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Sample records for nodulated alfalfa plants

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

    OpenAIRE

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

    2014-01-01

    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 characteri...

  2. 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.

  3. CO/sub 2/ fixation in alfalfa and birdsfoot trefoil root nodules and partitioning of /sup 14/C to the plant

    Energy Technology Data Exchange (ETDEWEB)

    Maxwell, C.A.; Vance, C.P.; Heichel, G.H.; Stade, S.

    The objectives of this study were to determine if nonphotosynthetic CO/sub 2/ fixation by root nodules contributes carbon for the assimilation of fixed N/sub 2/ in alfalfa (Medicago sativa L.) and birdsfoot trefoil (Lotus corniculatus L.) and if assimilation products are partitioned to different plant organs. Effective alfalfa nodules excised from or attached to roots had apparent /sup 14/CO/sub 2/ fixation rates of 50 to 80 ..mu..g CO/sub 2/ kg/sup -1/s/sup -1/ (dry weight) at 0.0012 to 0.0038 mole fraction CO/sub 2/. Nodule CO/sub 2/ fixation rates increased six- to seven-fold as ambient CO/sub 2/ was raised from 0.0038 to 0.0663 mole fraction. Respiration rates of nodules (3 to 4 mg CO/sub 2/ kg/sup -1/s/sup -1/) were 10 to 100-fold higher than /sup 14/CO/sub 2/ fixation rates of nodules. Pulse chase experiments with /sup 14/CO/sub 2/ combined with nodule and xylem sap analysis demonstrated the initial products of root and nodule CO/sub 2/ fixation were organic acids. However, the export of fixed /sup 14/C from effective nodules was primarily in the form of amino acids. In contrast, nodule and/or root fixed /sup 14/C in ineffectively nodulated alfalfa and denodulated effective alfalfa and birdsfoot trefoil was transported primarily as organic acids. Aspartate, asparagine, alanine, glutamate, and glutamine were the most heavily labeled compounds in the amino acid fraction of both effective alfalfa and birdsfoot trefoil nodules exposed to /sup 14/CO/sub 2/. By contrast, asparate, asparagine, and glutamine were the predominantly labeled amino acids in xylem sap collected from nodulated effective roots exposed to /sup 14/CO/sub 2/. The occurrence of nodule CO/sub 2/ fixation in alfalfa and birdsfoot trefoil and the export of fixed carbon as asparagine and aspartate to roots and shoots is consistent with a role for CO/sub 2/ fixation by nodules in providing carbon skeletons for assimilation and transport of symbiotically fixed N/sub 2/.

  4. Diversity of field isolates of sinorhizobium meliloti nodulating alfalfa

    Science.gov (United States)

    Most alfalfa seed is treated with a rhizobial inoculant consisting of one or more strains of Sinorhizobium meliloti before planting to enhance nodulation of seedlings. However, little is known about the persistence of inoculated strains later in the season. There is also a paucity of information on ...

  5. Lignin modification leads to increased nodule numbers in alfalfa.

    Science.gov (United States)

    Gallego-Giraldo, Lina; Bhattarai, Kishor; Pislariu, Catalina I; Nakashima, Jin; Jikumaru, Yusuke; Kamiya, Yuji; Udvardi, Michael K; Monteros, Maria J; Dixon, Richard A

    2014-03-01

    Reduction of lignin levels in the forage legume alfalfa (Medicago sativa) by down-regulation of the monolignol biosynthetic enzyme hydroxycinnamoyl coenzyme A:shikimate hydroxycinnamoyl transferase (HCT) results in strongly increased digestibility and processing ability of lignocellulose. However, these modifications are often also associated with dwarfing and other changes in plant growth. Given the importance of nitrogen fixation for legume growth, we evaluated the impact of constitutively targeted lignin modification on the belowground organs (roots and nodules) of alfalfa plants. HCT down-regulated alfalfa plants exhibit a striking reduction in root growth accompanied by an unexpected increase in nodule numbers when grown in the greenhouse or in the field. This phenotype is associated with increased levels of gibberellins and certain flavonoid compounds in roots. Although HCT down-regulation reduced biomass yields in both the greenhouse and field experiments, the impact on the allocation of nitrogen to shoots or roots was minimal. It is unlikely, therefore, that the altered growth phenotype of reduced-lignin alfalfa is a direct result of changes in nodulation or nitrogen fixation efficiency. Furthermore, HCT down-regulation has no measurable effect on carbon allocation to roots in either greenhouse or 3-year field trials.

  6. The tep1 gene of Sinorhizobium meliloti coding for a putative transmembrane efflux protein and N-acetyl glucosamine affect nod gene expression and nodulation of alfalfa plants

    Directory of Open Access Journals (Sweden)

    Soto María

    2009-01-01

    Full Text Available Abstract Background Soil bacteria collectively known as Rhizobium, characterized by their ability to establish beneficial symbiosis with legumes, share several common characteristics with pathogenic bacteria when infecting the host plant. Recently, it was demonstrated that a fadD mutant of Sinorhizobium meliloti is altered in the control of swarming, a type of co-ordinated movement previously associated with pathogenicity, and is also impaired in nodulation efficiency on alfalfa roots. In the phytopathogen Xanthomonas campestris, a fadD homolog (rpfB forms part of a cluster of genes involved in the regulation of pathogenicity factors. In this work, we have investigated the role in swarming and symbiosis of SMc02161, a S. meliloti fadD-linked gene. Results The SMc02161 locus in S. meliloti shows similarities with members of the Major Facilitator Superfamily (MFS of transporters. A S. meliloti null-mutant shows increased sensitivity to chloramphenicol. This indication led us to rename the locus tep1 for transmembrane efflux protein. The lack of tep1 does not affect the appearance of swarming motility. Interestingly, nodule formation efficiency on alfalfa plants is improved in the tep1 mutant during the first days of the interaction though nod gene expression is lower than in the wild type strain. Curiously, a nodC mutation or the addition of N-acetyl glucosamine to the wild type strain lead to similar reductions in nod gene expression as in the tep1 mutant. Moreover, aminosugar precursors of Nod factors inhibit nodulation. Conclusion tep1 putatively encodes a transmembrane protein which can confer chloramphenicol resistance in S. meliloti by expelling the antibiotic outside the bacteria. The improved nodulation of alfalfa but reduced nod gene expression observed in the tep1 mutant suggests that Tep1 transports compounds which influence nodulation. In contrast to Bradyrhizobium japonicum, we show that in S. meliloti there is no feedback regulation

  7. The tep1 gene of Sinorhizobium meliloti coding for a putative transmembrane efflux protein and N-acetyl glucosamine affect nod gene expression and nodulation of alfalfa plants.

    Science.gov (United States)

    van Dillewijn, Pieter; Sanjuán, Juan; Olivares, José; Soto, María José

    2009-01-27

    Soil bacteria collectively known as Rhizobium, characterized by their ability to establish beneficial symbiosis with legumes, share several common characteristics with pathogenic bacteria when infecting the host plant. Recently, it was demonstrated that a fadD mutant of Sinorhizobium meliloti is altered in the control of swarming, a type of co-ordinated movement previously associated with pathogenicity, and is also impaired in nodulation efficiency on alfalfa roots. In the phytopathogen Xanthomonas campestris, a fadD homolog (rpfB) forms part of a cluster of genes involved in the regulation of pathogenicity factors. In this work, we have investigated the role in swarming and symbiosis of SMc02161, a S. meliloti fadD-linked gene. The SMc02161 locus in S. meliloti shows similarities with members of the Major Facilitator Superfamily (MFS) of transporters. A S. meliloti null-mutant shows increased sensitivity to chloramphenicol. This indication led us to rename the locus tep1 for transmembrane efflux protein. The lack of tep1 does not affect the appearance of swarming motility. Interestingly, nodule formation efficiency on alfalfa plants is improved in the tep1 mutant during the first days of the interaction though nod gene expression is lower than in the wild type strain. Curiously, a nodC mutation or the addition of N-acetyl glucosamine to the wild type strain lead to similar reductions in nod gene expression as in the tep1 mutant. Moreover, aminosugar precursors of Nod factors inhibit nodulation. tep1 putatively encodes a transmembrane protein which can confer chloramphenicol resistance in S. meliloti by expelling the antibiotic outside the bacteria. The improved nodulation of alfalfa but reduced nod gene expression observed in the tep1 mutant suggests that Tep1 transports compounds which influence nodulation. In contrast to Bradyrhizobium japonicum, we show that in S. meliloti there is no feedback regulation of nodulation genes. Moreover, the Nod factor precursor

  8. Genetic diversity of indigenous Rhizobium leguminosarum bv. viciae isolates nodulating two different host plants during soil restoration with alfalfa.

    Science.gov (United States)

    Zhang, X X; Kosier, B; Priefer, U B

    2001-09-01

    A total of 360 Rhizobium leguminosarum bv. viciae strains was isolated from three brown-coal mining restoration fields of different age and plant cover (without and in the first and second year of alfalfa, Medicago sativa, cultivation) using two host species (Vicia hirsuta and Pisum sativum) as capture plants. The strains were genetically typed by restriction fragment length polymorphism analysis of polymerase chain reaction (PCR)-generated 16S-23S ribosomal DNA intergenic spacer regions (IGS-RFLP) and characterized by plasmid profiles and RFLP analysis of amplified nodABC genes. The R. leguminosarum bv. viciae population was dominated by the same group of strains (irrespective of the trap plant used). According to type richness, the genetic diversity of indigenous R. leguminosarum in the second year of restoration was lower than in the first year and it resembled that of the fallow field, except for plasmid types, in which it was higher than that of the fallow field. Some of the less frequent nodABC genotypes were associated with distinct chromosomal IGS genotypes and symbiotic plasmids (pSyms) of different sizes, indicating that horizontal transfer and rearrangements of pSym can occur in natural environments. However, the dominant pSym and chromosomal genotypes were strictly correlated suggesting a genetically stable persistence of the prevailing R. leguminosarum bv. viciae genotypes in the absence of its host plant.

  9. Genetic diversity of indigenous Rhizobium leguminosarum bv. viciae isolates nodulating two different host plants during soil restoration with alfalfa

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, X.X.; Kosier, B.; Priefer, U.B. [Rheinisch-Westfaelische TH Aachen, Aachen (Germany)

    2001-09-01

    A total of 360 Rhizobium leguminosarum bv. viciae strains was isolated from three brown-coal mining restoration fields of different age and plant cover (without and in the first and second year of alfalfa, Medicago sativa, cultivation) using two host species (Vicia hirsuta and Pisum sativum) as capture plants. The strains were genetically typed by restriction fragment length polymorphism analysis of polymerase chain reaction (PCR)-generated 16S-23S ribosomal DNA intergenic spacer regions (IGS-RFLP) and characterized by plasmid profiles and RFLP analysis of amplified nodABC genes. The R. leguminosarum bv. viciae population was dominated by the same group of strains (irrespective of the trap plant used). According to type richness, the genetic diversity of indigenous R. leguminosarum in the second year of restoration was lower than in the first year and it resembled that of the fallow field, except for plasmid types, in which it was higher than that of the fallow field. Some of the less frequent nodABC genotypes were associated with distinct chromosomal IGS genotypes and symbiotic plasmids (pSyms) of different sizes, indicating that horizontal transfer and rearrangements of pSym can occur in natural environments. However, the dominant pSym and chromosomal genotypes were strictly correlated suggesting a genetically stable persistence of the prevailing R. leguminosarum bv, viciae genotypes in the absence of its host plant.

  10. Persistence and diversity of rhizobial bacteria nodulating alfalfa

    Science.gov (United States)

    Most alfalfa seed is treated with an inoculant consisting of several strains of the nitrogen fixing bacterium Sinorhizobium meliloti to enhance nodulation of seedlings. One strategy for increasing alfalfa forage yields, particularly in less fertile sites, is selection and use of highly competitive a...

  11. [Alfalfa Planting as weed control

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — This document is a letter to farming cooperators regarding the stipulations surrounding alfalfa plantings in lieu of small grain plantings to provide weed control,...

  12. Nodulin gene expression and ENOD2 localization in effective, nitrogen fixing and ineffective, bacteria-free nodules of alfalfa.

    NARCIS (Netherlands)

    Wiel, van de C.C.M.; Nurris, J.H.; Bocheneck, B.; Dickstein, R.; Bisseling, T.; Hirsch, A.M.

    1990-01-01

    Alfalfa plants form bacteria-free nodules in response to a number of agents, including Rhizobium meliloti exo mutants, Agrobacterium tumefaciens transconjugants carrying cloned R. meliloti nodulation genes, and compounds that function as auxin transport inhibitors, N-( 1-naphthyl)phthalamic acid or

  13. Nodulin gene expression and ENOD2 localization in effective, nitrogen fixing and ineffective, bacteria-free nodules of alfalfa.

    NARCIS (Netherlands)

    Wiel, van de C.C.M.; Nurris, J.H.; Bocheneck, B.; Dickstein, R.; Bisseling, T.; Hirsch, A.M.

    1990-01-01

    Alfalfa plants form bacteria-free nodules in response to a number of agents, including Rhizobium meliloti exo mutants, Agrobacterium tumefaciens transconjugants carrying cloned R. meliloti nodulation genes, and compounds that function as auxin transport inhibitors, N-( 1-naphthyl)phthalamic acid or

  14. Alfalfa Enod12 genes are differentially regulated during nodule development by Nod factors and Rhizobium invasion.

    Science.gov (United States)

    Bauer, P; Crespi, M D; Szécsi, J; Allison, L A; Schultze, M; Ratet, P; Kondorosi, E; Kondorosi, A

    1994-01-01

    MsEnod12A and MsEnod12B are two early nodulin genes from alfalfa (Medicago sativa). Differential expression of these genes was demonstrated using a reverse transcription-polymerase chain reaction approach. MsEnod12A RNA was detected only in nodules and not in other plant tissues. In contrast, MsEnod12B transcripts were found in nodules and also at low levels in roots, flowers, stems, and leaves. MsEnod12B expression was enhanced in the root early after inoculation with the microsymbiont Rhizobium meliloti and after treatment with purified Nod factors, whereas MsEnod12A induction was detected only when developing nodules were visible. In situ hybridization showed that in nodules, MsEnod12 expression occurred in the infection zone. In empty Fix- nodules the MsEnod12A transcript level was much reduced, and in spontaneous nodules it was not detectable. These data indicate that MsEnod12B expression in roots is related to the action of Nod factors, whereas MsEnod12A expression is associated with the invasion process in nodules. Therefore, alfalfa possesses different mechanisms regulating MsEnod12A and MsEnod12B expression. PMID:8066132

  15. Screening of Highly Effective Sinorhizobium meliloti Strains for 'Vector' Alfalfa and Testing of Its Competitive Nodulation Ability in the Field

    Institute of Scientific and Technical Information of China (English)

    ZENG Zhao-Hai; CHEN Wen-Xin; HU Yue-Gao; SUI Xin-Hua; CHEN Dan-Ming

    2007-01-01

    Seventeen Sinorhizobium meliloti strains from seven provinces in China were used to screen highly effective strains for alfalfa cultivar in a greenhouse study and their symbiotic relationship and competitive ability were studied in the field.CCBAU30138 was the most effective strain,as evidenced by increase in dry weights.A field experiment showed that the inoculation of alfalfa with CCBAU30138 resulted in increases of 11.9%and 19.6%of dry matter production and crude protein production,respectively,in forage of monocultured plants.The total dry matter yields of alfalfa and tall fescue in binary culture were increased by 16.3%by inoculation of alfalfa with this strain.These results showed that S.Meliloti strain CCBAU30138 was an effective inoculant both in the greenhouse and in the field.The analysis of randomly amplified polymorphic DNA(RAPD)by polymerase chain reaction(PCR)from nodule extracts showed that the strain CCBAU30138 had high competitiveness in the field.It occupied 47.5%of nodules in alfalfa monoculture and 44.4% of nodules in alfalfa-tall fescue binary culture after 20 weeks of growth.In conclusion,a simple system to select highly effective and competitive symbiotic strains specific to alfalfa was established.Using this system.A strain suitable for the alfalfa cultivar'Vector’grown in Wuqiao County of Hebei Province was obtained.

  16. Molybdate in Rhizobial Seed-Coat Formulations Improves the Production and Nodulation of Alfalfa.

    Science.gov (United States)

    Zhou, Jiqiong; Deng, Bo; Zhang, Yingjun; Cobb, Adam B; Zhang, Zhao

    2017-01-01

    Rhizobia-legume symbiosis is the most well researched biological nitrogen fixation system. Coating legume seeds with rhizobia is now a recognized practical measure for improving the production of legume corp. However, the efficacy of some commercial rhizobia inoculants cannot be guaranteed in China due to the low rate of live rhizobia in these products. A greenhouse experiment was conducted to assess the effects of different rhizobial inoculant formulations on alfalfa productivity and nitrogen fixation. Two rhizobia strains, (ACCC17631 and ACCC17676), that are effective partners with alfalfa variety Zhongmu No. 1 were assessed with different concentrations of ammonium molybdate in seed-coat formulations with two different coating adhesives. Our study showed that the growth, nodulation, and nitrogen fixation ability of the plants inoculated with the ACCC17631 rhizobial strain were greatest when the ammonium molybdate application was0.2% of the formulation. An ammonium molybdate concentration of 0.1% was most beneficial to the growth of the plants inoculated with the ACCC17676 rhizobial strain. The sodium carboxymethyl cellulose and sodium alginate, used as coating adhesives, did not have a significant effect on alfalfa biomass and nitrogen fixation. However, the addition of skimmed milk to the adhesive improved nitrogenase activity. These results demonstrate that a new rhizobial seed-coat formulation benefitted alfalfa nodulation and yield.

  17. Molybdate in Rhizobial Seed-Coat Formulations Improves the Production and Nodulation of Alfalfa

    Science.gov (United States)

    Zhou, Jiqiong; Deng, Bo; Zhang, Yingjun; Cobb, Adam B.; Zhang, Zhao

    2017-01-01

    Rhizobia-legume symbiosis is the most well researched biological nitrogen fixation system. Coating legume seeds with rhizobia is now a recognized practical measure for improving the production of legume corp. However, the efficacy of some commercial rhizobia inoculants cannot be guaranteed in China due to the low rate of live rhizobia in these products. A greenhouse experiment was conducted to assess the effects of different rhizobial inoculant formulations on alfalfa productivity and nitrogen fixation. Two rhizobia strains, (ACCC17631 and ACCC17676), that are effective partners with alfalfa variety Zhongmu No. 1 were assessed with different concentrations of ammonium molybdate in seed-coat formulations with two different coating adhesives. Our study showed that the growth, nodulation, and nitrogen fixation ability of the plants inoculated with the ACCC17631 rhizobial strain were greatest when the ammonium molybdate application was0.2% of the formulation. An ammonium molybdate concentration of 0.1% was most beneficial to the growth of the plants inoculated with the ACCC17676 rhizobial strain. The sodium carboxymethyl cellulose and sodium alginate, used as coating adhesives, did not have a significant effect on alfalfa biomass and nitrogen fixation. However, the addition of skimmed milk to the adhesive improved nitrogenase activity. These results demonstrate that a new rhizobial seed-coat formulation benefitted alfalfa nodulation and yield. PMID:28099471

  18. Expression of nodule-specific genes in alfalfa root nodules blocked at an early stage of development.

    NARCIS (Netherlands)

    Dickstein, R.; Bisseling, T.; Reinhold, V.N.; Ausubel, F.M.

    1988-01-01

    To help dissect the molecular basis of the Rhizobium-legume symbiosis, we used in vitro translation and Northern blot analysis of nodule RNA to examine alfalfa-specific genes (nodulins) expressed in two types of developmentally defective root nodules elicited by Rhizobium meliloti. Fix- nodules were

  19. Expression of nodule-specific genes in alfalfa root nodules blocked at an early stage of development.

    NARCIS (Netherlands)

    Dickstein, R.; Bisseling, T.; Reinhold, V.N.; Ausubel, F.M.

    1988-01-01

    To help dissect the molecular basis of the Rhizobium-legume symbiosis, we used in vitro translation and Northern blot analysis of nodule RNA to examine alfalfa-specific genes (nodulins) expressed in two types of developmentally defective root nodules elicited by Rhizobium meliloti. Fix- nodules were

  20. Nodulation competitiveness of Ensifer meliloti alfalfa nodule isolates and their potential for application as inoculants.

