tetraploid plants in order to compare them with the observations of diploid plants. As a result of the study one tetraploid plant of Clementine 22D mandarin was recovered from the 0.4% colchicine treatment for 6 hours. Besides, mixoploid (2x+4x forms of W. Murcott mandarin and Moro blood orange were recovered.

  11. Breeding and improvement of quality traits in alfalfa (Medicago sativa ssp. sativa L.

    Directory of Open Access Journals (Sweden)

    Milić Dragan

    2014-01-01

    Full Text Available An important objective in alfalfa breeding is the improvement of forage quality. The aim of this experiment was to determine the genetic control of the most important quality traits in whole plant leaves and stems of alfalfa genotypes. Quantitative genetic parameters were estimated for crude protein (CP, neutral detergent fiber (NDF, acid detergent fiber (ADF and acid detergent lignin (ADL content. A 5x5 diallel design with reciprocals among parents from geographically different populations were studied. In the diallel study, effects due to general combining ability (GCA were higher than those due to specific combining ability (SCA for majority of the tested characters in analyses of whole plant. The F1 progeny effect was significant for almost all characters (except ADF content. The highest values of narrow sense heritability’s were found for NDF (h =0.19 and CP (h =0.17 in whole plant, and in leaves (0.11 and 0.08, while in stems most heritable traits are ADF (h = 0.29 and ADL (h = 0.16 content. As a consequence, CP, NDF and ADF content would be more efficient selection criteria in whole plant analyses, while ADF and ADL content would be more interesting to investigate in stems, in a breeding program for improved nutritive value of alfalfa. [Projekat Ministarstva nauke Republike Srbije, br. TR 31024

  12. Embryo rescue of crosses between diploid and tetraploid grape ...

    African Journals Online (AJOL)

    Five cross combinations Jumeigui×Xinghua No.1, 87-1×Kyoho, Kyoho×Muscat Hamburg, Jumeigui× Hongqitezao and Red globle×Kyoho were used as the testing materials. Factors that affect embryo rescue from crossed seeds between diploid and tetraploid grape were studied applying L25(55) orthogonal experiment ...

  13. Description of a tetraploid Tomopterna (Anura: Ranidae) from South ...

    African Journals Online (AJOL)

    1995-08-14

    Aug 14, 1995 ... alleles for lCD, lDH, G6PDH, and AAT. T tandyi and T. cryptotis shared alleles, different from T. delalandil for OPI and MDH. T tandyi shared alleles with T. delalandli, differ- ent from T. cryptotis, for FUMH, GDH, and LDH. This is evidence of hybridization as the tetraploid shares alleles with both putative ...

  14. Abnormal mitosis triggers p53-dependent cell cycle arrest in human tetraploid cells.

    Science.gov (United States)

    Kuffer, Christian; Kuznetsova, Anastasia Yurievna; Storchová, Zuzana

    2013-08-01

    Erroneously arising tetraploid mammalian cells are chromosomally instable and may facilitate cell transformation. An increasing body of evidence shows that the propagation of mammalian tetraploid cells is limited by a p53-dependent arrest. The trigger of this arrest has not been identified so far. Here we show by live cell imaging of tetraploid cells generated by an induced cytokinesis failure that most tetraploids arrest and die in a p53-dependent manner after the first tetraploid mitosis. Furthermore, we found that the main trigger is a mitotic defect, in particular, chromosome missegregation during bipolar mitosis or spindle multipolarity. Both a transient multipolar spindle followed by efficient clustering in anaphase as well as a multipolar spindle followed by multipolar mitosis inhibited subsequent proliferation to a similar degree. We found that the tetraploid cells did not accumulate double-strand breaks that could cause the cell cycle arrest after tetraploid mitosis. In contrast, tetraploid cells showed increased levels of oxidative DNA damage coinciding with the p53 activation. To further elucidate the pathways involved in the proliferation control of tetraploid cells, we knocked down specific kinases that had been previously linked to the cell cycle arrest and p53 phosphorylation. Our results suggest that the checkpoint kinase ATM phosphorylates p53 in tetraploid cells after abnormal mitosis and thus contributes to proliferation control of human aberrantly arising tetraploids.

  15. Sapogenin content variation in Medicago inter-specific hybrid derivatives highlights some aspects of saponin synthesis and control.

    Science.gov (United States)

    Carelli, Maria; Biazzi, Elisa; Tava, Aldo; Losini, Ilaria; Abbruscato, Pamela; Depedro, Claudia; Scotti, Carla

    2015-04-01

    In the Medicago genus, saponins are a complex mixture of triterpene glycosides showing a broad spectrum of biological properties. Here we analyzed the variation in the sapogenin content and composition of inter-specific hybrid Medicago sativa × Medicago arborea derivatives to highlight the pattern of this variation in plant organs (leaves/roots) and the possible mechanisms underlying it. In Sativa Arborea Cross (SAC) leaves and roots, saponins and sapogenins were evaluated using chromatographic methods. Phenotypic correlations between sapogenin content and bio-agronomic traits were examined. Expression studies on β-amyrin synthase and four cytochromes P450 (CYPs) involved in sapogenin biosynthesis and sequence analysis of the key gene of the hemolytic sapogenin pathway (CYP716A12) were performed. Chromatographic analyses revealed a different pattern of among-family variation for hemolytic and nonhemolytic sapogenins and saponins and for the two organs/tissues. Different correlation patterns of gene expression in roots and leaves were found. Diachronic analysis revealed a relationship between sapogenin content and gene transcriptional levels in the early stages of the productive cycle. The results suggest that there are different control mechanisms acting on sapogenin biosynthesis for leaves and roots, which are discussed. A key role for medicagenic acid in the control of sapogenin content in both the tissues is proposed and discussed. © 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.

  16. Relationships of the Woody Medicago Species (Section Dendrotelis) Assessed by Molecular Cytogenetic Analyses

    Science.gov (United States)

    Rosato, Marcela; Castro, Mercedes; Rosselló, Josep A.

    2008-01-01

    Background and Aims The organization of rDNA genes in the woody medic species from the agronomically important Medicago section Dendrotelis was analysed to gain insight into their taxonomic relationships, to assess the levels of infraspecific variation concerning ribosomal loci in a restricted and fragmented insular species (M. citrina) and to assess the nature of its polyploidy. Methods Fluorescence in situ hybridization (FISH) was used for physical mapping of 5S and 45S ribosomal DNA genes in the three species of section Dendrotelis (M. arborea, M. citrina, M. strasseri) and the related M. marina from section Medicago. Genomic in situ hybridization (GISH) was used to assess the genomic relationships of the polyploid M. citrina with the putatively related species from section Dendrotelis. Key Results The diploid (2n = 16) M. marina has a single 45S and two 5S rDNA loci, a pattern usually detected in previous studies of Medicago diploid species. However, polyploid species from section Dendrotelis depart from expectations. The tetraploid species (2n = 32) M. arborea and M. strasseri have one 45S rDNA locus and two 5S rDNA loci, whereas in the hexaploid (2n = 48) M. citrina four 45S rDNA and five 5S rDNA loci have been detected. No single chromosome of M. citrina was uniformly labelled after using genomic probes from M. arborea and M. strasseri. Instead, cross-hybridization signals in M. citrina were restricted to terminal chromosome arms and NOR regions. Conclusions FISH results support the close taxonomic interrelationship between M. arborea and M. strasseri. In these tetraploid species, NOR loci have experienced a diploidization event through physical loss of sequences, a cytogenetic feature so far not reported in other species of the genus. The high number of rDNA loci and GISH results support the specific status for the hexaploid M. citrina, and it is suggested that this species is not an autopolyploid derivative of M. arborea or M. strasseri. Further, molecular

  17. Medicago PhosphoProtein Database: a repository for Medicago truncatula phosphoprotein data

    OpenAIRE

    Rose, Christopher M.; Muthusubramanian eVenkateshwaran; Grimsrud, Paul A.; Westphall, Michael S.; Sussman, Michael R.; Coon, Joshua J.; Jean-Michel eAné

    2012-01-01

    The ability of legume crops to fix atmospheric nitrogen via a symbiotic association with soil rhizobia makes them an essential component of many agricultural systems. Initiation of this symbiosis requires protein phosphorylation-mediated signaling in response to rhizobial signals named Nod factors. Medicago truncatula (Medicago) is the model system for studying legume biology, making the study of its phosphoproteome essential. Here, we describe the Medicago Phosphoprotein Database (http://pho...

  18. Medicago PhosphoProtein Database: a repository for Medicago truncatula phosphoprotein data

    OpenAIRE

    Rose, Christopher M.; Venkateshwaran, Muthusubramanian; Grimsrud, Paul A.; Westphall, Michael S.; Sussman, Michael R.; Coon, Joshua J.; Ané, Jean-Michel

    2012-01-01

    The ability of legume crops to fix atmospheric nitrogen via a symbiotic association with soil rhizobia makes them an essential component of many agricultural systems. Initiation of this symbiosis requires protein phosphorylation-mediated signaling in response to rhizobial signals named Nod factors. Medicago truncatula (Medicago) is the model system for studying legume biology, making the study of its phosphoproteome essential. Here, we describe the Medicago PhosphoProtein Database (MPPD; http...

  19. Meiotic analysis in induced tetraploids of Brachiaria decumbens Stapf

    Directory of Open Access Journals (Sweden)

    Carine Simioni

    2011-01-01

    Full Text Available The meiotic behavior of three tetraploid plants (2n=4x=36 originated from somatic chromosome duplication ofsexually reproducing diploid plants of Brachiaria decumbens was evaluated. All the analyzed plants presented abnormalities relatedto polyploidy, such as irregular chromosome segregation, leading to precocious chromosome migration to the poles and micronucleiduring both meiotic divisions. However, the abnormalities observed did not compromise the meiotic products which were characterizedby regular tetrads and satisfactory pollen fertility varying from 61.36 to 64.86%. Chromosomes paired mostly as bivalents indiakinesis but univalents to tetravalents were also observed. These studies contributed to the choice of compatible fertile sexualgenitors to be crossed to natural tetraploid apomicts in the B. decumbens by identifying abnormalities and verifying pollen fertility.Intraespecific crosses should reduce sterility in the hybrids produced in the breeding program of Brachiaria, a problem observedwith the interspecific hybrids produced so far.

  20. Induksi Tetraploid Tunas Pucuk Jeruk Siam Simadu (Citrus Nobilis Lour) Menggunakan Kolkisin secara In Vitro

    OpenAIRE

    Yulianti, Fitri; Purwito, Agus; Husni, Ali; Dinarti, Diny

    2015-01-01

    Seedless fruit is one of the criteria (necessary) to improve the quality of Simadu tangerine. The most effective method to obtain seedless triploid cultivars is hybridisation between tetraploid and diploid parents. Simadu tangerine is a diploid plant. Tetraploid Simadu tangerine can be obtained with doubling chromosome using colchicines.The aim of this research was to obtain tetraploid Simadu tangerine shoot which would serve as parent to produced seedless Simadu tangerine. Shoot-tips of Sima...

  1. Induksi Tetraploid Tunas Pucuk Jeruk Siam Simadu (Citrus nobilis Lour) Menggunakan Kolkisin secara In Vitro

    OpenAIRE

    Fitri Yulianti; Agus Purwito; Ali Husni; Diny Dinarti

    2015-01-01

    Seedless fruit is one of the criteria (necessary) to improve the quality of Simadu tangerine. The most effective method to obtain seedless triploid cultivars is hybridisation between tetraploid and diploid parents. Simadu tangerine is a diploid plant. Tetraploid Simadu tangerine can be obtained with doubling chromosome using colchicines.The aim of this research was to obtain tetraploid Simadu tangerine shoot which would serve as parent to produced seedless Simadu tangerine. Shoot-tips of Sima...

  2. Induksi Tetraploid Tunas Pucuk Jeruk Siam Simadu (Citrus Nobilis Lour) Menggunakan Kolkisin secara In Vitro

    OpenAIRE

    Dinarti, Diny; Husni, Ali; Purwito, Agus; Yulianti, Fitri

    2014-01-01

    Seedless fruit is one of the criteria necessary to improve the quality of Simadu tangerine. The most effective method to obtain seedless triploid cultivars is hybridisation between tetraploid and diploid parents. Simadu tangerine is diploid plant. Tetraploid Simadu tangerine can be obtained with doubling chromosome using colchicines. The aim of this research was to obtain tetraploid Simadu tangerine shoot which would serve as parent to produced seedless Simadu tangerine. Shoot-tips of Simadu ...

  3. Alkaloid spectrum in diploid and tetraploid hairy root cultures of Datura stramonium.

    Science.gov (United States)

    Berkov, Strahil; Pavlov, Atanas; Kovatcheva, Petia; Stanimirova, Pepa; Philipov, Stefan

    2003-01-01

    Hairy root cultures were obtained from diploid and induced tetraploid plants of Datura stramonium and analyzed by gas chromatography/mass spectrometry. Twenty alkaloids (19 for diploid and 9 for tetraploid hairy root cultures) were identified. A new tropane ester 3-tigloyloxy-6-propionyloxy-7-hydroxytropane was identified on the basis of mass spectral data. Hyoscyamine was the main alkaloid in both diploid and tetraploid cultures. In contrast to diploid hairy roots, the percentage contributions of the alkaloids, with exceptions for hyoscyamine and apoatropine, were higher in the total alkaloid mixture of tetraploid hairy roots.

  4. Fermentation characteristics and nutritive value of low moisture silage made from mature bermudagrass (C. dactylon) and switchgrass (P. virgatum) in mixture with alfalfa (M. sativa) or treated with urea and plantain (Musa AAB

    Science.gov (United States)

    Two experiments were conducted at the University of Kentucky Spindletop Farm in Lexington, Kentucky between October and November, 2009 to evaluate the effect of different percentages of alfalfa (Medicago sativa) as mixtures in switchgrass (Panicum virgatus) and bermudagrass (Cynodon dactylon) silage...

  5. Allozyme Diversity in the Tetraploid Endemic Thymus loscosii (Lamiaceae)

    Science.gov (United States)

    LÓPEZ‐PUJOL, JORDI; BOSCH, MARIA; SIMON, JOAN; BLANCHÉ, CÈSAR

    2004-01-01

    • Background and Aims Thymus loscosii (Lamiaceae) is a tetraploid perennial species endemic to the Ebro river basin (north‐eastern Spain), which is included in the National Catalogue of Endangered Species. It is a tetraploid species (2n = 54), presumably an autotetraploid originated by the duplication of a 2n = 28 genome and the subsequent loss of two chromosomes. Allozyme electrophoresis was conducted to survey the levels and distribution of genetic diversity and to test the previous autopolyploid hypothesis for its origin. In addition, both in situ and ex situ conservation measures are proposed. • Methods Eight populations were sampled for analysis by standard methods of starch gel electrophoresis, and six putative enzymatic loci were resolved (five consistently and one only partially). • Key Results Banding patterns exhibited no evidence of fixed heterozygosity and showed both balanced and unbalanced heterozygotes. In addition, most individuals showed a pattern consistent with the presence of three or four alleles at a single locus. High levels of genetic variability were found at population level (P = 85 %, A = 3·0, He = 0·422), in addition to a trend of an excess of heterozygotes. • Conclusions Allozyme data support the hypothesis that T. loscosii is an autotetraploid, and the high number of alleles at some loci may be due to repeated polyploidization events. The high values of genetic variation found in this species agree with those expected for tetraploids. The excess of heterozygotes may be due to some barriers to inbreeding (e.g. occurrence of gynodioecy) and/or selection for heterozygosity. PMID:14742353

  6. SILENCIAMIENTO EPIGENETICO EN EL ROEDOR TETRAPLOIDE TYMPANOCTOMYS BARRERAE (DODENTIA, OCTODONTIDAE)

    OpenAIRE

    BACQUET PEREZ, CAROLINE NICOLE ORIANA

    2008-01-01

    El roedor Tympanoctomys barrerae (4n=102) es el primer mamífero tetraploide descrito. En esta tesis se estudió la contribución de los mecanismos epigenéticos en la diploidización funcional de su genoma duplicado. Para este efecto, se analizó la inactivación del cromosoma X(XCI), la impronta genómica y los cambios potenciales de la estructura croma única gatillados por la duplicación genómica. El cromosoma X inactivo se detectó mediante la unción inmunofluorescente de proteínas típica...

  7. Localized chiasmata and meiotic nodules in the tetraploid onion Allium porrum

    NARCIS (Netherlands)

    Stack, S M; Roelofs, D

    Allium porrum L. (cultivated leek) (2n = 4x = 32) is a fertile tetraploid that forms bivalents with pericentric chiasmata at metaphase I. To investigate the basis of this unusual behavior for a tetraploid, we describe the karyotype, axial cores, synaptonemal complexes (SCs), and meiotic nodules of

  8. Tetraploid Lolium perenne genotypes identified in Danish semi-natural habitats

    DEFF Research Database (Denmark)

    Damgaard, Christian; Simonsen, Vibeke; Sørensen, Jesper Givskov

    2014-01-01

    The ploidy of perennial ryegrass, Lolium perenne, was studied in order to assess the potential of tetraploidity as a means of preventing transgenes from spreading to natural populations.In contradiction to earlier observations, we found that tetraploid L. perenne genotypes was present in a semi...

  9. Aspects of gene regulation in the diploid and tetraploid Odontophrynus americanus (Amphibia, Anura, Leptodactylidae

    Directory of Open Access Journals (Sweden)

    Cianciarullo Aurora M.

    2000-01-01

    Full Text Available Erythropoietic and hemoglobin DNA transcriptional activities were analyzed in the diploid and the tetraploid Odontophrynus americanus. Flow cytometric analyses of DNA, RNA and mitochondrial contents showed increased genic activity in both diploid and tetraploid animals during erythropoiesis in vivo elicited by pretreatment phenylhydrazine. Generally, higher values were seen in immature tetraploid erythroid cells. On the 10th day of recovery from anemia, large amounts of messenger RNA were found in both specimens. Based on the mitochondrial content, the tetraploid cells had more intense energy metabolism than the diploid cells. Diploid O. americanus had about three times more erythroid cells than tetraploid specimens, indicating that there were differences in the regulatory mechanisms of erythroid cells. Hematological parameters showed that tetraploid cells had 30% more hemoglobin than the diploid, suggesting a regulatory mechanism of hemoglobin synthesis at the transcriptional level. Cytoplasmic inclusions resembling Heinz bodies were found in both types of cells. In the tetraploid cells they were previously found associated with RNA or RNP, suggesting that other regulatory system which controls the accumulation of nontranslated RNA transcribed in excess must be present. These differences at the physiological and molecular levels during erythropoiesis reinforce the hypothesis that speciation is occurring between diploid and tetraploid O. americanus.

  10. Early seedling development of Medicago truncatula genotypes ...

    African Journals Online (AJOL)

    adel

    2014-01-08

    Jan 8, 2014 ... dry weight and storage protein content ... Key words: Medicago truncatula, salt stress, seed weight, storage protein content, seed vigour, aged seeds. ..... My Forest. 29:225-230. Amouri AA, Fyad-Lamèche FZ (2012). Comparative analysis of salinity tolerance of the male gametophyte and the sporophyte in ...

  11. Genomic characterization of the LEED..PEEDs, a gene family unique to the medicago lineage.

    Science.gov (United States)

    Trujillo, Diana I; Silverstein, Kevin A T; Young, Nevin D

    2014-08-25

    The LEED..PEED (LP) gene family in Medicago truncatula (A17) is composed of 13 genes coding small putatively secreted peptides with one to two conserved domains of negatively charged residues. This family is not present in the genomes of Glycine max, Lotus japonicus, or the IRLC species Cicer arietinum. LP genes were also not detected in a Trifolium pratense draft genome or Pisum sativum nodule transcriptome, which were sequenced de novo in this study, suggesting that the LP gene family arose within the past 25 million years. M. truncatula accession HM056 has 13 LP genes with high similarity to those in A17, whereas M. truncatula ssp. tricycla (R108) and M. sativa have 11 and 10 LP gene copies, respectively. In M. truncatula A17, 12 LP genes are located on chromosome 7 within a 93-kb window, whereas one LP gene copy is located on chromosome 4. A phylogenetic analysis of the gene family is consistent with most gene duplications occurring prior to Medicago speciation events, mainly through local tandem duplications and one distant duplication across chromosomes. Synteny comparisons between R108 and A17 confirm that gene order is conserved between the two subspecies, although a further duplication occurred solely in A17. In M. truncatula A17, all 13 LPs are exclusively transcribed in nodules and absent from other plant tissues, including roots, leaves, flowers, seeds, seed shells, and pods. The recent expansion of LP genes in Medicago spp. and their timing and location of expression suggest a novel function in nodulation, possibly as an aftermath of the evolution of bacteroid terminal differentiation or potentially associated with rhizobial-host specificity. Copyright © 2014 Trujillo et al.

  12. Characterization and genetic dissection of resistance to spotted alfalfa aphid (Therioaphis trifolii) in Medicago truncatula

    Science.gov (United States)

    Kamphuis, Lars G.; Lichtenzveig, Judith; Peng, Kefan; Guo, Su-Min; Klingler, John P.

    2013-01-01

    Aphids cause significant yield losses in agricultural crops worldwide. Medicago truncatula, a model legume, cultivated pasture species in Australia and close relative of alfalfa (Medicago sativa), was used to study the defence response against Therioaphis trifolii f. maculate [spotted alfalfa aphid (SAA)]. Aphid performance and plant damage were compared among three accessions. A20 is highly susceptible, A17 has moderate resistance, and Jester is strongly resistant. Subsequent analyses using A17 and A20, reciprocal F1s and an A17×A20 recombinant inbred line (RIL) population revealed that this moderate resistance is phloem mediated and involves antibiosis and tolerance but not antixenosis. Electrical penetration graph analysis also identified a novel waveform termed extended potential drop, which occurred following SAA infestation of M. truncatula. Genetic dissection using the RIL population revealed three quantitative trait loci on chromosomes 3, 6, and 7 involved in distinct modes of aphid defence including antibiosis and tolerance. An antibiosis locus resides on linkage group 3 (LG3) and is derived from A17, whereas a plant tolerance and antibiosis locus resides on LG6 and is derived from A20, which exhibits strong temporary tolerance. The loci identified reside in regions harbouring classical resistance genes, and introgression of these loci in current medic cultivars may help provide durable resistance to SAA, while elucidation of their molecular mechanisms may provide valuable insight into other aphid–plant interactions. PMID:24058162

  13. Characterization and genetic dissection of resistance to spotted alfalfa aphid (Therioaphis trifolii) in Medicago truncatula

    KAUST Repository

    Kamphuis, L. G.

    2013-09-21

    Aphids cause significant yield losses in agricultural crops worldwide. Medicago truncatula, a model legume, cultivated pasture species in Australia and close relative of alfalfa (Medicago sativa), was used to study the defence response against Therioaphis trifolii f. maculate [spotted alfalfa aphid (SAA)]. Aphid performance and plant damage were compared among three accessions. A20 is highly susceptible, A17 has moderate resistance, and Jester is strongly resistant. Subsequent analyses using A17 and A20, reciprocal F1s and an A17×A20 recombinant inbred line (RIL) population revealed that this moderate resistance is phloem mediated and involves antibiosis and tolerance but not antixenosis. Electrical penetration graph analysis also identified a novel waveform termed extended potential drop, which occurred following SAA infestation of M. truncatula. Genetic dissection using the RIL population revealed three quantitative trait loci on chromosomes 3, 6, and 7 involved in distinct modes of aphid defence including antibiosis and tolerance. An antibiosis locus resides on linkage group 3 (LG3) and is derived from A17, whereas a plant tolerance and antibiosis locus resides on LG6 and is derived from A20, which exhibits strong temporary tolerance. The loci identified reside in regions harbouring classical resistance genes, and introgression of these loci in current medic cultivars may help provide durable resistance to SAA, while elucidation of their molecular mechanisms may provide valuable insight into other aphid–plant interactions.

  14. p53 Dependent Centrosome Clustering Prevents Multipolar Mitosis in Tetraploid Cells

    Science.gov (United States)

    Yi, Qiyi; Zhao, Xiaoyu; Huang, Yun; Ma, Tieliang; Zhang, Yingyin; Hou, Heli; Cooke, Howard J.; Yang, Da-Qing; Wu, Mian; Shi, Qinghua

    2011-01-01

    Background p53 abnormality and aneuploidy often coexist in human tumors, and tetraploidy is considered as an intermediate between normal diploidy and aneuploidy. The purpose of this study was to investigate whether and how p53 influences the transformation from tetraploidy to aneuploidy. Principal Findings Live cell imaging was performed to determine the fates and mitotic behaviors of several human and mouse tetraploid cells with different p53 status, and centrosome and spindle immunostaining was used to investigate centrosome behaviors. We found that p53 dominant-negative mutation, point mutation, or knockout led to a 2∼ 33-fold increase of multipolar mitosis in N/TERT1, 3T3 and mouse embryonic fibroblasts (MEFs), while mitotic entry and cell death were not significantly affected. In p53-/- tetraploid MEFs, the ability of centrosome clustering was compromised, while centrosome inactivation was not affected. Suppression of RhoA/ROCK activity by specific inhibitors in p53-/- tetraploid MEFs enhanced centrosome clustering, decreased multipolar mitosis from 38% to 20% and 16% for RhoA and ROCK, respectively, while expression of constitutively active RhoA in p53+/+ tetraploid 3T3 cells increased the frequency of multipolar mitosis from 15% to 35%. Conclusions p53 could not prevent tetraploid cells entering mitosis or induce tetraploid cell death. However, p53 abnormality impaired centrosome clustering and lead to multipolar mitosis in tetraploid cells by modulating the RhoA/ROCK signaling pathway. PMID:22076149

  15. Proteomic Analysis of Etiolated Juvenile Tetraploid Robinia pseudoacacia Branches during Different Cutting Periods

    Directory of Open Access Journals (Sweden)

    Nan Lu

    2014-04-01

    Full Text Available The propagation of hard-branch cuttings of tetraploid Robinia pseudoacacia (black locust is restricted by the low rooting rate; however, etiolated juvenile tetraploid black locust branches result in a significantly higher rooting rate of cuttings compared with non-etiolated juvenile tetraploid branches. To identify proteins that influence the juvenile tetraploid branch rooting process, two-dimensional electrophoresis (2-DE and matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectra (MALDI-TOF/TOF-MS were used to analyze proteomic differences in the phloem of tetraploid R. pseudoacacia etiolated and non-etiolated juvenile branches during different cutting periods. A total of 58 protein spots differed in expression level, and 16 protein spots were only expressed in etiolated branches or non-etiolated ones. A total of 40 highly expressed protein spots were identified by mass spectrometry, 14 of which were accurately retrieved. They include nucleoglucoprotein metabolic proteins, signaling proteins, lignin synthesis proteins and phyllochlorin. These results help to reveal the mechanism of juvenile tetraploid R. pseudoacacia etiolated branch rooting and provide a valuable reference for the improvement of tetraploid R. pseudoacacia cutting techniques.

  16. Characterization of two TT2-type MYB transcription factors regulating proanthocyanidin biosynthesis in tetraploid cotton, Gossypium hirsutum.

    Science.gov (United States)

    Lu, Nan; Roldan, Marissa; Dixon, Richard A

    2017-08-01

    Two TT2-type MYB transcription factors identified from tetraploid cotton are involved in regulating proanthocyanidin biosynthesis, providing new strategies for engineering condensed tannins in crops. Proanthocyanidins (PAs), also known as condensed tannins, are important secondary metabolites involved in stress resistance in plants, and are health supplements that help to reduce cholesterol levels. As one of the most widely grown crops in the world, cotton provides the majority of natural fabrics and is a supplemental food for ruminant animals. The previous studies have suggested that PAs present in cotton are a major contributor to fiber color. However, the biosynthesis of PAs in cotton still remains to be elucidated. AtTT2 (transparent testa 2) is a MYB family transcription factor from Arabidopsis that initiates the biosynthesis of PAs by inducing the expression of multiple genes in the pathway. In this study, we isolated two R2R3-type MYB transcription factors from Gossypium hirsutum that are homologous to AtTT2. Expression analysis showed that both genes were expressed at different levels in various cotton tissues, including leaf, seed coat, and fiber. Protoplast transactivation assays revealed that these two GhMYBs were able to activate promoters of genes encoding enzymes in the PA biosynthesis pathway, namely anthocyanidin reductase and leucoanthocyanidin reductase. Complementation experiments showed that both of the GhMYBs were able to recover the transparent testa seed coat phenotype of the Arabidopsis tt2 mutant by restoring PA biosynthesis. Ectopic expression of either of the two GhMYBs in Medicago truncatula hairy roots increased the contents of anthocyanins and PAs compared to control lines expressing the GUS gene, and expression levels of MtDFR, MtLAR, and MtANR were also elevated in lines expressing GhMYBs. Together, these data provide new insights into engineering condensed tannins in cotton.

  17. Selection on meiosis genes in diploid and tetraploid Arabidopsis arenosa.

    Science.gov (United States)

    Wright, Kevin M; Arnold, Brian; Xue, Katherine; Šurinová, Maria; O'Connell, Jeremy; Bomblies, Kirsten

    2015-04-01

    Meiotic chromosome segregation is critical for fertility across eukaryotes, and core meiotic processes are well conserved even between kingdoms. Nevertheless, recent work in animals has shown that at least some meiosis genes are highly diverse or strongly differentiated among populations. What drives this remains largely unknown. We previously showed that autotetraploid Arabidopsis arenosa evolved stable meiosis, likely through reduced crossover rates, and that associated with this there is strong evidence for selection in a subset of meiosis genes known to affect axis formation, synapsis, and crossover frequency. Here, we use genome-wide data to study the molecular evolution of 70 meiosis genes in a much wider sample of A. arenosa. We sample the polyploid lineage, a diploid lineage from the Carpathian Mountains, and a more distantly related diploid lineage from the adjacent, but biogeographically distinct Pannonian Basin. We find that not only did selection act on meiosis genes in the polyploid lineage but also independently on a smaller subset of meiosis genes in Pannonian diploids. Functionally related genes are targeted by selection in these distinct contexts, and in two cases, independent sweeps occurred in the same loci. The tetraploid lineage has sustained selection on more genes, has more amino acid changes in each, and these more often affect conserved or potentially functional sites. We hypothesize that Pannonian diploid and tetraploid A. arenosa experienced selection on structural proteins that mediate sister chromatid cohesion, the formation of meiotic chromosome axes, and synapsis, likely for different underlying reasons. © The Author 2014. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  18. Models Analyses for Allelopathic Effects of Chicory at Equivalent Coupling of Nitrogen Supply and pH Level on F. arundinacea, T. repens and M. sativa

    OpenAIRE

    Quanzhen Wang; Bao Xie; Chunhui Wu; Guo Chen; Zhengwei Wang; Jian Cui; Tianming Hu; Pawel Wiatrak

    2012-01-01

    Alllelopathic potential of chicory was investigated by evaluating its effect on seed germination, soluble sugar, malondialdehyde (MDA) and the chlorophyll content of three target plants species (Festuca arundinacea, Trifolium repens and Medicago sativa). The secretion of allelochemicals was regulated by keeping the donor plant (chicory) separate from the three target plant species and using different pH and nitrogen levels. Leachates from donor pots with different pH levels and nitrogen conce...

  19. Flavonolignans from Avena sativa.

    Science.gov (United States)

    Wenzig, Eva; Kunert, Olaf; Ferreira, Daneel; Schmid, Martin; Schühly, Wolfgang; Bauer, Rudolf; Hiermann, Alois

    2005-02-01

    Three flavonolignans derived from the flavone tricin were isolated from Avena sativa herb. This is the first report of the presence of flavonolignans in A. sativa. In the known compounds 1a and 1b, a coniferyl alcohol moiety is linked to the flavone by an ether bond; in the new natural product 2, it is linked by C-C bonds. Structure elucidation of compound 2 was performed by 1D and 2D NMR experiments, and the absolute configuration was determined from circular dichroic data.

  20. Tetraploidization events by chromosome doubling of nucellar cells are frequent in apomictic citrus and are dependent on genotype and environment

    Science.gov (United States)

    Aleza, Pablo; Froelicher, Yann; Schwarz, Sergio; Agustí, Manuel; Hernández, María; Juárez, José; Luro, François; Morillon, Raphael; Navarro, Luis; Ollitrault, Patrick

    2011-01-01

    Background and Aims Polyploidy is a major component of plant evolution. The citrus gene pool is essentially diploid but tetraploid plants are frequently encountered in seedlings of diploid apomictic genotypes. The main objectives of the present study were to establish the origin of these tetraploid plants and to ascertain the importance of genotypic and environmental factors on tetraploid formation. Methods Tetraploid seedlings from 30 diploid apomictic genotypes were selected by flow cytometry and genotyped with 24 single sequence repeat (SSR) markers to analyse their genetic origin. Embryo rescue was used to grow all embryos contained in polyembryonic seeds of ‘Tardivo di Ciaculli’ mandarin, followed by characterization of the plantlets obtained by flow cytometry and SSR markers to accurately establish the rate of tetraploidization events and their potential tissue location. Inter-annual variations in tetraploid seedling rates were analysed for seven genotypes. Variation in tetraploid plantlet rates was analysed between different seedlings of the same genotype (‘Carrizo’ citrange; Citrus sinensis × Poncirus trifoliata) from seeds collected in different tropical, subtropical and Mediterranean countries. Key Results Tetraploid plants were obtained for all the studied diploid genotypes, except for four mandarins. All tetraploid plants were identical to their diploid maternal line for SSR markers and were not cytochimeric. Significant genotypic and environmental effects were observed, as well as negative correlation between mean temperature during the flowering period and tetraploidy seedling rates. The higher frequencies (20 %) of tetraploids were observed for citranges cultivated in the Mediterranean area. Conclusions Tetraploidization by chromosome doubling of nucellar cells are frequent events in apomictic citrus, and are affected by both genotypic and environmental factors. Colder conditions in marginal climatic areas appear to favour the expression of

  1. The Mitochondrial Complexome of Medicago truncatula

    Science.gov (United States)

    Kiirika, Leonard Muriithi; Behrens, Christof; Braun, Hans-Peter; Colditz, Frank

    2013-01-01

    Legumes (Fabaceae, Leguminosae) are unique in their ability to carry out an elaborate endosymbiotic nitrogen fixation process with rhizobia proteobacteria. The symbiotic nitrogen fixation enables the host plants to grow almost independently of any other nitrogen source. Establishment of symbiosis requires adaptations of the host cellular metabolism, here foremost of the energy metabolism mainly taking place in mitochondria. Since the early 1990s, the galegoid legume Medicago truncatula Gaertn. is a well-established model for studying legume biology, but little is known about the protein complement of mitochondria from this species. An initial characterization of the mitochondrial proteome of M. truncatula (Jemalong A17) was published recently. In the frame of this study, mitochondrial protein complexes were characterized using Two-dimensional (2D) Blue native (BN)/SDS-PAGE. From 139 detected spots, the “first hit” (=most abundant) proteins of 59 spots were identified by mass spectrometry. Here, we present a comprehensive analysis of the mitochondrial “complexome” (the “protein complex proteome”) of M. truncatula via 2D BN/SDS-PAGE in combination with highly sensitive MS protein identification. In total, 1,485 proteins were identified within 158 gel spots, representing 467 unique proteins. Data evaluation by the novel GelMap annotation tool allowed recognition of protein complexes of low abundance. Overall, at least 36 mitochondrial protein complexes were found. To our knowledge several of these complexes were described for the first time in Medicago. The data set is accessible under http://www.gelmap.de/medicago/. The mitochondrial protein complex proteomes of Arabidopsis available at http://www.gelmap.de/arabidopsis/ and Medicago are compared. PMID:23596449

  2. Correspondence of trichome mutations in diploid and tetraploid cottons.

    Science.gov (United States)

    Desai, Aparna; Chee, Peng W; May, O Lloyd; Paterson, Andrew H

    2008-01-01

    Quantitative variation for leaf trichome number is observed within and among Gossypium species, varying from glabrous to densely pubescent phenotypes. Moreover, economically important cotton lint fibers are modified trichomes. Earlier studies have mapped quantitative trait loci (QTLs) affecting leaf pubescence in Gossypium using allotetraploids. In this study, we mapped genes responsible for leaf trichome density in a diploid A genome cross. We were able to map 3 QTLs affecting leaf pubescence based on trichome counts obtained from young leaves (YL) and mature leaves (ML). When the F(2) progeny were classified as pubescent versus glabrous, their ratio did not deviate significantly from a 3:1 model, suggesting that glabrousness is inherited in a simple Mendelian fashion. The glabrous mutation mapped to linkage group A3 at the position of major QTL YL1 and ML1 and appeared orthologous to the t1 locus of the allotetraploids. Interestingly, a fiber mutation, sma-4(ha), observed in the same F(2) population cosegregated with the glabrous marker, which indicates either close linkage or common genetic control of lint fiber and leaf trichomes. Studies of A genome diploids may help to clarify the genetic control of trichomes and fiber in both diploid and tetraploid cottons.

  3. Increase phosphorus availability from the use of alfalfa (Medicago sativa L) green manure in rice (Oryza sativa L.) agroecosystem.

    Science.gov (United States)

    Gao, Xiaoye; Shi, Dongyan; Lv, Aimin; Wang, Shengyin; Yuan, Shili; Zhou, Peng; An, Yuan

    2016-11-11

    Alfalfa is a good green manure source, but its effect on rice growth has not been fully elucidated. Two green manure species, alfalfa and broad bean (Vicia faba L.), and two N fertilizer levels, alone or combination, were applied to a rice field. The results indicated that alfalfa had more pronounced effects on increasing soil labile phosphorus (P) fractions (including NaHCO3-Pi, NaOH-Pi), P uptake and soil enzyme activities (dehydrogenase, urease, acid phosphatase and β-glucosidase) than broad bean and N fertilizer. The transformation of NaHCO3-Po to labile P regulated by alfalfa played a significant direct and indirect effect on grain yield. Although a much lower N input from alfalfa addition, a similar grain yield with N fertilizer treatment was achieved, and the integration of alfalfa with N fertilizer produced the highest grain yield and P availability, which was associated with the highest urease, acid phosphatase and β-glucosidase activity in soil. These results indicate that alfalfa green manure had a great ability of increasing grain yield through enhancing P availability in rice paddy, which could give us a way to reduce N fertilizer application by enhancing P availability.

  4. Induction of Tetraploids from Petiole Explants through Colchicine Treatments in Echinacea purpurea L.

    Directory of Open Access Journals (Sweden)

    Dahanayake Nilanthi

    2009-01-01

    Full Text Available Petiole explants were obtained from in vitro grown diploid (2x=22 Echinacea purpurea plantlets. Shoots were regenerated by culturing the explants on MS basal medium containing 0.3 mg/L benzyladenine (BA, 0.01 mg/L naphthaleneacetic acid (NAA and four concentrations (30, 60, 120, and 240 mg/L of colchicine for 30 days, or 120 mg/L of colchicine for various durations (7, 14, 21, and 28 days. The regenerated shoots were induced to root on MS basal medium with 0.01 mg/L NAA, and then the root-tips of the regenerated shoots were sampled for count of chromosome number. It was found that a treatment duration of >7 days was necessary for induction of tetraploid (4x=44 shoots, and treatment with 120 mg/L colchicine for 28 days was the most efficient for induction of tetraploids, yielding 23.5% of tetraploids among all the regenerated shoots. Chimeras were observed in almost all the treatments. However, the ratio of tetraploid to diploid cells in a chimeric plant was usually low. In comparison with diploid plants, tetraploid plants in vitro had larger stomata and thicker roots with more root branches, and had prominently shorter inflorescence stalk when mature.

  5. The role of tetraploids in the sexual-asexual cycle in dandelions (Taraxacum).

    Science.gov (United States)

    Verduijn, M H; Van Dijk, P J; Van Damme, J M M

    2004-10-01

    Apomictic plants often produce pollen that can function in crosses with related sexuals. Moreover, facultative apomicts can produce some sexual offspring. In dandelions, Taraxacum, a sexual-asexual cycle between diploid sexuals and triploid apomicts, has been described, based on experimental crosses and population genetic studies. Little is known about the actual hybridization processes in nature. We therefore studied the sexual-asexual cycle in a mixed dandelion population in the Netherlands. In this population, the frequencies of sexual diploids and triploids were 0.31 and 0.68, respectively. In addition, less than 1% tetraploids were detected. Diploids were strict sexuals, triploids were obligate apomicts, but tetraploids were most often only partly apomictic, lacking certain elements of apomixis. Tetraploid seed fertility in the field was significantly lower than that of apomictic triploids. Field-pollinated sexual diploids produced on average less than 2% polyploid offspring, implying that the effect of hybridization in the 2x-3x cycle in Taraxacum will be low. Until now, 2x-3x crosses were assumed to be the main pathway of new formation of triploid apomicts in the sexual-asexual cycle in Taraxacum. However, tetraploid pollen donors produced 28 times more triploid offspring in experimental crosses with diploid sexuals than triploid pollen donors. Rare tetraploids may therefore act as an important bridge in the formation of new triploid apomicts.

  6. Partial preferential chromosome pairing is genotype dependent in tetraploid rose.

    Science.gov (United States)

    Bourke, Peter M; Arens, Paul; Voorrips, Roeland E; Esselink, G Danny; Koning-Boucoiran, Carole F S; Van't Westende, Wendy P C; Santos Leonardo, Tiago; Wissink, Patrick; Zheng, Chaozhi; van Geest, Geert; Visser, Richard G F; Krens, Frans A; Smulders, Marinus J M; Maliepaard, Chris

    2017-04-01

    It has long been recognised that polyploid species do not always neatly fall into the categories of auto- or allopolyploid, leading to the term 'segmental allopolyploid' to describe everything in between. The meiotic behaviour of such intermediate species is not fully understood, nor is there consensus as to how to model their inheritance patterns. In this study we used a tetraploid cut rose (Rosa hybrida) population, genotyped using the 68K WagRhSNP array, to construct an ultra-high-density linkage map of all homologous chromosomes using methods previously developed for autotetraploids. Using the predicted bivalent configurations in this population we quantified differences in pairing behaviour among and along homologous chromosomes, leading us to correct our estimates of recombination frequency to account for this behaviour. This resulted in the re-mapping of 25 695 SNP markers across all homologues of the seven rose chromosomes, tailored to the pairing behaviour of each chromosome in each parent. We confirmed the inferred differences in pairing behaviour among chromosomes by examining repulsion-phase linkage estimates, which also carry information about preferential pairing and recombination. Currently, the closest sequenced relative to rose is Fragaria vesca. Aligning the integrated ultra-dense rose map with the strawberry genome sequence provided a detailed picture of the synteny, confirming overall co-linearity but also revealing new genomic rearrangements. Our results suggest that pairing affinities may vary along chromosome arms, which broadens our current understanding of segmental allopolyploidy. © 2017 The Authors The Plant Journal published by John Wiley & Sons Ltd and Society for Experimental Biology.

  7. High-quality permanent draft genome sequence of Ensifer medicae strain WSM244, a microsymbiont isolated from Medicago polymorpha growing in alkaline soil.

    Science.gov (United States)

    Ardley, Julie; Tian, Rui; O'Hara, Graham; Seshadri, Rekha; Reddy, T B K; Pati, Amrita; Woyke, Tanja; Markowitz, Victor; Ivanova, Natalia; Kyrpides, Nikos; Howieson, John; Reeve, Wayne

    2015-01-01

    Ensifer medicae WSM244 is an aerobic, motile, Gram-negative, non-spore-forming rod that can exist as a soil saprophyte or as a legume microsymbiont of Medicago species. WSM244 was isolated in 1979 from a nodule recovered from the roots of the annual Medicago polymorpha L. growing in alkaline soil (pH 8.0) in Tel Afer, Iraq. WSM244 is the only acid-sensitive E. medicae strain that has been sequenced to date. It is effective at fixing nitrogen with M. polymorpha L., as well as with more alkaline-adapted Medicago spp. such as M. littoralis Loisel., M. scutellata (L.) Mill., M. tornata (L.) Mill. and M. truncatula Gaertn. This strain is also effective with the perennial M. sativa L. Here we describe the features of E. medicae WSM244, together with genome sequence information and its annotation. The 6,650,282 bp high-quality permanent draft genome is arranged into 91 scaffolds of 91 contigs containing 6,427 protein-coding genes and 68 RNA-only encoding genes, and is one of the rhizobial genomes sequenced as part of the DOE Joint Genome Institute 2010 Genomic Encyclopedia for Bacteria and Archaea-Root Nodule Bacteria (GEBA-RNB) project proposal.

  8. and black cumin (Nigella sativa)

    African Journals Online (AJOL)

    USER

    2016-03-30

    Mar 30, 2016 ... Lipid composition of egg yolk and serum in laying hens fed diets containing black cumin (Nigella sativa). Int. J. Poult. Sci. 5, 574-578. El-Dakhakhny, M., Mady, N.I. & Halim, M.A., 2000. Nigella sativa L. oil protects against induced hepatotoxicity and improves serum lipid profile in rats. Arzneimittelforschung ...

  9. Genetic effects of chromosomes 01, 04, and 18 from three tetraploid gossypium species in topcrosses with five elite cultivars

    Science.gov (United States)

    Chromosome substitution lines (CSL) have been developed for selected chromosomes from two tetraploid species of Gossypium and have been shown to be effective ways to target introgression of useful alleles from exotic tetraploid species into Upland cotton, G. hirsutum L. Genetic effects of chromosome...

  10. “New” lectins for the identification of erythrocyte cryptantigens and the classification of erythrocyte polyagglutinability: Medicago disciformis and Medicago turbinata

    OpenAIRE

    Bird, GWG; Wingham, June

    1983-01-01

    The seeds of Medicago disciformis and Medicago turbinata contain lectins for T or Th but not Tk or Tx red cell cryptantigens. Extracts of Medicago disciformis seeds are a useful addition to a panel of lectins used in the classification of red cell cryptantigens and of red cell polyagglutinability.