    Science.gov (United States)

    Marek-Kozaczuk, Monika; Wielbo, Jerzy; Pawlik, Anna; Skorupska, Anna

    2014-01-01

    Alfalfa (Medicago sativa) is a widely cultivated legume, which enters into nitrogen-fixing symbiosis with Ensifer (Sinorhizobium) spp. In this study, an autochthonous rhizobial population of Ensifer sp. occupying alfalfa nodules grown in arable soil was used as the basis for selection of potential inoculants. Alfalfa nodule isolates were identified as Ensifer meliloti by partial 16S rDNA, recA, atpD and nodC nucleotide sequencing. The sampled isolates displayed different symbiotic performance and diversity in the number of plasmids and molecular weight. Isolates that were the most efficient in symbiotic nitrogen fixation were tagged with a constitutively expressed gusA gene carried by a stable plasmid vector pJBA21Tc and used in competition experiments in soil under greenhouse conditions. Two E. meliloti strains LU09 and LU12, which effectively competed with indigenous soil rhizobia, were selected. The metabolic profiles of these selected strains showed differences in the use of carbon and energy sources. In addition, the LU09 strain exhibited bacteriocin production and LU12 mineral phosphate solubilization, which are valuable traits for soil survival. These strains may be considered as potential biofertilizers for alfalfa cultivation.

  1. Rhizobial strains exert a major effect on the amino acid composition of alfalfa nodules under NaCl stress.

    Science.gov (United States)

    Bertrand, Annick; Bipfubusa, Marie; Dhont, Catherine; Chalifour, François-P; Drouin, Pascal; Beauchamp, Chantal J

    2016-11-01

    Specific amino acids have protective functions in plants under stress conditions. This study assessed the effects of rhizobial strains on the amino acid composition in alfalfa under salt stress. Two alfalfa cultivars (Medicago sativa L. cv Apica and salt-tolerant cv Halo) in association with two Sinorhizobium meliloti strains with contrasting growth under salt stress (strain A2 and salt-tolerant strain Rm1521) were exposed to different levels of NaCl (0, 20, 40, 80 or 160 mM NaCl) under controlled conditions. We compared root and shoot biomasses, as well as root:shoot ratio for each association under these conditions as indicators of the salt tolerance of the symbiosis. Amino acid concentrations were analyzed in nodules, leaves and roots. The total concentration of free amino acids in nodules was mostly rhizobial-strain dependent while in leaves and roots it was mostly responsive to salt stress. For both cultivars, total and individual concentrations of amino acids including asparagine, proline, glutamine, aspartate, glutamate, γ-aminobutyric acid (GABA), histidine and ornithine were higher in Rm1521 nodules than in A2 nodules. Conversely, lysine and methionine were more abundant in A2 nodules than in Rm1521 nodules. Proline, glutamine, arginine, GABA and histidine substantially accumulated in salt-stressed nodules, suggesting an enhanced production of amino acids associated with osmoregulation, N storage or energy metabolism to counteract salt stress. Combining the salt-tolerant strain Rm1521 and the salt-tolerant cultivar Halo enhanced the root:shoot ratios and amino acid concentrations involved in plant protection which could be in part responsible for the enhancement of salt tolerance in alfalfa. Crown Copyright © 2016. Published by Elsevier Masson SAS. All rights reserved.

  2. Antibacterial activity of plant defensins against alfalfa crown rot pathogens

    Science.gov (United States)

    Alfalfa (Medicago sativa) is the fourth most widely grown crop in the United States. Alfalfa crown rot is a disease complex that severely decreases alfalfa stand density and productivity in all alfalfa-producing areas. Currently, there are no viable methods of disease control. Plant defensins are sm...

  3. ALFALFA

    OpenAIRE

    Mirko Stjepanović; Robert Zimmer; Marijana Tucak; Gordana Bukvić; Svetislav Popović; Zvonimir Štafa

    2009-01-01

    The academic textbook Alfalfa contains the following chapters: Review outline, Foreword, Origin and spread of alfalfa, Economic importance of alfalfa, Biological properties and breeding alfalfa, Morphological traits of alfalfa, Growth and development of alfalfa, Ecological conditions for alfalfa growth, Agronomy of alfalfa for fodder production, Pests and diseases of alfalfa and their control, Usage of alfalfa and hay, silage and haylage production, Alfalfa seed production, Prospects for alfa...

  4. Relationship between photosynthetic capacity, nitrogen assimilation and nodule metabolism in alfalfa (Medicago sativa) grown with sewage sludge.

    Science.gov (United States)

    Antolín, M Carmen; Fiasconaro, M Laura; Sánchez-Díaz, Manuel

    2010-10-15

    Sewage sludge has been used as N fertilizer because it contains some of inorganic N, principally as nitrate and ammonium ions. However, sewage sludge addition to legumes could result in impaired nodule metabolism due to the presence of inorganic N from sludge. A greenhouse experiment was conducted to examine the effects of sewage sludge on growth, photosynthesis, nitrogen assimilation and nodule metabolism in alfalfa (Medicago sativa L. cv. Aragón). Plants were grown in pots with a mixture of perlite and vermiculite (2:1, v/v). The experiment included three treatments: (1) plants inoculated with rhizobia and amended with sewage sludge at rate of 10% (w/w) (RS); (2) plants inoculated with rhizobia without any amendment (R); and (3) non-inoculated plants fed with ammonium nitrate (N). N(2)-fixing plants had lower growth and sucrose phosphate synthase activity but higher photosynthesis than nitrate-fed plants because they compensated the carbon cost of the rhizobia. However, sewage sludge-treated plants evidenced a loss of carbon sink strength due to N(2) fixation by means of decreased photosynthetic capacity, leaf chlorophylls and N concentration in comparison to untreated plants. Sewage sludge did no affect nodulation but decreased nodule enzyme activities involved in carbon and N metabolisms that may lead to accumulation of toxic N-compounds.

  5. Composted organic wastes from the pharmaceutical and agro-food industries induce soil bioactivity and nodulation in alfalfa.

    Science.gov (United States)

    Fornes, Fernando; Jaramillo, Claudia X; García-de-la-Fuente, Rosana; Belda, Rosa M; Lidón, Antonio

    2014-11-01

    Environmentally friendly agriculture needs to reduce the use of synthetic fertilizers and to reclaim nutrients from organic wastes. In this study the effect of five doses (0, 12, 24, 48 and 96 t ha(-1) ) of two two-phase olive mill waste (TPOMW)-based composts on the bioactivity and chemical characteristics of an agricultural soil and their potential to fertilize alfalfa (Medicago sativa) and stimulate nodulation were assessed during a two-year incubation experiment. The two composts were prepared either with the olive mill waste alone (compost A), which served as control, or mixed with a liquid fatty-proteinaceous hydrolyzate waste (FPH) from the pharmaceutical industry (compost AH). Compost AH resulted in greater N immobilization than compost A because the former supplied the soil with easily degradable C and N, which increased microbial biomass and activity. Both compost mineralizations during the first year of incubation supplied the soil with more nutrients (mainly N), more so with A than with AH. Nevertheless, plant growth was similar in soils amended with either A or AH. Both composts induced nodulation similarly and the highest dose (96 t ha(-1) ) increased the formation of nodules by a factor of 11 compared with the four lower doses. TPOMW serves as an effective ground material for co-composting with liquid wastes such as FPH. TPOMW supplies key nutrients and stimulates nodulation in alfalfa. © 2014 Society of Chemical Industry.

  6. Effects of water stress on antioxidant enzymes of leaves and nodules of transgenic alfalfa overexpressing superoxide dismutases.

    Science.gov (United States)

    Rubio, Maria C; González, Esther M; Minchin, Frank R; Webb, K. Judith; Arrese-Igor, Cesar; Ramos, Javier; Becana, Manuel

    2002-08-01

    The antioxidant composition and relative water stress tolerance of nodulated alfalfa plants (Medicago sativa L. x Sinorhizobium meliloti 102F78) of the elite genotype N4 and three derived transgenic lines have been studied in detail. These transgenic lines overproduced, respectively, Mn-containing superoxide dismutase (SOD) in the mitochondria of leaves and nodules, MnSOD in the chloroplasts, and FeSOD in the chloroplasts. In general for all lines, water stress caused moderate decreases in MnSOD and FeSOD activities in both leaves and nodules, but had distinct tissue-dependent effects on the activities of the peroxide-scavenging enzymes. During water stress, with a few exceptions, ascorbate peroxidase and catalase activities increased moderately in leaves but decreased in nodules. At mild water stress, transgenic lines showed, on average, 20% higher photosynthetic activity than the parental line, which suggests a superior tolerance of transgenic plants under these conditions. However, the untransformed and the transgenic plants performed similarly during moderate and severe water stress and recovery with respect to important markers of metabolic activity and of oxidative stress in leaves and nodules. We conclude that the base genotype used for transformation and the background SOD isozymic composition may have a profound effect on the relative tolerance of the transgenic lines to abiotic stress.

  7. Oxygen Regulation of a Nodule-Located Carbonic Anhydrase in Alfalfa1

    Science.gov (United States)

    Gálvez, Susana; Hirsch, Ann M.; Wycoff, Keith L.; Hunt, Stephen; Layzell, David B.; Kondorosi, Adam; Crespi, Martin

    2000-01-01

    Control of the permeability to oxygen is critical for the function of symbiotic nitrogen fixation in legume nodules. The inner cortex (IC) seems to be a primary site for this regulation. In alfalfa (Medicago sativa) nodules, expression of the Msca1 gene encoding a carbonic anhydrase (CA) was previously found to be restricted to the IC. We have now raised antibodies against recombinant Msca1 protein and used them, together with antibodies raised against potato leaf CA, to demonstrate the presence of two forms of CA in mature nodules. Each antibody recognizes a different CA isoform in nodule tissues. Immunolocalization revealed that leaf-related CAs were localized primarily in the nitrogen-fixing zone, whereas the Msca1 protein was restricted exclusively to the IC region, in indeterminate and determinate nodules. In alfalfa nodules grown at various O2 concentrations, an inverse correlation was observed between the external oxygen pressure and Msca1 protein content in the IC, the site of the putative diffusion barrier. Thus Msca1 is a molecular target of physiological processes occurring in the IC cells involved in gas exchange in the nodule. PMID:11080283

  8. Synthesis of an opine-like compound, a rhizopine, in alfalfa nodules is symbiotically regulated.

    OpenAIRE

    Murphy, P J; Heycke, N; Trenz, S P; Ratet, P.; de Bruijn, F J; Schell, J.

    1988-01-01

    We show that the promoter of the mos locus, which encodes genes required for the synthesis of a nodule-specific, opine-like compound, a rhizopine, in alfalfa nodules is regulated by the symbiotic nitrogen-fixation regulatory gene nifA. The 5'-regulatory region and amino-terminal end of the first open reading frame of the mos locus are highly homologous to the 5'-regulatory region and amino-terminal portion of the Rhizobium meliloti nifH gene. The coordinate regulation of mos and nif genes sug...

  9. 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.

  10. Requirement of succinate dehydrogenase activity for symbiotic bacteroid differentiation of Rhizobium meliloti in alfalfa nodules.

    OpenAIRE

    Gardiol, A E; Truchet, G L; Dazzo, F. B.

    1987-01-01

    Transmission electron microscopy was used to study the cellular morphologies of a wild-type Rhizobium meliloti strain (L5-30), a nitrogen fixation-ineffective (Fix-) succinate dehydrogenase mutant (Sdh-) strain, and a Fix+ Sdh+ revertant strain within alfalfa nodules and after free-living growth in a minimal medium containing 27 mM mannitol plus 20 mM succinate. The results showed a requirement of succinate dehydrogenase activity for symbiotic differentiation and maintenance of R. meliloti ba...

  11. Construction and environmental release of a Sinorhizobium meliloti strain genetically modified to be more competitive for alfalfa nodulation.

    Science.gov (United States)

    van Dillewijn, P; Soto, M J; Villadas, P J; Toro, N

    2001-09-01

    Highly efficient nitrogen-fixing strains selected in the laboratory often fail to increase legume production in agricultural soils containing indigenous rhizobial populations because they cannot compete against these populations for nodule formation. We have previously demonstrated, with a Sinorhizobium meliloti PutA- mutant strain, that proline dehydrogenase activity is required for colonization and therefore for the nodulation efficiency and competitiveness of S. meliloti on alfalfa roots (J. I. Jiménez-Zurdo, P. van Dillewijn, M. J. Soto, M. R. de Felipe, J. Olivares, and N. Toro, Mol. Plant-Microbe Interact. 8:492-498, 1995). In this work, we investigated whether the putA gene could be used as a means of increasing the competitiveness of S. meliloti strains. We produced a construct in which a constitutive promoter was placed 190 nucleotides upstream from the start codon of the putA gene. This resulted in an increase in the basal expression of this gene, with this increase being even greater in the presence of the substrate proline. We found that the presence of multicopy plasmids containing this putA gene construct increased the competitiveness of S. meliloti in microcosm experiments in nonsterile soil planted with alfalfa plants subjected to drought stress only during the first month. We investigated whether this construct also increased the competitiveness of S. meliloti strains under agricultural conditions by using it as the inoculum in a contained field experiment at León, Spain. We found that the frequency of nodule occupancy was higher with inoculum containing the modified putA gene for samples that were analyzed after 34 days but not for samples that were analyzed later.

  12. The Sinorhizobium meliloti ntrX gene is involved in succinoglycan production, motility, and symbiotic nodulation on alfalfa.

    Science.gov (United States)

    Wang, Dong; Xue, Haiying; Wang, Yiwen; Yin, Ruochun; Xie, Fang; Luo, Li

    2013-12-01

    Rhizobia establish a symbiotic relationship with their host legumes to induce the formation of nitrogen-fixing nodules. This process is regulated by many rhizobium regulators, including some two-component regulatory systems (TCSs). NtrY/NtrX, a TCS that was first identified in Azorhizobium caulinodans, is required for free-living nitrogen metabolism and symbiotic nodulation on Sesbania rostrata. However, its functions in a typical rhizobium such as Sinorhizobium meliloti remain unclear. Here we found that the S. meliloti response regulator NtrX but not the histidine kinase NtrY is involved in the regulation of exopolysaccharide production, motility, and symbiosis with alfalfa. A plasmid insertion mutant of ntrX formed mucous colonies, which overproduced succinoglycan, an exopolysaccharide, by upregulating its biosynthesis genes. This mutant also exhibited motility defects due to reduced flagella and decreased expression of flagellins and regulatory genes. The regulation is independent of the known regulatory systems of ExoR/ExoS/ChvI, EmmABC, and ExpR. Alfalfa plants inoculated with the ntrX mutant were small and displayed symptoms of nitrogen starvation. Interestingly, the deletion mutant of ntrY showed a phenotype similar to that of the parent strain. These findings demonstrate that the S. meliloti NtrX is a new regulator of succinoglycan production and motility that is not genetically coupled with NtrY.

  13. Alfalfa

    Science.gov (United States)

    Alfalfa is an herb. People use the leaves, sprouts, and seeds to make medicine. Alfalfa is used for kidney conditions, bladder and prostate ... bleeding disorder called thrombocytopenic purpura. People also take alfalfa as a source of vitamins A, C, E, ...

  14. The Nodulation of Alfalfa by the Acid-Tolerant Rhizobium sp. Strain LPU83 Does Not Require Sulfated Forms of Lipochitooligosaccharide Nodulation Signals▿

    Science.gov (United States)

    Torres Tejerizo, Gonzalo; Del Papa, María Florencia; Soria-Diaz, M. Eugenia; Draghi, Walter; Lozano, Mauricio; Giusti, María de los Ángeles; Manyani, Hamid; Megías, Manuel; Gil Serrano, Antonio; Pühler, Alfred; Niehaus, Karsten; Lagares, Antonio; Pistorio, Mariano

    2011-01-01

    The induction of root nodules by the majority of rhizobia has a strict requirement for the secretion of symbiosis-specific lipochitooligosaccharides (nodulation factors [NFs]). The nature of the chemical substitution on the NFs depends on the particular rhizobium and contributes to the host specificity imparted by the NFs. We present here a description of the genetic organization of the nod gene cluster and the characterization of the chemical structure of the NFs associated with the broad-host-range Rhizobium sp. strain LPU83, a bacterium capable of nodulating at least alfalfa, bean, and Leucena leucocephala. The nod gene cluster was located on the plasmid pLPU83b. The organization of the cluster showed synteny with those of the alfalfa-nodulating rhizobia, Sinorhizobium meliloti and Sinorhizobium medicae. Interestingly, the strongest sequence similarity observed was between the partial nod sequences of Rhizobium mongolense USDA 1844 and the corresponding LPU83 nod genes sequences. The phylogenetic analysis of the intergenic region nodEG positions strain LPU83 and the type strain R. mongolense 1844 in the same branch, which indicates that Rhizobium sp. strain LPU83 might represent an early alfalfa-nodulating genotype. The NF chemical structures obtained for the wild-type strain consist of a trimeric, tetrameric, and pentameric chitin backbone that shares some substitutions with both alfalfa- and bean-nodulating rhizobia. Remarkably, while in strain LPU83 most of the NFs were sulfated in their reducing terminal residue, none of the NFs isolated from the nodH mutant LPU83-H were sulfated. The evidence obtained supports the notion that the sulfate decoration of NFs in LPU83 is not necessary for alfalfa nodulation. PMID:20971905

  15. Isolation and Characterization of Alfalfa-Nodulating Rhizobia Present in Acidic Soils of Central Argentina and Uruguay

    OpenAIRE

    1999-01-01

    We describe the isolation and characterization of alfalfa-nodulating rhizobia from acid soils of different locations in Central Argentina and Uruguay. A collection of 465 isolates was assembled, and the rhizobia were characterized for acid tolerance. Growth tests revealed the existence of 15 acid-tolerant (AT) isolates which were able to grow at pH 5.0 and formed nodules in alfalfa with a low rate of nitrogen fixation. Analysis of those isolates, including partial sequencing of the genes enco...

  16. Biogeography of Sinorhizobium meliloti nodulating alfalfa in different Croatian regions.

    Science.gov (United States)

    Donnarumma, Francesca; Bazzicalupo, Marco; Blažinkov, Mihaela; Mengoni, Alessio; Sikora, Sanjia; Babić, Katarina Huić

    2014-09-01

    Sinorhizobium meliloti is a nitrogen-fixing rhizobium symbiont of legumes, widespread in many temperate environments the high genetic diversity of which enables it to thrive as a symbiont of host legumes and free-living in soil. Soil type, together with geographic differences and host plant genotype, seem to be prominent factors in shaping rhizobial genetic diversity. While a large body of research supports the idea that the genetic structure of free-living microbial taxa exhibits a clear biogeographic pattern, few investigations have been performed on the biogeographic pattern of S. meliloti genotypes in a restricted geographic range. In the present study, a collection of 128 S. meliloti isolates from three different regions in Croatia was investigated to analyze the relationship between genetic diversity, geographic distribution, soil features and isolate phenotypes by using amplified fragment length polymorphism (AFLP) as a genome-wide scanning method. Results obtained led to the conclusion that the genotypes of isolates cluster according to the region of origin and that the differentiation of S. meliloti populations can be mainly ascribed to geographic isolation following an isolation-by-distance model, with a strong distance-decay relationship of genetic similarity with distance, in which local soil conditions are not the major component influencing the isolate phenotypes or their genomic differentiation.

  17. 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.

  18. N abundance of nodules as an indicator of N metabolism in n(2)-fixing plants.

    Science.gov (United States)

    Shearer, G; Feldman, L; Bryan, B A; Skeeters, J L; Kohl, D H; Amarger, N; Mariotti, F; Mariotti, A

    1982-08-01

    This paper expands upon previous reports of (15)N elevation in nodules (compared to other tissues) of N(2)-fixing plants. N(2)-Fixing nodules of Glycine max (soybeans), Vigna unguiculata (cowpea), Phaseolus vulgaris (common bean), Phaseolus coccineus (scarlet runner bean), Prosopis glandulosa (mesquite), and Olneya tesota (desert ironwood) were enriched in (15)N. Nodules of Vicia faba (fava beans), Arachis hypogaea (peanut), Trifolium pratense (red clover), Pisum sativum (pea), Lathyrus sativus (grass pea), Medicago sativa (alfalfa), and Lupinus mutabilis (South American lupine) were not; nor were the nodules of nine species of N(2)-fixing nonlegumes. The nitrogen of ineffective nodules of soybeans and cowpeas was not enriched in (15)N. Thus, (15)N elevation in nodules of these plants depends on active N(2)-fixation. Results obtained so far on the generality of (15)N enrichment in N(2)-fixing nodules suggest that only the nodules of plants which actively fix N(2) and which transport allantoin or allantoic acid exhibit (15)N enrichment.