  11. Arbuscular Mycorrhiza Stimulates Biological Nitrogen Fixation in Two Medicago spp. through Improved Phosphorus Acquisition

    Science.gov (United States)

    Püschel, David; Janoušková, Martina; Voříšková, Alena; Gryndlerová, Hana; Vosátka, Miroslav; Jansa, Jan

    2017-01-01

    Legumes establish root symbioses with rhizobia that provide plants with nitrogen (N) through biological N fixation (BNF), as well as with arbuscular mycorrhizal (AM) fungi that mediate improved plant phosphorus (P) uptake. Such complex relationships complicate our understanding of nutrient acquisition by legumes and how they reward their symbiotic partners with carbon along gradients of environmental conditions. In order to disentangle the interplay between BNF and AM symbioses in two Medicago species (Medicago truncatula and M. sativa) along a P-fertilization gradient, we conducted a pot experiment where the rhizobia-treated plants were either inoculated or not inoculated with AM fungus Rhizophagus irregularis ‘PH5’ and grown in two nutrient-poor substrates subjected to one of three different P-supply levels. Throughout the experiment, all plants were fertilized with 15N-enriched liquid N-fertilizer to allow for assessment of BNF efficiency in terms of the fraction of N in the plants derived from the BNF (%NBNF). We hypothesized (1) higher %NBNF coinciding with higher P supply, and (2) higher %NBNF in mycorrhizal as compared to non-mycorrhizal plants under P deficiency due to mycorrhiza-mediated improvement in P nutrition. We found a strongly positive correlation between total plant P content and %NBNF, clearly documenting the importance of plant P nutrition for BNF efficiency. The AM symbiosis generally improved P uptake by plants and considerably stimulated the efficiency of BNF under low P availability (below 10 mg kg-1 water extractable P). Under high P availability (above 10 mg kg-1 water extractable P), the AM symbiosis brought no further benefits to the plants with respect to P nutrition even as the effects of P availability on N acquisition via BNF were further modulated by the environmental context (plant and substrate combinations). As a response to elevated P availability in the substrate, the extent of root length colonization by AM fungi was

  12. Effects of drought stress on germination characters of some alfalfa (Medicago sativa L. varieties

    Directory of Open Access Journals (Sweden)

    Emine BUDAKLI ÇARPICI

    2015-12-01

    Full Text Available This study was conducted to determine the responses of some alfalfa varieties to different drought stress at germination stage. Alfalfa varieties Bilensoy-80, Alsancak, Gözlü-1, Prosementi and Isıde were used as plant materials. Four different drought stress levels (0, -2.95, -4.91 and -10.27 bar were handled in this study and were used Polyethylene glycol-6000 (PEG-6000 to create of drought stress at different levels. The experiment was conducted as completely randomized design with two factors and four replications. In the experiment, some characters such as germination percentage, plumula length, radicle length and vigour index were examined. The results revealed that drought stress had a significant effect on all characters. Germination percentage, plumula length, radicle length, and vigour index ranged 0.00-99.50 %, 0.00-43.24 mm, 0.00-54.45 mm and 0.00-43.79, respectively. Considering germination percentage, the cultivars tolerated the drought stress up to-4.91 bar, but after this level germination percentage decreased significantly. In terms of plumula length and vigour index, the cultivars tolerated the drought stress up to -2.95 bar and in terms of radicle length the cultivars tolerated the drought stress up to -4.91 bar and were observed a significant decreases or deaths after this levels. Bilensoy-80 and Alsancak had been more successful for drought than the other varieties in experiment.

  13. Alfalfa (Medicago sativa L. seed yield in relation to phosphorus fertilization and honeybee pollination

    Directory of Open Access Journals (Sweden)

    Saad Naser Al-Kahtani

    2017-07-01

    Full Text Available This investigation was conducted at the Agricultural and Veterinary Training and Research Station, King Faisal University, Al-Ahsa, Saudi Arabia, during the alfalfa growing season in 2014. The study aimed to evaluate the impact of phosphorus fertilization and honeybee pollination on alfalfa seed production. The experiment was divided into 9 treatments of open pollination, honeybee pollination, and non-pollination with three different levels (0, 300 or 600 kg P2O5/ha/year of triple super phosphate. All vegetative growth attributes of Hassawi alfalfa were significantly higher in the non-insect pollination plots, while the yield and yield component traits were significantly higher with either open pollination or honeybee pollination in parallel with the increasing level of phosphorus fertilizer up to 600 kg P2O5/ha/year in light salt-affected loamy sand soils. There was no seed yield in Hassawi alfalfa without insect pollination. Therefore, placing honeybee colonies near the fields of Hassawi alfalfa and adding 600 kg P2O5/ha/year can increase seed production.

  14. Alfalfa (Medicago sativa L.) seed yield in relation to phosphorus fertilization and honeybee pollination.

    Science.gov (United States)

    Al-Kahtani, Saad Naser; Taha, El-Kazafy Abdou; Al-Abdulsalam, Mohammed

    2017-07-01

    This investigation was conducted at the Agricultural and Veterinary Training and Research Station, King Faisal University, Al-Ahsa, Saudi Arabia, during the alfalfa growing season in 2014. The study aimed to evaluate the impact of phosphorus fertilization and honeybee pollination on alfalfa seed production. The experiment was divided into 9 treatments of open pollination, honeybee pollination, and non-pollination with three different levels (0, 300 or 600 kg P2O5/ha/year) of triple super phosphate. All vegetative growth attributes of Hassawi alfalfa were significantly higher in the non-insect pollination plots, while the yield and yield component traits were significantly higher with either open pollination or honeybee pollination in parallel with the increasing level of phosphorus fertilizer up to 600 kg P2O5/ha/year in light salt-affected loamy sand soils. There was no seed yield in Hassawi alfalfa without insect pollination. Therefore, placing honeybee colonies near the fields of Hassawi alfalfa and adding 600 kg P2O5/ha/year can increase seed production.

  15. Addition of alfalfa (Medicago sativa L.) to lamb diets enhances production and profits in northern China

    Science.gov (United States)

    The agro-pastoral transitional zone of northern China is an ecologically fragile area with a short growing season and extensive livestock production, where feed shortage is a key constraint to ruminant production, especially in winter. This study was conducted to evaluate the effects of various rat...

  16. Isolation and functional characterization of a Medicago sativa L. gene, MsLEA3-1.

    Science.gov (United States)

    Bai, Yongqin; Yang, Qingchuan; Kang, Junmei; Sun, Yan; Gruber, Margaret; Chao, Yuehui

    2012-03-01

    A full-length cDNA of 1,728 nt, called MsLEA3-1, was cloned from alfalfa by rapid amplification of cDNA ends from an expressed sequence tag homologous to soybean pGmPM10 (accession No. AAA91965.1). MsLEA3-1, encodes a deduced protein of 436 amino acids, a calculated molecular weight of 47.0 kDa, a theoretical isoelectric point of 5.18, and closest homology with late embryogenesis abundant proteins in soybean. Sequence homology suggested a signal peptide in the N terminus, and subcellular localization with GFP revealed that MsLEA3-1 was localized preferentially to the nucleolus. The transcript titre of MsLEA3-1 was strongly enriched in leaves compared with roots and stems of mature alfalfa plants. Gene expression of MsLEA3-1 was strongly induced when seedlings were treated with NaCl and ABA. Expression of the MsLEA3-1 transgenic was detected in transgenic tobacco. Malondialdehyde content and, electrical conductivity content were reduced and electrical conductivity and proline content were increased in transgenic tobacco compared with non-transgenic tobacco under salt stress. The results showed that accumulation of the MsLEA3-1 protein in the vegetative tissues of transgenic plants enhanced their tolerance to salt stress. These results demonstrate a role for the MsLEA3-1 protein in stress protection and suggest the potential of the MsLEA3-1 gene for genetic engineering of salt tolerance.

  17. Extraction, composition, and functional properties of dried alfalfa (Medicago sativa L.) leaf protein.

    Science.gov (United States)

    Hojilla-Evangelista, Mila P; Selling, Gordon W; Hatfield, Ronald; Digman, Matthew

    2017-02-01

    Alfalfa is considered a potential feedstock for biofuels; co-products with value-added uses would enhance process viability. This work evaluated dried alfalfa leaves for protein production and describes the functional properties of the protein. Dried alfalfa leaves contained 260 g kg-1 dry basis (DB) crude protein, with albumins being the major fraction (260 g kg-1 of total protein). Alkali solubilization for 2 h at 50 °C, acid precipitation, dialysis, and freeze-drying produced a protein concentrate (600 g kg-1 DB crude protein). Alfalfa leaf protein concentrate showed moderate solubility (maximum 500 g kg-1 soluble protein from pH 5.5 to 10), excellent emulsifying properties (activity 158-219 m2  g-1 protein, stability 17-49 min) and minimal loss of solubility during heating at pH ≥ 7.0. It is technically feasible to extract protein with desirable emulsifying and heat stability properties from dried alfalfa leaves; however, the dried form may not be a practical starting material for protein production, given the difficulty of achieving high yields and high-purity protein product. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  18. Expected Genetic Gain for several Quantitative Traits in Alfalfa (Medicago sativa L.

    Directory of Open Access Journals (Sweden)

    Hassan MONIRIFAR

    2011-03-01

    Full Text Available In order to determine genetic gain for some quantitative traits in alfalfa ecotypes an experiment was conducted during cropping seasons of 2001-2007 at East Azarbaijan Agricultural and Natural Resources Research Center, Tabriz, Iran. Twenty nine native ecotypes collected from northwest of Iran and an improved variety were used in a polycross nursery. A randomized complete block design was used with 12 replications to ensure the random mating in the polycross nursery. The 30 half-sib families resulted from polycross nursery were planted individually in pots and 30 day old seedlings transplanted in the field and various traits were measured for three cropping seasons in a polycross test. The results of data analysis showed large variations among ecotypes for the traits studied. This indicates that successful selection for desired traits among their progenies is possible. Based on general combining ability, especially for fresh and dry yield, several ecotypes, including �Satellou�, �Gara-yonje�, �Almard�, �Legan�, �Baftan�, �Khaje�, �Sivan�, �Ilan jouj�, �Dizaj Safar Ali�, �Khosrovang� and �Garababa� were selected as promising parents for developing synthetic variety. The narrow�sense heritability values for fresh yield, dry matter, plant height, fresh leaf to stem and dry leaf to stem ratios were about 60%, 59%, 50%, 11% and 19%, respectively. Using selection intensity of 30%, an increase in fresh yield and dry matter yields were estimated to be 3.2 and 1.58 t/ha, respectively.

  19. Role of Silicon Counteracting Cadmium Toxicity in Alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Kabir, Ahmad H; Hossain, Mohammad M; Khatun, Most A; Mandal, Abul; Haider, Syed A

    2016-01-01

    Cadmium (Cd) is one of the most phytotoxic elements causing an agricultural problem and human health hazards. This work investigates whether and how silicon (Si) ameliorates Cd toxicity in Alfalfa. The addition of Si in Cd-stressed plants caused significant improvement in morpho-physiological features as well as total protein and membrane stability, indicating that Si does have critical roles in Cd detoxification in Alfalfa. Furthermore, Si supplementation in Cd-stressed plants showed a significant decrease in Cd and Fe concentrations in both roots and shoots compared with Cd-stressed plants, revealing that Si-mediated tolerance to Cd stress is associated with Cd inhibition in Alfalfa. Results also showed no significant changes in the expression of two metal chelators [MsPCS1 (phytochelatin synthase) and MsMT2 (metallothionein)] and PC (phytochelatin) accumulation, indicating that there may be no metal sequestration or change in metal sequestration following Si application under Cd stress in Alfalfa. We further performed a targeted study on the effect of Si on Fe uptake mechanisms. We observed the consistent reduction in Fe reductase activity, expression of Fe-related genes [MsIRT1 (Fe transporter), MsNramp1 (metal transporter) and OsFRO1 (ferric chelate reductase] and Fe chelators (citrate and malate) by Si application to Cd stress in roots of Alfalfa. These results support that limiting Fe uptake through the down-regulation of Fe acquisition mechanisms confers Si-mediated alleviation of Cd toxicity in Alfalfa. Finally, an increase of catalase, ascorbate peroxidase, and superoxide dismutase activities along with elevated methionine and proline subjected to Si application might play roles, at least in part, to reduce H2O2 and to provide antioxidant defense against Cd stress in Alfalfa. The study shows evidence of the effect of Si on alleviating Cd toxicity in Alfalfa and can be further extended for phytoremediation of Cd toxicity in plants.

  20. Integrative analysis of transgenic alfalfa (Medicago sativa L. suggests new metabolic control mechanisms for monolignol biosynthesis.

    Directory of Open Access Journals (Sweden)

    Yun Lee

    2011-05-01

    Full Text Available The entanglement of lignin polymers with cellulose and hemicellulose in plant cell walls is a major biological barrier to the economically viable production of biofuels from woody biomass. Recent efforts of reducing this recalcitrance with transgenic techniques have been showing promise for ameliorating or even obviating the need for costly pretreatments that are otherwise required to remove lignin from cellulose and hemicelluloses. At the same time, genetic manipulations of lignin biosynthetic enzymes have sometimes yielded unforeseen consequences on lignin composition, thus raising the question of whether the current understanding of the pathway is indeed correct. To address this question systemically, we developed and applied a novel modeling approach that, instead of analyzing the pathway within a single target context, permits a comprehensive, simultaneous investigation of different datasets in wild type and transgenic plants. Specifically, the proposed approach combines static flux-based analysis with a Monte Carlo simulation in which very many randomly chosen sets of parameter values are evaluated against kinetic models of lignin biosynthesis in different stem internodes of wild type and lignin-modified alfalfa plants. In addition to four new postulates that address the reversibility of some key reactions, the modeling effort led to two novel postulates regarding the control of the lignin biosynthetic pathway. The first posits functionally independent pathways toward the synthesis of different lignin monomers, while the second postulate proposes a novel feedforward regulatory mechanism. Subsequent laboratory experiments have identified the signaling molecule salicylic acid as a potential mediator of the postulated control mechanism. Overall, the results demonstrate that mathematical modeling can be a valuable complement to conventional transgenic approaches and that it can provide biological insights that are otherwise difficult to obtain.

  1. Isolation, taxonomic analysis, and phenotypic characterization of bacterial endophytes present in alfalfa (Medicago sativa) seeds.

    Science.gov (United States)

    López, José Luis; Alvarez, Florencia; Príncipe, Analía; Salas, María Eugenia; Lozano, Mauricio Javier; Draghi, Walter Omar; Jofré, Edgardo; Lagares, Antonio

    2017-12-29

    A growing body of evidence has reinforced the central role of microbiomes in the life of sound multicellular eukaryotes, thus more properly described as true holobionts. Though soil was considered a main source of plant microbiomes, seeds have been shown to be endophytically colonized by microorganisms thus representing natural carriers of a selected microbial inoculum to the young seedlings. In this work we have investigated the type of culturable endophytic bacteria that are carried within surface-sterilized alfalfa seeds. MALDI-TOF analysis revealed the presence of bacteria that belonged to 40 separate genera, distributed within four taxa (Proteobacteria, Actinobacteria, Firmicutes, and Bacteroidetes). Nonsymbiotic members of the Rhizobiaceae family were also found. The evaluation of nine different in-vitro biochemical activities demonstrated isolates with complex combinations of traits that, upon a Principal-Component-Analysis, could be classified into four phenotypic groups. That isolates from nearly half of the genera identified had been able to colonize alfalfa plants grown under axenic conditions was remarkable. Further analyses should be addressed to investigating the colonization mechanisms of the alfalfa seeds, the evolutionary significance of the alfalfa-seed endophytes, and also how after germination the seed microbiome competes with spermospheric and rhizospheric soil bacteria to colonize newly emerging seedlings. Copyright © 2017. Published by Elsevier B.V.

  2. Bacterial population dynamics during the ensiling of Medicago sativa (alfalfa) and subsequent exposure to air.

    Science.gov (United States)

    McGarvey, J A; Franco, R B; Palumbo, J D; Hnasko, R; Stanker, L; Mitloehner, F M

    2013-06-01

    To describe, at high resolution, the bacterial population dynamics and chemical transformations during the ensiling of alfalfa and subsequent exposure to air. Samples of alfalfa, ensiled alfalfa and silage exposed to air were collected and their bacterial population structures compared using 16S rRNA gene libraries containing approximately 1900 sequences each. Cultural and chemical analyses were also performed to complement the 16S gene sequence data. Sequence analysis revealed significant differences (P alfalfa-derived library contained mostly sequences associated with the Gammaproteobacteria (including the genera: Enterobacter, Erwinia and Pantoea); the ensiled material contained mostly sequences associated with the lactic acid bacteria (LAB) (including the genera: Lactobacillus, Pediococcus and Lactococcus). Exposure to air resulted in even greater percentages of LAB, especially among the genus Lactobacillus, and a significant drop in bacterial diversity. In-depth 16S rRNA gene sequence analysis revealed significant bacterial population structure changes during ensiling and again during exposure to air. This in-depth description of the bacterial population dynamics that occurred during ensiling and simulated feed out expands our knowledge of these processes. © 2013 The Society for Applied Microbiology No claim to US Government works.

  3. Lucerne transient streak virus; a Recently Detected Virus Infecting Alfafa (Medicago sativa in Central Saudi Arabia

    Directory of Open Access Journals (Sweden)

    Ahmed Raza

    2017-02-01

    Full Text Available A survey was conducted to determine the status of Lucerne transient streak virus (LTSV in three high-yielding alfalfa regions in central Saudi Arabia (Riyadh, Qassim, and Hail during 2014. Three hundred and eight symptomatic alfalfa, and seven Sonchus oleraceus samples were collected. DAS-ELISA indicated that 59 of these samples were positive to LTSV. Two isolates of LTSV from each region were selected for molecular studies. RT-PCR confirmed the presence of LTSV in the selected samples using a specific primer pair. Percentage identity and homology tree comparisons revealed that all Saudi isolates were more closely related to each other but also closely related to the Canadian isolate-JQ782213 (97.1–97.6% and the New Zealand isolate-U31286 (95.8–97.1%. Comparing Saudi isolates of LTSV with ten other sobemoviruses based on the coat protein gene sequences confirmed the distant relationship between them. Eleven out of fourteen plant species used in host range study were positive to LTSV. This is the first time to document that Trifolium alexandrinum, Nicotiana occidentalis, Chenopodium glaucum, and Lathyrus sativus are new host plant species for LTSV and that N. occidentalis being a good propagative host for it.

  4. Alleviation of cadmium toxicity in Medicago sativa by hydrogen-rich water

    Energy Technology Data Exchange (ETDEWEB)

    Cui, Weiti; Gao, Cunyi; Fang, Peng [College of Life Sciences, Nanjing Agricultural University, Nanjing 210095 (China); Lin, Guoqing [Laboratory Center of Life Sciences, Co. Laboratory of Nanjing Agricultural University and Carl Zeiss Far East, Nanjing Agricultural University, Nanjing 210095 (China); Shen, Wenbiao, E-mail: wbshenh@njau.edu.cn [College of Life Sciences, Nanjing Agricultural University, Nanjing 210095 (China)

    2013-09-15

    Highlights: • HRW can alleviate Cd-induced alfalfa seedling growth inhibition and DNA laddering. • HRW alleviates Cd-induced oxidative stress by activating antioxidant enzymes. • Cd uptake in alfalfa seedling roots was decreased by HRW. • HRW can re-establish glutathione homeostasis under Cd stress. -- Abstract: Hydrogen gas (H{sub 2}) induces plant tolerance to several abiotic stresses, including salinity and paraquat exposure. However, the role of H{sub 2} in cadmium (Cd)-induced stress amelioration is largely unknown. Here, pretreatment with hydrogen-rich water (HRW) was used to characterize physiological roles and molecular mechanisms of H{sub 2} in the alleviation of Cd toxicity in alfalfa plants. Our results showed that the addition of HRW at 10% saturation significantly decreased contents of thiobarbituric acid reactive substances (TBARS) caused by Cd, and inhibited the appearance of Cd toxicity symptoms, including the improvement of root elongation and seedling growth. These responses were related to a significant increase in the total or isozymatic activities of representative antioxidant enzymes, or their corresponding transcripts. In vivo imaging of reactive oxygen species (ROS), and the detection of lipid peroxidation and the loss of plasma membrane integrity provided further evidence for the ability of HRW to improve Cd tolerance significantly, which was consistent with a significant enhancement of the ratio of reduced/oxidized (homo)glutathione ((h)GSH). Additionally, plants pretreated with HRW accumulated less amounts of Cd. Together, this study suggested that the usage of HRW could be an effective approach for Cd detoxification and could be explored in agricultural production systems.

  5. Uso del agua en la alfalfa (Medicago sativa con riego por goteo subsuperficial

    Directory of Open Access Journals (Sweden)

    José Alfredo Montemayor Trejo

    2010-01-01

    Full Text Available Con el objetivo de analizar la eficiencia en el uso del agua con el sistema de riego por goteo subsuperficial y por gravedad, durante el año 2004 se realizó la investigación en la Comarca Lagunera de los estados de Coahuila y Durango, México. Se estableció un diseño de bloques al azar con cuatro repeticiones y tres tratamientos de separación de cintas de riego de 0.8, 0.9 y 1.0 m, y como testigo se evaluó el método de riego por gravedad. Se utilizó el método de regresión lineal simple entre las variables días después de siembra y lámina de agua aplicada, para obtener modelos de aplicación de agua con riego por goteo subsuperficial (RGS y riego por gravedad. La variedad de alfalfa establecida fue la WL 712 con una densidad de siembra de 40 kg ha-1, el gasto de la cinta fue de 3 L/h por metro lineal con orificios espaciados a 30 cm y espesor de 10,000 micrones. La lámina de riego aplicada fue de 1.39 m en el sistema por gravedad y 1.16 m con riego por goteo subsuperficial. Los consumos de agua promedio diario de los modelos presentaron diferencias t(a=0.05 y fueron 0.417 y 0.299 cm dia-1 en gravedad y RGS. El rendimiento de materia seca en siete cortes y la eficiencia en el uso del agua fueron de 24.08 t ha-1, 2.04 kg m-3 en subsuperficial y 18.46 t ha-1, 1.33 kg m-3 en gravedad (P<0.05.

  6. Replacement of alfalfa hay (Medicago sativa) with maralfalfahay (Pennisetum sp.) in diets of lactating dairy goats

    DEFF Research Database (Denmark)

    Criscioni, P.; Marti, J.V.; Pérez-Baena, I.

    2016-01-01

    .5 vs. 15.6 mol/100 mol, respectively)and milk C15:0 (0.81 vs. 0.62 g/100 g), C17:0 (0.33 vs. 0.24 g/100 g) fatty acids, indicatingpredominance of amylolytic fermentation. Milk uric acid, a potential biomarker of rumennitrogen flow and feed efficiency, was higher in A than M (133 vs. 102 mol/L). Higher......-circuit indirect calorimetry system using a head box. Daily dry matter intakewas higher on A than on M diets (1.8 vs. 1.6, for A and M respectively), but no differences wasobserved in metabolizable energy intake (1089 kJ/kg of BW0.75, on average). Greater valuesin M than A were found in rumen propionic acid (17...

  7. Physiological and molecular characterization of lucerne (Medicago sativa L.) germplasm with improved seedling freezing tolerance

    Science.gov (United States)

    We conducted greenhouse experiments to compare 14 alfalfa germplasms for their survival following freezing. Among these germplasms are collections adapted to the Grand River National Grasslands in South Dakota. Our hypothesis was that these collections developed tolerance to survive the frigid gro...

  8. Replacement of alfalfa hay (Medicago sativa) with maralfalfahay (Pennisetum sp.) in diets of lactating dairy goats

    DEFF Research Database (Denmark)

    Criscioni, P.; Marti, J.V.; Pérez-Baena, I.

    2016-01-01

    tThe objective of this experiment was to study the effects of substituting alfalfa (Medicagosativa) with maralfalfa (Pennisetum sp.) on energy, nitrogen and carbon balance, methaneemission, and milk performance in dairy goats. Ten Murciano-Granadina dairy goats in latelactation (45.7 ± 2.96 kg...... of 40/60. Diets were isoen-ergetic and isoproteic. The goats were allocated to individual metabolism cages. After 17d of adaptation, feed intake, total faecal and urine output and milk yield were recordeddaily over a 5 d period. Then, gas exchange measurements were recorded individually by amobile open...... of body weight [BW]) were selected in a 2-treatment and crossoverdesign experiment where each goat received both treatments in 2 periods. One group offive goats was fed a mixed ration with alfalfa as forage (A diet) and the other diet replacedalfalfa with maralfalfa (M diet) in a forage concentrate ratio...

  9. Phosphate Rock Application on Alfalfa (Medicago sativa L. Production and Macronutrients in Latosol Soil

    Directory of Open Access Journals (Sweden)

    Y. Liani

    2011-12-01

    Full Text Available The objective of this study was to evaluate the effectiveness of phosphate rocks (PRs fertilizer compared to chemical P fertilizer for the best crop production and macronutrients of alfalfa. A completely randomized design under 3x3 factorial patterns was used in this research. The first factor was different sources of P fertilizer: Guizhou Phosphate Rock (GPR, Jingxiang Phosphate Rock (JPR, and Single Super Phosphate (SSP. The second factor was level of P fertilizer: 75, 100, and 125 mg P2O5/kg soil. A control treatment (without addition of P fertilizer, CK was added as a comparison with the treatments. The results showed that JPR was the best for alfalfa production, whereas GPR and SSP were better for nutrient content in the alfalfa tissue than JPR. On the whole, phosphate rocks had similar effect on alfalfa growth compared to SSP at the experimental conditions.

  10. Effect of Humic Acid on some Agronomic Characters of some Varieties of Alfalfa (Medicago sativa L.

    Directory of Open Access Journals (Sweden)

    Ali Tadayyon

    2016-10-01

    Full Text Available To evaluate the effect of humic acid on some cultural characters of some alfalfa varieties, a field experiment was conducted as factorial split plot based on RCBD design with three replications in 2014. In this experiment, 5 varieties of alfalfa (Yazdi, Hamedani, Baghdadi, Bami and Ranger and three levels of humic acid (5 and 10 lha-1 and without humic acid, as control were considered as the first factor and harvest time was as the second factor. Result indicated that except plant height, other characteristics such as dry weight yield, dry weight per plant, root and shoot dry weight per plant, number of leaves and stem diameter were affected by triple interaction effect of harvest time×variety×humic acid. Plant height was affected by double interaction effect of variety×humic acid and also harvest time×variety. Yazdi and Bami had the highest and Ranger had the least values of traits under study in two cuttings, respectively. Also in the between of humic acid treatments, 10 lha-1 treatment produced highest traits measured as compared to control and 5 lha-1 treatment. Bami, Ranger and Yazdi varieties with 10 lha-1of humic acid in two cuttings produced the highest dry weight yield (3.64, 3.55 and 3.26 t.ha-1 respectively in this experiment.

  11. Rhizobium favelukesii sp. nov., isolated from the root nodules of alfalfa (Medicago sativa L).

    Science.gov (United States)

    Torres Tejerizo, Gonzalo; Rogel, Marco Antonio; Ormeño-Orrillo, Ernesto; Althabegoiti, María Julia; Nilsson, Juliet Fernanda; Niehaus, Karsten; Schlüter, Andreas; Pühler, Alfred; Del Papa, María Florencia; Lagares, Antonio; Martínez-Romero, Esperanza; Pistorio, Mariano

    2016-11-01

    Strains LPU83T and Or191 of the genus Rhizobium were isolated from the root nodules of alfalfa, grown in acid soils from Argentina and the USA. These two strains, which shared the same plasmid pattern, lipopolysaccharide profile, insertion-sequence fingerprint, 16S rRNA gene sequence and PCR-fingerprinting pattern, were different from reference strains representing species of the genus Rhizobium with validly published names. On the basis of previously reported data and from new DNA-DNA hybridization results, phenotypic characterization and phylogenetic analyses, strains LPU83T and Or191 can be considered to be representatives of a novel species of the genus Rhizobium, for which the name Rhizobium favelukesii sp. nov. is proposed. The type strain of this species is LPU83T (=CECT 9014T=LMG 29160T), for which an improved draft-genome sequence is available.

  12. Evaluación cuantitativa de diez variedades de alfalfa (Medicago sativa L.).

    OpenAIRE

    Álvarez Vázquez, Perpetuo

    2013-01-01

    En México, la alfalfa es la leguminosa más utilizada en la alimentación del ganado lechero, debido a su alto rendimiento y valor nutritivo. El objetivo del presente estudio fue evaluar las variables que determinan el crecimiento y rendimiento de diez variedades de alfalfa a intervalos de corte definidos estacionalmente. El estudio se realizó de septiembre de 2011 a septiembre de 2012 en el Colegio de Postgraduados, Campus Montecillo, Texcoco, México. Las variedades de alfalfa estudiadas fuero...

  13. Alleviation of cadmium toxicity in Medicago sativa by hydrogen-rich water.

    Science.gov (United States)

    Cui, Weiti; Gao, Cunyi; Fang, Peng; Lin, Guoqing; Shen, Wenbiao

    2013-09-15

    Hydrogen gas (H₂) induces plant tolerance to several abiotic stresses, including salinity and paraquat exposure. However, the role of H₂ in cadmium (Cd)-induced stress amelioration is largely unknown. Here, pretreatment with hydrogen-rich water (HRW) was used to characterize physiological roles and molecular mechanisms of H₂ in the alleviation of Cd toxicity in alfalfa plants. Our results showed that the addition of HRW at 10% saturation significantly decreased contents of thiobarbituric acid reactive substances (TBARS) caused by Cd, and inhibited the appearance of Cd toxicity symptoms, including the improvement of root elongation and seedling growth. These responses were related to a significant increase in the total or isozymatic activities of representative antioxidant enzymes, or their corresponding transcripts. In vivo imaging of reactive oxygen species (ROS), and the detection of lipid peroxidation and the loss of plasma membrane integrity provided further evidence for the ability of HRW to improve Cd tolerance significantly, which was consistent with a significant enhancement of the ratio of reduced/oxidized (homo)glutathione ((h)GSH). Additionally, plants pretreated with HRW accumulated less amounts of Cd. Together, this study suggested that the usage of HRW could be an effective approach for Cd detoxification and could be explored in agricultural production systems. Copyright © 2013 Elsevier B.V. All rights reserved.

  14. Evaluation of alfalfa (Medicago sativa L.) populations' response to salinity stress

    Science.gov (United States)

    Alfalfa is a moderately salt tolerant crop with high economic return, therefore more suitable for production with lower quality water than most high value crops. This study was conducted to examine the effects of the irrigation water salt type (ST=Cl- or SO4 2-) and five salinity levels (ECiw= 0.85,...

  15. Low Level Gamma Spectroscopy Measurements of Radium and Cesium in Lucerne (Medicago Sativa)

    Science.gov (United States)

    Fokapić, S.; Bikit, I.; Mrđa, D.; Vesković, M.; Slivka, J.; Mihaljev, Ž.; Ćupić, Ž.

    2007-04-01

    Nineteen years after Chernobyl nuclear accident, activity concentration of 137Cs still could be detected in food and soil samples in Central and Eastern Europe. In this paper radiation levels of radium and cesium in Lucerne will be presented. It is a perennial plant with a deep root system and it is widely grown throughout the world as forage for cattle. The samples of Lucerne were taken from twelve different locations in Vojvodina in the summer period July-September 2004. The samples were specially dried on the air and after that ground, powdered and mineralized by method of dry burning on the temperature of 450°C. Gamma spectrometry measurements of the ash were performed by means of actively shielded germanium detector with maximal background reduction. For cesium 137Cs 10 mBq/kg order of magnitude detection limits were achieved.

  16. A CLIPS expert system for maximizing alfalfa (Medicago Sativa L.) production

    Science.gov (United States)

    Engel, B. A.; Jones, D. D.; Rhykerd, R. L.; Rhykerd, L. M.; Rhykerd, C. L., Jr.; Rhykerd, C. L.

    1990-01-01

    An alfalfa management expert system originally developed by Purdue University agricultural scientists on the PC Plus expert system shell from Texas Instrument has been updated and successfully converted to CLIPS (C Language Integrated Production System). This reduces the cost and legal restrictions associated with making the expert system available to agribusiness industries, extension personnel and farm managers and operators. The expert system includes recommendations concerning soil drainage, liming, P and K fertilization, weed control, variety selection and seeding rate including pure live seeds.

  17. Extraction, composition, and functional properties of dried alfalfa (Medicago sativa L.) leaf protein

    Science.gov (United States)

    Alfalfa, traditionally used for animal feed, has attracted attention as a potential feedstock for biofuels and the viability of the process would be enhanced by co-products with value-added uses. This study describes extraction of protein from dried alfalfa leaves and the functional properties of th...

  18. Root characters of Lucerne (Medicago sativa L.) under rain-fed and irrigated conditions

    Science.gov (United States)

    Raza, Amir; Moghaddam, Ali; Loiskandl, Willibald; Friedel, Jürgen K.; Himmelbauer, Margaritta; Bodner, Gernot

    2010-05-01

    In organic farming, only limited use of selected fertilizers is allowed and plants have to meet their nutritional requirements through mobilization of nutrients provided by organic amendments, crop residue input within the rotation, and released from the soil reservoir. The crop varieties used in such systems shall be efficient in nutrient and water uptake. Root length, surface area and depth distribution are important root characters that demonstrate a potential for nutrient and water uptake. Detailed information on these root characters is lacking for Lucerne, one of the most important legume crop widely used in organic farming. A study was designed to compare three lucerne cultivars from different geographical origin viz. Niva, Mohajaren and Sitel for their root characters in two different sets of experiments planted under rain-fed and irrigated conditions in 2007. The irrigated experiment should provide root traits under potential growth conditions while the rain-fed experiment should highlight root characteristics under water limited conditions. The experiments were conducted on fields of the research station Groß Enzersdorf of the University or Natural Resources and Applied Life Sciences(BOKU), Vienna, Austria. Both experiments were laid out in α-lattice design with two replicates. At the end of vegetation period, root samples were taken for every 30 cm soil profile depth till the depth of 90 cm. From each plot, one sample was taken on the row and two samples between the rows using a cylindrical auger for sampling depth of 0 - 30, 30 - 60 and 60 - 90 cm. Root samples were washed out and analyzed with WinRhizo software to determine root length, surface area, root volume and average diameter. Results revealed that cultivars under rain-fed conditions had higher root length density and surface area than under irrigated conditions. The differences in root parameters estimated for each of the Lucerne cultivars are discussed.

  19. Detoxification of Atrazine by Low Molecular Weight Thiols in Alfalfa (Medicago sativa).

    Science.gov (United States)

    Zhang, Jing Jing; Xu, Jiang Yan; Lu, Feng Fan; Jin, She Feng; Yang, Hong

    2017-10-16

    Low molecular weight (LMW) thiols in higher plants are a group of sulfur-rich nonprotein compounds and play primary and multiple roles in cellular redox homeostasis, enzyme activities, and xenobiotics detoxification. This study focused on identifying thiols-related protein genes from the legume alfalfa exposed to the herbicide atrazine (ATZ) residues in environment. Using high-throughput RNA-sequencing, a set of ATZ-responsive thiols-related protein genes highly up-regulated and differentially expressed in alfalfa was identified. Most of the differentially expressed genes (DEGs) were involved in regulation of biotic and abiotic stress responses. By analyzing the genes involved in thiols-mediated redox homeostasis, we found that many of them were thiols-synthetic enzymes such as γ-glutamylcysteine synthase (γECS), homoglutathione synthetase (hGSHS), and glutathione synthetase (GSHS). Using liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS), we further characterized a group of ATZ-thiols conjugates, which are the detoxified forms of ATZ in plants. Cysteine S-conjugate ATZ-HCl+Cys was the most important metabolite detected by MS. Several other ATZ-conjugates were also examined as ATZ-detoxified metabolites. Such results were validated by characterizing their analogs in rice. Our data showed that some conjugates under ATZ stress were detected in both plants, indicating that some detoxified mechanisms and pathways can be shared by the two plant species. Overall, these results indicate that LMW thiols play critical roles in detoxification of ATZ in the plants.

  20. Identification and Selection for Salt Tolerance in Alfalfa (Medicago sativa L. Ecotypes via Physiological Traits

    Directory of Open Access Journals (Sweden)

    Hassan MONIRIFAR

    2009-12-01

    Full Text Available Salt stress is a serious environmental problem throughout the world which may be partially relieved by breeding cultivars that can tolerate salt stress. Plant breeding may provide a relatively cost effective short-term solution to the salinity problem by producing cultivars able to remain productive at low to moderate levels of salinity. Five alfalfa cultivars, �Seyah-Roud�, �Ahar-Hourand�, �Oskou�, �Malekan� and �Sefida-Khan� were assessed for salt tolerance at mature plant stage. A greenhouse screening system was used to evaluate individual alfalfa plants grown in perlit medium, and irrigated with water containing different amounts of NaCl. Three salt levels were achieved by adding 0, 100 and 200 mM NaCl to Hoagland nutrient solution, respectively. Forage yield, sodium and potassium contents and K/Na ratio was determined. Also, leaf samples were analyzed for proline and chlorophyll contents. The ecotypes Seyha-Roud and �Sefida-Khan� had comparatively less sodium contents than �Oskou�, �Ahar-Hourand� and �Malekan� ecotypes, also potassium content increased under saline condition. Forage yield of different alfalfa ecotypes was significantly influenced by the salinity. The ecotypes �Malekan�, Ahar- Hourand and �Oskou� were successful in maintaining forage yield under salinity stress. Sodium contents increased due to salinity in all alfalfa ecotypes however ecotypes �Ahar-Hourand� and �Malekan� maintained the highest leaf Na concentration. They showed higher content of K than other ecotypes but had lower K/Na ratio. It was concluded that, two ecotypes �Malekan� and �Ahar-Hourand� were better.

  1. Methane alleviates copper-induced seed germination inhibition and oxidative stress in Medicago sativa.

    Science.gov (United States)

    Samma, Muhammad Kaleem; Zhou, Heng; Cui, Weiti; Zhu, Kaikai; Zhang, Jing; Shen, Wenbiao

    2017-02-01

    Recent results discovered the protective roles of methane (CH4) against oxidative stress in animals. However, the possible physiological roles of CH4 in plants are still unknown. By using physiological, histochemical and molecular approaches, the beneficial role of CH4 in germinating alfalfa seeds upon copper (Cu) stress was evaluated. Endogenous production of CH4 was significantly increased in Cu-stressed alfalfa seeds, which was mimicked by 0.39 mM CH4. The pretreatment with CH4 significantly alleviated the inhibition of seed germination and seedling growth induced by Cu stress. Cu accumulation was obviously blocked as well. Meanwhile, α/β amylase activities and sugar contents were increased, all of which were consistent with the alleviation of seed germination inhibition triggered by CH4. The Cu-triggered oxidative stress was also mitigated, which was confirmed by the decrease of lipid peroxidation and reduction of Cu-induced loss of plasma membrane integrity in CH4-pretreated alfalfa seedlings. The results of antioxidant enzymes, including ascorbate peroxidase (APX), superoxide dismutase (SOD), catalase (CAT), and guaiacol peroxidase (POD) total or isozymatic activities, and corresponding transcripts (APX1/2, Cu/Zn SOD and Mn-SOD), indicated that CH4 reestablished cellular redox homeostasis. Further, Cu-induced proline accumulation was partly impaired by CH4, which was supported by the alternation of proline metabolism. Together, these results indicated that CH4 performs an advantageous effect on the alleviation of seed germination inhibition caused by Cu stress, and reestablishment of redox homeostasis mainly via increasing antioxidant defence.

  2. Combining -Omics to Unravel the Impact of Copper Nutrition on Alfalfa (Medicago sativa) Stem Metabolism.

    Science.gov (United States)

    Printz, Bruno; Guerriero, Gea; Sergeant, Kjell; Audinot, Jean-Nicolas; Guignard, Cédric; Renaut, Jenny; Lutts, Stanley; Hausman, Jean-Francois

    2016-02-01

    Copper can be found in the environment at concentrations ranging from a shortage up to the threshold of toxicity for plants, with optimal growth conditions situated in between. The plant stem plays a central role in transferring and distributing minerals, water and other solutes throughout the plant. In this study, alfalfa is exposed to different levels of copper availability, from deficiency to slight excess, and the impact on the metabolism of the stem is assessed by a non-targeted proteomics study and by the expression analysis of key genes controlling plant stem development. Under copper deficiency, the plant stem accumulates specific copper chaperones, the expression of genes involved in stem development is decreased and the concentrations of zinc and molybdenum are increased in comparison with the optimum copper level. At the optimal copper level, the expression of cell wall-related genes increases and proteins playing a role in cell wall deposition and in methionine metabolism accumulate, whereas copper excess imposes a reduction in the concentration of iron in the stem and a reduced abundance of ferritins. Secondary ion mass spectrometry (SIMS) analysis suggests a role for the apoplasm as a copper storage site in the case of copper toxicity. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists.

  3. Complete nucleotide sequence and genome organization of a novel allexivirus from alfalfa (Medicago sativa)

    Science.gov (United States)

    A new species of the family Alphaflexiviridae provisionally named Alfalfa virus S (AVS) was diagnosed in alfalfa samples originating from Sudan. A complete nucleotide sequence of the viral genome consisting of 8,349 nucleotides excluding the 3’ poly(A) tail was determined by Illumina NGS technology ...

  4. Induksi Tetraploid Tunas Pucuk Jeruk Siam Simadu (Citrus nobilis Lour Menggunakan Kolkisin secara In Vitro

    Directory of Open Access Journals (Sweden)

    Fitri Yulianti

    2015-06-01

    Full Text Available Seedless fruit is one of the criteria (necessary to improve the quality of Simadu tangerine. The most effective method to obtain seedless triploid cultivars is hybridisation between tetraploid and diploid parents. Simadu tangerine is a diploid plant. Tetraploid Simadu tangerine can be obtained with doubling chromosome using colchicines.The aim of this research was to obtain tetraploid Simadu tangerine shoot which would serve as parent to produced seedless Simadu tangerine. Shoot-tips of Simadu tangerine without leaves were treated with colchicines at four different concentrations (0, 0.1, 0.2, and 0.3% for 3 hours. The results showed that the high concentration of 0.3% reduced survival rate. The colchicine treatments reduced growth of shoot-tip of Simadu Tangerine.The leaves of colchicines treated shoots were thicker than control. Leaves from control (0% colchicine and 0.1% colchicine treated shoots had 8.67 and 18.25 chloroplast per pair of guard cells. Compared to those of control, leaves with 0.1% colchicine had lower stomatal density, and larger stomatal size. It appeared that 0.1% colchicine treatment resulted in tetraploid Simadu Tangerine Shoot.Keywords:chloroplasts, doubling chromosomes, stomatal size, stomatal density

  5. Genotype calling in tetraploid species from bi-allelic marker data using mixture models

    NARCIS (Netherlands)

    Voorrips, R.E.; Gort, G.; Vosman, B.

    2011-01-01

    Background: Automated genotype calling in tetraploid species was until recently not possible, which hampered genetic analysis. Modern genotyping assays often produce two signals, one for each allele of a bi-allelic marker. While ample software is available to obtain genotypes (homozygous for either

  6. In vitro isoflavonoid production and analysis in natural tetraploid Trifolium pratense (red clover calluses

    Directory of Open Access Journals (Sweden)

    Tugba Ercetin

    2012-10-01

    Full Text Available Isoflavones are polyphenolic phytoestrogens, predominantly found in leguminous plants. Trifolium pratense L., Fabaceae (red clover, is rich in isoflavones that possess estrogenic activity due to their similar molecular structure and effectiveness in preventing health conditions such as menopause, osteoporosis, cardiovascular disease, hypertension and hormone-dependent cancers. In this study, presence and amount of various phytoestrogens in the tetraploid plant and in the calluses derived from the plants were investigated. Calluses were generated from explants obtained from natural tetraploid T. pratense seedlings. The best callus formation was obtained from hypocotyl explants cultured in Phillips Collins and Gamborg B5 media containing different plant growth regulators. Flowers of plants and calluses were analysed for formononetin, biochanin A, genistein and daidzein contents using HPLC. In HPLC analysis, high levels of formononetin (0.249 µg/mg were determined in natural tetraploid T. pratense flowers in addition to genistein and biochanin A. In calluses, highest isoflavone content (1.15 µg/mg formononetin was observed in modified Gamborg B5 medium. Biochanin A content of calluses and the plant were found to be nearly the same. But formononetin and genistein contents of the calluses in this medium were found to be respectively 4.62 and 21.39 folds higher than the tetraploid plant.

  7. In vitro isoflavonoid production and analysis in natural tetraploid Trifolium pratense (red clover calluses

    Directory of Open Access Journals (Sweden)

    Tugba Ercetin

    2012-09-01

    Full Text Available Isoflavones are polyphenolic phytoestrogens, predominantly found in leguminous plants. Trifolium pratense L., Fabaceae (red clover, is rich in isoflavones that possess estrogenic activity due to their similar molecular structure and effectiveness in preventing health conditions such as menopause, osteoporosis, cardiovascular disease, hypertension and hormone-dependent cancers. In this study, presence and amount of various phytoestrogens in the tetraploid plant and in the calluses derived from the plants were investigated. Calluses were generated from explants obtained from natural tetraploid T. pratense seedlings. The best callus formation was obtained from hypocotyl explants cultured in Phillips Collins and Gamborg B5 media containing different plant growth regulators. Flowers of plants and calluses were analysed for formononetin, biochanin A, genistein and daidzein contents using HPLC. In HPLC analysis, high levels of formononetin (0.249 µg/mg were determined in natural tetraploid T. pratense flowers in addition to genistein and biochanin A. In calluses, highest isoflavone content (1.15 µg/mg formononetin was observed in modified Gamborg B5 medium. Biochanin A content of calluses and the plant were found to be nearly the same. But formononetin and genistein contents of the calluses in this medium were found to be respectively 4.62 and 21.39 folds higher than the tetraploid plant.

  8. Identification of agronomically important QTL in tetraploid potato cultivars using a marker-trait association analysis

    NARCIS (Netherlands)

    hoop, D' B.B.; Keizer, L.C.P.; Paulo, M.J.; Visser, R.G.F.; Eeuwijk, van F.A.; Eck, van H.J.

    2014-01-01

    Two association mapping panels were analysed for marker–trait associations to identify quantitative trait loci (QTL). The first panel comprised 205 historical and contemporary tetraploid potato cultivars that were phenotyped in field trials at two locations with two replicates (the academic panel).

  9. The simulation of meiosis in diploid and tetraploid organisms using various genetic models

    NARCIS (Netherlands)

    Voorrips, R.E.; Maliepaard, C.A.