  19. 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.

  20. Elevated CO{sub 2} enhances plant growth in droughted N{sub 2}-fixing alfalfa without improving water status

    Energy Technology Data Exchange (ETDEWEB)

    Luis, I. De; Irigoyen, J.J.; Sanchez-Diaz, M. [Univ. de Navarra, Dept. de Fisiologia Vegetal, Pamplona (Spain)

    1999-07-01

    The long-term interaction between elevated CO{sub 2} and soil water deficit was analysed in N{sub 2}-fixing alfalfa plants in order to assess the possible drought tolerance effect of CO{sub 2}. Elevated CO{sub 2} could delay the onset of drought stress by decreasing transpiration rates, but this effect was avoided by subjecting plants to the same soil water content. Nodulated alfalfa plants subjected to ambient (400 {mu}mol mol{sup -1}) or elevated (700 {mu}mol mol{sup -1}) CO{sub 2} were either well watered or partially watered by restricting water to obtain 30% of the water content at field capacity (approximately 0.55 g water cm{sup -3}). The negative effects of soil water deficit on plant growth were counterbalanced by elevated CO{sub 2}. In droughted plants, elevated CO{sub 2} stimulated carbon fixation and, as a result, biomass production was even greater than in well-watered plants grown in ambient CO{sub 2}. Below-ground production was preferentially stimulated by elevated CO{sub 2} in droughted plants, increasing nodule biomass production and the availability of photosynthates to the nodules. As a result, total nitrogen content in droughted plants was higher than in well-watered plants grown in ambient CO{sub 2}. The beneficial effect of elevated CO{sub 2} was not correlated with a better plant water status. It is concluded that elevated CO{sub 2} enhances growth of droughted plants by stimulating carbon fixation, preferentially increasing the availability of photosynthates to below-ground production (roots and nodules) without improving water status. This means that elevated CO{sub 2} enhances the ability to produce more biomass in N{sub 2}-fixing alfalfa under given soil water stress, improving drought tolerance. (au)

  1. Regulation of symbiotic nitrogen fixation in root nodules of alfalfa (Medicago sativa) infected with Rhizobium meliloti.

    Science.gov (United States)

    Kamberger, W

    1977-10-24

    Symbiotic nitrogen fixation of Rhizobium meliloti bacteroids in Medicago sativa root nodules was suppressed by several inorganic nitrogen sources. Amino acids like glutamine, glutamic acid and aspartic acid, which can serve as sole nitrogen sources for the unnodulated plant did not influence nitrogenase activity of effective nodules, even at high concetrations. Ammonia and nitrate suppressed symbiotic nitrogen fixation in vivo only at concentrations much higher than those needed for suppression of nitrogenase activity in free living nitrogen fixing bacteria. The kinetics of suppression were slow compared with that of free living nitrogen fixing bacteria. On the other hand, nitrite, which acts as a direct inhibitor of nitrogenase, suppressed very quickly and at low concentrations. Glutamic acid and glutamine enhanced the effect of ammonia dramatically, while the suppression by nitrate was enhanced only slightly.

  2. Genetically engineered alfalfa and feral alfalfa plants: What should growers know?

    Science.gov (United States)

    Alfalfa (Medicago sativa subsp. sativa L) is the world’s most important forage crop. The western United States is the most important production area for both alfalfa forage and alfalfa seed. Alfalfa was the first major perennial genetically-engineered (GE)crop and a GE trait for resistance to glypho...

  3. Activity screening of plant growth promoting rhizobacteria isolated from alfalfa rhizosphere

    Directory of Open Access Journals (Sweden)

    shahla pashapour

    2016-03-01

    Full Text Available Introduction: Some rhizobacteria by various mechanisms influence plant growth as they are called plant growth promoting rhizobacteria (PGPR. Scientists identified some PGPR characters involved in promoting plant growth, while all these characters are not able to study. The aim of this study was to evaluate PGP activities of bacterial isolates, (45 isolates belonged to rhizobium and 2 bacterial isolates belonged to Pseudomonas fluorescens, which were isolated from alfalfa (Medicago sativa rhizosphere and root nodules grown around Zanjan. Materials and methods: These bacteria were isolated from alfalfa roots grown around Zinc industries in Zanjan province. After bacterial isolation and purification from root and soil samples, isolates were screened in vitro for plant growth promoting traits such as IAA (Indole Acetic Acid, ACC- deaminase (Amino Cyclopropan Carboxylate, HCN (Hydrogen Cyanide, siderophore, chitinase production and mineral and organic phosphate solubilization activities. Results: The results indicated that 43 bacterial isolates produced IAA (4.04- 4.95 μg/ml and 15 isolates produced ACC- deaminase (0.23- 1.05 μg/ml. Only one isolate (Rm66 produced high amount of HCN. Qualitative siderophore production was observed in 9 isolates. None of the isolates produced chitinase. Solubilization of mineral phosphate was commonly detected in 19 isolates (4.33- 5.86 μg/ml, and 15 isolates solubilized organic phosphate (1.66- 144.28 μg/ml. Discussion and conclusion: This study shows that most of the bacterial strains which isolated from alfalfa cultivated lands had PGP activities and also a good potential to increase plant growth after inoculation with to seeds as eco- friendly fertilizers.

  4. Potentiality Of Stressed Alfalfa Plants To Modify Soil Traits

    Directory of Open Access Journals (Sweden)

    Bardees M. Mickky

    2015-08-01

    Full Text Available Leguminous plants play a key role in the sustainability of agricultural systems. Nevertheless it is to somewhat scarce to find investigations about the interaction of such plants with the soil in which they grow under stress. A pot experiment was conducted to investigate the influence of graduated water regimes on the chemical features of different soil types varying in sand proportion before and after their cultivation with alfalfa Medicago sativa plants. The results indicated that soil cultivation with the studied plants whether water-satisfied or not generally increased the amount of soil ammonia nitrate peptide total soluble and total nitrogen. On contrary the amount of soil amino amide nitrite and protein nitrogen generally decreased after cultivation. Concerning soil ionic composition the post-planting values of soil sodium calcium magnesium and sodium potassium ratio were significantly lower than their pre-cultivation synonyms. Furthermore the percentages of decline caused by cultivating well-watered plants were higher than those caused by their moderately-droughted relatives which were in turn higher than those caused by severely-droughted ones. For soil chloride potassium sodium adsorption ratio and potassium adsorption ratio culturing water-unstressed plants reduced these ionic fractions while droughted plants markedly favored such values. As a general feature the maximum titers of all soil nitrogenous and ionic constituents were recorded for soil with the least sand proportion. The other chemical soil characteristics pH electric conductivity organic carbon organic matter calcium carbonate bicarbonates sulphates and total soluble salts were all fluctuated in a random fashion among the various soil types before and after planting water-stressed or control alfalfa plants. Thus the results obtained herein recommend alfalfa as a pioneer plant that can be introduced to infertile and or dry lands with a paramount efficacy to enhance soil

  5. Photosynthetic and Molecular Markers of CO2-mediated Photosynthetic Downregulation in Nodulated Alfalfa

    Institute of Scientific and Technical Information of China (English)

    (A)lvaro Sanz-Sáez; Gorka Erice; Iker Aranjuelo; Ricardo Aroca; Juan Manuel Ruíz-Lozano; Jone Aguirreolea; Juan José Irigoyen

    2013-01-01

    Elevated CO2 leads to a decrease in potential net photosynthesis in long-term experiments and thus to a reduction in potential growth.This process is known as photosynthetic downregulation.There is no agreement on the definition of which parameters are the most sensitive for detecting CO2 acclimation.In order to investigate the most sensitive photosynthetic and molecular markers of CO2 acclimation,the effects of elevated CO2,and associated elevated temperature were analyzed in alfalfa plants inoculated with different Sinorhizobium meliloti strains.Plants (Medicago sativa L.cv.Aragón) were grown in summer or autumn in temperature gradient greenhouses (TGG).At the end of the experiment,all plants showed acclimation in both seasons,especially under elevated summer temperatures.This was probably due to the lower nitrogen (N) availability caused by decreased N2-fixation under higher temperatures.Photosynthesis measured at growth CO2 concentration,rubisco in vitro activity and maximum rate of carboxylation were the most sensitive parameters for detecting downregulation.Severe acclimation was also related with decreases in leaf nitrogen content associated with declines in rubisco content (large and small subunits) and activity that resulted in a drop in photosynthesis.Despite the sensitivity of rubisco content as a marker of acclimation,it was not coordinated with gene expression,possibly due to a lag between gene transcription and protein translation.

  6. Nitrogen use efficiency and life cycle of nodules in alfalfa after different mineral fertilization and soil cultivation

    Directory of Open Access Journals (Sweden)

    V. Vasileva

    2015-09-01

    Full Text Available Nitrogen use efficiency and life cycle of root nodules in alfalfa after different mineral fertilization and soil cultivation practices were studied. Field trial was conducted in the Institute of Forage Crops, Pleven, Bulgaria on leached chernozem subsoil type and no irrigation. The next treatments were tested: i for fertilization as follows: N0P0K0 (control; N60P100K80 (an accepted technology; N23P100K35 (nitrogen was applied 1/2 in first year of growing and 1/2 in third year; N23P100K35 (nitrogen was supplied pre-sowing; N35P80K50, and Аmophose - 250 kg/ha, calculated at fertilizing rates N27P120K0; ii for soil cultivation as follows: soil loosing 10-12 cm, plough at depth 12-15 cm, 22-24 cm (an accepted technology, 18-22 cm and 30-35 cm. It was found that soil cultivation and mineral fertilization had effect on nitrogen use efficiency and life cycle of root nodules in Alfalfa. Nitrogen use efficiency was found to be highest at N23P100K35 and plough at the depth of 22-24 cm. Life cycle of root nodules was the longest at N35P80K50 and plough at the depth of 18-22 cm. The better root mass to nodule number ratio was found at N23P100K35 and plough at the depth of 22-24 cm.

  7. 15N Abundance of Nodules as an Indicator of N Metabolism in N2-Fixing Plants 1

    Science.gov (United States)

    Shearer, Georgia; Feldman, Lori; Bryan, Barbara A.; Skeeters, Jerri L.; Kohl, Daniel H.; Amarger, Nöelle; Mariotti, Françoise; Mariotti, André

    1982-01-01

    This paper expands upon previous reports of 15N elevation in nodules (compared to other tissues) of N2-fixing plants. N2-Fixing nodules of Glycine max (soybeans), Vigna unguiculata (cowpea), Phaseolus vulgaris (common bean), Phaseolus coccineus (scarlet runner bean), Prosopis glandulosa (mesquite), and Olneya tesota (desert ironwood) were enriched in 15N. Nodules of Vicia faba (fava beans), Arachis hypogaea (peanut), Trifolium pratense (red clover), Pisum sativum (pea), Lathyrus sativus (grass pea), Medicago sativa (alfalfa), and Lupinus mutabilis (South American lupine) were not; nor were the nodules of nine species of N2-fixing nonlegumes. The nitrogen of ineffective nodules of soybeans and cowpeas was not enriched in 15N. Thus, 15N elevation in nodules of these plants depends on active N2-fixation. Results obtained so far on the generality of 15N enrichment in N2-fixing nodules suggest that only the nodules of plants which actively fix N2 and which transport allantoin or allantoic acid exhibit 15N enrichment. PMID:16662517

  8. Alfalfa nodules elicited by a flavodoxin-overexpressing Ensifer meliloti strain display nitrogen-fixing activity with enhanced tolerance to salinity stress.

    Science.gov (United States)

    Redondo, Francisco J; Coba de la Peña, Teodoro; Lucas, M Mercedes; Pueyo, José J

    2012-12-01

    Nitrogen fixation by legumes is very sensitive to salinity stress, which can severely reduce the productivity of legume crops and their soil-enriching capacity. Salinity is known to cause oxidative stress in the nodule by generating reactive oxygen species (ROS). Flavodoxins are involved in the response to oxidative stress in bacteria and cyanobacteria. Prevention of ROS production by flavodoxin overexpression in bacteroids might lead to a protective effect on nodule functioning under salinity stress. Tolerance to salinity stress was evaluated in alfalfa nodules elicited by an Ensifer meliloti strain that overexpressed a cyanobacterial flavodoxin compared with nodules produced by the wild-type bacteria. Nitrogen fixation, antioxidant and carbon metabolism enzyme activities were determined. The decline in nitrogenase activity associated to salinity stress was significantly less in flavodoxin-expressing than in wild-type nodules. We detected small but significant changes in nodule antioxidant metabolism involving the ascorbate-glutathione cycle enzymes and metabolites, as well as differences in activity of the carbon metabolism enzyme sucrose synthase, and an atypical starch accumulation pattern in flavodoxin-containing nodules. Salt-induced structural and ultrastructural alterations were examined in detail in alfalfa wild-type nodules by light and electron microscopy and compared to flavodoxin-containing nodules. Flavodoxin reduced salt-induced structural damage, which primarily affected young infected tissues and not fully differentiated bacteroids. The results indicate that overexpression of flavodoxin in bacteroids has a protective effect on the function and structure of alfalfa nodules subjected to salinity stress conditions. Putative protection mechanisms are discussed.

  9. Plant regeneration from proroplasts of alfalfa (Medicago sativa) via somatic embryogenesis

    OpenAIRE

    Monteiro, Mariza; Appezzato-da-Glória, Beatriz; Valarini,Maria José; Oliveira, Carlos Alberto; Vieira, Maria Lucia Carneiro

    2003-01-01

    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 pro...

  10. Ensifer, Phyllobacterium and Rhizobium species occupy nodules of Medicago sativa (alfalfa) and Melilotus alba (sweet clover) grown at a Canadian site without a history of cultivation

    Science.gov (United States)

    Phage resistant and susceptible bacteria from nodules of alfalfa and sweet clover grown at a site without a known history of cultivation, were identified as Ensifer, Rhizobium and Phyllobacterium species based on sequence analysis of ribosomal (16S and 23S rRNA)and protein encoding (atpD and recA) g...

  11. Leguminous plants: inventors of root nodules to accommodate symbiotic bacteria.

    Science.gov (United States)

    Suzaki, Takuya; Yoro, Emiko; Kawaguchi, Masayoshi

    2015-01-01

    Legumes and a few other plant species can establish a symbiotic relationship with nitrogen-fixing rhizobia, which enables them to survive in a nitrogen-deficient environment. During the course of nodulation, infection with rhizobia induces the dedifferentiation of host cells to form primordia of a symbiotic organ, the nodule, which prepares plants to accommodate rhizobia in host cells. While these nodulation processes are known to be genetically controlled by both plants and rhizobia, recent advances in studies on two model legumes, Lotus japonicus and Medicago truncatula, have provided great insight into the underlying plant-side molecular mechanism. In this chapter, we review such knowledge, with particular emphasis on two key processes of nodulation, nodule development and rhizobial invasion. Copyright © 2015 Elsevier Inc. All rights reserved.

  12. The elicitor-inducible alfalfa isoflavone reductase promoter confers different patterns of developmental expression in homologous and heterologous transgenic plants.

    Science.gov (United States)

    Oommen, A; Dixon, R A; Paiva, N L

    1994-01-01

    In legumes, the synthesis of infection- and elicitor-inducible antimicrobial phytoalexins occurs via the isoflavonoid branch of the phenylpropanoid pathway. To study transcriptional regulation of isoflavonoid pathway-specific genes, we have isolated the gene encoding isoflavone reductase (IFR), which is the enzyme that catalyzes the penultimate step in the synthesis of the phytoalexin medicarpin in alfalfa. Chimeric gene fusions were constructed between 765- and 436-bp promoter fragments of the IFR gene and the beta-glucuronidase reporter gene and transferred to alfalfa and tobacco by Agrobacterium-mediated transformation. Both promoter fragments conferred elicitor-mediated expression in cell suspension cultures derived from transgenic plants of both species and fungal infection-mediated expression in leaves of transgenic alfalfa. Developmental expression directed by both promoter fragments in transgenic alfalfa was observed only in the root meristem, cortex, and nodules, which is consistent with the accumulation of endogenous IFR transcripts. However, in transgenic tobacco, expression from the 765-bp promoter was observed in vegetative tissues (root meristem and cortex, inner vascular tissue of stems and petioles, leaf tips, and stem peripheries adjacent to petioles) and in reproductive tissues (stigma, placenta, base of the ovary, receptacle, seed, tapetal layer, and pollen grains), whereas the 436-bp promoter was expressed only in fruits, seed, and pollen. These data indicate that infection/elicitor inducibility of the IFR promoter in both species and developmental expression in alfalfa are determined by sequences downstream of position -436, whereas sequences between -436 and -765 confer a complex pattern of strong ectopic developmental expression in the heterologous species that lacks the isoflavonoid pathway. PMID:7866024

  13. Ensifer, Phyllobacterium and Rhizobium species occupy nodules of Medicago sativa (alfalfa) and Melilotus alba (sweet clover) grown at a Canadian site without a history of cultivation.

    Science.gov (United States)

    Bromfield, E S P; Tambong, J T; Cloutier, S; Prévost, D; Laguerre, G; van Berkum, P; Thi, T V Tran; Assabgui, R; Barran, L R

    2010-02-01

    Phage-resistant and -susceptible bacteria from nodules of alfalfa and sweet clover, grown at a site without a known history of cultivation, were identified as diverse genotypes of Ensifer, Rhizobium and Phyllobacterium species based on sequence analysis of ribosomal (16S and 23S rRNA) and protein-encoding (atpD and recA) genes, Southern hybridization/RFLP and a range of phenotypic characteristics. Among phage-resistant bacteria, one genotype of Rhizobium sp. predominated on alfalfa (frequency approximately 68 %) but was recovered infrequently ( approximately 1 %) from sweet clover. A second genotype was isolated infrequently only from alfalfa. These genotypes fixed nitrogen poorly in association with sweet clover and Phaseolus vulgaris, but were moderately effective with alfalfa. They produced a near-neutral reaction on mineral salts agar containing mannitol, which is atypical of the genus Rhizobium. A single isolate of Ensifer sp. and two of Phyllobacterium sp. were recovered only from sweet clover. All were highly resistant to multiple antibiotics. Phylogenetic analysis indicated that Ensifer sp. strain T173 is closely related to, but separate from, the non-symbiotic species 'Sinorhizobium morelense'. Strain T173 is unique in that it possesses a 175 kb symbiotic plasmid and elicits ineffective nodules on alfalfa, sweet clover, Medicago lupulina and Macroptilium atropurpureum. The two Phyllobacterium spp. were non-symbiotic and probably represent bacterial opportunists. Three genotypes of E. meliloti that were symbiotically effective with alfalfa and sweet clover were encountered infrequently. Among phage-susceptible isolates, two genotypes of E. medicae were encountered infrequently and were highly effective with alfalfa, sweet clover and Medicago polymorpha. The ecological and practical implications of the findings are discussed.

  14. 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.

  15. Easy synthesis of graphene sheets from alfalfa plants by treatment of nitric acid

    Energy Technology Data Exchange (ETDEWEB)

    Qu, Jiao, E-mail: qujiao@bhu.edu.cn [School of Chemistry and Chemical Engineering, Bohai University, Jinzhou 121013 (China); School of Urban and Environmental Sciences, Northeast Normal University, Changchun 130024 (China); Luo, Chunqiu, E-mail: fplj_lcq@163.com [School of Chemistry and Chemical Engineering, Bohai University, Jinzhou 121013 (China); Zhang, Qian; Cong, Qiao [School of Chemistry and Chemical Engineering, Bohai University, Jinzhou 121013 (China); Yuan, Xing [School of Urban and Environmental Sciences, Northeast Normal University, Changchun 130024 (China)

    2013-04-01

    Highlights: ► An easy method for synthesis of graphene sheets using alfalfa plants was introduced. ► An novelty formation mechanism of graphene sheets using alfalfa plants was proposed. ► This method exploits a new carbon source and provides a novel idea to synthesize graphene sheets. -- Abstract: This letter focuses on synthesis of graphene sheets from alfalfa plants by treatment of nitric acid. The transmission electron microscopy image (TEM) demonstrates that the graphene sheets are agglomerated and overlapped, the energy dispersive spectrum (EDS) indicates that the products are pure, and the Raman spectrum shows the graphene sheets are well graphitized. In addition, the formation mechanism of the graphene sheets from alfalfa plants by treatment nitric acid is discussed. These findings inspire the search for a new strategy for synthesis of graphene sheets from renewable natural products, and the lower cost of this new process and carbon source may facilitate industrial production.