    2012-01-01

    Background: While the genetics of diploid inheritance are well studied and software for linkage mapping, haplotyping and QTL analysis are available, for tetraploids the available tools are limited. In order to develop such tools it would be helpful if simulated populations based on a variety of

  10. Discrimination of the closely related A and D genomes of the hexaploid oat Avena sativa L.

    Science.gov (United States)

    Linares, Concha; Ferrer, Esther; Fominaya, Araceli

    1998-01-01

    A satellite DNA sequence, As120a, specific to the A-genome chromosomes in the hexaploid oat, Avena sativa L., was isolated by subcloning a fragment with internal tandem repeats from a plasmid, pAs120, that had been obtained from an Avena strigosa (As genome) genomic library. Southern and in situ hybridization showed that sequences with homology to sequences within pAs120 were dispersed throughout the genome of diploid (A and C genomes), tetraploid (AC genomes), and hexaploid (ACD genomes) Avena species. In contrast, sequences homologous to As120a were found in two A-genome species (A. strigosa and Avena longiglumis) and in the hexaploid A. sativa whereas this sequence was little amplified in the tetraploid Avena murphyi and was absent in the remaining A- and C-genome diploid species. In situ hybridization of pAs120a to hexaploid oat species revealed the distribution of elements of the As120a repeated family over both arms of 14 of 42 chromosomes of this species. By using double in situ hybridization with pAs120a and a C genome-specific probe, three sets of 14 chromosomes were revealed corresponding to the A, C, and D genomes of the hexaploid species. Simultaneous in situ hybridizations with pAs120a and ribosomal probes were used to assign the SAT chromosomes of hexaploid species to their correct genomes. This work reports a sequence able to distinguish between the closely related A and D genomes of hexaploid oats. This sequence offers new opportunities to analyze the relationships of Avena species and to explore the possible evolution of various polyploid oat species. PMID:9770506

  11. Overexpression of Medicago SVP genes causes floral defects and delayed flowering in Arabidopsis but only affects floral development in Medicago.

    Science.gov (United States)

    Jaudal, Mauren; Monash, Jacob; Zhang, Lulu; Wen, Jiangqi; Mysore, Kirankumar S; Macknight, Richard; Putterill, Joanna

    2014-02-01

    The MADS-domain transcription factor SHORT VEGETATIVE PHASE plays a key role as a repressor of the transition to flowering and as a regulator of early floral development in Arabidopsis thaliana (Arabidopsis). However, no flowering-time repressors have been functionally identified in the model legume Medicago truncatula (Medicago). In this study, phylogenetic analysis of two closely-related MtSVP-like sequences, MtSVP1 and MtSVP2, showed that their predicted proteins clustered together within the eudicot SVP clade. To determine if the MtSVP-like genes have a role in flowering, they were functionally characterized in Medicago and Arabidopsis. Transcripts of both MtSVP genes were abundant and broadly expressed in vegetative tissues but were detected at much lower levels in flowers in Medicago. Over-expression of the MtSVP genes in Arabidopsis resulted in delayed flowering and flowers with many abnormal phenotypes such as leafy sepals, changes to floral organ number and longer pedicels than the wild type. By contrast, in transgenic Medicago, over-expression of MtSVP1 resulted in alterations to flower development, but did not alter flowering time, suggesting that MtSVP1 may not function to repress the transition to flowering in Medicago.

  12. Losmapimod Overcomes Gefitinib Resistance in Non-small Cell Lung Cancer by Preventing Tetraploidization.

    Science.gov (United States)

    Yeung, Yiu To; Yin, Shuying; Lu, Bingbing; Fan, Suyu; Yang, Ran; Bai, Ruihua; Zhang, Chengjuan; Bode, Ann M; Liu, Kangdong; Dong, Zigang

    2018-02-01

    The epidermal growth factor receptor (EGFR) is known to play a critical role in non-small cell lung cancer (NSCLC). Constitutively active EGFR mutations, including in-frame deletion in exon 19 and L858R point mutation in exon 21, contribute about 90% of all EGFR-activating mutations in NSCLC. Although oral EGFR-tyrosine kinase inhibitors (TKIs), gefitinib and erlotinib, show dramatic clinical efficacy with significantly prolonged progression-free survival in patients harboring these EGFR-activating mutations, most of these patients will eventually develop acquired resistance. Researchers have recently named genomic instability as one of the hallmarks of cancer. Genomic instability usually involves a transient phase of polyploidization, in particular tetraploidization. Tetraploid cells can undergo asymmetric cell division or chromosome loss, leading to tumor heterogeneity and multidrug resistance. Therefore, identification of signaling pathways involved in tetraploidization is crucial in overcoming drug resistance. In our present study, we found that gefitinib could activate YAP-MKK3/6-p38 MAPK-STAT3 signaling and induce tetraploidization in gefitinib-resistance cells. Using p38 MAPK inhibitors, SB203580 and losmapimod, we could eliminate gefitinib-induced tetraploidization and overcome gefitinib-resistance. In addition, shRNA approach to knockdown p38α MAPK could prevent tetraploidy formation and showed significant inhibition of cancer cell growth. Finally, in an in vivo study, losmapimod could successfully overcome gefitinib resistance using an in-house established patient-derived xenograft (PDX) mouse model. Overall, these findings suggest that losmapimod could be a potential clinical agent to overcome gefitinib resistance in NSCLC. Copyright © 2018 The Author(s). Published by Elsevier B.V. All rights reserved.

  13. Hemp (Cannabis sativa L.).

    Science.gov (United States)

    Feeney, Mistianne; Punja, Zamir K

    2015-01-01

    Hemp (Cannabis sativa L.) suspension culture cells were transformed with Agrobacterium tumefaciens strain EHA101 carrying the binary plasmid pNOV3635. The plasmid contains a phosphomannose isomerase (PMI) selectable marker gene. Cells transformed with PMI are capable of metabolizing the selective agent mannose, whereas cells not expressing the gene are incapable of using the carbon source and will stop growing. Callus masses proliferating on selection medium were screened for PMI expression using a chlorophenol red assay. Genomic DNA was extracted from putatively transformed callus lines, and the presence of the PMI gene was confirmed using PCR and Southern hybridization. Using this method, an average transformation frequency of 31.23% ± 0.14 was obtained for all transformation experiments, with a range of 15.1-55.3%.

  14. Chestnut, European (Castanea sativa).

    Science.gov (United States)

    Corredoira, Elena; Valladares, Silvia; Vieitez, Ana M; Ballester, Antonio

    2015-01-01

    Development of a system for direct transfer of antifungal candidate genes into European chestnut (Castanea sativa) would provide an alternative approach to conventional breeding for production of chestnut trees that are tolerant to ink disease caused by Phytophthora spp. Overexpression of genes encoding PR proteins (such as thaumatin-like proteins), which display antifungal activity, may represent an important advance in control of the disease. We have used a chestnut thaumatin-like protein gene (CsTL1) isolated from European chestnut cotyledons and have achieved overexpression of the gene in chestnut somatic embryogenic lines used as target material. We have also acclimatized the transgenic plants and grown them on in the greenhouse. Here, we describe the various steps of the process, from the induction of somatic embryogenesis to the production of transgenic plants.

  15. A cold responsive galactinol synthase gene from Medicago falcata (MfGolS1) is induced by myo-inositol and confers multiple tolerances to abiotic stresses.

    Science.gov (United States)

    Zhuo, Chunliu; Wang, Ting; Lu, Shaoyun; Zhao, Yaqing; Li, Xiaoguang; Guo, Zhenfei

    2013-09-01

    Galactinol synthase (GolS, EC 2.4.1.123) catalyzes formation of galactinol and the subsequent synthesis of raffinose family oligosaccharides. The relationship of GolS to drought and salt tolerance has been well documented, however, little information is available about the role of GolS gene in cold tolerance. A coding sequence of MfGolS1 cDNA was cloned from Medicago sativa spp falcata (i.e. M. falcata), a species that exhibits greater cold tolerance than alfalfa (M. sativa). MfGolS1 transcript was not detected in untreated vegetative tissues using RNA blot hybridization; however, it was greatly induced in leaves, but not in stem and petiole, after cold treatment. Higher levels of MfGolS1 transcript were induced and maintained in M. falcata than in M. sativa during cold acclimation. Accordingly, more sugars including sucrose, galactinol, raffinose and stachyose were accumulated in M. falcata than in M. sativa. The data indicated that MfGolS1 transcript and its resultant sugar accumulation were associated with the differential cold tolerance between M. falcata and M. sativa. MfGolS1 transcript was weakly induced by dehydration and salt stresses, but not responsive to abscisic acid. MfGolS1 could be induced by myo-inositol, which is proposed to participate in cold-induced MfGolS1 expression. Overexpression of MfGolS1 in tobacco resulted in elevated tolerance to freezing and chilling in transgenic plants as a result of enhanced levels of galactinol, raffinose and stachyose. Tolerance to drought and salt stresses was also increased in the transgenic tobacco plants. It is suggested that MfGolS1 plays an important role in plant tolerance to abiotic stresses. © 2012 Scandinavian Plant Physiology Society.

  16. The Medicago Genome Provides Insight into the Evolution of Rhizobial Symbioses

    Science.gov (United States)

    Young, Nevin D.; Debellé, Frédéric; Oldroyd, Giles E. D.; Geurts, Rene; Cannon, Steven B.; Udvardi, Michael K.; Benedito, Vagner A.; Mayer, Klaus F. X.; Gouzy, Jérôme; Schoof, Heiko; Van de Peer, Yves; Proost, Sebastian; Cook, Douglas R.; Meyers, Blake C.; Spannagl, Manuel; Cheung, Foo; De Mita, Stéphane; Krishnakumar, Vivek; Gundlach, Heidrun; Zhou, Shiguo; Mudge, Joann; Bharti, Arvind K.; Murray, Jeremy D.; Naoumkina, Marina A.; Rosen, Benjamin; Silverstein, Kevin A. T.; Tang, Haibao; Rombauts, Stephane; Zhao, Patrick X.; Zhou, Peng; Barbe, Valérie; Bardou, Philippe; Bechner, Michael; Bellec, Arnaud; Berger, Anne; Bergès, Hélène; Bidwell, Shelby; Bisseling, Ton; Choisne, Nathalie; Couloux, Arnaud; Denny, Roxanne; Deshpande, Shweta; Dai, Xinbin; Doyle, Jeff; Dudez, Anne-Marie; Farmer, Andrew D.; Fouteau, Stéphanie; Franken, Carolien; Gibelin, Chrystel; Gish, John; Goldstein, Steven; González, Alvaro J.; Green, Pamela J.; Hallab, Asis; Hartog, Marijke; Hua, Axin; Humphray, Sean; Jeong, Dong-Hoon; Jing, Yi; Jöcker, Anika; Kenton, Steve M.; Kim, Dong-Jin; Klee, Kathrin; Lai, Hongshing; Lang, Chunting; Lin, Shaoping; Macmil, Simone L; Magdelenat, Ghislaine; Matthews, Lucy; McCorrison, Jamison; Monaghan, Erin L.; Mun, Jeong-Hwan; Najar, Fares Z.; Nicholson, Christine; Noirot, Céline; O’Bleness, Majesta; Paule, Charles R.; Poulain, Julie; Prion, Florent; Qin, Baifang; Qu, Chunmei; Retzel, Ernest F.; Riddle, Claire; Sallet, Erika; Samain, Sylvie; Samson, Nicolas; Sanders, Iryna; Saurat, Olivier; Scarpelli, Claude; Schiex, Thomas; Segurens, Béatrice; Severin, Andrew J.; Sherrier, D. Janine; Shi, Ruihua; Sims, Sarah; Singer, Susan R.; Sinharoy, Senjuti; Sterck, Lieven; Viollet, Agnès; Wang, Bing-Bing; Wang, Keqin; Wang, Mingyi; Wang, Xiaohong; Warfsmann, Jens; Weissenbach, Jean; White, Doug D.; White, Jim D.; Wiley, Graham B.; Wincker, Patrick; Xing, Yanbo; Yang, Limei; Yao, Ziyun; Ying, Fu; Zhai, Jixian; Zhou, Liping; Zuber, Antoine; Dénarié, Jean; Dixon, Richard A.; May, Gregory D.; Schwartz, David C.; Rogers, Jane; Quétier, Francis; Town, Christopher D.; Roe, Bruce A.

    2011-01-01

    Legumes (Fabaceae or Leguminosae) are unique among cultivated plants for their ability to carry out endosymbiotic nitrogen fixation with rhizobial bacteria, a process that takes place in a specialized structure known as the nodule. Legumes belong to one of the two main groups of eurosids, the Fabidae, which includes most species capable of endosymbiotic nitrogen fixation 1. Legumes comprise several evolutionary lineages derived from a common ancestor 60 million years ago (Mya). Papilionoids are the largest clade, dating nearly to the origin of legumes and containing most cultivated species 2. Medicago truncatula (Mt) is a long-established model for the study of legume biology. Here we describe the draft sequence of the Mt euchromatin based on a recently completed BAC-assembly supplemented with Illumina-shotgun sequence, together capturing ~94% of all Mt genes. A whole-genome duplication (WGD) approximately 58 Mya played a major role in shaping the Mt genome and thereby contributed to the evolution of endosymbiotic nitrogen fixation. Subsequent to the WGD, the Mt genome experienced higher levels of rearrangement than two other sequenced legumes, Glycine max (Gm) and Lotus japonicus (Lj). Mt is a close relative of alfalfa (M. sativa), a widely cultivated crop with limited genomics tools and complex autotetraploid genetics. As such, the Mt genome sequence provides significant opportunities to expand alfalfa’s genomic toolbox. PMID:22089132

  17. The Medicago truncatula gene expression atlas web server

    Directory of Open Access Journals (Sweden)

    Tang Yuhong

    2009-12-01

    Full Text Available Abstract Background Legumes (Leguminosae or Fabaceae play a major role in agriculture. Transcriptomics studies in the model legume species, Medicago truncatula, are instrumental in helping to formulate hypotheses about the role of legume genes. With the rapid growth of publically available Affymetrix GeneChip Medicago Genome Array GeneChip data from a great range of tissues, cell types, growth conditions, and stress treatments, the legume research community desires an effective bioinformatics system to aid efforts to interpret the Medicago genome through functional genomics. We developed the Medicago truncatula Gene Expression Atlas (MtGEA web server for this purpose. Description The Medicago truncatula Gene Expression Atlas (MtGEA web server is a centralized platform for analyzing the Medicago transcriptome. Currently, the web server hosts gene expression data from 156 Affymetrix GeneChip® Medicago genome arrays in 64 different experiments, covering a broad range of developmental and environmental conditions. The server enables flexible, multifaceted analyses of transcript data and provides a range of additional information about genes, including different types of annotation and links to the genome sequence, which help users formulate hypotheses about gene function. Transcript data can be accessed using Affymetrix probe identification number, DNA sequence, gene name, functional description in natural language, GO and KEGG annotation terms, and InterPro domain number. Transcripts can also be discovered through co-expression or differential expression analysis. Flexible tools to select a subset of experiments and to visualize and compare expression profiles of multiple genes have been implemented. Data can be downloaded, in part or full, in a tabular form compatible with common analytical and visualization software. The web server will be updated on a regular basis to incorporate new gene expression data and genome annotation, and is accessible

  18. A comparative study on rooting of in vitro regenerated shoots in haploid, diploid and tetraploid purple coneflower (Echinacea purpurea L.

    Directory of Open Access Journals (Sweden)

    Rong Chen

    2016-01-01

    Full Text Available Haploid, diploid and tetraploid shoots of Echinacea purpurea L. sharing the same genome were cultured in medium and their rooting response to the composition of the culture medium was investigated. It was found that in medium without growth regulators, haploid shoots could initiate roots quite efficiently with the shortest time required for the emergence of roots and with the highest rooting rate; the response of the diploids was similar to that of the haploids and largely different from that of the tetraploids. The tetraploids obviously required longer time for the initiation of roots and had the lowest rooting rate. Supplementing the medium with 0.05 and 0.15 mg/L naphthaleneacetic acid or 0.1 and 0.3 mg/L indole-3-butyric acid (IBA had little positive effect on the rooting of diploid shoots and, in some cases, had even negative effect on the rooting of haploid shoots, but enhanced effectively the rooting of the tetraploid shoots. By supplementing the medium with 0.3 mg/L IBA, the time required for the emergence of roots from the tetraploid shoots was shortened and the rooting rate was increased largely. As a result, healthy tetraploid plantlets with fully developed root system could be efficiently propagated.

  19. Genomic prediction of starch content and chipping quality in tetraploid potato using genotyping-by-sequencing

    DEFF Research Database (Denmark)

    Sverrisdóttir, Elsa; Byrne, Stephen; Nielsen, Ea Høegh Riis

    2017-01-01

    potato necessitates large training populations to efficiently capture the genetic diversity of elite potato germplasm and enable accurate prediction across the entire spectrum of elite potatoes. Nonetheless, our results demonstrate that GS is a promising breeding strategy for tetraploid potato.......Genomic selection uses genome-wide molecular markers to predict performance of individuals and allows selections in the absence of direct phenotyping. It is regarded as a useful tool to accelerate genetic gain in breeding programs, and is becoming increasingly viable for crops as genotyping costs...... continue to fall. In this study, we have generated genomic prediction models for starch content and chipping quality in tetraploid potato to facilitate varietal development. Chipping quality was evaluated as the colour of a potato chip after frying following cold induced sweetening. We used genotyping...

  20. Genomic prediction for dry matter and chipping quality in tetraploid potato

    DEFF Research Database (Denmark)

    Sverrisdóttir, Elsa; Nielsen, Ea Høegh Riis; Johnsen, Heidi Øllegaard

    Potato is one of the most space-efficient food crops and is of vital importance for global food security. The traditional “mate and phenotype” breeding approach is costly and time-consuming; however, genomic selection using genome-wide molecular markers is becoming increasingly applicable to crops...... across-population predictions were low or moderate. Overall, the results suggest that genomic prediction and hence selection of breeding material can be obtained with good accuracies within tetraploid potato. Although the most optimal prediction accuracies were obtained when predicting within the same...... population, the results from combining training populations with genotypes from different populations suggest a promising approach for establishing a broad-application prediction model for the implementation of genomic selection in tetraploid potato breeding programmes....

  1. Disomic segregation of microsatellites in the tetraploid Prunus serotina Ehrh. (Rosaceae)

    OpenAIRE

    Pairon, Marie; Jacquemart, Anne-Laure

    2005-01-01

    Tetraploid black cherry (Prunus serotina) is the only Prunus L. species that has commercial importance as a timber tree in North America and is well known in Europe for its invasive behavior. Inheritance studies have never been performed and it is not known whether the species is allo or autotetraploid. Six microsatellite nuclear markers were used to test the inheritance in progenies of controlled crosses. Inheritance was proven to be disomic at all loci and a typical diploid mendelian inheri...

  2. Assessment of genetic variability of haploids extracted from tetraploid (2n = 4x = 48) Solanum tuberosum.

    Science.gov (United States)

    Ercolano, M R; Carputo, D; Li, J; Monti, L; Barone, A; Frusciante, L

    2004-08-01

    The objectives of this study were to assess the genetic variability of haploids (2n = 2x = 24) extracted from tetraploid Solanum tuberosum through 4x x 2x crosses with Solanum phureja. Molecular and phenotypic analyses were performed to fingerprint the genotypes used and to evaluate their potential use in breeding programs. AFLP analysis revealed the presence of specific bands derived from the tetraploid seed parent S. phureja, as well as ex novo originated bands. On average, 210 bands were visualized per genotype, 149 (70%) of which were common to both parental genotypes. The percentage of S. tuberosum specific bands ranged from 25.1% to 18.6%, with an average of 22%. The fraction of genome coming from S. phureja ranged from 1.9% to 6.5%, with an average value of 4%. The percentage of ex novo bands varied from 1.9% to 9.0%. The presence of S. phureja DNA is very interesting because it indicated that S. phureja pollinator is involved in the mechanism of haploid formation. The characterization for resistance to Erwinia carotovora subsp. carotovora and potato virus X (PVX) provided evidence that haploids may express traits that are lacking in the tetraploids they come from, which can be useful for both genetic studies and breeding purposes. It is noteworthy that genotypes combining resistance to both diseases and good pollen stainability were identified. Other possible breeding implications owing to the presence of S. phureja genome in the haploids analyzed are discussed.

  3. Field evaluation of in vitro-induced tetraploid and diploid Centella asiatica (L.) Urban.

    Science.gov (United States)

    Thong-On, Wachiraporn; Arimatsu, Panida; Pitiporn, Supaporn; Soonthornchareonnon, Noppamas; Prathanturarug, Sompop

    2014-04-01

    Centella asiatica-a medicinal plant that produces high-value active triterpenoids-is in increasing demand by the pharmaceutical and cosmetic industries. The aim of this study was to field-test one induced tetraploid and three diploid C. asiatica lines for the selection of high-quality plants with high phytomass and triterpenoid content and to determine their optimal harvesting time. All tested C. asiatica were micropropagated using an established protocol. One-month-old plantlets were acclimatized for the field experiment. The plants were grown in a randomized complete block design (RCBD) with three replications, ten plantlets per replication, and the experimental bed site was 0.6 × 1.0 m. Growth parameters, phytomass and the amounts of four active triterpenoids were evaluated. All lines exhibited the highest growth, yields and triterpenoids at 4 months after cultivation. The tetraploid line showed significantly better characteristics, i.e., larger leaf area, leaf width, petiole length, and greater yields, than diploid lines. Dry weight per cultivated area (77.53 ± 3.07 g/m(2)) and total triterpenoids (15.38 ± 0.76 % dry weight) were increased significantly in tetraploid plants of C. asiatica. Furthermore, the harvesting time had an effect on the yield and triterpenoid content (P asiatica.

  4. Heterogeneity in sarcoma cell lines reveals enhanced motility of tetraploid versus diploid cells

    Science.gov (United States)

    Jemaà, Mohamed; Abdallah, Samer; Lledo, Gwendaline; Perrot, Gaelle; Lesluyes, Tom; Teyssier, Catherine; Roux, Pierre; van Dijk, Juliette; Chibon, Frederic; Abrieu, Ariane; Morin, Nathalie

    2017-01-01

    Soft tissue sarcomas with complex genomics are very heterogeneous tumors lacking simple prognosis markers or targeted therapies. Overexpression of a subset of mitotic genes from a signature called CINSARC is of bad prognosis, but the significance of this signature remains elusive. Here we precisely measure the cell cycle and mitosis duration of sarcoma cell lines and we found that the mitotic gene products overexpression does not reflect variation in the time spent during mitosis or G2/M. We also found that the CINSARC cell lines, we studied, are composed of a mixture of aneuploid, diploid, and tetraploid cells that are highly motile in vitro. After sorting diploid and tetraploid cells, we showed that the tetraploid cell clones do not possess a proliferative advantage, but are strikingly more motile and invasive than their diploid counterparts. This is correlated with higher levels of mitotic proteins overexpression. Owing that mitotic proteins are almost systematically degraded at the end of mitosis, we propose that it is the abnormal activity of the mitotic proteins during interphase that boosts the sarcoma cells migratory properties by affecting their cytoskeleton. To test this hypothesis, we designed a screen for mitotic or cytoskeleton protein inhibitors affecting the sarcoma cell migration potential independently of cytotoxic activities. We found that inhibition of several mitotic kinases drastically impairs the CINSARC cell invasive and migratory properties. This finding could provide a handle by which to selectively inhibit the most invasive cells. PMID:28035071

  5. Induction and flow cytometry identification of tetraploids from seed-derived explants through colchicine treatments in Catharanthus roseus (L.) G. Don.

    Science.gov (United States)

    Xing, Shi-Hai; Guo, Xin-Bo; Wang, Quan; Pan, Qi-Fang; Tian, Yue-Sheng; Liu, Pin; Zhao, Jing-Ya; Wang, Guo-Feng; Sun, Xiao-Fen; Tang, Ke-Xuan

    2011-01-01

    The tetraploid plants of Catharanthus roseus (L.) G. Don was obtained by colchicine induction from seeds explants, and the ploidy of the plants was identified by flow cytometry. The optimal treatment is 0.2% colchicine solution treated for 24 hours, and the induction rate reaches up to 30%. Comparing with morphological characteristics and growth habits between tetraploids and the control, we found that tetraploids of C. roseus had larger stoma and more branches and leaves. HPLC analysis showed tetraploidization could increase the contents of terpenoid indole alkaloids in C. roseus. Thus, tetraploidization could be used to produce higher alkaloids lines for commercial use. QRT-PCR results showed that the expression of enzymes involved in terpenoid indole alkaloids biosynthesis pathway had increased in the tetraploid plants. To our knowledge, this was the first paper to explore the secondary metabolism in autotetraploid C. roseus induced by colchicine.

  6. Induction and Flow Cytometry Identification of Tetraploids from Seed-Derived Explants through Colchicine Treatments in Catharanthus roseus (L. G. Don

    Directory of Open Access Journals (Sweden)

    Shi-Hai Xing

    2011-01-01

    Full Text Available The tetraploid plants of Catharanthus roseus (L. G. Don was obtained by colchicine induction from seeds explants, and the ploidy of the plants was identified by flow cytometry. The optimal treatment is 0.2% colchicine solution treated for 24 hours, and the induction rate reaches up to 30%. Comparing with morphological characteristics and growth habits between tetraploids and the control, we found that tetraploids of C. roseus had larger stoma and more branches and leaves. HPLC analysis showed tetraploidization could increase the contents of terpenoid indole alkaloids in C. roseus. Thus, tetraploidization could be used to produce higher alkaloids lines for commercial use. QRT-PCR results showed that the expression of enzymes involved in terpenoid indole alkaloids biosynthesis pathway had increased in the tetraploid plants. To our knowledge, this was the first paper to explore the secondary metabolism in autotetraploid C. roseus induced by colchicine.

  7. Induction and Flow Cytometry Identification of Tetraploids from Seed-Derived Explants through Colchicine Treatments in Catharanthus roseus (L.) G. Don

    Science.gov (United States)

    Xing, Shi-Hai; Guo, Xin-Bo; Wang, Quan; Pan, Qi-Fang; Tian, Yue-Sheng; Liu, Pin; Zhao, Jing-Ya; Wang, Guo-Feng; Sun, Xiao-Fen; Tang, Ke-Xuan

    2011-01-01

    The tetraploid plants of Catharanthus roseus (L.) G. Don was obtained by colchicine induction from seeds explants, and the ploidy of the plants was identified by flow cytometry. The optimal treatment is 0.2% colchicine solution treated for 24 hours, and the induction rate reaches up to 30%. Comparing with morphological characteristics and growth habits between tetraploids and the control, we found that tetraploids of C. roseus had larger stoma and more branches and leaves. HPLC analysis showed tetraploidization could increase the contents of terpenoid indole alkaloids in C. roseus. Thus, tetraploidization could be used to produce higher alkaloids lines for commercial use. QRT-PCR results showed that the expression of enzymes involved in terpenoid indole alkaloids biosynthesis pathway had increased in the tetraploid plants. To our knowledge, this was the first paper to explore the secondary metabolism in autotetraploid C. roseus induced by colchicine. PMID:21660143

  8. Induction of Tetraploid Male Sterile Tagetes erecta by Colchicine Treatment and Its Application for Interspecific Hybridization

    Directory of Open Access Journals (Sweden)

    Yanhong He

    2016-09-01

    Full Text Available Tagetes erecta is an annual multifunctional plant which can be cultivated under a broad range of climatic conditions. Polyploidization and interspecific hybridization are applied to facilitate breeding cultivars of T. erecta with improved ornamental qualities. Colchicine treatment to the germinating seeds was proved to be a useful tool for chromosome doubling of the male sterile two-type line ‘M525AB’, with the resulting frequency of polyploid seedlings ranging from 88.89% (following 0.05% w/v colchicine applied for a 3–6 h exposure period to a maximum of 100.00% (following 0.1% for 3–6 h, or 0.2% for 3 h. Morphological observation, stomatal size and density analysis, flow cytometric analysis and chromosome counting were conducted to identify the tetraploid plants. Distinctive morphological changes were observed in a notable proportion of polyploid plants. The colchicine-treated polyploid T. erecta plants showed dwarfed and more robust growth, thicker, larger and greener leaves, larger inflorescences and florets. The mutant plants identified through morphological observation all aligned as polyploid plants, thus morphological observation could be an effective method for the detection of polyploidy. The polyploid plants had significant larger stomata size over the abaxial leaf surface, whereas the density of stomata distribution was remarkably reduced. The survival rate of tetraploid cuttings (i.e. 38% was greatly reduced compared to that of diploid plants. The fertility of tetraploid plants was also decreased, as shown by cross-pollination yields. Interspecific hybridizations between colchicine-induced tetraploid plants of a male sterile T. erecta line and the naturally tetraploid fully fertile Tagetes patula species resulted in hybrid progeny. Most of these hybrids displayed the dwarfed growth stature and compact, larger-flower morphology which is the typical ideotype of herbaceous flowers. Thus, polyploidization may be employed

  9. A temperature induced lipocalin gene from Medicago falcata (MfTIL1) confers tolerance to cold and oxidative stress.

    Science.gov (United States)

    He, Xueying; Sambe, Mame Abdou Nahr; Zhuo, Chunliu; Tu, Qinghua; Guo, Zhenfei

    2015-04-01

    Temperature-induced lipocalins (TIL) are plasmalemma-localized proteins and responsive to environmental stresses. Physiological functions of MfTIL1 from Medicago sativa subsp. falcata (L.) Arcang. (hereafter falcata), a forage legume with cold and drought tolerance, were investigated in this study. MfTIL1 expression was greatly induced by 4-96 h of cold treatment, while transcript levels of the orthologs in Medicago truncatula, a model legume plant with lower cold tolerance than falcata, were reduced or not altered within 48-96 h. MfTIL1 expression was not responsive to dehydration and salinity. Compared to the wild type, transgenic tobacco plants overexpressing MfTIL1 had lower temperature (LT50) that resulted in 50 % lethal and elevated survival rate in response to freezing, elevated F v/F m and decreased ion leakage after treatments with chilling, high light and methyl viologen (MV). H2O2 and O2 (-) were less accumulated in transgenic plants than in the wild type after treatments with chilling, high light and MV, while antioxidant enzyme activities showed no difference between the two types of plants prior to or following treatments. Higher transcript levels of NtDREB3 and NtDREB4 genes were observed in transgenic plants than in the wild type under non-stressed conditions, but higher transcript levels of NtDREB1, NtDREB2, NtDREB4 and NtCOR15a genes under chilling conditions. It is suggested that MfTIL1 plays an important role in plant tolerance to cold and oxidative stress through promoted scavenging of reactive oxygen species and up-regulating expression of multiple cold responsive genes.

  10. Characterization of genetic structure of alfalfa (Medicago sp.) from ...

    African Journals Online (AJOL)

    Jane

    2011-08-08

    Aug 8, 2011 ... Slatkin M (1987). Gene flow and the geographic structure of natural populations. Science, 236: 787-792. Small E Jomphe M (1989). A synopsis of the genus Medicago. (Leguminosae). Can. J. Bot. 67: 3260-3294. Soltis DE, Soltis PS, Tate JA (2003). Advances in the study of polyploidy since plant speciation ...

  11. Phylogenetic signal variation in the genomes of Medicago (Fabaceae).

    Science.gov (United States)

    Yoder, Jeremy B; Briskine, Roman; Mudge, Joann; Farmer, Andrew; Paape, Timothy; Steele, Kelly; Weiblen, George D; Bharti, Arvind K; Zhou, Peng; May, Gregory D; Young, Nevin D; Tiffin, Peter

    2013-05-01

    Genome-scale data offer the opportunity to clarify phylogenetic relationships that are difficult to resolve with few loci, but they can also identify genomic regions with evolutionary history distinct from that of the species history. We collected whole-genome sequence data from 29 taxa in the legume genus Medicago, then aligned these sequences to the Medicago truncatula reference genome to confidently identify 87 596 variable homologous sites. We used this data set to estimate phylogenetic relationships among Medicago species, to investigate the number of sites needed to provide robust phylogenetic estimates and to identify specific genomic regions supporting topologies in conflict with the genome-wide phylogeny. Our full genomic data set resolves relationships within the genus that were previously intractable. Subsampling the data reveals considerable variation in phylogenetic signal and power in smaller subsets of the data. Even when sampling 5000 sites, no random sample of the data supports a topology identical to that of the genome-wide phylogeny. Phylogenetic relationships estimated from 500-site sliding windows revealed genome regions supporting several alternative species relationships among recently diverged taxa, consistent with the expected effects of deep coalescence or introgression in the recent history of Medicago.

  12. Genetic variation patterns of Medicago ruthenica populations from ...

    African Journals Online (AJOL)

    According to GST, gene flow at population level was 1.4427 for allozymes and 1.2040 for SSR markers, which indicated there was definite gene flow between populations. Low differentiations among ... in M. ruthenica populations. Key words: Medicago ruthenica, genetic diversity, allozyme, micro-satellite, North China.

  13. Characterization of Medicago populations under cold acclimation by ...

    African Journals Online (AJOL)

    SAM

    2014-07-02

    Jul 2, 2014 ... Moreover, there were differences between populations of the same species of cold acclimation, which will encourage for a study of association between cold acclimation and molecular polymorphism. Key words: Cold acclimation, root: shoot ratios, molecular polymorphism, annuals populations, Medicago.

  14. Identification of genes induced by salt stress from Medicago ...

    African Journals Online (AJOL)

    In order to identify genes induced during the salt stress response in barrel medic (Medicago truncatula L) seedlings, a cDNA library by salt stress was constructed by suppression subtractive hybridization (SSH). Total RNA from 15-day-old seedlings was used as a 'driver', and total RNA from seedlings induced by salt was ...

  15. Characterization of Medicago populations under cold acclimation by ...

    African Journals Online (AJOL)

    Characterization of Medicago populations under cold acclimation by morphological traits and microsatellite (SSR) markers. ... altitude (lower ability to cold acclimation). Tests differentiation between species by fisher pair indicates that all species were different from each other. Results show the highest level of homozygosity ...

  16. Pharmacology of Marihuana (Cannabis sativa)

    Science.gov (United States)

    Maickel, Roger P.

    1973-01-01

    A detailed discussion of marihuana (Cannabis sativa) providing the modes of use, history, chemistry, and physiologic properties of the drug. Cites research results relating to the pharmacologic effects of marihuana. These effects are categorized into five areas: behavioral, cardiovascular-respiratory, central nervous system, toxicity-toxicology,…

  17. [Effect of gas-turbine green discoloring and drying processing methods on herbal quality of tetraploid Lonicerae Japonicae Flos].

    Science.gov (United States)

    Hu, Xuan; Li, Wei-dong; Li, Ou; Hao, Jiang-bo; Liu, Jia-kun

    2012-09-01

    To study the effect of gas-turbine green discoloring and drying processing method on the quality of various Lonicerae Japonicae Flos herbs. DIKMA DiamonsilTM-C18 column (4.6 mm x 250 mm, 5 microm) was adopted using HPLC Waters 1525 and eluted with acetonitrile and 0.1% phosphate acid as the mobile phase. The flow rate was 1.0 mL x min(-1) , the column temperature was 25 degrees C the detection wavelength was 355 nm. After being processed by the gas-turbine green discoloring and drying method, tetraploid Lonicerae Japonicae Flos showed a green color. The contents of chlorogenic acid and galuteolin were 5.31% and 0.105% , both significantly higher by 18.0% and 32.1% than those of diploid Lonicerae Japonicae Flos processed by the same method. The content of chlorogenic acid in tetraploid Lonicerae Japonicae Flos processed the gas-turbine green discoloring and drying method were also remarkably higher than that of tetraploid and diploid Lonicerae Japonicae Flos processed by traditional processing method of natural drying. The gas-turbine green discoloring and drying processing method is a new-type drying method suitable for tetraploid Lonicerae Japonicae Flos. Under the condition of gas-turbine green discoloring and drying processing, tetraploid Lonicerae Japonicae Flos shows much higher quality than Lonicerae Japonicae Flos, suggesting that it is a good variety worth popularizing and applying.

  18. Genetic structure and origin of a tetraploid toad species Bufo danatensis Pisanetz, 1978 (Amphibia, Bufonidae) from central Asia: Description of biochemical polymorphism and comparison of heterozygosity levels in diploid and tetraploid species

    Energy Technology Data Exchange (ETDEWEB)

    Mezhzherin, S.V.; Pisanets, E.M. [Schmalhausen Institute of Zoology, Kiev (Ukraine)

    1995-01-01

    Comparison of individual variation at 24 biochemical loci in members of the species complex of Palearctic green toads showed that the heterozygosity of the tetraploid species Bufo danatensis (H{sub obs} = 0.45) was significantly higher than that of the diploid species B. viridis, B. sp., and B. raddei (H{sub obs} = 0.009 - 0.103). Such difference can be explained only by a hybrid origin of the tetraploid species. Individual electrophoretic variability of the polyploid toad species is associated with an allelic variation that is manifested in constantly heterozygous spectra as the gene dosage effect. At the population level, this phenomenon found in Pamir toads is caused by irregular meiosis in founders of the population or by directional changes in gene regulation. Genotypic distributions in zones of contact of the diploid and tetraploid taxons demonstrate the possibility of restricted introgressive hybridization.

  19. Aggressiveness and host range of Phoma medicaginis isolated from Medicago species growing in Tunisia

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    Naceur DJEBALI

    2013-05-01

    Full Text Available Aggressiveness of 14 Phoma medicaginis isolates obtained from Medicago truncatula (barrel medic and M. ciliaris (ciliate medic growing in Tunisia was measured after inoculation on leaves and roots of M. truncatula. The ability of one isolate to cause disease on M. sativa (alfalfa, Cicer arietinum (chickpea, Pisum sativum (pea, Lens culinaris (lentil and Phaseolus vulgaris (common bean was also tested. The pathogen caused dark lesions that enlarged and coalesced causing yellowing and premature abscission of leaves, resulting in decreased shoot fresh weight in barrel medic plants. All P. medicaginis isolates infected barrel medic roots causing collar rot, brown root discoloration, yellowing of cotyledons and reduced shoot and root development. The pathogen colonized the cortex and the stele of plants and produced fertile pycnidia on infected roots. Symptoms on leaves allowed for greater discrimination in aggressiveness among isolates in comparison to symptoms on roots. No correlations were observed between the parameters measured on leaves and roots suggesting organ specialization in this pathogen. Phoma medicaginis infected leaves of alfalfa, pea, common bean and chickpea causing necrosis and tissue yellowing at 15 d post inoculation (dpi. Pycnidium production was observed on dead and dying foliar tissues of alfalfa, pea and common bean, but not on chickpea. The pathogen caused symptoms of collar rot and brown root discoloration on alfalfa, chickpea, pea and common bean, but did not cause symptoms on leaves or roots of lentil at 15 dpi. Phoma medicaginis was more pathogenic on barrel medic, the host of origin, in comparison to the other legumes, suggesting that these species are likely to be secondary hosts for this pathogen.

  20. Inducing triploids and tetraploids with high temperatures in Populus sect. Tacamahaca.

    Science.gov (United States)

    Guo, Liqin; Xu, Wenting; Zhang, Yan; Zhang, Jinfeng; Wei, Zunzheng

    2017-02-01

    This study is the first to report that triploids and tetraploids have been successfully produced through embryo sac and zygotic embryo chromosome doubling with high temperatures in P. simonii Carr. and its hybrid. A new synthetic polyploid induced by hybridization with unreduced gametes and heterozygotic embryo chromosome doubling can effectively combine polyploidy and heterosis, which can provide two major breeding advantages. In Populus, successfully creating and cultivating new polyploid varieties have economic and ecological production value. This was the first successful study in which embryo sac and zygotic embryo chromosome doubling was induced using high temperatures to produce triploids and tetraploids in Populus simonii Carr. and its hybrid, P. simonii × P. nigra var. Italica, of Populus sect. Tacamahaca. The relationship between flower bud morphological characteristics (time after pollination) and female meiotic stage (embryo sac and zygotic embryo development) was established to guide the induction treatment period. In the resulting progeny, 37 triploids and 12 tetraploids were obtained and identified using flow cytometry. The optimal temperatures for embryo sac and zygotic embryo chromosome doubling were 38 and 41 °C, respectively. Cytogenetic analysis revealed that 66-72 h after pollination (HAP), a period characterized by a high proportion of one-nucleate and two-nucleate embryo sacs, was the optimal period for embryo sac chromosome doubling. For zygotic embryo chromosome doubling, 168 HAP was the optimal induction period, as there was a high proportion of two-cell and four-cell proembryos. The results indicate that inducing embryo sac and zygotic embryo chromosome doubling is an ideal method for producing polyploids. The methods for inducing polyploids and for evaluating ploidy and offspring with different ploidies and heterozygosity in this study will be useful for genetic research and Populus breeding programmes.

  1. A new tetraploid species of Solanum L. sect. Solanum (Solanaceae) from Tanzania.

    Science.gov (United States)

    Manoko, Mkabwa L K; van Weerden, Gerard M; van Berg, Ronald G; Mariani, Celestina

    2012-01-01

    Solanum umalilaense Manoko sp. nov. (Solanaceae) is described from the Umalila area, in the southern highlands of Tanzania. Its novelty is supported with both morphological and AFLP data. Phenetic and phylogenetic analyses place Solanum umalilaense as a unique and well-supported taxon among tetraploid species of Solanum sect. Solanum from Africa. It can be distinguished from other African species by its extremely developed branching, each branch producing many multi-flowered inflorescences, flowers with short calyx lobes and its persistent, small, light yellowish brown fruits.

  2. Volatile compound in cut and un-cut flowers of tetraploid Freesia hybrida.

    Science.gov (United States)

    Ao, Man; Liu, Baofeng; Wang, Li

    2013-01-01

    The flower volatile compounds (FVCs) of two tetraploid Freesia hybrida (pink-yellow and yellow) cultivars and their cut flowers were analysed by headspace solid-phase microextraction combined with gas chromatography-mass spectrometry. Twelve FVCs were identified in the pink-yellow cultivar, with linalool as the major compound; 30 FVCs were identified in the yellow cultivar, with linalool and terpineol as the two major compounds. The FVCs (>1%) of the two cut flower cultivars were very similar to that of the un-cut flowers, and no significant difference was observed.

  3. 2n megagametophyte formed via SDR contributes to tetraploidization in polyembryonic 'Nadorcott' tangor crossed by citrus allotetraploids.

    Science.gov (United States)

    Xie, Kai-Dong; Wang, Xiao-Pei; Biswas, Manosh Kumar; Liang, Wu-Jun; Xu, Qiang; Grosser, Jude W; Guo, Wen-Wu

    2014-10-01

    2 n megagametophyte formation plays an important role in polyploidization in polyembryonic citrus and is valuable for plant improvement. Tetraploid plants are frequently observed in the seedlings of diploid polyembryonic citrus genotypes. However, the mechanisms underlying the formation of tetraploids are still indistinct when apomictic citrus genotypes are used as female parent to cross with tetraploids. Herein, 54 tetraploid progenies, which were unexpectedly obtained previously from four 2x × 4x crosses using polyembryonic 'Nadorcott' tangor as seed parent, were analyzed by 22 simple sequence repeat (SSR) markers, aiming to reveal their genetic origin and the mechanism underlying 2n megagametophyte formation. The results showed that 13 tetraploids from all these four crosses were doubled diploids as indicated by their identical SSR allelic profile with their female parent; while the remaining 41 tetraploids apparently exhibited paternally derived alleles, which confirmed their zygotic origin. Furthermore, the genotyping of all hybrids indicated that all of them arose from 2n megagametophytes. Based on the genotypes of 2n megagametophytes, the analysis of maternal heterozygosity restitution (HR) for each marker showed that it varied from 0.00 to 87.80 % with a mean value of 40.89 %. In addition, it was observed that 13 markers displayed a lower rate than 50 %. On the basis of the above results, it can be speculated that the second division restitution (SDR) is the mechanism underlying the 2n megagametophyte formation in 'Nadorcott' tangor. The elucidation of the mechanism of 2n megagametophyte formation will be of great help to optimize further sexual hybridization for polyploids in citrus.

  4. Mixoploidía Diploide - Tetraploide: Primer Reporte en Nuestro Medio

    Directory of Open Access Journals (Sweden)

    Héctor Pimentel Benítez

    1999-09-01

    Full Text Available Se presenta un caso de mixoploidía diploide-tetraploide, con ligeros signos clínicos pertenecientes a esta condición de la poliploidía en humanos. Estudios citogenéticos a partir de cultivos de sangre periférica y fibroblastos de piel permitieron establecer la fórmula cromosómica 46,XX/92XXXX en la paciente. Se realiza correlación fenotipocariotipo, así como su presentación clínica. Este estudio de casos ayuda a enriquecer el conocimiento sobre esta condición, contribuye a su mejor diagnóstico y a dilucidar la etiología de síndromes malformativos inespecíficos.A case of diploid-tetraploid mixoploidy with mild clinical signs correspondoing to this condition of polyploidy in humans is presented. Cytogenetic studies based on cultures of peripheral blood and fibroblasts of skin allowed to obtain the 46,XX/92XXXX chromosomic formula in the patient. The phenotype-kariotype correlation was stablished and its clinical presentation was made. This study of cases helps to improve the knowledge about this condition, and it also contributes to have a better diagnosis and to dilucidate the etiology of unspecific syndromes of malformation.

  5. Localized chiasmata and meiotic nodules in the tetraploid onion Allium porrum.

    Science.gov (United States)

    Stack, S M; Roelofs, D

    1996-08-01

    Allium porrum L. (cultivated leek) (2n = 4x = 32) is a fertile tetraploid that forms bivalents with pericentric chiasmata at metaphase I. To investigate the basis of this unusual behavior for a tetraploid, we describe the karyotype, axial cores, synaptonemal complexes (SCs), and meiotic nodules of A. porrum. The karyotype appears to be autotetraploid. This conclusion is also supported by presynaptic alignment of axial cores in groups of four and partner trades between pairs of SCs. Numerous early nodules are distributed all along axial cores and SCs during zygonema, but they are lost by late zygonema - early pachynema. Late (recombination) nodules (RNs) are present on SCs near kinetochores throughout the remainder of pachynema. This pattern of RNs corresponds to the pattern of pericentric chiasmata. Pachytene quadrivalents usually are resolved into bivalents because partner trades between SC lateral elements rarely occur between RNs on the same segment of SC. Thus, the patterns of crossing-over and partner trades promote balanced disjunction and high fertility in autotetraploid A. porrum. Rare quadrivalents observed at metaphase I must be due to infrequent partner trades between RNs. Polycomplexes, unusual in their number and size, were observed during zygonema. Key words : synaptonemal complex, recombination nodules, localized chiasmata, polycomplex, Allium porrum.