  16. Transgenic Alfalfa Plants Expressing the Sweetpotato Orange Gene Exhibit Enhanced Abiotic Stress Tolerance

    Science.gov (United States)

    Wang, Zhi; Ke, Qingbo; Kim, Myoung Duck; Kim, Sun Ha; Ji, Chang Yoon; Jeong, Jae Cheol; Lee, Haeng-Soon; Park, Woo Sung; Ahn, Mi-Jeong; Li, Hongbing; Xu, Bingcheng; Deng, Xiping; Lee, Sang-Hoon; Lim, Yong Pyo; Kwak, Sang-Soo

    2015-01-01

    Alfalfa (Medicago sativa L.), a perennial forage crop with high nutritional content, is widely distributed in various environments worldwide. We recently demonstrated that the sweetpotato Orange gene (IbOr) is involved in increasing carotenoid accumulation and enhancing resistance to multiple abiotic stresses. In this study, in an effort to improve the nutritional quality and environmental stress tolerance of alfalfa, we transferred the IbOr gene into alfalfa (cv. Xinjiang Daye) under the control of an oxidative stress-inducible peroxidase (SWPA2) promoter through Agrobacterium tumefaciens-mediated transformation. Among the 11 transgenic alfalfa lines (referred to as SOR plants), three lines (SOR2, SOR3, and SOR8) selected based on their IbOr transcript levels were examined for their tolerance to methyl viologen (MV)-induced oxidative stress in a leaf disc assay. The SOR plants exhibited less damage in response to MV-mediated oxidative stress and salt stress than non-transgenic plants. The SOR plants also exhibited enhanced tolerance to drought stress, along with higher total carotenoid levels. The results suggest that SOR alfalfa plants would be useful as forage crops with improved nutritional value and increased tolerance to multiple abiotic stresses, which would enhance the development of sustainable agriculture on marginal lands. PMID:25946429

  17. Transgenic alfalfa plants expressing the sweetpotato Orange gene exhibit enhanced abiotic stress tolerance.

    Directory of Open Access Journals (Sweden)

    Zhi Wang

    Full Text Available Alfalfa (Medicago sativa L., a perennial forage crop with high nutritional content, is widely distributed in various environments worldwide. We recently demonstrated that the sweetpotato Orange gene (IbOr is involved in increasing carotenoid accumulation and enhancing resistance to multiple abiotic stresses. In this study, in an effort to improve the nutritional quality and environmental stress tolerance of alfalfa, we transferred the IbOr gene into alfalfa (cv. Xinjiang Daye under the control of an oxidative stress-inducible peroxidase (SWPA2 promoter through Agrobacterium tumefaciens-mediated transformation. Among the 11 transgenic alfalfa lines (referred to as SOR plants, three lines (SOR2, SOR3, and SOR8 selected based on their IbOr transcript levels were examined for their tolerance to methyl viologen (MV-induced oxidative stress in a leaf disc assay. The SOR plants exhibited less damage in response to MV-mediated oxidative stress and salt stress than non-transgenic plants. The SOR plants also exhibited enhanced tolerance to drought stress, along with higher total carotenoid levels. The results suggest that SOR alfalfa plants would be useful as forage crops with improved nutritional value and increased tolerance to multiple abiotic stresses, which would enhance the development of sustainable agriculture on marginal lands.

  18. [Genetic resources of nodule bacteria].

    Science.gov (United States)

    Rumiantseva, M L

    2009-09-01

    Nodule bacteria (rhizobia) form highly specific symbiosis with leguminous plants. The efficiency of accumulation of biological nitrogen depends on molecular-genetic interaction between the host plant and rhizobia. Genetic characteristics of microsymbiotic strains are crucial in developing highly productive and stress-resistant symbiotic pairs: rhizobium strain-host plant cultivar (species). The present review considers the issue of studying genetic resources of nodule bacteria to identify genes and their blocks, responsible for the ability of rhizobia to form highly effective symbiosis in various agroecological conditions. The main approaches to investigation of intraspecific and interspecific genetic and genomic diversity of nodule bacteria are considered, from MLEE analysis to the recent methods of genomic DNA analysis using biochips. The data are presented showing that gene centers of host plants are centers of genetic diversification of nodule bacteria, because the intraspecific polymorphism of genetic markers of the core and the accessory rhizobial genomes is extremely high in them. Genotypic features of trapped and nodule subpopulations of alfalfa nodule bacteria are discussed. A survey of literature showed that the genomes of natural strains in alfalfa gene centers exhibit significant differences in genes involved in control of metabolism, replication, recombination, and the formation of defense response (hsd genes). Natural populations of rhizobia are regarded as a huge gene pool serving as a source of evolutionary innovations.

  19. USDA-ARS Plant Science Research Unit, St. Paul Alfalfa/Forage Research Program

    Science.gov (United States)

    The Plant Science Research Unit (PSRU) located at the University of Minnesota in St. Paul receives approximately $1.5 million to fund the research of six scientists who direct their research efforts toward developing new uses and improved traits for alfalfa. Our overarching goal is to develop alfalf...

  20. Efficacy of a BVDV subunit vaccine produced in alfalfa transgenic plants.

    Science.gov (United States)

    Peréz Aguirreburualde, María Sol; Gómez, María Cristina; Ostachuk, Agustín; Wolman, Federico; Albanesi, Guillermo; Pecora, Andrea; Odeon, Anselmo; Ardila, Fernando; Escribano, José M; Dus Santos, María José; Wigdorovitz, Andrés

    2013-02-15

    Bovine viral diarrhea virus (BVDV) is considered an important cause of economic loss within bovine herds worldwide. In Argentina, only the use of inactivated vaccines is allowed, however, the efficacy of inactivated BVDV vaccines is variable due to its low immunogenicity. The use of recombinant subunit vaccines has been proposed as an alternative to overcome this difficulty. Different studies on protection against BVDV infection have focused the E2 protein, supporting its putative use in subunit vaccines. Utilization of transgenic plants expressing recombinant antigens for the formulation of experimental vaccines represents an innovative and cost effective alternative to the classical fermentation systems. The aim of this work was to develop transgenic alfalfa plants (Medicago sativa, L.) expressing a truncated version of the structural protein E2 from BVDV fused to a molecule named APCH, that target to antigen presenting cells (APCH-tE2). The concentration of recombinant APCH-tE2 in alfalfa leaves was 1 μg/g at fresh weight and its expression remained stable after vegetative propagation. A methodology based an aqueous two phases system was standardized for concentration and partial purification of APCH-tE2 from alfalfa. Guinea pigs parentally immunized with leaf extracts developed high titers of neutralizing antibodies. In bovine, the APCH-tE2 subunit vaccine was able to induce BVDV-specific neutralizing antibodies. After challenge, bovines inoculated with 3 μg of APCH-tE2 produced in alfalfa transgenic plants showed complete virological protection.

  1. Eficiência de inoculantes de rizóbio na nodulação de alfafa em solo de cerrado Efficiency of rhizobia inoculants on nodulation of alfalfa in a "cerrado" soil

    Directory of Open Access Journals (Sweden)

    Deise Ferreira Xavier

    2005-06-01

    harvest was performed at 5 cm height from ground level, to determine dry matter (DM weight and total-N in the above ground parts. At ninety days, the whole plants were sampled, to measure the DM yield of both above ground parts and roots, and N content of above ground parts. The presence of nodules in the roots was examined. Results of DM yield of above ground parts and roots, and of total-N in the above ground parts, indicated that the two commercial inoculants used were efficient for all the three alfalfa cultivars in the soil, which had its pH corrected. Regarding nodulation, the checks with and without nitrogen did not nodulate, indicating that Rhizobium meliloti was not present in the soil. These observations reinforce the need of inoculation of alfalfa with appropriate rhizobia in tropical regions.

  2. Symbiotic leghemoglobins are crucial for nitrogen fixation in legume root nodules but not for general plant growth and development

    DEFF Research Database (Denmark)

    Ott, Thomas; van Dongen, Joost T; Günther, Catrin

    2005-01-01

    fixation (SNF). In legumes, SNF occurs in specialized organs, called nodules, which contain millions of nitrogen-fixing rhizobia, called bacteroids. The induction of nodule-specific plant genes, including those encoding symbiotic leghemoglobins (Lb), accompanies nodule development. Leghemoglobins...

  3. Effects of plant enzyme inactivation or sterilization on lipolysis and proteolysis in alfalfa silage.

    Science.gov (United States)

    Ding, W R; Long, R J; Guo, X S

    2013-04-01

    This experiment studied the contribution of plant enzymes and microbial activity on lipolysis and proteolysis in ensiled alfalfa. Before ensiling, the wilted alfalfa was treated with plant enzyme inactivation by autoclaving or with sterilization by γ-ray irradiation. The treated alfalfa was then inoculated with commercial lactic acid bacteria inoculants and ensiled for 40 d. Alfalfa without treatment was ensiled as the control. The content of total fatty acid (FA) after ensiling decreased 43% in the control silage and 28% in the γ-ray-treated silage, but did not change in the autoclave-treated silage. Among the major FA (C16:0, C18:2n-6, C18:3n-3), a considerable increase was observed in proportion of C16:0 in the control silage as compared with fresh alfalfa; conversely, decreases in proportions of C18:2n-6 and C18:3n-3 occurred during ensilage. Silage treated with γ-ray radiation at ensiling had a smaller proportion of C16:0 and greater proportions of C18:2n-6 and C18:3n-3 than control silage. Autoclave treatment further decreased proportions of C16:0 and most of the other FA, and increased C18:2n-6 and C18:3n-3 proportions in comparison with γ-ray treatment. Proportions of C16:0, C18:2n-6, C18:3n-3 and other detected FA (except for the proportion of C15:0) did not differ between fresh forage and autoclave-treated silage. Remarkably, smaller nonprotein nitrogen content was observed in the autoclave-treated silage compared with the γ ray-treated silage or the control silage. These results indicated that an extensive lipolysis occurred during ensiling of alfalfa, and plant enzymes played a major role in lipolysis and proteolysis. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  4. Effect of foliar feeding on nitrogen assimilation in alfalfa plants at insufficient molybdenum supply.

    Science.gov (United States)

    Hristozkova, Marieta; Geneva, Maria; Stancheva, Ira

    2009-06-01

    The influence of foliar feeding on the nitrogen assimilation in alfalfa plants under conditions of Mo shortage was studied. It was established that foliar fertilization with 0.3% solution of Agroleaf® resulted in increase of nitrogen fixation and nitrogen assimilation in the absence of Mo. Insufficient molybdenum supply leads to significant reduction of plant Mo content and nitrogen-fixing activity, while stress induced amino acids as alanine, GABA, threonine, proline and serine increased repeatedly. The negative effect of Mo deficiency on the enzyme activities related to the primary nitrogen assimilation (NR, GS, GOGAT) and plant growth diminished due to the foliar absorbed nutrients.

  5. Excessive ammonia inhibited transcription of MsU2 gene and furthermore affected accumulation distribution of allantoin and amino acids in alfalfa Medicago sativa

    Institute of Scientific and Technical Information of China (English)

    WANG Li; JIANG Lin-lin; Nomura Mika; Tajima Shigeyuki; CHENG Xian-guo

    2015-01-01

    In legume plants, uricase gene (Nodulin-35) plays a positive role in metabolism of ureide and amide compounds in symbiotic nitrogen-ifxing in the nodules. In this study, a pot experiment was performed to examine the effects of ammonium application on the transcription of MsU2 gene and distribution of major nitrogen compounds in alfalfa Medicago sativa. Data showed that alfalfa plant has a signiifcant difference in contents of nitrogen compounds in xylem saps compared with soybean plant, and belongs to typical amide type legume plants with little ureide accumulation, and the accumulation of asparagines and ureide in the tissues of alfalfa is mainly gathered in the nodules. Northern blotting showed that excessive ammonium signiifcantly inhibited the transcription of MsU2 gene in the nodules and roots, and mRNA accumulation of MsU2 gene in the plants exposed to excessive ammonium decreased gradual y with culture time extension, indicating that application of ammonium signiifcantly inhibited the transcription of MsU2 gene in the alfalfa plants. Although the application of exces-sive ammonium increased the contents of amino acids in various tissues of alfalfa, the accumulation of al antoin relfecting the strength of uricase activity is remarkably reduced in the xylem saps, stems and nodules when alfalfa plants exposed to excessive ammonium, suggesting that application of excessive ammonium generated a negative effect on symbiosis ifxing-nitrogen system due to inhibition of ammonium ion on uricase activity in the nodules of alfalfa. This result seems to imply that application of excessive ammonium in legume plants should not be proposed to avoid affecting the ability of ifxing nitrogen in the nodules of legume plants, and reasonable dose of ammonium should be recommended to effectively utilize the ifxed N from atmosphere in legume plant production.

  6. Transgenic alfalfa plants expressing AtNDPK2 exhibit increased growth and tolerance to abiotic stresses.

    Science.gov (United States)

    Wang, Zhi; Li, Hongbing; Ke, Qingbo; Jeong, Jae Cheol; Lee, Haeng-Soon; Xu, Bingcheng; Deng, Xi-Ping; Lim, Yong Pyo; Kwak, Sang-Soo

    2014-11-01

    In this study, we generated and evaluated transgenic alfalfa plants (Medicago sativa L. cv. Xinjiang Daye) expressing the Arabidopsis nucleoside diphosphate kinase 2 (AtNDPK2) gene under the control of the oxidative stress-inducible SWPA2 promoter (referred to as SN plants) to develop plants with enhanced tolerance to various abiotic stresses. We selected two SN plants (SN4 and SN7) according to the expression levels of AtNDPK2 and the enzyme activity of NDPK in response to methyl viologen (MV)-mediated oxidative stress treatment using leaf discs for further characterization. SN plants showed enhanced tolerance to high temperature, NaCl, and drought stress on the whole-plant level. When the plants were subjected to high temperature treatment (42 °C for 24 h), the non-transgenic (NT) plants were severely wilted, whereas the SN plants were not affected because they maintained high relative water and chlorophyll contents. The SN plants also showed significantly higher tolerance to 250 mM NaCl and water stress treatment than the NT plants. In addition, the SN plants exhibited better plant growth through increased expression of auxin-related indole acetic acid (IAA) genes (MsIAA3, MsIAA5, MsIAA6, MsIAA7, and MsIAA16) under normal growth conditions compared to NT plants. The results suggest that induced overexpression of AtNDPK2 in alfalfa will be useful for increasing biomass production under various abiotic stress conditions.

  7. Comparison of the nodule vs. root transcriptome of the actinorhizal plant Datisca glomerata: actinorhizal nodules contain a specific class of defensins.

    Directory of Open Access Journals (Sweden)

    Irina V Demina

    Full Text Available Actinorhizal root nodule symbioses are very diverse, and the symbiosis of Datisca glomerata has previously been shown to have many unusual aspects. In order to gain molecular information on the infection mechanism, nodule development and nodule metabolism, we compared the transcriptomes of D. glomerata roots and nodules. Root and nodule libraries representing the 3'-ends of cDNAs were subjected to high-throughput parallel 454 sequencing. To identify the corresponding genes and to improve the assembly, Illumina sequencing of the nodule transcriptome was performed as well. The evaluation revealed 406 differentially regulated genes, 295 of which (72.7% could be assigned a function based on homology. Analysis of the nodule transcriptome showed that genes encoding components of the common symbiosis signaling pathway were present in nodules of D. glomerata, which in combination with the previously established function of SymRK in D. glomerata nodulation suggests that this pathway is also active in actinorhizal Cucurbitales. Furthermore, comparison of the D. glomerata nodule transcriptome with nodule transcriptomes from actinorhizal Fagales revealed a new subgroup of nodule-specific defensins that might play a role specific to actinorhizal symbioses. The D. glomerata members of this defensin subgroup contain an acidic C-terminal domain that was never found in plant defensins before.

  8. Alfalfa endophytes as novel sources of antimicrobial compounds that inhibit the growth of human and plant pathogens

    Science.gov (United States)

    Fungal endophytes may contribute to plant health and disease protection, yet little is known about their various roles in alfalfa. Also, endophytes from several plant species produce novel antimicrobial compounds that may be useful clinically. We isolated endophytic fungi from over 50 samples from s...

  9. Characterization of the plant growth promoting bacterium, Enterobacter cloacae MSR1, isolated from roots of non-nodulating Medicago sativa.

    Science.gov (United States)

    Khalifa, Ashraf Y Z; Alsyeeh, Abdel-Moneium; Almalki, Mohammed A; Saleh, Farag A

    2016-01-01

    The aim of the present study was to characterize the endophytic bacterial strain designated MSR1 that was isolated from inside the non-nodulating roots of Medicago sativa after surface-sterilization. MSR1 was identified as Enterobacter cloacae using both 16S rDNA gene sequence analysis and API20E biochemical identification system (Biomerieux, France). Furthermore, this bacterium was characterized using API50CH kit (Biomerieux, France) and tested for antibacterial activities against some food borne pathogens. The results showed that E. cloacae consumed certain carbohydrates such as glycerol, d-xylose, d-maltose and esculin melibiose as a sole carbon source and certain amino acids such as arginine, tryptophan ornithine as nitrogen source. Furthermore, MSR1 possessed multiple plant-growth promoting characteristics; phosphate solubility, production of phytohormones acetoin and bioactive compounds. Inoculation of Pisum sativum with MSR1 significantly improved the growth parameters (the length and dry weight) of this economically important grain legume compared to the non-treated plants. To our knowledge, this is the first report addressing E. cloacae which exist in roots of alfalfa growing in Al-Ahsaa region. The results confirmed that E. cloacae exhibited traits for plant growth promoting and could be developed as an eco-friendly biofertilizer for P. sativum and probably for other important plant species in future.

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

    OpenAIRE

    Aranjuelo, Iker; Tcherkez, Guillaume,; Molero, Gemma; Gilard, Françoise; AVICE, JEAN-CHRISTOPHE; Nogués, Salvador

    2013-01-01

    Although the mechanisms of nodule N2 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 N2-fixing alfalfa (Medicago sativa L.) plants was investigated, and proteomic, metabolomic, and physiological analyses were ...

  11. Nodulation of leguminous plants as affected by root secretions and red light

    NARCIS (Netherlands)

    Lie, T.A.

    1964-01-01

    Nodulation of bean plants, Phaseolus vulgaris L., in water culture was poor during hot sunny weather in the greenhouse. It did not improve when indoleacetic acid, kinetin, gibberellic acid, purines and pyrimidines, yeast and soil extract were added. Nodulation was enhanced by adding used

  12. Medicago truncatula root nodule proteome analysis reveals differential plant and bacteroid responses to drought stress.

    Science.gov (United States)

    Larrainzar, Estíbaliz; Wienkoop, Stefanie; Weckwerth, Wolfram; Ladrera, Rubén; Arrese-Igor, Cesar; González, Esther M

    2007-07-01

    Drought is one of the environmental factors most affecting crop production. Under drought, symbiotic nitrogen fixation is one of the physiological processes to first show stress responses in nodulated legumes. This inhibition process involves a number of factors whose interactions are not yet understood. This work aims to further understand changes occurring in nodules under drought stress from a proteomic perspective. Drought was imposed on Medicago truncatula 'Jemalong A17' plants grown in symbiosis with Sinorhizobium meliloti strain 2011. Changes at the protein level were analyzed using a nongel approach based on liquid chromatography coupled to tandem mass spectrometry. Due to the complexity of nodule tissue, the separation of plant and bacteroid fractions in M. truncatula root nodules was first checked with the aim of minimizing cross contamination between the fractions. Second, the protein plant fraction of M. truncatula nodules was profiled, leading to the identification of 377 plant proteins, the largest description of the plant nodule proteome so far. Third, both symbiotic partners were independently analyzed for quantitative differences at the protein level during drought stress. Multivariate data mining allowed for the classification of proteins sets that were involved in drought stress responses. The isolation of the nodule plant and bacteroid protein fractions enabled the independent analysis of the response of both counterparts, gaining further understanding of how each symbiotic member is distinctly affected at the protein level under a water-deficit situation.