  6. A tetraploid intermediate precedes aneuploid formation in yeasts exposed to fluconazole.

    Directory of Open Access Journals (Sweden)

    Benjamin D Harrison

    2014-03-01

    Full Text Available Candida albicans, the most prevalent human fungal pathogen, is generally diploid. However, 50% of isolates that are resistant to fluconazole (FLC, the most widely used antifungal, are aneuploid and some aneuploidies can confer FLC resistance. To ask if FLC exposure causes or only selects for aneuploidy, we analyzed diploid strains during exposure to FLC using flow cytometry and epifluorescence microscopy. FLC exposure caused a consistent deviation from normal cell cycle regulation: nuclear and spindle cycles initiated prior to bud emergence, leading to "trimeras," three connected cells composed of a mother, daughter, and granddaughter bud. Initially binucleate, trimeras underwent coordinated nuclear division yielding four daughter nuclei, two of which underwent mitotic collapse to form a tetraploid cell with extra spindle components. In subsequent cell cycles, the abnormal number of spindles resulted in unequal DNA segregation and viable aneuploid progeny. The process of aneuploid formation in C. albicans is highly reminiscent of early stages in human tumorigenesis in that aneuploidy arises through a tetraploid intermediate and subsequent unequal DNA segregation driven by multiple spindles coupled with a subsequent selective advantage conferred by at least some aneuploidies during growth under stress. Finally, trimera formation was detected in response to other azole antifungals, in related Candida species, and in an in vivo model for Candida infection, suggesting that aneuploids arise due to azole treatment of several pathogenic yeasts and that this can occur during the infection process.

  7. The Medicago truncatula lysin [corrected] motif-receptor-like kinase gene family includes NFP and new nodule-expressed genes.

    Science.gov (United States)

    Arrighi, Jean-François; Barre, Annick; Ben Amor, Besma; Bersoult, Anne; Soriano, Lidia Campos; Mirabella, Rossana; de Carvalho-Niebel, Fernanda; Journet, Etienne-Pascal; Ghérardi, Michèle; Huguet, Thierry; Geurts, René; Dénarié, Jean; Rougé, Pierre; Gough, Clare

    2006-09-01

    Rhizobial Nod factors are key symbiotic signals responsible for starting the nodulation process in host legume plants. Of the six Medicago truncatula genes controlling a Nod factor signaling pathway, Nod Factor Perception (NFP) was reported as a candidate Nod factor receptor gene. Here, we provide further evidence for this by showing that NFP is a lysin [corrected] motif (LysM)-receptor-like kinase (RLK). NFP was shown both to be expressed in association with infection thread development and to be involved in the infection process. Consistent with deviations from conserved kinase domain sequences, NFP did not show autophosphorylation activity, suggesting that NFP needs to associate with an active kinase or has unusual functional characteristics different from classical kinases. Identification of nine new M. truncatula LysM-RLK genes revealed a larger family than in the nonlegumes Arabidopsis (Arabidopsis thaliana) or rice (Oryza sativa) of at least 17 members that can be divided into three subfamilies. Three LysM domains could be structurally predicted for all M. truncatula LysM-RLK proteins, whereas one subfamily, which includes NFP, was characterized by deviations from conserved kinase sequences. Most of the newly identified genes were found to be expressed in roots and nodules, suggesting this class of receptors may be more extensively involved in nodulation than was previously known.

  8. Differential antifungal activity of alfalfa (Medicago santva L. saponins originated from roots and aerial parts for some ornamental plant pathogens

    Directory of Open Access Journals (Sweden)

    Alicja Saniewska

    2013-12-01

    Full Text Available The total saponins isolated from aerial parts and roots of alfalfa (Medicago sativa L. at the concentration of 0,01%, 0,05% and 0,1% showed differential influence on the mycelium growth of Alternaria zinniae, Botrytis cinerea, Botrytis tulipae, Phoma narcissi, Phoma poolensis and Rhizoctonia solani. A higher inhibitory effect on in vitro growth of mycelium of all tested pathogens indicated saponins from roots of alfalfa than from aerial parts. Tested saponins from roots at lhe concentration of 0,1% totally inhibited linear growth of Phoma narcissi, and linear growth of Alternaria zinniae was limited about 67%, Botrytis cinerea about 74%, Botrytis tulipae about 68%, Phoma poolensis about 38%, and Rhizoctonia solani about 74% in comparison to the control. The saponins of alfalfa from roots at the concentration of 0,1% and 0,2% applied as preinoculation sprays evidently inhibited the development of Phoma narcissi on Hippeastrum leaves. This dose of aerial saponins of alfalfa did not effect the development of necrotic spots on the leaves.

  9. Study of epidermis characters and venation of certain species of Medicago L. in Leguminosae (Fabaceae family in Iraq

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    Noor J.J. Al-Tememmi

    2017-09-01

    Full Text Available The anatomical study of the epidermis leaflet for seven species and variety wild belonging to the genus Medicago L. species are: M. constricta Dur., M. coronata L., M. intertexta L., M. intertexta.var. ciliaris L., M. laciniata L., M. lupulina L., M. minima L. and M. sativa L. were studied, The search included epidermis characters and stomatal complexes addition to venation system in leaflets. It is revealed through the study, epidermis leaflet type Amphistomatic (the stomata spread on the upper and lower surface as well as the presence of three types of stomatal complexes namely: Anisocytic (the guard cells surrounded by three unequal cell size, Anomocytic (not differential from subsidiary cells in epidermis and Anomotetracytic (four cells surrounding the guard cell is irregular and varied. The results showed variations in dimensions and forms stomata and nature vertical walls and tangent to the upper and lower surfaces. The venation system distinguish by all the species studied as one of the type Simple craspedodromous in this system all secondary veins and branching ends at the margin of the blade leaflets, this found in all species. But there are clear differences in the Areoles shape and type of vienlets and at the ends, where these characters taxonomic and diagnostic value of the task in the species under study.

  10. Sinorhizobium meliloti can protect Medicago truncatula against Phoma medicaginis attack

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    Moncef MRABET

    2011-09-01

    Full Text Available The Sinorhizobium meliloti microsymbiont of Medicago spp. was used in an antibiosis test against Phoma medicaginis and in bioprotection assays of Medicago truncatula JA17 from the pathogen. Among 17 S. meliloti strains isolated from root nodules of M. truncatula and Medicago laciniata grown in Tunisian soils, six showed up to 60% growth inhibition of five P. medicaginis strains isolated from infected field-grown M. truncatula. Two S. meliloti strains with differing in vitro effects on P. medicaginis, 10.16/R6 antagonist and 5M6 non antagonist, were used in a bioprotection assay of M. truncatula JA17 from the pathogen. The inoculation of P. medicaginis caused complete root and stem rotting, and the mortality of all treated plantlets. Inoculation of the antagonist S. meliloti strain 10.16/R6 to M. truncatula JA17 infected with P. medicaginis was associated with a significant 65% decrease of vegetative rotting length, an 80% decrease of plant mortality, an increase of root length, and enhancement of root and shoot biomass comparatively to control plantlets treated with P. medicaginis. The inoculation of the non antagonistic S. meliloti strain 5M6 slightly decreased disease and slightly increased plant growth parameters.

  11. Seasonal variations of artemisinin and its biosynthetic precursors in tetraploid Artemisia annua plants compared with the diploid wild-type

    NARCIS (Netherlands)

    Wallaart, T.E.; Pras, N.; Quax, Wim

    1999-01-01

    Using colchicine we induced tetraploidy in Artemisia annua L. plants. During a vegetation period we monitored the time course of the levels of artemisinin, its direct precursors, the biosynthetically related sesquiterpenes and the essential oil content in the diploid (wild-type) and tetraploid A.

  12. Frequency of local, regional, and long-distance dispersal of diploid and tetraploid Saxifraga oppositifolia (Saxifragaceae) to Arctic glacier forelands.

    Science.gov (United States)

    Müller, Eike; Eidesen, Pernille Bronken; Ehrich, Dorothee; Alsos, Inger Greve

    2012-03-01

    Climate change forces many species to migrate. Empirical small-scale data on migration and colonization in the Arctic are scarce. Retreating glaciers provide new territory for cold-adapted plant species, but the genetic consequences depend on dispersal distances and frequencies. We estimated local, regional, and long-distance dispersal frequencies, as well as their effect on levels of genetic diversity, in diploid and tetraploid individuals of Saxifraga oppositifolia. Samples were collected in four aged moraines in each of three glacier forelands, in surrounding areas and reference populations in the Arctic archipelago Svalbard. These samples were analyzed for neutral amplified fragment length polymorphisms (AFLPs, n = 707) and ploidy levels (n = 30). Genetic clustering and ploidy analyses revealed two distinct genetic groups representing diploids and tetraploids, with few intermediate triploids. The groups were intermixed in most sampled populations. No differences in genetic diversity were found between tetraploids and diploids, or between established and glacier foreland populations. Seeds were dispersed over local, regional, and long distances, with the highest proportions of seeds originating from close sources. A minimum of 4-15 founding individuals from several source populations had initially established in each glacier foreland. Our data suggest that S. oppositifolia can rapidly colonize new deglaciated areas without losing genetic diversity. Thus, glacier forelands can be alternative habitats for cold-adapted vascular plants tracking their climatic niche. Our data show no difference in colonization success between diploid and tetraploid individuals.

  13. INITIAL ACYTOKINESIS DURING LEAF PROTOPLAST CULTURE OF DIHAPLOID AND TETRAPLOID SOLANUM-TUBEROSUM AND DIPLOID S-BULBOCASTANUM

    NARCIS (Netherlands)

    VANEVERDINK, WJ; PIJNACKER, LP

    1994-01-01

    Leaf protoplasts of dihaploid (2n=2x=24) and tetraploid (2n=4x=48) Solanum tuberosum, rr, and diploid S. bulbocastanum (2n=2x=24) were cultured in liquid medium. The cultures were studied for early karyological changes during their development. Giemsa staining of spread preparations revealed

  14. Two major groups of chloroplast DNA haplotypes in diploid and tetraploid Aconitum subgen. Aconitum (Ranunculaceae in the Carpathians

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    J. Mitka

    2016-04-01

    Full Text Available Aconitum in Europe is represented by ca. 10% of the total number of species and the Carpathian Mts. are the center of the genus variability in the subcontinent. We studied the chloroplast DNA intergenic spacer trnL(UAG-rpl32- ndhF (cpDNA variability of the Aconitum subgen. Aconitum in the Carpathians: diploids (2n=16, sect. Cammarum, tetraploids (2n=32, sect. Aconitum and triploids (2n=24, nothosect. Acomarum. Altogether 25 Aconitum accessions representing the whole taxonomic variability of the subgenus were sequenced and subjected to phylogenetic analyses. Both parsimony, Bayesian and character network analyses showed the two distinct types of the cpDNA chloroplast, one typical of the diploid and the second of the tetraploid groups. Some specimens had identical cpDNA sequences (haplotypes and scattered across the whole mountain arch. In the sect. Aconitum 9 specimens shared one haplotype, while in the sect. Camarum one haplotype represents 4 accessions and the second – 5 accessions. The diploids and tetraploids were diverged by 6 mutations, while the intrasectional variability amounted maximally to 3 polymorphisms. Taking into consideration different types of cpDNA haplotypes and ecological profiles of the sections (tetraploids – high‑mountain species, diploids – species from forest montane belt we speculate on the different and independent history of the sections in the Carpathians.

  15. Cultivation of diploid and tetraploid hairy roots of Datura stramonium L. in stirred tank bioreactor for tropane alkaloids production

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    ATANAS PAVLOV

    2012-01-01

    Full Text Available Biomass accumulation and tropane alkaloids production by diploid and tetraploid hairy root cultures of Datura stramonium L. cultivated in stirred tank bioreactor at different aeration rates were investigated. The maximal growth for both hairy root cultures (ADB = 8.3 g/L and 6.8 g/L for diploid and tetraploid line, respectively was achieved at aeration rate of 15.0 L/(L.h. The corresponding growth indexes were remarkably high (GIDW = 9.0 and 7.8 for diploid and tetraploid line, respectively compared to the values, usually reported for other hairy root cultures. The optimal aeration rate for biomass accumulation was also optimal for alkaloids biosynthesis. According to our survey, the achieved maximal amounts of accumulated hyoscyamine (35.0 mg/L and 27.0 mg/L for diploid and tetraploid line were the highest reported in the scientific literature for D. stramonium L. hairy roots. During the cultivation in stirred tank bioreactor, the hairy roots biosynthesized pharmaceutically important alkaloid scopolamine in minor concentrations. This is an important observation since scopolamine was not detected during submerged cultivation of these hairy root lines in other bioreactor types. However, the ploidy level was found to be the most important factor concerning scopolamine production by D. stramonium L. hairy roots cultures. The present work demonstrated the effect of ploidity levels on biomass accumulation and tropane alkaloids production by D. stramonium L. hairy roots cultivated in stirred tank bioreactor. This investigation show that the stirred tank bioreactor could be successfully applied for both maximal biomass accumulations, as well as for manipulation of tropane alkaloids production by diploid and tetraploid D. stramonium L. hairy root cultures.

  16. Changes in motility, ATP content, morphology and fertilisation capacity during the movement phase of tetraploid Pacific oyster (Crassostrea gigas) sperm.

    Science.gov (United States)

    Suquet, M; Labbe, C; Brizard, R; Donval, A; Le Coz, J R; Quere, C; Haffray, P

    2010-07-01

    Changes in sperm features during the movement phase are especially interesting to study in external fertilization species whose sperm duration movement is long because this implies a significant adaptation of moving cells to the external medium. This study describes the changes in tetraploid Pacific oyster sperm characteristics in relation to time post activation. Sperm individually collected on three tetraploid males were activated in seawater. Their features were analysed over a 24h period and compared to a sperm pool collected on three diploid males as a reference. The percentage of motile spermatozoa, the intracellular ATP content, and the fine structure of spermatozoa were studied in relation to time post activation. Furthermore, the fertilisation capacity of sperm individually collected on five diploid males was assessed after 1 and 24h post activation. A forward progressive movement was maintained for at least a 20h duration. Compared to diploid males, the percentage of motile spermatozoa was lower in tetraploid males. The intracellular ATP concentration was higher in spermatozoa from tetraploid males than in spermatozoa from diploid males. A decrease in ATP content was observed in the first 6h post activation and severe alterations were observed in sperm morphology after 24h. Then, a lower fertilisation capacity of sperm from diploid males was observed at the end of the movement phase. The cessation of Pacific oyster sperm motility was unlikely caused by ATP consumption as ATP concentration was still high at the end of sperm movement but rather caused by drastic changes in sperm morphology. Compared to sperm collected on diploid males, the lower quality of sperm from tetraploid males was emphasized by a shorter movement duration and deeper morphological alterations at the end of the movement phase.

  17. Hyperhaploid and tetraploid sperm detected in men who ingested ultra-high doses of diazepam

    Energy Technology Data Exchange (ETDEWEB)

    Baumgartner, A. [Lawrence Livermore National Lab., CA (United States)]|[GSF-Inst. Saeugetiergenetik, Neuherberg (Germany); Adler, I.D.; Schmid, T.E. [GSF-Inst. Saeugetiergenetik, Neuherberg (Germany)] [and others

    1997-10-01

    Diazepam is widely administered as a sedative, muscle relaxant and anxiolytic drug. Five young non-smoking men who were hospitalized after their suicide attempt using diazepam, {approximately}1-7 mg/kg (oral intake), provided semen samples 40-50 days and {approximately}100 days after exposure to assess drug effects on meiotic cells and to evaluate persistence. Five healthy men served as local clinical controls. A multicolor FISH assay was applied to detect aneuploidy for chromosome X, Y, and 21 in sperm. Sex ratios were not significantly different from 1:1 among 133,143 cells analyzed. The 40-day samples showed an increase in several sperm aneuploidy groups: disomy 21 (1.5 fold, p=0.04); disomy X (2.7 fold, p=0.0006), and XY aneuploidy (1.6 folk, p=0 0.017). The results for {approximately}100 days after exposure were similar to controls suggesting that hyperhaploidy effects may not persist. Phase contrast microscopy was used to identify flagellated tetraploid sperm, i.e., X-X-Y-Y-21-21-21-21. Tetraploid sperm were found among 8 semen samples provided by five patients (1.4 {+-} 1.2 per 10,000 cells; >80,000 cells) while none were detected among >50,000 cells from healthy men. Our findings are consistent with the possible aneuploidy-inducing effect of diazepam during male meiosis but further studies are needed before these results can be extrapolated to therapeutic dosing because suicide patients are a highly exposed cohort and other confounding factors (alcohol, drugs, antidotes) cannot be ruled out.

  18. Bioactive spirans and other constituents from the leaves of Cannabis sativa f. sativa.

    Science.gov (United States)

    Guo, Tian-Tian; Zhang, Jian-Chun; Zhang, Hai; Liu, Qing-Chao; Zhao, Yong; Hou, Yu-Fei; Bai, Lu; Zhang, Li; Liu, Xue-Qiang; Liu, Xue-Ying; Zhang, Sheng-Yong; Bai, Nai-Sheng

    2017-08-01

    In this paper, 17 compounds (1-17) were isolated from the leaves of Hemp (Cannabis sativa f. sativa). Among the isolates, two were determined to be new spirans: cannabispirketal (1), and α-cannabispiranol 4'-O-β-D-glucopyranose (2) by 1D and 2D NMR spectroscopy, LC-MS, and HRESIMS. The known compounds 7, 8, 10, 13, 15, and 16 were isolated from Hemp (C. sativa f. sativa) for the first time. Furthermore, compounds 8 and 13 were isolated from the nature for the first time. All isolated compounds were evaluated for cytotoxicity on different tissue-derived passage cancer cell lines through cell viability and apoptosis assay. Among these compounds, compounds 5, 9 and 16 exhibited a broad-spectrum antitumor effect via inhibiting cell proliferation and promoting apoptosis. These results obtained have provided valuable clues to the understanding of the cytotoxic profile for these isolated compounds from Hemp (C. sativa f. sativa).

  19. Differential proteomics: a study in Medicago truncatula somatic embryogenesis

    OpenAIRE

    Ventosa, Miguel André Lourenço da Luz, 1987

    2010-01-01

    Tese de mestrado. Biologia (Biologia Celular e Biotecnologia). Universidade de Lisboa, Faculdade de Ciências, 2010 As leguminosas são um importante grupo de cultivares, utilizadas como alimento humano e animal, principalmente devido ao seu alto conteúdo proteico. A informação disponível sobre leguminosas modelo como Medicago truncatula e Lotus japonicus nos últimos anos tem melhorado consideravelmente a nossa compreensão sobre a estrutura do genoma, função de gene e de proteínas, das legum...

  20. Plant regeneration from proroplasts of alfalfa (Medicago sativa via somatic embryogenesis Regeneração de plantas a partir de protoplastos de alfafa (Medicago sativa via embriogênese somática

    Directory of Open Access Journals (Sweden)

    Mariza Monteiro

    2003-12-01

    Full Text Available Alfalfa is one of the most frequently studied species from the production of tissue culture-derived embryos point of view. In this study, five alfalfa cultivars were analyzed with reference to their ability to regenerate plants from protoplast cultures via somatic embryogenesis. Plant regeneration from leaf-derived protoplasts isolated from the cultivar Rangelander was achieved using a protocol defined for alfalfa cell suspension-derived embryogenesis. Because of its high efficiency, this procedure is recommended for protoplast electroporation-mediated genetic transformation of alfalfa.A alfafa é uma das espécies mais freqüentemente estudadas do ponto de vista da produção de embriões somáticos derivados da cultura de tecidos. Neste trabalho, cinco cultivares de alfafa foram analisados com referência à capacidade de regenerar plantas a partir de culturas de protoplastos via embriogênese somática. Regeneração de plantas a partir de protoplastos isolados de folhas da cultivar Rangelander foi obtida usando-se um protocolo definido para embriogênese somática derivada de suspensões celulares de alfafa. Em função da sua alta eficiência, recomenda-se o uso deste procedimento para transformação genética de alfafa mediada por eletroporação de protoplastos.

  1. Origin and domestication of Lactuca sativa L.

    NARCIS (Netherlands)

    Vries, de I.M.

    1997-01-01

    The domestication of lettuce, Lactuca sativa L. is described on the basis of literature study. The centre of origin is discussed. A historical survey is made of the distribution of the groups of Lactuca cultivars over the world.

  2. Nigella sativa provides protection against metabolic syndrome ...

    African Journals Online (AJOL)

    Nigella sativa provides protection against metabolic syndrome. Abdul Saboor Shah, Gul Majid Khan, Amir Badshah, Shefaat Ullah Shah, Kifayat Ullah Shah, Shakeel Ahmad Mirza, Kifayat Ullah Shah, Kamran Ahmad Khan ...

  3. The molecular genetic linkage map of the model legume Medicago truncatula: an essential tool for comparative legume genomics and the isolation of agronomically important genes

    Science.gov (United States)

    Thoquet, Philippe; Ghérardi, Michele; Journet, Etienne-Pascal; Kereszt, Attila; Ané, Jean-Michel; Prosperi, Jean-Marie; Huguet, Thierry

    2002-01-01

    Background The legume Medicago truncatula has emerged as a model plant for the molecular and genetic dissection of various plant processes involved in rhizobial, mycorrhizal and pathogenic plant-microbe interactions. Aiming to develop essential tools for such genetic approaches, we have established the first genetic map of this species. Two parental homozygous lines were selected from the cultivar Jemalong and from the Algerian natural population (DZA315) on the basis of their molecular and phenotypic polymorphism. Results An F2 segregating population of 124 individuals between these two lines was obtained using an efficient manual crossing technique established for M. truncatula and was used to construct a genetic map. This map spans 1225 cM (average 470 kb/cM) and comprises 289 markers including RAPD, AFLP, known genes and isoenzymes arranged in 8 linkage groups (2n = 16). Markers are uniformly distributed throughout the map and segregation distortion is limited to only 3 linkage groups. By mapping a number of common markers, the eight linkage groups are shown to be homologous to those of diploid alfalfa (M. sativa), implying a good level of macrosynteny between the two genomes. Using this M. truncatula map and the derived F3 populations, we were able to map the Mtsym6 symbiotic gene on linkage group 8 and the SPC gene, responsible for the direction of pod coiling, on linkage group 7. Conclusions These results demonstrate that Medicago truncatula is amenable to diploid genetic analysis and they open the way to map-based cloning of symbiotic or other agronomically-important genes using this model plant. PMID:11825338

  4. From model to crop: functional analysis of a STAY-GREEN gene in the model legume Medicago truncatula and effective use of the gene for alfalfa improvement.