  13. Do glyphosate resistant feral plants and hay fields spread the transgene to conventional alfalfa seed fields?

    Science.gov (United States)

    In addition to meeting domestic needs, large amounts of alfalfa seed and hay produced in the US are being exported overseas. Because alfalfa is an insect pollinated crop, gene flow is a concern. Adding to this alfalfa readily naturalizes along roadsides, irrigation ditches, and unmanaged habitats; a...

  14. 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)

    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 alfalfa accessions for mapping loci associated with plant growth and forage production under salt stress using genome-wide association stud...

  15. 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.

  16. 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.

  17. Enhancement of lead uptake by alfalfa (Medicago sativa) using EDTA and a plant growth promoter.

    Science.gov (United States)

    López, Martha L; Peralta-Videa, Jose R; Benitez, Tenoch; Gardea-Torresdey, Jorge L

    2005-10-01

    Phytoremediation is a novel cleanup technology for the removal of contaminants from polluted waters and soils. In phytoremediation, the plant uptake capability and the availability of the pollutant in the media are important. Here we show the results of a study on the combined effects of ethylenediaminetetraacetic acid (EDTA) and the phytohormone indole-3-acetic acid (IAA) on Pb uptake by Medicago sativa (alfalfa). Plants were grown in hydroponics media containing a nutrient solution amended with Pb at 0.2mM and different combinations of EDTA, and IAA. After 10d of treatment, the Pb content in plant tissues was quantified using an Inductively Coupled Plasma Optical Emission Spectrometer (ICP/OES). The results showed that the combination of 100 microM IAA/0.2 mM EDTA increased the Pb accumulation in leaves by about 2800% and by about 600%, as compared to Pb content in leaves of plants exposed to Pb alone and those cultivated with Pb/EDTA, respectively. These results indicate that non-metal hyperaccumulator plants could increase their hyperaccumulating potential without genetic manipulation.

  18. Leaf d15N as a physiological indicator of the responsiveness of N2-fixing alfalfa plants to elevated [CO2], temperature and low water availability

    Directory of Open Access Journals (Sweden)

    Idoia eAriz

    2015-08-01

    Full Text Available The natural 15N/14N isotope composition (δ15N of a tissue is a consequence of its N source and N physiological mechanisms in response to the environment. It could potentially be used as a tracer of N metabolism in plants under changing environmental conditions, where primary N metabolism may be complex, and losses and gains of N fluctuate over time. In order to test the utility of δ15N as an indicator of plant N status in N2-fixing plants grown under various environmental conditions, alfalfa (Medicago sativa L. plants were subjected to distinct conditions of [CO2] (400 versus 700 mol mol-1, temperature (ambient versus ambient + 4ºC and water availability (fully watered versus water deficiency - WD. As expected, increased [CO2] and temperature stimulated photosynthetic rates and plant growth, whereas these parameters were negatively affected by WD. The determination of δ15N in leaves, stems, roots and nodules showed that leaves were the most representative organs of the plant response to increased [CO2] and WD. Depletion of heavier N isotopes in plants grown under higher [CO2] and WD conditions reflected decreased transpiration rates, but could also be related to a higher N demand in leaves, as suggested by the decreased leaf N and total soluble protein (TSP contents detected at 700 mol mol-1 [CO2] and WD conditions. In summary, leaf δ15N provides relevant information integrating parameters which condition plant responsiveness (e.g. photosynthesis, TSP, N demand and water transpiration to environmental conditions.

  19. Assessment of heat shock protein 70 induction by heat in alfalfa varieties and constitutive overexpression in transgenic plants.

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    Nicoletta Ferradini

    Full Text Available Heat shock proteins (HSPs are molecular chaperones involved in many cellular functions. It has been shown that mammalian cytosolic HSP70 binds antigenic peptides mediating the activation of the immune system, and that it plays a determining role in tumour immunogenicity. This suggests that HSP70 may be used for the production of conjugated vaccines. Human and plant HSPs share high sequence similarity and some important biological functions in vitro. In addition, plant HSPs have no endotoxic side effects. Extraction of HSP70 from plants for use as vaccine adjuvant requires enhancing its concentration in plant tissues. In this work, we explored the possibility to produce HSP70 in both transgenic and non-transgenic plants, using alfalfa as a model species. First, a transcriptional analysis of a constitutive and an inducible HSP70 genes was conducted in Arabidopsis thaliana. Then the coding sequence of the inducible form was cloned and introduced into alfalfa by Agrobacterium-mediated transformation, and the accumulation of the protein in leaf tissue of transgenic plants was demonstrated. We also tested diverse alfalfa varieties for heat-inducible expression of endogenous HSP70, revealing variety-specific responses to heat shock.

  20. PII Overexpression in Lotus japonicus Affects Nodule Activity in Permissive Low-Nitrogen Conditions and Increases Nodule Numbers in High Nitrogen Treated Plants.

    Science.gov (United States)

    D'Apuzzo, Enrica; Valkov, Vladimir Totev; Parlati, Aurora; Omrane, Selim; Barbulova, Ani; Sainz, Maria Martha; Lentini, Marco; Esposito, Sergio; Rogato, Alessandra; Chiurazzi, Maurizio

    2015-04-01

    We report here the first characterization of a GLNB1 gene coding for the PII protein in leguminous plants. The main purpose of this work was the investigation of the possible roles played by this multifunctional protein in nodulation pathways. The Lotus japonicus LjGLB1 gene shows a significant transcriptional regulation during the light-dark cycle and different nitrogen availability, conditions that strongly affect nodule formation, development, and functioning. We also report analysis of the spatial profile of expression of LjGLB1 in root and nodule tissues and of the protein's subcellular localization. Transgenic L. japonicus lines overexpressing the PII protein were obtained and tested for the analysis of the symbiotic responses in different conditions. The uncoupling of PII from its native regulation affects nitrogenase activity and nodule polyamine content. Furthermore, our results suggest the involvement of PII in the signaling of the nitrogen nutritional status affecting the legumes' predisposition for nodule formation.

  1. Fixação do nitrogênio em alfafa nodulada sob supressão e ressuprimento de fósforo Nitrogen fixation in alfalfa nodulated under phosphorus supression and resupply

    Directory of Open Access Journals (Sweden)

    Fernando Teixeira Gomes

    2002-12-01

    Full Text Available Estudaram-se os efeitos da supressão e do ressuprimento de fósforo (Pi sobre a fixação biológica do nitrogênio atmosférico (N2 em plantas de alfafa cv. Flórida 77 inoculadas com Sinorhizobium meliloti, em diferentes estádios do desenvolvimento vegetativo (V3 e V4 e reprodutivo (R6 e R8. O ensaio foi conduzido em casa de vegetação e as plantas cultivadas em solução nutritiva. O período de supressão de Pi por dez dias reduziu os teores de Pi nas folhas e nos nódulos em todos os estádios do desenvolvimento, enquanto nas raízes essa redução somente foi observada nos estádios vegetativos. Após o ressuprimento de Pi à solução nutritiva por igual período, dez dias, foi observada a recuperação nos teores de Pi nos estádios R6 e R8 para folhas, V3 e V4 para raízes e V3 para os nódulos. A supressão de Pi alterou o processo de fixação do N2, estimado pela concentração de aminoácidos totais nas folhas e nas raízes. Os teores de aminoácidos nas folhas e nas raízes foram significativamente menores nas plantas sob supressão de Pi, em relação aos das plantas do tratamento controle. Após o ressuprimento os teores de aminoácidos totais nas raízes, em todos os estádios do desenvolvimento, alcançaram valores similares àqueles das plantas do tratamento controle, enquanto nas folhas isso só ocorreu nos estádios vegetativos. A supressão de Pi não influenciou a proporção de aminoácidos livres na seiva do xilema.Phosphorus (Pi suppression and resupply effects were studied on nitrogen biological fixation (N2 in alfalfa cv. Florida 77 inoculated with Sinorhizobium meliloti in different vegetative (V3 and V4 and reproductive (R6 and R8 stages. The experiment was carried in greenhouse and the plants cultivated in nutritive solution. The inorganic phosphorus (Pi ten days suppression period decreased Pi levels in leaves and nodules in all growth stages, whereas in the roots this decrease was observed only in the

  2. 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...

  3. Morphological, physiological and plant infectivity characterization of Frankia strains isolated from Casuarina’s nodules

    OpenAIRE

    Patrick Moritz; Kelly Campos Guerra P. de Goes; Souza,José Roberto P de; Letícia Trindade Ataíde; Diva Souza Andrade

    2007-01-01

    Frankia are soil microorganisms that form symbiosis with roots of tree species called actinorhizal plants and are capable of fixing atmospheric N2. This study was carried out to characterize morphologically, physiologically and to assess the nodulation of four Frankia reference strains (HFPCcI3, JCT287, KB5 and F59) and 12 (IPRF) isolated from root nodules of Casuarina plants. All strains (Reference and IPRF) were characterized as Gram-positive and 50% as acid-fast. The Frankia strains produc...

  4. [Analysis of Symbiotic Genes of Leguminous Plants Nodule Bacteria Grown in the Southern Urals].

    Science.gov (United States)

    Baymiev, An Kh; Ivanova, E S; Gumenko, R S; Chubukova, O V; Baymiev, Al Kh

    2015-12-01

    Bacterial strains isolated from the nodules, tissues, and root surface of wild legumes growing in the Southern Urals related to the tribes Galegeae, Hedysareae, Genisteae, Trifolieae, and Loteae were examined for the presence in their genomes of symbiotic (sym) genes. It was found that the sym-genes are present in microorganisms isolated only from the nodules of the analyzed plants (sym+ -strains). Phylogenetic analysis of sym+ -strains on the basis of a comparative analysis of 16S rRNA gene sequences showed that sym+ -strains belong to five families of nodule bacteria: Mesorhizobium, Bradyrhizobium, Sinorhizobium, Rhizobium, and Phyllobacterium. A study the phylogeny of the sym-genes showed that the nodule bacteria of leguminous plants of the Southern Urals at the genus level are mainly characterized by a parallel evolution of symbiotic genes and the 16S rRNA gene. Thus, cases of horizontal transfer of sym genes, which sometimes leads to the formation of certain types of atypical rhizobial strains ofleguminous plants, are detected in nodule bacteria populations.

  5. Diversity of frankia strains in root nodules of plants from the families elaeagnaceae and rhamnaceae

    Science.gov (United States)

    Clawson; Caru; Benson

    1998-09-01

    Partial 16S ribosomal DNAs (rDNAs) were PCR amplified and sequenced from Frankia strains living in root nodules of plants belonging to the families Elaeagnaceae and Rhamnaceae, including Colletia hystrix, Elaeagnus angustifolia, an unidentified Elaeagnus sp., Talguenea quinquenervia, and Trevoa trinervis. Nearly full-length 16S rDNAs were sequenced from strains of Frankia living in nodules of Ceanothus americanus, C. hystrix, Coriaria arborea, and Trevoa trinervis. Partial sequences also were obtained from Frankia strains isolated and cultured from the nodules of C. hystrix, Discaria serratifolia, D. trinervis, Retanilla ephedra, T. quinquenervia, and T. trinervis (Rhamnaceae). Comparison of these sequences and other published sequences of Frankia 16S rDNA reveals that the microsymbionts and isolated strains from the two plant families form a distinct phylogenetic clade, except for those from C. americanus. All sequences in the clade have a common 2-base deletion compared with other Frankia strains. Sequences from C. americanus nodules lack the deletion and cluster with Frankia strains infecting plants of the family Rosaceae. Published plant phylogenies (based on chloroplast rbcL sequences) group the members of the families Elaeagnaceae and Rhamnaceae together in the same clade. Thus, with the exception of C. americanus, actinorhizal plants of these families and their Frankia microsymbionts share a common symbiotic origin.

  6. Comparative phytotoxicity of ZnO NPs, bulk ZnO, and ionic zinc onto the alfalfa plants symbiotically associated with Sinorhizobium meliloti in soil.

    Science.gov (United States)

    Bandyopadhyay, Susmita; Plascencia-Villa, Germán; Mukherjee, Arnab; Rico, Cyren M; José-Yacamán, Miguel; Peralta-Videa, Jose R; Gardea-Torresdey, Jorge L

    2015-05-15

    ZnO nanoparticles (NPs) are reported as potentially phytotoxic in hydroponic and soil media. However, studies on ZnO NPs toxicity in a plant inoculated with bacterium in soil are limited. In this study, ZnO NPs, bulk ZnO, and ZnCl₂ were exposed to the symbiotic alfalfa (Medicago sativa L.)-Sinorhizobium meliloti association at concentrations ranging from 0 to 750 mg/kg soil. Plant growth, Zn bioaccumulation, dry biomass, leaf area, total protein, and catalase (CAT) activity were measured in 30 day-old plants. Results showed 50% germination reduction by bulk ZnO at 500 and 750 mg/kg and all ZnCl₂ concentrations. ZnO NPs and ionic Zn reduced root and shoot biomass by 80% and 25%, respectively. Conversely, bulk ZnO at 750 mg/kg increased shoot and root biomass by 225% and 10%, respectively, compared to control. At 500 and 750 mg/kg, ZnCl₂ reduced CAT activity in stems and leaves. Total leaf protein significantly decreased as external ZnCl₂ concentration increased. STEM-EDX imaging revealed the presence of ZnO particles in the root, stem, leaf, and nodule tissues. ZnO NPs showed less toxicity compared to ZnCl₂ and bulk ZnO found to be growth enhancing on measured traits. These findings are significant to reveal the toxicity effects of different Zn species (NPs, bulk, and ionic Zn) into environmentally important plant-bacterial system in soil.

  7. Common and divergent roles of plant hormones in nodulation and arbuscular mycorrhizal symbioses.

    Science.gov (United States)

    Foo, Eloise; Ferguson, Brett J; Reid, James B

    2014-01-01

    All of the classical plant hormones have been suggested to influence nodulation, including some that interact with the Autoregulation of Nodulation (AON) pathway. Leguminous plants strictly regulate the number of nodules formed through this AON pathway via a root-shoot-root loop that acts to suppress excessive nodulation. A related pathway, the Autoregulation of Mycorrhization (AOM) pathway controls the more ancient, arbuscular mycorrhizal (AM) symbiosis. A comparison of the published responses to the classical hormones in these 2 symbioses shows that most influence the symbioses in the same direction. This may be expected if they affect the symbioses via common components of these symbiotic regulatory pathways. However, some hormones influence these symbioses in opposite directions, suggesting a more complex relationship, and probably one that is not via the common components of these pathways. In a recent paper we showed, using a genetic approach, that strigolactones and brassinosteroids do not act downstream of the AON genes examined and argued that they probably act independently to promote nodule formation. Recently it has been shown that the control of nodulation via the AON pathway involves mobile CLE peptide signals. It is therefore suggested that a more direct avenue to determine if the classical hormones play a direct role in the autoregulatory pathways is to further examine whether CLE peptides and other components of these processes can influence, or be influenced by, the classical hormones. Such studies and other comparisons between the nodulation and mycorrhizal symbioses should allow the role of the classical hormones in these critical symbioses to be rapidly advanced.

  8. MicroRNA156 as a promising tool for alfalfa improvement.

    Science.gov (United States)

    Aung, Banyar; Gruber, Margaret Y; Amyot, Lisa; Omari, Khaled; Bertrand, Annick; Hannoufa, Abdelali

    2015-08-01

    A precursor of miR156 (MsmiR156d) was cloned and overexpressed in alfalfa (Medicago sativa L.) as a means to enhance alfalfa biomass yield. Of the five predicted SPL genes encoded by the alfalfa genome, three (SPL6, SPL12 and SPL13) contain miR156 cleavage sites and their expression was down-regulated in transgenic alfalfa plants overexpressing miR156. These transgenic plants had reduced internode length and stem thickness, enhanced shoot branching, increased trichome density, a delay in flowering time and elevated biomass production. Minor effects on sugar, starch, lignin and cellulose contents were also observed. Moreover, transgenic alfalfa plants had increased root length, while nodulation was maintained. The multitude of traits affected by miR156 may be due to the network of genes regulated by the three target SPLs. Our results show that the miR156/SPL system has strong potential as a tool to substantially improve quality and yield traits in alfalfa. © 2014 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.

  9. Sinorhizobium meliloti nifA gene exerts a pleiotropic effect on nodulation through the enhanced plant defense response

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Sinorhizobium meliloti nifA gene is required for the expression of a bunch of nif and fix genes. Here, we report its pleiotropic effects on the nodule formation. Compared with wild type strain, nifA mutant significantly reduced nodule suppression rate in split-root system. The plants inoculated with mutant strain produced lower amount of daidzein and less necrotic cells on their roots. In addition, the defense genes failed to be evoked by nifA mutant at the early nodulation stage. These findings indicated that host defense response was one of the mechanisms mediated by nifA gene to regulate nodule formation during symbiosis. Even though nifA mutant could increase the number of nodules in host plant, it synthesized lower Nod factors than wild type. This suggested that nifA gene mediated multiple and diverse instances in nodulation formation.

  10. Establishing alfalfa in silage corn

    Science.gov (United States)

    According to recent agricultural statistics, alfalfa was planted on 0.44 million acres and harvested from 2.2 million acres and silage corn was planted and harvested from 1.0 million acres per year in Wisconsin. Because both crops are often grown in rotation, alfalfa could be interseeded at corn pla...

  11. Establishing alfalfa in corn silage

    Science.gov (United States)

    According to recent agricultural statistics, alfalfa was planted on 0.44 million acres and harvested from 2.2 million acres, and corn silage was planted and harvested from 1.0 million acres per year in Wisconsin. Because both crops are often grown in rotation, alfalfa could be interseeded at corn pl...

  12. Metabolomic Profiling of Bradyrhizobium diazoefficiens-Induced Root Nodules Reveals Both Host Plant-Specific and Developmental Signatures

    OpenAIRE

    Lardi, Martina; Murset, Valérie; Fischer, Hans-Martin; Mesa, Socorro; Ahrens, Christian H.; Zamboni, Nicola; Pessi, Gabriella

    2016-01-01

    Bradyrhizobium diazoefficiens is a nitrogen-fixing endosymbiont, which can grow inside root-nodule cells of the agriculturally important soybean and other host plants. Our previous studies described B. diazoefficiens host-specific global expression changes occurring during legume infection at the transcript and protein level. In order to further characterize nodule metabolism, we here determine by flow injection–time-of-flight mass spectrometry analysis the metabolome of (i) nodules and roots...

  13. 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

    OpenAIRE

    Mariza Monteiro; Beatriz Appezzato-da-Glória; Maria José Valarini; Carlos Alberto de Oliveira; Maria Lucia Carneiro Vieira

    2003-01-01

    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 pro...

  14. Nitrogen assimilation by nodulate plants of Phaseolus vulgaris l. and Vigna unguiculata (l. ) walp

    Energy Technology Data Exchange (ETDEWEB)

    Neves, M.C.P.; Fernandes, M.S.; Sa, M.F.M. (Universidade Federal Rural do Rio de Janeiro (Brazil). Dept. de Solos)

    1982-05-01

    Under field conditions, the processes of nitrogen assimilation via nitrogenase and nitrate-reductase, the transport and the accumulation of nitrogen in nodulated plants of Phaseolus vulgaris cv. Rio Tibagi and Vigna unguiculata cv. Vita 34 were compared and contrasted. V. unguiculata showed better nodulation than P. vulgaris and consequently had higher rates of nitrogenase activity. The small nodulation of P. vulgaris resulted in greater dependence on soil mineral nitrogen as indicated by the higher rates of nitrate-reductase acitivty compared with V. unguiculata, especially during reproductive stage of growth. The superiority of V. unguiculata in terms of assimilation and remobilization of stored nitrogen resulted in a seed yield 28% greater than that of P. vulgaris. P. vulgaris showed a negative correlation between the nitrate-reductase activity and the ureide content of the sap indicating that the metabolic pathways leading to ureide production operates alternatively to nitrate assimilation.

  15. Effects of simulated acid precipitation on growth and nodulation of leguminous plants

    Energy Technology Data Exchange (ETDEWEB)

    Chang, F.H.; Alexander, M.

    1983-04-01

    Soil samples were collected from the Panther Lake, Sagamore Lake and Woods Lake watersheds of the Adirondack region of New York. The first two soils were from under coniferous canopy and the second from under a deciduous canopy. Agricultural soil was tested for comparative purposes. Simulated acid rain was applied to the soil samples for 14 days before seeds of Arlington red clover (Trifolium pratense L.) and Viking birdsfoot trefoil (Lotus corniculatus L.) were sown. Results show that, based on nodule numbers and depth, nodulation of birdsfoot trefoil was more tolerant to acidity than that of red clover. The depth to which nodulation occurred was reduced with decreasing pH of the simulated rain. Yield and nitrogen content of both plant species were significantly reduced in two soils. Under the test conditions, the legumes were stunted and neither species nodulated in Sagamore soil. The enhancement of plant growth and nitrogen uptake in both plants in Howard soil (pH 6.7) may be a reflection of improved nutrient availability arising from chemical action of the more acid solutions. (JMT)

  16. nip, a symbiotic Medicago truncatula mutant that forms root nodules with aberrant infection threads and plant defense-like response.

    Science.gov (United States)

    Veereshlingam, Harita; Haynes, Janine G; Penmetsa, R Varma; Cook, Douglas R; Sherrier, D Janine; Dickstein, Rebecca

    2004-11-01

    To investigate the legume-Rhizobium symbiosis, we isolated and studied a novel symbiotic mutant of the model legume Medicago truncatula, designated nip (numerous infections and polyphenolics). When grown on nitrogen-free media in the presence of the compatible bacterium Sinorhizobium meliloti, the nip mutant showed nitrogen deficiency symptoms. The mutant failed to form pink nitrogen-fixing nodules that occur in the wild-type symbiosis, but instead developed small bump-like nodules on its roots that were blocked at an early stage of development. Examination of the nip nodules by light microscopy after staining with X-Gal for S. meliloti expressing a constitutive GUS gene, by confocal microscopy following staining with SYTO-13, and by electron microscopy revealed that nip initiated symbiotic interactions and formed nodule primordia and infection threads. The infection threads in nip proliferated abnormally and very rarely deposited rhizobia into plant host cells; rhizobia failed to differentiate further in these cases. nip nodules contained autofluorescent cells and accumulated a brown pigment. Histochemical staining of nip nodules revealed this pigment to be polyphenolic accumulation. RNA blot analyses demonstrated that nip nodules expressed only a subset of genes associated with nodule organogenesis, as well as elevated expression of a host defense-associated phenylalanine ammonia lyase gene. nip plants were observed to have abnormal lateral roots. nip plant root growth and nodulation responded normally to ethylene inhibitors and precursors. Allelism tests showed that nip complements 14 other M. truncatula nodulation mutants but not latd, a mutant with a more severe nodulation phenotype as well as primary and lateral root defects. Thus, the nip mutant defines a new locus, NIP, required for appropriate infection thread development during invasion of the nascent nodule by rhizobia, normal lateral root elongation, and normal regulation of host defense-like responses

  17. Co-expression of tonoplast Cation/H(+) antiporter and H(+)-pyrophosphatase from xerophyte Zygophyllum xanthoxylum improves alfalfa plant growth under salinity, drought and field conditions.

    Science.gov (United States)

    Bao, Ai-Ke; Du, Bao-Qiang; Touil, Leila; Kang, Peng; Wang, Qiang-Long; Wang, Suo-Min

    2016-03-01

    Salinity and drought are major environmental factors limiting the growth and productivity of alfalfa worldwide as this economically important legume forage is sensitive to these kinds of abiotic stress. In this study, transgenic alfalfa lines expressing both tonoplast NXH and H(+)-PPase genes, ZxNHX and ZxVP1-1 from the xerophyte Zygophyllum xanthoxylum L., were produced via Agrobacterium tumefaciens-mediated transformation. Compared with wild-type (WT) plants, transgenic alfalfa plants co-expressing ZxNHX and ZxVP1-1 grew better with greater plant height and dry mass under normal or stress conditions (NaCl or water-deficit) in the greenhouse. The growth performance of transgenic alfalfa plants was associated with more Na(+), K(+) and Ca(2+) accumulation in leaves and roots, as a result of co-expression of ZxNHX and ZxVP1-1. Cation accumulation contributed to maintaining intracellular ions homoeostasis and osmoregulation of plants and thus conferred higher leaf relative water content and greater photosynthesis capacity in transgenic plants compared to WT when subjected to NaCl or water-deficit stress. Furthermore, the transgenic alfalfa co-expressing ZxNHX and ZxVP1-1 also grew faster than WT plants under field conditions, and most importantly, exhibited enhanced photosynthesis capacity by maintaining higher net photosynthetic rate, stomatal conductance, and water-use efficiency than WT plants. Our results indicate that co-expression of tonoplast NHX and H(+)-PPase genes from a xerophyte significantly improved the growth of alfalfa, and enhanced its tolerance to high salinity and drought. This study laid a solid basis for reclaiming and restoring saline and arid marginal lands as well as improving forage yield in northern China.

  18. Soil factors exhibit greater influence than bacterial inoculation on alfalfa growth and nitrogen fixation.

    Science.gov (United States)

    Neumann, Ute; Kosier, Bob; Jahnke, Joachim; Priefer, Ursula B; Al-Halbouni, Djamila

    2011-09-01

    In order to study the effects of soil factors and bacterial inoculation on alfalfa (Medicago sativa), plants were inoculated with Ensifer meliloti L33 and Azospirillum brasilense Sp7 in pot experiments using two different soils separately as well as in a mixture. One soil was contaminated with chemical waste products; the other was an arable soil. Soil factors, including the availability of macro- and micronutrients as well as carbon and nitrogen contents, were found to exhibit a much greater influence on the growth of alfalfa than any of the inoculations. In contaminated soil, the shoot and root growth of alfalfa was decreased and nodules were diminished and ineffective. Bacterial inoculations did not significantly improve this hostile growth environment. However, in a mixture (44% arable, 22% contaminated soil, 34% vermiculite), growth conditions for alfalfa were improved so that shoot dry weight and nodule numbers increased up to 100- and 20-fold, respectively, compared with the contaminated soil. For the strain L33, its persistence in the rhizosphere was correlated to the presence of its host plant, but its dynamics were influenced by competition with indigenous rhizobia. The strain Sp7, once provided with a suitable soil, was not dependent on the plant's rhizosphere, but it enhanced the performance of L33 and native rhizobia.

  19. The effects of selenate and sulphate supply on the accumulation and volatilization of Se by cabbage, kohlrabi and alfalfa plants grown hydroponically

    Directory of Open Access Journals (Sweden)

    R. HAJIBOLAND

    2008-12-01

    Full Text Available The effect of Selenium (Se supplementation at five levels of 0 (control, 5, 10, 15, 20 ìM in plants supplied with one of four concentrations of sulphur (S including 0.05, 0.25, 0.5 and 1.0 mM was investigated in two varieties of Brassica oleracea (cabbage and kohlrabi and alfalfa (Medicago sativa L. in a hydroponic experiment. In severely S deficient plants (0.05 mM, Se acted as a toxic element, alfalfa was the most susceptible plant that died at this treatment. However, in plants supplied with near adequate (0.5 mM or adequate (1.0 mM S, Se acted as a growth promoting element. The most pronounced stimulation of growth was observed in cabbage and the lowest in alfalfa. Increasing S concentration in the medium, reduced Se uptake and transport. In contrast, S uptake and transport increased in response to Se addition. Se volatilization was higher in alfalfa than cabbage and kohlrabi when expressed on unit shoot dry weight or leaf area basis, but not when expressed per plant. Results suggested that Se supplementation of plants supplied with adequate S, not only had beneficial effects on plants growth but also can have application in enrichment of livestock fodder and human food.;

  20. Fungi isolated from the initial industrial soil planted with a mixture of alfalfa and grasses

    Directory of Open Access Journals (Sweden)

    Maria Kowalik

    2014-08-01

    Full Text Available Research were conducted during the years 1988-1990 on the "Machów" industrial waste dump. It was determined, that a mixture of alfalfa with grasses. introduced as first culture on the raw substrate of industrial wastes. was helpful for the development of soil fungi communities.

  1. 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.

  2. Transgenic alfalfa plants co-expressing glutathione S-transferase (GST) and human CYP2E1 show enhanced resistance to mixed contaminates of heavy metals and organic pollutants

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Yuanyuan [Department of Pharmaceutics, Qingdao University of Science and Technology, 53 Zhengzhou Road, P.O. Box 70, Qingdao 266042 (China); Liu, Junhong, E-mail: liujh@qust.edu.cn [Department of Pharmaceutics, Qingdao University of Science and Technology, 53 Zhengzhou Road, P.O. Box 70, Qingdao 266042 (China)

    2011-05-15

    Transgenic alfalfa plants simultaneously expressing human CYP2E1 and glutathione S-transferase (GST) were generated from hypocotyl segments by the use of an Agrobacterium transformation system for the phytoremediation of the mixed contaminated soil with heavy metals and organic pollutants. The transgenic alfalfa plants were screened by a combination of kanamycin resistance, PCR, GST and CYP2E1 activity and Western blot analysis. The capabilities of mixed contaminants (heavy metals-organic compounds) resistance of pKHCG transgenic alfalfa plants became markedly increased compared with the transgenic alfalfa plants expressing single gene (GST or CYP2E1) and the non-transgenic control plants. The pKHCG alfalfa plants exhibited strong resistance towards the mixtures of cadmium (Cd) and trichloroethylene (TCE) that were metabolized by the introduced GST and CYP2E1 in combination. Our results show that the pKHCG transgenic alfalfa plants have good potential for phytoremediation because they have cross-tolerance towards the complex contaminants of heavy metals and organic pollutants. Therefore, these transgenic alfalfa plants co-expressing GST and human P450 CDNAs may have a great potential for phytoremediation of mixed environmental contaminants.

  3. Both plant and bacterial nitrate reductases contribute to nitric oxide production in Medicago truncatula nitrogen-fixing nodules.

    Science.gov (United States)

    Horchani, Faouzi; Prévot, Marianne; Boscari, Alexandre; Evangelisti, Edouard; Meilhoc, Eliane; Bruand, Claude; Raymond, Philippe; Boncompagni, Eric; Aschi-Smiti, Samira; Puppo, Alain; Brouquisse, Renaud

    2011-02-01

    Nitric oxide (NO) is a signaling and defense molecule of major importance in living organisms. In the model legume Medicago truncatula, NO production has been detected in the nitrogen fixation zone of the nodule, but the systems responsible for its synthesis are yet unknown and its role in symbiosis is far from being elucidated. In this work, using pharmacological and genetic approaches, we explored the enzymatic source of NO production in M. truncatula-Sinorhizobium meliloti nodules under normoxic and hypoxic conditions. When transferred from normoxia to hypoxia, nodule NO production was rapidly increased, indicating that NO production capacity is present in functioning nodules and may be promptly up-regulated in response to decreased oxygen availability. Contrary to roots and leaves, nodule NO production was stimulated by nitrate and nitrite and inhibited by tungstate, a nitrate reductase inhibitor. Nodules obtained with either plant nitrate reductase RNA interference double knockdown (MtNR1/2) or bacterial nitrate reductase-deficient (napA) and nitrite reductase-deficient (nirK) mutants, or both, exhibited reduced nitrate or nitrite reductase activities and NO production levels. Moreover, NO production in nodules was found to be inhibited by electron transfer chain inhibitors, and nodule energy state (ATP-ADP ratio) was significantly reduced when nodules were incubated in the presence of tungstate. Our data indicate that both plant and bacterial nitrate reductase and electron transfer chains are involved in NO synthesis. We propose the existence of a nitrate-NO respiration process in nodules that could play a role in the maintenance of the energy status required for nitrogen fixation under oxygen-limiting conditions.

  4. Proteomic insights into intra- and intercellular plant-bacteria symbiotic association during root nodule formation

    Directory of Open Access Journals (Sweden)

    Afshin eSalavati

    2013-02-01

    Full Text Available Over the last several decades, there have been a large number of studies done on the all aspects of legumes and bacteria which participate in nitrogen-fixing symbiosis. The analysis of legume-bacteria interaction is not just a matter of numerical complexity in terms of variants of gene products that can arise from a single gene. Bacteria regulate their quorum-sensing genes to enhance their ability to induce conjugation of plasmids and symbiotic islands, and various protein secretion mechanisms; that can stimulate a collection of chain reactions including species-specific combinations of plant-secretion isoflavonoids, complicated calcium signaling pathways and autoregulation of nodulation mechanisms. Quorum-sensing systems are introduced by the intra- and intercellular organization of gene products lead to protein–protein interactions or targeting of proteins to specific cellular structures. In this study, an attempt has been made to review significant contributions related to nodule formation and development and their impacts on cell proteome for better understanding of plant-bacterium interaction mechanism at protein level. This review would not only provide new insights into the plant-bacteria symbiosis response mechanisms but would also highlights the importance of studying changes in protein abundance inside and outside of cells in response to symbiosis. Furthermore, the application to agriculture programe of plant-bacteria interaction will be discussed.

  5. A Nodule-Specific Lipid Transfer Protein AsE246 Participates in Transport of Plant-Synthesized Lipids to Symbiosome Membrane and Is Essential for Nodule Organogenesis in Chinese Milk Vetch1[C][W][OPEN

    Science.gov (United States)

    Lei, Lei; Chen, Ling; Shi, Xiaofeng; Li, Yixing; Wang, Jianyun; Chen, Dasong; Xie, Fuli; Li, Youguo

    2014-01-01

    Rhizobia in legume root nodules fix nitrogen in symbiosomes, organelle-like structures in which a membrane from the host plant surrounds the symbiotic bacteria. However, the components that transport plant-synthesized lipids to the symbiosome membrane remain unknown. This study identified and functionally characterized the Chinese milk vetch (Astragalus sinicus) lipid transfer protein AsE246, which is specifically expressed in nodules. It was found that AsE246 can bind lipids in vitro. More importantly, AsE246 can bind the plant-synthesized membrane lipid digalactosyldiacylglycerol in vivo. Immunofluorescence and immunoelectron microscopy showed that AsE246 and digalactosyldiacylglycerol localize in the symbiosome membrane and are present in infection threads. Overexpression of AsE246 resulted in increased nodule numbers; knockdown of AsE246 resulted in reduced nodule numbers, decreased lipids contents in nodules, diminished nitrogen fixation activity, and abnormal development of symbiosomes. AsE246 knockdown also resulted in fewer infection threads, nodule primordia, and nodules, while AsE246 overexpression resulted in more infection threads and nodule primordia, suggesting that AsE246 affects nodule organogenesis associated with infection thread formation. Taken together, these results indicate that AsE246 contributes to lipids transport to the symbiosome membrane, and this transport is required for effective legume-rhizobium symbiosis. PMID:24367021

  6. enod40, a gene expressed during nodule organogenesis, codes for a non-translatable RNA involved in plant growth.

    Science.gov (United States)

    Crespi, M D; Jurkevitch, E; Poiret, M; d'Aubenton-Carafa, Y; Petrovics, G; Kondorosi, E; Kondorosi, A

    1994-01-01

    Rhizobium meliloti can interact symbiotically with Medicago plants, thereby inducing root nodules. However, certain Medicago plants can form nodules spontaneously, in the absence of rhizobia. A differential screening was performed using spontaneous nodule versus root cDNAs from Medicago sativa ssp. varia. Transcripts of a differentially expressed clone, Msenod40, were detected in all differentiating cells of nodule primordia and spontaneous nodules, but were absent in fully differentiated cells. Msenod40 showed homology to a soybean early nodulin gene, Gmenod40, although no significant open reading frame (ORF) or coding capacity was found in the Medicago sequence. Furthermore, in the sequences of cDNAs and a genomic clone (Mtenod40) isolated from Medicago truncatula, a species containing a unique copy of this gene, no ORFs were found either. In vitro translation of purified Mtenod40 transcripts did not reveal any protein product. Evaluation of the RNA secondary structure indicated that both msenod40 and Gmenod40 transcripts showed a high degree of stability, a property shared with known non-coding RNAs. The Mtenod40 RNA was localized in the cytoplasm of cells in the nodule primordium. Infection with Agrobacterium tumefaciens strains bearing antisense constructs of Mtenod40 arrested callus growth of Medicago explants, while overexpressing Mtenod40 embryos developed into teratomas. These data suggest that the enod40 genes might have a role in plant development, acting as 'riboregulators', a novel class of untranslated RNAs associated with growth control and differentiation. Images PMID:7957074

  7. Effect of shoot removal on remobilization of carbon and nitrogen during regrowth of nitrogen-fixing alfalfa.

    Science.gov (United States)

    Aranjuelo, Iker; Molero, Gemma; Erice, Gorka; Aldasoro, Joseba; Arrese-Igor, Cesar; Nogués, Salvador

    2015-01-01

    The contribution of carbon and nitrogen reserves to regrowth following shoot removal has been studied in the past. However, important gaps remain in understanding the effect of shoot cutting on nodule performance and its relevance during regrowth. In this study, isotopic labelling was conducted at root and canopy levels with both (15) N2 and (13) C-depleted CO2 on exclusively nitrogen-fixing alfalfa plants. As expected, our results indicate that the roots were the main sink organs before shoots were removed. Seven days after regrowth the carbon and nitrogen stored in the roots was invested in shoot biomass formation and partitioned to the nodules. The large depletion in nodule carbohydrate availability suggests that root-derived carbon compounds were delivered towards nodules in order to sustain respiratory activity. In addition to the limited carbohydrate availability, the upregulation of nodule peroxidases showed that oxidative stress was also involved during poor nodule performance. Fourteen days after cutting, and as a consequence of the stimulated photosynthetic and N2 -fixing machinery, availability of Cnew and Nnew strongly diminished in the plants due to their replacement by C and N assimilated during the post-labelling period. In summary, our study indicated that during the first week of regrowth, root-derived C and N remobilization did not overcome C- and N-limitation in nodules and leaves. However, 14 days after cutting, leaf and nodule performance were re-established.

  8. A comparative genomics screen identifies a Sinorhizobium meliloti 1021 sodM-like gene strongly expressed within host plant nodules

    Directory of Open Access Journals (Sweden)

    Queiroux Clothilde

    2012-05-01

    Full Text Available Abstract Background We have used the genomic data in the Integrated Microbial Genomes system of the Department of Energy’s Joint Genome Institute to make predictions about rhizobial open reading frames that play a role in nodulation of host plants. The genomic data was screened by searching for ORFs conserved in α-proteobacterial rhizobia, but not conserved in closely-related non-nitrogen-fixing α-proteobacteria. Results Using this approach, we identified many genes known to be involved in nodulation or nitrogen fixation, as well as several new candidate genes. We knocked out selected new genes and assayed for the presence of nodulation phenotypes and/or nodule-specific expression. One of these genes, SMc00911, is strongly expressed by bacterial cells within host plant nodules, but is expressed minimally by free-living bacterial cells. A strain carrying an insertion mutation in SMc00911 is not defective in the symbiosis with host plants, but in contrast to expectations, this mutant strain is able to out-compete the S. meliloti 1021 wild type strain for nodule occupancy in co-inoculation experiments. The SMc00911 ORF is predicted to encode a “SodM-like” (superoxide dismutase-like protein containing a rhodanese sulfurtransferase domain at the N-terminus and a chromate-resistance superfamily domain at the C-terminus. Several other ORFs (SMb20360, SMc01562, SMc01266, SMc03964, and the SMc01424-22 operon identified in the screen are expressed at a moderate level by bacteria within nodules, but not by free-living bacteria. Conclusions Based on the analysis of ORFs identified in this study, we conclude that this comparative genomics approach can identify rhizobial genes involved in the nitrogen-fixing symbiosis with host plants, although none of the newly identified genes were found to be essential for this process.

  9. 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.

  10. Nod Factor-Independent Nodulation in Aeschynomene evenia Required the Common Plant-Microbe Symbiotic Toolkit.

    Science.gov (United States)

    Fabre, Sandrine; Gully, Djamel; Poitout, Arthur; Patrel, Delphine; Arrighi, Jean-François; Giraud, Eric; Czernic, Pierre; Cartieaux, Fabienne

    2015-12-01

    Nitrogen fixation in the legume-rhizobium symbiosis is a crucial area of research for more sustainable agriculture. Our knowledge of the plant cascade in response to the perception of bacterial Nod factors has increased in recent years. However, the discovery that Nod factors are not involved in the Aeschynomene-Bradyrhizobium spp. interaction suggests that alternative molecular dialogues may exist in the legume family. We evaluated the conservation of the signaling pathway common to other endosymbioses using three candidate genes: Ca(2+)/Calmodulin-Dependent Kinase (CCaMK), which plays a central role in cross signaling between nodule organogenesis and infection processes; and Symbiosis Receptor Kinase (SYMRK) and Histidine Kinase1 (HK1), which act upstream and downstream of CCaMK, respectively. We showed that CCaMK, SYMRK, and HK1 are required for efficient nodulation in Aeschynomene evenia. Our results demonstrate that CCaMK and SYMRK are recruited in Nod factor-independent symbiosis and, hence, may be conserved in all vascular plant endosymbioses described so far.