    Science.gov (United States)

    Zhou, Chuanen; Han, Lu; Pislariu, Catalina; Nakashima, Jin; Fu, Chunxiang; Jiang, Qingzhen; Quan, Li; Blancaflor, Elison B; Tang, Yuhong; Bouton, Joseph H; Udvardi, Michael; Xia, Guangmin; Wang, Zeng-Yu

    2011-11-01

    Medicago truncatula has been developed into a model legume. Its close relative alfalfa (Medicago sativa) is the most widely grown forage legume crop in the United States. By screening a large population of M. truncatula mutants tagged with the transposable element of tobacco (Nicotiana tabacum) cell type1 (Tnt1), we identified a mutant line (NF2089) that maintained green leaves and showed green anthers, central carpels, mature pods, and seeds during senescence. Genetic and molecular analyses revealed that the mutation was caused by Tnt1 insertion in a STAY-GREEN (MtSGR) gene. Transcript profiling analysis of the mutant showed that loss of the MtSGR function affected the expression of a large number of genes involved in different biological processes. Further analyses revealed that SGR is implicated in nodule development and senescence. MtSGR expression was detected across all nodule developmental zones and was higher in the senescence zone. The number of young nodules on the mutant roots was higher than in the wild type. Expression levels of several nodule senescence markers were reduced in the sgr mutant. Based on the MtSGR sequence, an alfalfa SGR gene (MsSGR) was cloned, and transgenic alfalfa lines were produced by RNA interference. Silencing of MsSGR led to the production of stay-green transgenic alfalfa. This beneficial trait offers the opportunity to produce premium alfalfa hay with a more greenish appearance. In addition, most of the transgenic alfalfa lines retained more than 50% of chlorophylls during senescence and had increased crude protein content. This study illustrates the effective use of knowledge gained from a model system for the genetic improvement of an important commercial crop.

  5. The molecular genetic linkage map of the model legume Medicago truncatula: an essential tool for comparative legume genomics and the isolation of agronomically important genes

    Directory of Open Access Journals (Sweden)

    Ané Jean-Michel

    2002-01-01

    Full Text Available Abstract Background The legume Medicago truncatula has emerged as a model plant for the molecular and genetic dissection of various plant processes involved in rhizobial, mycorrhizal and pathogenic plant-microbe interactions. Aiming to develop essential tools for such genetic approaches, we have established the first genetic map of this species. Two parental homozygous lines were selected from the cultivar Jemalong and from the Algerian natural population (DZA315 on the basis of their molecular and phenotypic polymorphism. Results An F2 segregating population of 124 individuals between these two lines was obtained using an efficient manual crossing technique established for M. truncatula and was used to construct a genetic map. This map spans 1225 cM (average 470 kb/cM and comprises 289 markers including RAPD, AFLP, known genes and isoenzymes arranged in 8 linkage groups (2n = 16. Markers are uniformly distributed throughout the map and segregation distortion is limited to only 3 linkage groups. By mapping a number of common markers, the eight linkage groups are shown to be homologous to those of diploid alfalfa (M. sativa, implying a good level of macrosynteny between the two genomes. Using this M. truncatula map and the derived F3 populations, we were able to map the Mtsym6 symbiotic gene on linkage group 8 and the SPC gene, responsible for the direction of pod coiling, on linkage group 7. Conclusions These results demonstrate that Medicago truncatula is amenable to diploid genetic analysis and they open the way to map-based cloning of symbiotic or other agronomically-important genes using this model plant.

  6. Phytochrome chromophore biosynthesis. [Avena sativa

    Energy Technology Data Exchange (ETDEWEB)

    Elich, T.D.; Lagarias, J.C.

    1987-06-01

    Etiolated Avena sativa L. seedlings grown in the presence of gabaculine (5-amino-1,3-cyclohexadienylcarboxylic acid) contained reduced levels of phytochrome as shown by spectrophotometric and immunochemical assays. Photochromic phytochrome levels in gabaculine-grown plants were estimated to be 20% of control plants, while immunoblot analysis showed that the phytochrome protein moiety was present at approximately 50% of control levels. Gabaculine-grown seedlings administered either 5-aminolevulinic acid or biliverdin exhibited a rapid increase of spectrophotometrically detectable phytochrome. Phytochrome concentrations estimated immunochemically did not similarly increase throughout treatment with either compound. Similar experiments with 5-amino(4-/sup 14/C) levulinic acid showed radiolabeling of phytochrome with kinetics that paralleled the spectrally detected increase. These results are consistent with (a) the intermediacy of both 5-aminolevulinic acid and biliverdin in the biosynthetic pathway of the phytochrome chromophore and (b) the lack of coordinate regulation of chromophore and apoprotein synthesis in Ayena seedlings.

  7. [Therapeutic potential of Cannabis sativa].

    Science.gov (United States)

    Avello L, Marcia; Pastene N, Edgar; Fernández R, Pola; Córdova M, Pia

    2017-03-01

    Cannabis sativa (marihuana) is considered an illicit drug due to its psychoactive properties. Recently, the Chilean government opened to the use cannabis in the symptomatic treatment of some patients. The biological effects of cannabis render it useful for the complementary treatment of specific clinical situations such as chronic pain. We retrieved scientific information about the analgesic properties of cannabis, using it as a safe drug. The drug may block or inhibit the transmission of nervous impulses at different levels, an effect associated with pain control. Within this context and using adequate doses, forms and administration pathways, it can be used for chronic pain management, considering its effectiveness and low cost. It could also be considered as an alternative in patients receiving prolonged analgesic therapies with multiple adverse effects.

  8. Review: Nigella sativa (Prophetic Medicine): A Review.

    Science.gov (United States)

    Ijaz, Hira; Tulain, Ume Ruqia; Qureshi, Junaid; Danish, Zeeshan; Musayab, Samina; Akhtar, Muhammad Furqan; Saleem, Ammara; Khan, Khanzada Khanzada; Zaman, Muhammad; Waheed, Imran; Khan, Imran; Abdel-Daim, Mohamed

    2017-01-01

    The present report is a significant effort to explore detail description of N. Sativa, its pharmacognostic characteristics, morphological characteristics, and mechanism of actions, doses and medicinal uses. Nigella sativa (N. Sativa) is greatest form of healing medicine. It is also known as Prophetic Medicine as its use has been mentioned in Prophetic Hadit, as natural remedy for all the diseases except death. It is recommended on daily basis in Tibb-e-Nabwi (Prophetic Medicine). Hazrat Abu Hurairah States ''I have heard from Rasool Allah (PBUH) that there is cure for every disease in black seeds except death and black seeds are shooneeze''. Salim Bin Abdullah narrates with reference to his father Hazrat Abdullah Bin Omar that Rasool Allah (PBUH) said, 'Let all the black seed upon you, these contain cure of all diseases except death'. N. sativa claimed to have anti-inflammatory, analgesic, hepato-protective, neuro-protective, gastro-protective and other useful properties. Biological and pharmacological effects are attributed to its two important constituents Thymoquinone (TQ) and Nigella sativa oil (NSO). TQ has interaction with human serum albumin. Seeds containing volatile oils mainly Melanthin showed toxicity at larger doses. This report is a reference for all pharmaceutical researchers, physicians and biologists researching on N. sativa and will open a door towards novel agent.

  9. Comunidad de arañas (Arachnida, Araneae del cultivo de alfalfa (Medicago sativa en Buenos Aires, Argentina

    Directory of Open Access Journals (Sweden)

    Andrea Armendano

    2010-06-01

    Full Text Available En las últimas décadas se ha dado un interés creciente en el uso de enemigos naturales para controlar plagas de insectos, como arañas. Se estudió una comunidad de arañas en Argentina mediante un muestreo cada dos semanas durante el periodo 2004-2006 en lotes de una hectárea. En el estrato del suelo las arañas fueron colectadas con redes de arrastre y trampas de caída. Se recolecto un total de 6 229 ejemplares (15 familias y 50 especies. Siete familias se encuentran en el estrato herbáceo, las más abundantes fueron: Thomisidae (n=2 012, 32.30%, Araneidae (n=1 516, 24.33% y Oxyopidae (n=604, 9.70%. El suelo habían 14 familias, principalmente: Lycosidae (n=629, 10.10% y Linyphiidae (n=427, 6.85%. Predominaron las arañas cazadoras: por emboscadas (32.99%, al acecho (11.77%, corredoras de suelo (10.84% y tejedoras orbiculares (27.56%. Los índices de diversidad fueron: H´=2.97, Dsp=0.11 y J=0.79, evidenciando una comunidad de arañas moderadamente diversa, con predominio de Misumenops pallidus, Oxyopes salticus, Lycosa poliostoma and L. erythrognatha. Las arañas estuvieron presentes durante el desarrollo fenológico del cultivo con picos de abundancia en primavera y verano.

  10. Cloning and function analysis of an alfalfa (Medicago sativa L.) zinc finger protein promoter MsZPP.

    Science.gov (United States)

    Li, Yan; Sun, Yan; Yang, Qingchuan; Kang, Junmei; Zhang, Tiejun; Gruber, Margaret Yvonne; Fang, Feng

    2012-08-01

    A 1272 bp upstream sequence of MsZFN gene was cloned from alfalfa, which was designed as MsZPP (Genbank accession number: FJ 161979.2) using an adaptor-mediated genome walking method. A sole transcription start site was located 69 bp upstream of the translation start site. Its pattern of expression included roots, stem vascular tissues, floral reproductive organs, and leaves, but the promoter did not express in seeds, petals or sepals. Transcription levels can be stimulated by dark, MeJA, and IAA. However, GUS fusion activities had no change by treatments of GA, ABA, drought and high salt for 3 days. Deletion analysis revealed that all sections of the promoter can drive gus gene expression in the root, stem, leaves and floral reproductive organs; however, only fragments longer than the -460 bp promoter can stimulate strong gus gene expression in these organs. In addition, the -460 bp promoter fragment can drive gus expression not only in the vascular tissue, but also in leaf guard cells. The results suggest that the promoter MsZPP plays roles in the regulation of transgene expression, particularly due to its darkness, MeJA, and IAA responsiveness.

  11. Exploitation of alfalfa seed (Medicago sativa L.) flour into gluten-free rice cookies: Nutritional, antioxidant and quality characteristics.

    Science.gov (United States)

    Giuberti, Gianluca; Rocchetti, Gabriele; Sigolo, Samantha; Fortunati, Paola; Lucini, Luigi; Gallo, Antonio

    2018-01-15

    In an effort to increase the nutritional value of common gluten-free (GF) cereal-based foods, GF cookies using alfalfa seed flour (ASF), at different substitution levels to common rice flour (0% as control, 15%, 30% and 45% w/w), were produced. Crude protein, total dietary fibre, total polyunsaturated, total n-3 and n-6 fatty acid contents increased linearly (p<0.05) by raising the substitution levels of rice with ASF. The hardness, the total phenolic content, the in vitro antioxidant capacity and the resistant starch increased linearly (p<0.05), whereas the starch hydrolysis index decreased linearly (p<0.05) by raising the substitution levels of rice flour with ASF. Despite the fact that ASF-substituted GF cookies had inferior sensory attributes compared to the control, the score given by the panellists remained at fairly good levels for all tested parameters, showing acceptability of the substituted GF cookies. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Farklı Yonca (Medicago sativa L.) Genotiplerinin Ege Bölgesi Koşullarına Adaptasyonu

    OpenAIRE

    DEMİROĞLU, Gülcan; GEREN, Hakan; AVCIOĞLU, Rıza

    2015-01-01

    B u çalışma; 2002-2003 ve 2003-2004 yılı yetiştirme dönemlerinde, Ege Üniversitesi Ziraat Fakültesi Tarla Bitkileri Bölümü’nün Bornova’da bulunan deneme tarlaları ile Ödemiş Meslek Yüksek Okulu'na ait deneme tarlalarında farklı yonca genotiplerinin (Tru-test, Bacana, FG8R612 ve Elçi) morfolojik ve bazı agronomik özelliklerinin farklı toprak bünyelerindeki lokasyonlarda adaptasyonunu belirlemek amacıyla yürütülmüştür. Denemede; çiçeklenme gün sayısı, ana sap uzunluğu, ana sap kalınlığı, b...

  13. Stress-related phenomena and detoxification mechanisms induced by common pharmaceuticals in alfalfa (Medicago sativa L.) plants.

    Science.gov (United States)

    Christou, Anastasis; Antoniou, Chrystalla; Christodoulou, Charalampia; Hapeshi, Evroula; Stavrou, Ioannis; Michael, Costas; Fatta-Kassinos, Despo; Fotopoulos, Vasileios

    2016-07-01

    Pharmaceutically active compounds (PhACs) have been recently shown to exert phytotoxic effects. The present study explores the uptake, systemic translocation, and abiotic stress responses and detoxification mechanisms induced by the exposure of alfalfa plants grown in sand under greenhouse conditions to four common, individually applied PhACs (10μgL(-1)) (diclofenac, sulfamethoxazole, trimethoprim, 17a-ethinylestradiol) and their mixture. Stress physiology markers (lipid peroxidation, proline, H2O2 and NO content, antioxidant activity assays) and gene expression levels of key plant detoxification components (including glutathione S-transferases, GST7, GST17; superoxide dismutases, CuZnSOD, FeSOD; proton pump, H(+)-ATP, and cytochrome c oxidase, CytcOx), were evaluated. PhACs were detected in significantly higher concentrations in roots compared with leaves. Stress related effects, manifested via membrane lipid peroxidation and oxidative burst, were local (roots) rather than systemic (leaves), and exacerbated when the tested PhACs were applied in mixture. Systemic accumulation of H2O2 in leaves suggests its involvement in signal transduction and detoxification responses. Increased antioxidant enzymatic activities, as well as upregulated transcript levels of GST7, GST17, H(+)-ATPase and CytcOx, propose their role in the detoxification of the selected PhACs in plants. The current findings provide novel biochemical and molecular evidence highlighting the studied PhACs as an emerging abiotic stress factor, and point the need for further research on wastewater flows under natural agricultural environments. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Effect of photoperiod prior to cold acclimation on freezing tolerance and carbohydrate metabolism in alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Bertrand, Annick; Bipfubusa, Marie; Claessens, Annie; Rocher, Solen; Castonguay, Yves

    2017-11-01

    Cold acclimation proceeds sequentially in response to decreases in photoperiod and temperature. This study aimed at assessing the impact of photoperiod prior to cold acclimation on freezing tolerance and related biochemical and molecular responses in two alfalfa cultivars. The fall dormant cultivar Evolution and semi-dormant cultivar 6010 were grown in growth chambers under different photoperiods (8, 10, 12, 14 or 16h) prior to cold acclimation. Freezing tolerance was evaluated as well as carbohydrate concentrations, levels of transcripts encoding enzymes of carbohydrate metabolism as well as a K-3dehydrin, before and after cold acclimation. The fall dormant cultivar Evolution had a better freezing tolerance than the semi-dormant cultivar 6010. The effect of photoperiod prior to cold acclimation on the level of freezing tolerance differed between the two cultivars: an 8h-photoperiod induced the highest level of freezing tolerance in Evolution and the lowest in 6010. In Evolution, the 8h-induced superior freezing tolerance was associated with higher concentration of raffinose-family oligosaccharides (RFO). The transcript levels of sucrose synthase (SuSy) decreased whereas those of sucrose phosphatase synthase (SPS) and galactinol synthase (GaS) increased in response to cold acclimation in both cultivars. Our results indicate that RFO metabolism could be involved in short photoperiod-induced freezing tolerance in dormant alfalfa cultivars. Crown Copyright © 2017. Published by Elsevier B.V. All rights reserved.

  15. Assessing of the contributions of pod photosynthesis to carbon acquisition of seed in alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Zhang, Wenxu; Mao, Peisheng; Li, Yuan; Wang, Mingya; Xia, Fangshan; Wang, Hui

    2017-02-07

    The distribution of carbon from a branch setting pod in alfalfa was investigated during the seed development of seeds to determine the relative contribution of pod and leaf photoassimilates to the total C balance and to investigate the partitioning of these photoassimilates to other plant organs. A 13Clabeling procedure was used to label C photoassimilates of pods and leaves in alfalfa, and the Δ13C values of a pod, leaves, a section of stem and roots were measured during seed development on day 10, 15, 20 and 25 after labeling of the pod. The results showed that the alfalfa pod had photosynthetic capacity early in the development of seeds, and that pod photosynthesis could provide carbon to alfalfa organs including seeds, pods, leaves, stems and roots, in addition to leaf photosynthesis. Photosynthesis in the pod affected the total C balance of the alfalfa branch with the redistribution of a portion of pod assimilates to other plant organs. The assimilated 13C of the pod was used for the growth requirements of plant seeds and pods. The requirements for assimilated C came primarily from the young pod in early seed development, with later requirements provided primarily from the leaf.

  16. Degradation of lignified secondary cell walls of lucerne (Medicago sativa L.) by rumen fungi growing in methanogenic co-culture.

    Science.gov (United States)

    Bootten, T J; Joblin, K N; McArdle, B H; Harris, P J

    2011-11-01

    To compare the abilities of the monocentric rumen fungi Neocallimastix frontalis, Piromyces communis and Caecomyces communis, growing in coculture with Methanobrevibacter smithii, to colonize and degrade lignified secondary cell walls of lucerne (alfalfa) hay. The cell walls of xylem cylinders isolated from stems of lucerne contained mostly xylans, cellulose and lignin together with a small proportion of pectic polysaccharides. All of these major components were removed during incubation with the three fungi, and differing cell wall polysaccharides were degraded to different extents. The greatest dry weight loss was found with N. frontalis and least with C. communis, and scanning electron microscopy revealed that these extensively colonized different cell types. C. communis specifically colonized secondary xylem fibres and showed much less degradation than N. frontalis and P. communis. Neocallimastix frontalis and P. communis were efficient degraders of the cell walls of lucerne xylem cylinders. Degradation occurred of pectic polysaccharides, xylan and cellulose. Loss of lignin from the xylem cylinders probably resulted from the cleavage of xylan releasing xylan-lignin complexes. Unlike rumen bacteria, the rumen fungi N. frontalis, P. communis and C. communis are able to degrade lignified secondary walls in lucerne stems. These fungi could improve forage utilization by ruminants and may have potential in the degradation of lignocellulosic biomass in the production of biofuels. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

  17. An alfalfa (Medicago sativa L.) ethylene response factor gene, MsERF11, enhances salt tolerance in transgenic Arabidopsis.

    Science.gov (United States)

    Chen, Tingting; Yang, Qingchuan; Zhang, Xinquan; Ding, Wang; Gruber, Margaret

    2012-09-01

    A novel orthologue of ethylene response factor gene, MsERF11, was isolated from alfalfa in this study. It has an open reading frame of 807 bp, encoding a predicted polypeptide of 268 amino acids. Sequence similarity analysis clearly suggested that MsERF11 encoded an ethylene response factor protein. The results of transient expression of MsERF11 in onion epidermal cells indicated that MsERF11 is a nuclear protein. The expression pattern of MsERF11 gene was analyzed by real-time quantitative PCR and a higher level of expression was observed in leaves than was observed in roots, stems, flower buds and flowers. Furthermore, the expression was induced by PEG6000, NaCl, Al2(SO4)3 and six different hormones. Over-expressing MsERF11 resulted in enhanced tolerances to salt stress in transgenic Arabidopsis plants. This research indicates that MsERF11 has the potential to be used for improving crop's salt tolerance in areas where salinity is a limiting factor for agricultural productivity. MsERF11 was isolated from alfalfa. Its expression was induced by different abiotic stresses and hormones. Over-expressing MsERF11 resulted in enhanced salt tolerance in transgenic Arabidopsis plants.

  18. Structural, thermal, and anti-inflammatory properties of a novel pectic polysaccharide from alfalfa (Medicago sativa L.) stem.

    Science.gov (United States)

    Chen, Lei; Liu, Jie; Zhang, Yaqiong; Dai, Bona; An, Yuan; Yu, Liangli Lucy

    2015-04-01

    A pectic polysaccharide (APPS) was purified from the cold alkali extract of alfalfa stem and characterized to be a rhamnogalacturonan I (RG-I) type pectin with the molecular weight of 2.38 × 10(3) kDa and a radius of 123 nm. The primary structural analysis indicated that APPS composed of a →2)-α-l-Rhap-(1→4)-α-d-GalpA-(1→ backbone with 12% branching point at C-4 of Rhap forming side chains by l-arabinosyl and d-galactosyl oligosaccharide units. Transmission electron microscopy (TEM) analysis revealed a primary linear-shaped structure with a few branches in its assembly microstructures. The thermal decomposition evaluation revealed the stability of APPS with an apparent activation energy (Ea) of 226.5 kJ/mol and a pre-exponential factor (A) of 2.10 × 10(25)/s, whereas its primary degradation occurred in the temperature range from 215.6 to 328.0 °C. In addition, APPS showed significant anti-inflammatory effect against mRNA expressions of the pro-inflammatory cytokine genes, especially for IL-1β, suggesting its potential utilization in functional foods and dietary supplement products.

  19. A novel alkaline hemicellulosic heteroxylan isolated from alfalfa (Medicago sativa L.) stem and its thermal and anti-inflammatory properties.

    Science.gov (United States)

    Chen, Lei; Liu, Jie; Zhang, Yaqiong; Niu, Yuge; Dai, Bona; Yu, Liangli Lucy

    2015-03-25

    A novel hemicellulosic polysaccharide (ACAP) was purified from the cold alkali extraction of alfalfa stems and characterized as a heteroxylan with a weight-average molecular weight of 7.94 × 10(3) kDa and a radius of 58 nm. Structural analysis indicated that ACAP consisted of a 1,4-linked β-D-Xylp backbone with 4-O-MeGlcpA and T-L-Araf substitutions at O-2 and O-3 positions, respectively. Transmission electron microscopy (TEM) examination revealed the entangled chain morphology of ACAP molecules. The evaluation of thermal degradation property revealed a primary decomposition temperature range of 238.8-314.0 °C with an apparent activation energy (Ea) and a pre-exponential factor (A) of 220.0 kJ/mol and 2.81 × 10(24)/s, respectively. ACAP also showed significant inhibitory activities on IL-1β, IL-6, and COX-2 gene expressions in cultured RAW 264.7 mouse macrophage cells. These results suggested the potential utilization of ACAP in functional foods and dietary supplement products.

  20. The effect of the stage of maturity of alfalfa (Medicago sativa L. on the chemical composition and in sacco digestibility

    Directory of Open Access Journals (Sweden)

    Petr Doležal

    2008-01-01

    Full Text Available The influence of the stage of maturity of alfalfa on the chemical composition and in sacco digestibility was studied in a laboratory experiment. Alfalfa plants were analysed in 8 vegetation stages harvested in regular intervals before the beginning of bud setting until the fall of blossoms. The contents of nitrogenous substances, fat, crude fibre, ADF, NDF, Ca, P, Na, K and Mg were determined in the samples, and using the in sacco method also the organic matter digestibility in the rumen. Further we assessed the energy content and calculated the N−free extractives (BNLV value. In some stages the chemical changes were monitored in the whole plant. It was discovered that during vegetation there was a statistically significant increase in the crude fibre content and in the ADF and NDF fractions and a reduction in nitrogenous substances. A strong negative correlation was detected between the stage of maturity of alfalfa and content of crude protein and net energy for lactation (r = −0.97 and r = −0.92, respectively. In the individual stages of vegetation the differences in the content of nitrogenous substances, fibre and fat were statistically significant (P < 0.05. The statistically highest content of crude protein (28.97% was detected in the first stage before bud setting and the lowest (11.97% in the stage after the fall of blossoms. Later vegetation also had a stronger depressive effect on the content of fat and NEL. The organic matter digestibility (P < 0.05 was the lowest in alfalfa in the last stage of sampling (62.6% when it reached a mere 85.99% of the digestibility at the beginning of bud setting. The highest organic matter digestibility (72.80% was detected in a sample of young alfalfa prior to bud setting. In the experiment we confirmed a strong and negative correlation (r = −0.97 between the crude fibre content and organic matter digestibility. The correlation was also strong between the stage of maturity of alfalfa and the content of magnesium (Mg.