  11. Plant nodulation inducers enhance horizontal gene transfer of Azorhizobium caulinodans symbiosis island.

    Science.gov (United States)

    Ling, Jun; Wang, Hui; Wu, Ping; Li, Tao; Tang, Yu; Naseer, Nawar; Zheng, Huiming; Masson-Boivin, Catherine; Zhong, Zengtao; Zhu, Jun

    2016-11-29

    Horizontal gene transfer (HGT) of genomic islands is a driving force of bacterial evolution. Many pathogens and symbionts use this mechanism to spread mobile genetic elements that carry genes important for interaction with their eukaryotic hosts. However, the role of the host in this process remains unclear. Here, we show that plant compounds inducing the nodulation process in the rhizobium-legume mutualistic symbiosis also enhance the transfer of symbiosis islands. We demonstrate that the symbiosis island of the Sesbania rostrata symbiont, Azorhizobium caulinodans, is an 87.6-kb integrative and conjugative element (ICE(Ac)) that is able to excise, form a circular DNA, and conjugatively transfer to a specific site of gly-tRNA gene of other rhizobial genera, expanding their host range. The HGT frequency was significantly increased in the rhizosphere. An ICE(Ac)-located LysR-family transcriptional regulatory protein AhaR triggered the HGT process in response to plant flavonoids that induce the expression of nodulation genes through another LysR-type protein, NodD. Our study suggests that rhizobia may sense rhizosphere environments and transfer their symbiosis gene contents to other genera of rhizobia, thereby broadening rhizobial host-range specificity.

  12. Expression of an alfalfa (Medicago sativa L.) ethylene response factor gene MsERF8 in tobacco plants enhances resistance to salinity.

    Science.gov (United States)

    Chen, Tingting; Yang, Qingchuan; Gruber, Margaret; Kang, Junmei; Sun, Yan; Ding, Wang; Zhang, Tiejun; Zhang, Xinquan

    2012-05-01

    Ethylene response factors (ERF) play crucial roles in plant development and response to stresses. Here, a novel cDNA fragment (MsERF8) encoding an ERF protein with an AP2 domain was isolated and characterized from alfalfa. The MsERF8 cDNA has an open reading frame of 603 bp and encodes a nuclear protein of 201 amino acids. Q-RT-PCR analysis revealed that MsERF8 was strongly enriched in roots and leaves compared with stems, flower buds and flowers of mature alfalfa plants. Bioinformatic analysis of the MsERF8 promoter indicated a number of elements associated with stress-related responses, and MsERF8 transcripts in alfalfa seedlings were induced by NaCl, PEG6000, Al(2)(SO(4))(3) and five different hormones. Expression of MsERF8 in transgenic tobacco plants resulted in higher tolerance to salinity than with non-transgenic plants. This data shows that MsERF8 is a gene which prevents or alleviates salinity damage and has strong potential to impart salt tolerance to other crop plants.

  13. Effect of salinity on some growth indices and total protein content of alfalfa genotypes inoculated with Sinorhizobium meliloti strains under greenhouse conditions

    Directory of Open Access Journals (Sweden)

    A. Fazaeli

    2012-06-01

    Full Text Available This greenhouse experiment was carried out to evaluate the effects of salinity and bacterial inoculation on some growth indices and total protein content of alfalfa (Medicago sativa using a factorial completely randomized design with three replications. The effect of three salinity levels (0, 6 and 12 dS/m induced by a mixture of NaCl, CaCl2 and MgCl2 salts on growth indices and protein content of three alfalfa genotypes (Hamadani, Gharahyonjeh and Gharghalogh at three levels of inoculation with Sinorhizobium meliloti bacteria (no inoculation, inoculation with salinity-tolerant strain and inoculation with salinity-sensitive strain was investigated. After the isolation and purification of alfalfa-symbiotic-bacteria from alfalfa fields in Tehran province, two isolates of S. meliloti, one salinity-resistant and the other one salinity-sensitive, which are effective in symbiosis with alfalfa, were selected. Analysis of the results showed that by increasing the salinity level, the shoot and root dry weight, number of active nodules, and nitrogen (N concentration were decreased significantly (P<0.01. Inoculation with salinity-resistant strain of S. meliloti caused significant increase in shoot and root dry weight, number of active nodules, and N concentration of plants. Moreover, under saline conditions, the salinity-resistant S. meliloti strain increased significantly most alfalfa growth-indices and yield compared to control (without inoculation and inoculation with salinity-sensitive bacteria. There was no significant difference among alfalfa genotypes in yield and other growth indices under saline conditions. In general, the R59 isolate seems to be the best isolate of S. meliloti for greenhouse-grown alfalfa in saline conditions.

  14. Effect of Space Flight Factors on Plant Biomass Developed from Alfalfa Seeds Carried by the Satellite%卫星搭载对紫花苜蓿当代植株生物量的影响

    Institute of Scientific and Technical Information of China (English)

    任卫波; 赵亮; 王蜜; 陈立波; 郭慧琴

    2008-01-01

    [Objective] The aim of this study was to investigate the effect of space flight factors on plant biomass in the generation of alfalfa carried by the satellite. [Method] Seeds from three lines of alfalfa were carried by the seed-breeding satellite Shijian-8. After the satellite returned to the ground, stem diameter, primary branch number and current-year individual biomass of alfalfa were studied. [Result] After space flight, primary branch number and current-year individual biomass of alfalfa increased significantly, while the stem diameter had no significant change. Using the value over(the mean value of control + three standard deviation) as a criterion to screen, the variants with enlarged stem diameter, increased primary branch number and individual biomass was two, five and twelve respectively. [Conclusion] The obtained variants can be used in the variety improvement of alfalfa and its new variety breeding, but whether its favorable variation can inherit stably to the progenies needs further study.

  15. Cowpea Nodules Harbor Non-rhizobial Bacterial Communities that Are Shaped by Soil Type Rather than Plant Genotype

    Science.gov (United States)

    Leite, Jakson; Fischer, Doreen; Rouws, Luc F. M.; Fernandes-Júnior, Paulo I.; Hofmann, Andreas; Kublik, Susanne; Schloter, Michael; Xavier, Gustavo R.; Radl, Viviane

    2017-01-01

    Many studies have been pointing to a high diversity of bacteria associated to legume root nodules. Even though most of these bacteria do not form nodules with legumes themselves, it was shown that they might enter infection threads when co-inoculated with rhizobial strains. The aim of this work was to describe the diversity of bacterial communities associated with cowpea (Vigna unguiculata L. Walp) root nodules using 16S rRNA gene amplicon sequencing, regarding the factors plant genotype and soil type. As expected, Bradyrhizobium was the most abundant genus of the detected genera. Furthermore, we found a high bacterial diversity associated to cowpea nodules; OTUs related to the genera Enterobacter, Chryseobacterium, Sphingobacterium, and unclassified Enterobacteriacea were the most abundant. The presence of these groups was significantly influenced by the soil type and, to a lesser extent, plant genotype. Interestingly, OTUs assigned to Chryseobacterium were highly abundant, particularly in samples obtained from an Ultisol soil. We confirmed their presence in root nodules and assessed their diversity using a target isolation approach. Though their functional role still needs to be addressed, we postulate that Chryseobacterium strains might help cowpea plant to cope with salt stress in semi-arid regions. PMID:28163711

  16. 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.

  17. 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.

  18. 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.

  19. Complete genome sequence and integrated protein localization and interaction map for alfalfa dwarf virus, which combines properties of both cytoplasmic and nuclear plant rhabdoviruses

    Energy Technology Data Exchange (ETDEWEB)

    Bejerman, Nicolás, E-mail: n.bejerman@uq.edu.au [Instituto de Patología Vegetal (IPAVE), Centro de Investigaciones Agropecuarias (CIAP), Instituto Nacional de Tecnología Agropecuaria INTA, Camino a 60 Cuadras k 5,5, Córdoba X5020ICA (Argentina); Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, St Lucia, QLD 4072 (Australia); Giolitti, Fabián; Breuil, Soledad de; Trucco, Verónica; Nome, Claudia; Lenardon, Sergio [Instituto de Patología Vegetal (IPAVE), Centro de Investigaciones Agropecuarias (CIAP), Instituto Nacional de Tecnología Agropecuaria INTA, Camino a 60 Cuadras k 5,5, Córdoba X5020ICA (Argentina); Dietzgen, Ralf G. [Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, St Lucia, QLD 4072 (Australia)

    2015-09-15

    Summary: We have determined the full-length 14,491-nucleotide genome sequence of a new plant rhabdovirus, alfalfa dwarf virus (ADV). Seven open reading frames (ORFs) were identified in the antigenomic orientation of the negative-sense, single-stranded viral RNA, in the order 3′-N-P-P3-M-G-P6-L-5′. The ORFs are separated by conserved intergenic regions and the genome coding region is flanked by complementary 3′ leader and 5′ trailer sequences. Phylogenetic analysis of the nucleoprotein amino acid sequence indicated that this alfalfa-infecting rhabdovirus is related to viruses in the genus Cytorhabdovirus. When transiently expressed as GFP fusions in Nicotiana benthamiana leaves, most ADV proteins accumulated in the cell periphery, but unexpectedly P protein was localized exclusively in the nucleus. ADV P protein was shown to have a homotypic, and heterotypic nuclear interactions with N, P3 and M proteins by bimolecular fluorescence complementation. ADV appears unique in that it combines properties of both cytoplasmic and nuclear plant rhabdoviruses. - Highlights: • The complete genome of alfalfa dwarf virus is obtained. • An integrated localization and interaction map for ADV is determined. • ADV has a genome sequence similarity and evolutionary links with cytorhabdoviruses. • ADV protein localization and interaction data show an association with the nucleus. • ADV combines properties of both cytoplasmic and nuclear plant rhabdoviruses.

  20. Increased photosynthetic acclimation in alfalfa associated with arbuscular mycorrhizal fungi (AMF) and cultivated in greenhouse under elevated CO2.

    Science.gov (United States)

    Goicoechea, Nieves; Baslam, Marouane; Erice, Gorka; Irigoyen, Juan José

    2014-11-15

    Medicago sativa L. (alfalfa) can exhibit photosynthetic down-regulation when grown in greenhouse conditions under elevated atmospheric CO2. This forage legume can establish a double symbiosis with nitrogen fixing bacteria and arbuscular mycorrhizal fungi (AMF), which may increase the carbon sink effect of roots. Our aim was to assess whether the association of alfalfa with AMF can avoid, diminish or delay the photosynthetic acclimation observed in previous studies performed with nodulated plants. The results, however, showed that mycorrhizal (M) alfalfa at the end of their vegetative period had lower carbon (C) discrimination than non-mycorrhizal (NM) controls, indicating photosynthetic acclimation under ECO2 in plants associated with AMF. Decreased C discrimination was due to the acclimation of conductance, since the amount of Rubisco and the expression of genes codifying both large and small subunits of Rubisco were similar or slightly higher in M than in NM plants. Moreover, M alfalfa accumulated a greater amount of soluble sugars in leaves than NM plants, thus favoring a down-regulation effect on photosynthetic rates. The enhanced contents of sugars in leaves coincided with a reduced percentage of arbuscules in roots, suggesting decreased sink of carbohydrates from shoots to roots in M plants. The shorter life cycle of alfalfa associated with AMF in comparison with the NM controls may also be related to the accelerated photosynthetic acclimation in M plants. Further research is needed to clarify to what extent this behavior could be extrapolated to alfalfa cultivated in the field and subjected to periodic cutting of shoots under climatic change scenarios.

  1. Overexpression of EsMcsu1 from the halophytic plant Eutrema salsugineum promotes abscisic acid biosynthesis and increases drought resistance in alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Zhou, C; Ma, Z Y; Zhu, L; Guo, J S; Zhu, J; Wang, J F

    2015-12-17

    The stress phytohormone abscisic acid (ABA) plays pivotal roles in plants' adaptive responses to adverse environments. Molybdenum cofactor sulfurases influence aldehyde oxidase activity and ABA biosynthesis. In this study, we isolated a novel EsMcsu1 gene encoding a molybdenum cofactor sulfurase from Eutrema salsugineum. EsMcus1 transcriptional patterns varied between organs, and its expression was significantly upregulated by abiotic stress or ABA treatment. Alfalfa plants that overexpressed EsMcsu1 had a higher ABA content than wild-type (WT) plants under drought stress conditions. Furthermore, levels of reactive oxygen species (ROS), ion leakage, and malondialdehyde were lower in the transgenic plants than in the WT plants after drought treatment, suggesting that the transgenic plants experienced less ROS-mediated damage. However, the expression of several stress-responsive genes, antioxidant enzyme activity, and osmolyte (proline and total soluble sugar) levels in the transgenic plants were higher than those in the WT plants after drought treatment. Therefore, EsMcsu1 overexpression improved drought tolerance in alfalfa plants by activating a series of ABA-mediated stress responses.

  2. The Sinorhizobium (Ensifer) fredii HH103 Nodulation Outer Protein NopI Is a Determinant for Efficient Nodulation of Soybean and Cowpea Plants.

    Science.gov (United States)

    Jiménez-Guerrero, Irene; Pérez-Montaño, Francisco; Medina, Carlos; Ollero, Francisco Javier; López-Baena, Francisco Javier

    2017-03-01

    The type III secretion system (T3SS) is a specialized secretion apparatus that is commonly used by many plant and animal pathogenic bacteria to deliver proteins, termed effectors, to the interior of the host cells. These effectors suppress host defenses and interfere with signal transduction pathways to promote infection. Some rhizobial strains possess a functional T3SS, which is involved in the suppression of host defense responses, host range determination, and symbiotic efficiency. The analysis of the genome of the broad-host-range rhizobial strain Sinorhizobium fredii HH103 identified eight genes that code for putative T3SS effectors. Three of these effectors, NopL, NopP, and NopI, are Rhizobium specific. In this work, we demonstrate that NopI, whose amino acid sequence shows a certain similarity with NopP, is secreted through the S. fredii HH103 T3SS in response to flavonoids. We also determined that NopL can be considered an effector since it is directly secreted to the interior of the host cell as demonstrated by adenylate cyclase assays. Finally, the symbiotic phenotype of single, double, and triple nopI, nopL, and nopP mutants in soybean and cowpea was assayed, showing that NopI plays an important role in determining the number of nodules formed in both legumes and that the absence of both NopL and NopP is highly detrimental for symbiosis.IMPORTANCE The paper is focused on three Rhizobium-specific T3SS effectors of Sinorhizobium fredii HH103, NopL, NopP, and NopI. We demonstrate that S. fredii HH103 is able to secrete through the T3SS in response to flavonoids the nodulation outer protein NopI. Additionally, we determined that NopL can be considered an effector since it is secreted to the interior of the host cell as demonstrated by adenylate cyclase assays. Finally, nodulation assays of soybean and cowpea indicated that NopI is important for the determination of the number of nodules formed and that the absence of both NopL and NopP negatively affected

  3. 钙磷对紫花苜蓿-根瘤菌体系酸铝胁迫的修复效应%Remediating Effect of Ca2+ and P on Growth and Nodulation of Alfalfa-Rhizobium After Acidity and Aluminum Stresses

    Institute of Scientific and Technical Information of China (English)

    王晓锋; 罗珍; 刘晓燕; 朱丹; 张磊

    2013-01-01

    为初步探究钙、磷在酸铝胁迫之后对豆科植物及其根瘤菌共生结瘤的修复效应,本试验采用水培培养法,以紫花苜蓿和耐酸苜蓿根瘤菌91522为材料,探讨了钙磷对酸铝胁迫后苜蓿根瘤菌生长、存活及苜蓿结瘤、初生根伸长的影响.结果显示,5 mmol/L Ca2+能够显著提高酸铝胁迫下耐酸根瘤菌的生长和存活,4 μmol/L P素在Ca2+的基础上进一步提高根瘤菌存活率.在钙、磷共同存在的条件下,苜蓿初生根的伸长量比酸铝胁迫对照提高了5~7倍;在铝毒害之后,通过补充Ca2+和P,苜蓿结瘤数、瘤鲜重、现瘤时间、农艺性状得到了显著的恢复;高浓度(10 mmol/L)Ca2+对苜蓿根瘤菌生长有更好效果,但对苜蓿结瘤效果则不如5 mmol/L Ca2+.初步认为,钙磷可以通过提高根瘤菌在酸性环境下的存活、苜蓿根系生长以及参与根瘤菌与豆科宿主识别过程来恢复酸铝毒害后的苜蓿-根瘤菌共生结瘤过程.%Alfalfa and its acid-tolerant rhizobium strain 91522 were used as materials in a water culture experiment to study the effects of calcium and phosphorus on the growth and nodulation of the alfalfa-rhizobium system under acidity and aluminum stresses.The research showed that 5 mmol/L Ca2+ significantly improved the growth and survival of rhizobium compared with the control and that under the same Ca2+ level the addition of 4 μmol/L P further enhanced rhizobium survival rate.By combining Ca2+ and P,the elongation of alfalfa primary root was raised by 5-7 times over that of the control under the acidity + aluminum stress condition.When Ca2+ and P were added in combination,the nodular number,nodule fresh weight,time of nodule occurrence and agronomic traits of alfalfa were satisfactorily recovered.Compared with the treatment 5 mmol/L Ca2+,10 mmol/L Ca2+ had better effect of improving nodule growth of alfalfa but poorer effect on nodulation.It was tentatively concluded that Ca2+ and P

  4. Altered susceptibility to infection by Sinorhizobium meliloti and Nectria haematococca in alfalfa roots with altered cell cycle.

    Science.gov (United States)

    Woo, H-H; Hirsch, A M; Hawes, M C

    2004-07-01

    Most infections of plant roots are initiated in the region of elongation; the mechanism for this tissue-specific localization pattern is unknown. In alfalfa expressing PsUGT1 antisense mRNA under the control of the cauliflower mosaic virus (CaMV) 35S promoter, the cell cycle in roots is completed in 48 h instead of 24 h, and border cell number is decreased by more than 99%. These plants were found to exhibit increased root-tip infection by a fungal pathogen and reduced nodule formation by a bacterial symbiont. Thus, the frequency of infection in the region of elongation by Nectria haematocca was unaffected, but infection of the root tip was increased by more than 90%; early stages of Sinorhizobium meliloti infection and nodule morphology were normal, but the frequency of nodulation was fourfold lower than in wild-type roots.

  5. TCP在种植苜蓿土壤中的降解研究%Degradation of TCP in Soil Planted with Alfalfa (Medicago sativa L.)

    Institute of Scientific and Technical Information of China (English)

    丁克强

    2010-01-01

    [目的]研究TCP在苜蓿种植土壤中的降解作用,为氯酚类物质污染土壤生物修复技术的实际应用提供依据.[方法]利用玻璃房盆栽试验,研究苜蓿对土壤中2,4,6-三氯酚(TCP)污染的修复作用以及苜蓿的生长情况和TCP对土壤多酚氧化酶、脱氢酶和过氧化氢酶活性的影响.[结果]苜蓿经过75 d的生长后,在低、中、高3个浓度处理中,土壤中TCP含量均在15 d内迅速降低,随后降低速度逐渐变缓;在苜蓿生长30d时,3个处理的苜蓿鲜重与对照间无显著差异(P<0.05),而在生长75 d时,各处理的苜蓿鲜重明显对照低(P<0.05),表明土壤中TCP对苜蓿的生长具有抑制作用;苜蓿能显著提高土壤中多酚氧化酶、脱氢酶和过氧化氢酶的活性,从而提高了土壤植物和微生物对污染物的降解能力.[结论]苜蓿能促进土壤酶活性的提高,促进土壤中有机物的降解,从而可以利用苜蓿进行TCP污染土壤的植物修复.%[Objective]The research aimed to study the degradation of 2,4,6-trichlorophenol(TCP)in soil planted with alfalfa(Medicago sativa L.),as well as to provide references for the Chlorophenols phytoremediation technology in the practical application.[Method]By the use of pot culture experiment in greenhouse,the phytoremediation effect of alfalfa on TCP-contaminated soil,the growth conditions of alfalfa,as well as the effect of TCP on the activity of soil polyphenol oxidase,dehydrogenase and catalase were studied.[Result]After the alfalfa was grown for 75 d,the TCP content in soil of three different concentrations treatments(low,middle and high)decreased dramatically within 15 d,and then the decreasing rate was gradually slow;on the 30th d of cultivation,the fresh weight of treated alfalfa showed no significant difference with the control(P<0.05),indicating that TCP in soil had inhibition effect on the growth of alfalfa;alfalfa could significantly enhance the activities of polyphenol oxidase

  6. Calcium-dependent regulation of genes for plant nodulation in Rhizobium leguminosarum detected by iTRAQ quantitative proteomic analysis.

    Science.gov (United States)

    Arrigoni, Giorgio; Tolin, Serena; Moscatiello, Roberto; Masi, Antonio; Navazio, Lorella; Squartini, Andrea

    2013-11-01

    Rhizobia, the nitrogen-fixing bacterial symbionts of legumes, represent an agricultural application of primary relevance and a model of plant-microbe molecular dialogues. We recently described rhizobium proteome alterations induced by plant flavonoids using iTRAQ. Herein, we further extend that experimentation, proving that the transient elevation in cytosolic calcium is a key signaling event necessary for the expression of the nodulation (nod) genes. Ca(2+) involvement in nodulation is a novel issue that we recently flagged with genetic and physiological approaches and that hereby we demonstrate also by proteomics. Exploiting the multiple combinations of 4-plex iTRAQ, we analyzed Rhizobium leguminosarum cultures grown with or without the nod gene-inducing plant flavonoid naringenin and in the presence or absence of the extracellular Ca(2+) chelator EGTA. We quantified over a thousand proteins, 189 of which significantly altered upon naringenin and/or EGTA stimulation. The expression of NodA, highly induced by naringenin, is strongly reduced when calcium availability is limited by EGTA. This confirms, from a proteomic perspective, that a Ca(2+) influx is a necessary early step in flavonoid-mediated legume nodulation by rhizobia. We also observed other proteins affected by the different treatments, whose identities and roles in nodulation and rhizobium physiology are likewise discussed.

  7. 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.

  8. 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.

  9. Characterization of root-nodule bacteria isolated from Vicia faba and selection of plant growth promoting isolates.

    Science.gov (United States)

    Saïdi, Sabrine; Chebil, Samir; Gtari, Maher; Mhamdi, Ridha

    2013-06-01

    A collection of 104 isolates from root-nodules of Vicia faba was submitted to 16S rRNA PCR-RFLP typing. A representative sample was further submitted to sequence analysis of 16S rRNA. Isolates were assigned to 12 genera. All the nodulating isolates (45 %) were closely related to Rhizobium leguminosarum USDA2370(T) (99.34 %). The remaining isolates, including potential human pathogens, failed to nodulate their original host. They were checked for presence of symbiotic genes, P-solubilization, phytohormone and siderophore production, and then tested for their growth promoting abilities. Results indicated that 9 strains could induce significant increase (41-71 %) in shoot dry yield of faba bean. A Pseudomonas strain was further assessed in on-farm trial in combination with a selected rhizobial strain. This work indicated that nodule-associated bacteria could be a valuable pool for selection of effective plant growth promoting isolates. Nevertheless, the possible involvement of nodules in increasing risks related to pathogenic bacteria should not be neglected and needs to be investigated further.

  10. Investigating the origin of the Chinese name for alfalfa

    Science.gov (United States)

    Sun, Q. Z.; Xu, L. J.; Tang, X. J.; Ma, J. T.; Wang, D.; Li, D.; Liu, Q.; Tao, Y.; Li, F.

    2017-02-01

    It is assured that alfalfa (Medicago sativa L.) was introduced in Han dynasty. There are cognitive differences on whether Zhang Qian introduced alfalfa. Based on the previous studies, research inductive method was used. The relationship between Zhang Qian and alfalfa introduction was analyzed from the motivation, experience and influence of Zhang Qian to the Western Regions and the image generation of Zhang Qian brought back alfalfa’s seeds. Till to now, there are four opinions about Zhang Qian introducing Alfalfa seeds, including : (1) Zhang Qian introduced alfalfa seeds;(2) Zhang Qian did not introduce alfalfa seeds;(3) the information of Zhang Qian transferring alfalfa;(4)for commemoration Zhang Qian to the Western Regions. Although there are not direct historical materials to support Zhang Qian brought alfalfa seed to Han dynasty, it believes and confirms that the introducing of alfalfa is inextricably interwoven with Zhang Qian’s western travel. Zhangqian brought relative information from western regions during the introduction, which was the basis of non-native theory, and after that, Chinese began to plant alfalfa in Han dynasty., According to historical literatures, it is clear that the Chinese diplomat brought alfalfa seeds back to China. Alfalfa, as the favorite forage to Ferghana horse, have been already planted in Dawan in Han dynasty. Despite the debate, Zhangqian played an important pioneering role in introducing alfalfa.

  11. Variation in Frankia populations of the Elaeagnus host infection group in nodules of six host plant species after inoculation with soil.

    Science.gov (United States)

    Mirza, Babur S; Welsh, Allana; Rasul, Ghulam; Rieder, Julie P; Paschke, Mark W; Hahn, Dittmar

    2009-08-01

    The potential role of host plant species in the selection of symbiotic, nitrogen-fixing Frankia strains belonging to the Elaeagnus host infection group was assessed in bioassays with two Morella, three Elaeagnus, and one Shepherdia species as capture plants, inoculated with soil slurries made with soil collected from a mixed pine/grassland area in central Wisconsin, USA. Comparative sequence analysis of nifH gene fragments amplified from homogenates of at least 20 individual lobes of root nodules harvested from capture plants of each species confirmed the more promiscuous character of Morella cerifera and Morella pensylvanica that formed nodules with frankiae of the Alnus and the Elaeagnus host infection groups, while frankiae in nodules formed on Elaeagnus umbellata, Elaeagnus angustifolia, Elaeagnus commutata, and Shepherdia argentea generally belonged to the Elaeagnus host infection group. Diversity of frankiae of the Elaeagnus host infection groups was larger in nodules on both Morella species than in nodules formed on the other plant species. None of the plants, however, captured the entire diversity of nodule-forming frankiae. The distribution of clusters of Frankia populations and their abundance in nodules was unique for each of the plant species, with only one cluster being ubiquitous and most abundant while the remaining clusters were only present in nodules of one (six clusters) or two (two clusters) host plant species. These results demonstrate large effects of the host plant species in the selection of Frankia strains from soil for potential nodule formation and thus the significant effect of the choice of capture plant species in bioassays on diversity estimates in soil.

  12. Impact of Long-Term Alfalfa Cropping on Soil Potassium Content and Clay Minerals in a Semi-Arid Loess Soil in China

    Institute of Scientific and Technical Information of China (English)

    LI De-Cheng; B. VELDE; LI Feng-Min; ZHANG Gan-Lin; ZHAO Ming-Song; HUANG Lai-Ming

    2011-01-01

    Alfalfa cropping has been considered an efficient method of increasing soil fertility.Usually nitrogen increase in root nodules is considered to be the major beneficial effect.A 21-year time series (five sampling periods) of alfalfa cultivation plots on a loess soil,initially containing illite and chlorite,in Lanzhou of northwestern China was selected to investigate the relationships among alfalfa cropping,soil potassium (K) content and soil clay minerals.The results indicated that soil K significantly accumulated after cropping,with a peak value at about 15 years,and decreased afterwards.The accumulated K was associated with the K increase in the well-crystallized illite,which was not extracted by the traditional laboratory K extraction methods in assessing bioavailability.The steep decline in soil K content after 15-year cropping was in accord with the observed fertility loss in the alfalfa soil.Plant biomass productivity peaked at near 9 years of culture,whereas soil K and clay minerals continued to increase until cropping for 15 years.This suggested that K increased in the topsoil came from the deep root zone.Thus alfalfa continued to store K in clays even after peak production occurred.Nitrogen did not follow these trends,showing a general decline compared with the native prairie soils that had not been cropped.Therefore,the traditional alfalfa cropping can increase K content in the topsoil.

  13. How membranes shape plant symbioses: Signaling and transport in nodulation and arbuscular mycorrhiza

    Directory of Open Access Journals (Sweden)

    Laure eBapaume

    2012-10-01

    Full Text Available As sessile organisms that cannot evade adverse environmental conditions, plants have evolved various adaptive strategies to cope with environmental stresses. One of the most successful adaptations is the formation of symbiotic associations with beneficial microbes. In these mutualistic interactions the partners exchange essential nutrients and improve their resistance to biotic and abiotic stresses. In arbuscular mycorrhiza (AM and in root nodule symbiosis (RNS, AM fungi and rhizobia, respectively, penetrate roots and accommodate within the cells of the plant host. In these endosymbiotic associations, both partners keep their plasma membranes intact and use them to control the bidirectional exchange of signaling molecules and nutrients. Intracellular accommodation requires the exchange of symbiotic signals and the reprogramming of both interacting partners. This involves fundamental changes at the level of gene expression and of the cytoskeleton, as well as of organelles such as plastids, endoplasmic reticulum (ER, and the central vacuole. Symbiotic cells are highly compartmentalized and have a complex membrane system specialized for the diverse functions in molecular communication and nutrient exchange. Here, we discuss the roles of the different cellular membrane systems and their symbiosis-related proteins in AM and RNS, and we review recent progress in the analysis of membrane proteins involved in endosymbiosis.

  14. How membranes shape plant symbioses: signaling and transport in nodulation and arbuscular mycorrhiza

    Science.gov (United States)

    Bapaume, Laure; Reinhardt, Didier

    2012-01-01

    As sessile organisms that cannot evade adverse environmental conditions, plants have evolved various adaptive strategies to cope with environmental stresses. One of the most successful adaptations is the formation of symbiotic associations with beneficial microbes. In these mutualistic interactions the partners exchange essential nutrients and improve their resistance to biotic and abiotic stresses. In arbuscular mycorrhiza (AM) and in root nodule symbiosis (RNS), AM fungi and rhizobia, respectively, penetrate roots and accommodate within the cells of the plant host. In these endosymbiotic associations, both partners keep their plasma membranes intact and use them to control the bidirectional exchange of signaling molecules and nutrients. Intracellular accommodation requires the exchange of symbiotic signals and the reprogramming of both interacting partners. This involves fundamental changes at the level of gene expression and of the cytoskeleton, as well as of organelles such as plastids, endoplasmic reticulum (ER), and the central vacuole. Symbiotic cells are highly compartmentalized and have a complex membrane system specialized for the diverse functions in molecular communication and nutrient exchange. Here, we discuss the roles of the different cellular membrane systems and their symbiosis-related proteins in AM and RNS, and we review recent progress in the analysis of membrane proteins involved in endosymbiosis. PMID:23060892

  15. AGROCHEMICALS GROWTH REGULATORS ON ALFALFA

    Directory of Open Access Journals (Sweden)

    Buldykova I. A.

    2015-04-01

    Full Text Available One of the methods of increasing the productivity of alfalfa is the use of growth regulators, and the use of polymeric compositions of growth regulators, binders with synthesized growth regulators of sim-triazine series is promising with the possibility of creating environmentally friendly technologies for growing alfalfa. Studies have shown that the application of the test on alfalfa growth regulators has a positive effect on growth, physiological and morphogenetic processes, plant resistance to adverse environmental conditions. Intensity impact on plant growth regulators depends on the type of plant growth regulators, concentration and method of application. Processing alfalfa seed growth regulators on germination increases energy 3,0-14,0 % germination on 8,0-17,0 %. Processing plant growth regulators to enhance the growth of plants in height (on the 7th day – 2,6-11,9 % , on the 14th 41,9-48,0 % , the growth of aboveground biomass , expands on the number of productive branches of the 1st ( 24,1-41,3 % and 2nd order (21,7-55,0 %. Pre-sowing seed treatment and plant growth regulators alfalfa sim- triazine series contributed to the increase in seed yield of 15,5 %. On average, the yield of green mass increased by 0,8-2,4 t /ha or 5,2-15,5 % and seed yield at 0,19-0,42 h/ha or 8,7-19, 3 %. Growth regulators of sim-triazine series increase the protein content of the vegetative mass of alfalfa at 3,2-4,6 %

  16. Effect of Plant Growth-Promoting Rhizobacteria on Growth,Nodulation and Nutrient Accumulation of Lentil Under Controlled Conditions

    Institute of Scientific and Technical Information of China (English)

    M.ZAFAR; M.K.ABBASI; M.A.KHAN; A.KHALIQ; T.SULTAN; M.ASLAM

    2012-01-01

    Application of plant growth-promoting rhizobacteria (PGPR) has been shown to increase legume growth and development under field and controlled environmental conditions.The present study was conducted to isolate plant growth-promoting rhizobacteria (PGPR) from the root nodules of lentil (Lens culinaris Medik.) grown in arid/semi-arid region of Punjab,Pakistan and examined their plant growth-promoting abilities.Five bacterial isolates were isolated,screened in vitro for plant growth-promoting (PGP)characteristics and their effects on the growth of lentil were assessed under in vitro,hydroponic and greenhouse (pot experiment)conditions.All the isolates were Gram negative,rod-shaped and circular in form and exhibited the plant growth-promoting attributes of phosphate solubilization and auxin (indole acetic acid,IAA) production.The IAA production capacity ranged in 0.5-11.0 μgmL-1and P solubilization ranged in 3 16 mg L-1.When tested for their effects on plant growth,the isolated strains had a stimulatory effect on growth,nodulation and nitrogen (N) and phosphorus (P) uptake in plants on nutrient-deficient soil.In the greenhouse pot experiment,application of PGPR significantly increased shoot length,fresh weight and dry weight by 65%,43% and 63% and the increases in root length,fresh weight and dry weight were 74%,54% and 92%,respectively,as compared with the uninoculated control.The relative increases in growth characteristics under in vitro and hydroponic conditions were even higher.PGPR also increased the number of pods per plant,1000-grain weight,dry matter yield and grain yield by 50%,13%,28% and 29%,respectively,over the control.The number of nodules and nodule dry mass increased by 170% and 136%,respectively.After inoculation with effective bacterial strains,the shoot,root and seed N and P contents increased,thereby increasing both N and P uptake in plants. The root elongation showed a positive correlation (R2 =0.67) with the IAA

  17. A Sinorhizobium meliloti-specific N-acyl homoserine lactone quorum-sensing signal increases nodule numbers in Medicago truncatula independent of autoregulation

    Directory of Open Access Journals (Sweden)

    Debora Fabiola Veliz Vallejos

    2014-10-01

    Full Text Available N-acyl homoserine lactones (AHLs act as quorum sensing signals that regulate cell-density dependent behaviors in many gram-negative bacteria, in particular those important for plant-microbe interactions. AHLs can also be recognized by plants, and this may influence their interactions with bacteria. Here we tested whether the exposure to AHLs affects the nodule-forming symbiosis between legume hosts and rhizobia. We treated roots of the model legume, Medicago truncatula, with a range of AHLs either from its specific symbiont, Sinorhizobium meliloti, or from the potential pathogens, Pseudomonas aeruginosa and Agrobacterium vitis. We found increased numbers of nodules formed on root systems treated with the S. meliloti-specific AHL, 3-oxo-C14-homoserine lactone, at a concentration of 1 μM, while the other AHLs did not result in significant changes to nodule numbers. We did not find any evidence for altered nodule invasion by the rhizobia. Quantification of flavonoids that could act as nod gene inducers in S. meliloti did not show any correlation with increased nodule numbers. The effects of AHLs were specific for an increase in nodule numbers, but not lateral root numbers or root length. Increased nodule numbers following 3-oxo-C14-homoserine lactone treatment were under control of autoregulation of nodulation and were still observed in the autoregulation mutant, sunn4 (super numeric nodules4. However, increases in nodule numbers by 3-oxo-C14-homoserine lactone were not found in the ethylene-insensitive sickle mutant. A comparison between M. truncatula with M. sativa (alfalfa and Trifolium repens (white clover showed that the observed effects of AHLs on nodule numbers were specific to M. truncatula, despite M. sativa nodulating with the same symbiont. We conclude that plant perception of the S. meliloti-specific 3-oxo-C14-homoserine lactone influences nodule numbers in M. truncatula via an ethylene-dependent, but autoregulation

  18. Leguminous plants nodulated by selected strains of Cupriavidus necator grow in heavy metal contaminated soils amended with calcium silicate.

    Science.gov (United States)

    Avelar Ferreira, Paulo Ademar; Lopes, Guilherme; Bomfeti, Cleide Aparecida; de Oliveira Longatti, Silvia Maria; de Sousa Soares, Cláudio Roberto Fonseca; Guimarães Guilherme, Luiz Roberto; de Souza Moreira, Fatima Maria

    2013-11-01

    Increasing concern regarding mining area environmental contamination with heavy metals has resulted in an emphasis of current research on phytoremediation. The aim of the present study was to assess the efficiency of symbiotic Cupriavidus necator strains on different leguminous plants in soil contaminated with heavy metals following the application of inorganic materials. The application of limestone and calcium silicate induced a significant increase in soil pH, with reductions in zinc and cadmium availability of 99 and 94 %, respectively. In addition, improved nodulation of Mimosa caesalpiniaefolia, Leucaena leucocephala and Mimosa pudica in soil with different levels of contamination was observed. Significant increases in the nitrogen content of the aerial parts of the plant were observed upon nodulation of the root system of Leucaena leucocephala and Mimosa pudica by strain UFLA01-659 (36 and 40 g kg(-1)) and by strain UFLA02-71 in Mimosa caesalpiniaefolia (39 g kg(-1)). The alleviating effect of calcium silicate resulted in higher production of dry matter from the aerial part of the plant, an increase in nodule number and an increase in the nitrogen fixation rate. The results of the present study demonstrate that the combination of rhizobia, leguminous plants and calcium silicate may represent a key factor in the remediation of areas contaminated by heavy metals.

  19. Stress-related phenomena and detoxification mechanisms induced by common pharmaceuticals in alfalfa (Medicago sativa L.) plants

    Energy Technology Data Exchange (ETDEWEB)

    Christou, Anastasis [Agricultural Research Institute, P.O. Box 22016, 1516 Nicosia (Cyprus); Antoniou, Chrystalla; Christodoulou, Charalampia [Department of Agricultural Sciences, Biotechnology and Food Science, Cyprus University of Technology, 3603 Lemesos (Cyprus); Hapeshi, Evroula; Stavrou, Ioannis; Michael, Costas [NIREAS-International Water Research Center, University of Cyprus, P.O. Box 20537, 1678 Nicosia (Cyprus); Fatta-Kassinos, Despo [Department of Civil and Environmental Engineering, University of Cyprus, P.O. Box 20537, 1678 Nicosia (Cyprus); NIREAS-International Water Research Center, University of Cyprus, P.O. Box 20537, 1678 Nicosia (Cyprus); Fotopoulos, Vasileios, E-mail: vassilis.fotopoulos@cut.ac.cy [Department of Agricultural Sciences, Biotechnology and Food Science, Cyprus University of Technology, 3603 Lemesos (Cyprus)

    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 (10 μg L{sup −1}) (diclofenac, sulfamethoxazole, trimethoprim, 17a-ethinylestradiol) and their mixture. Stress physiology markers (lipid peroxidation, proline, H{sub 2}O{sub 2} 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{sup +}-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 H{sub 2}O{sub 2} 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{sup +}-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. - Highlights: • PhACs were detected in higher concentrations in roots compared with leaves. • Stress effects were local and exacerbated when PhACs were applied in mixture. • H{sub 2}O{sub 2} may be involved in signal transduction and detoxification responses. • GSTs, H{sup +}-ATPase and CytcOx contribute to the

  20. DROUGHT-INDUCED EFFECTS AND RECOVERY OF NITRATE ASSIMILATION AND NODULE ACTIVITY IN COWPEA PLANTS INOCULATED WITH BRADYRHIZOBIUM SPP. UNDER MODERATE NITRATE LEVEL

    Directory of Open Access Journals (Sweden)

    Silveira Joaquim Albenísio Gomes da

    2001-01-01

    Full Text Available This study was carried out to establish comparative effects of drought and recovery on the nitrate assimilation and nodule activity related to N2 fixation in cowpea plants [Vigna unguiculata L. (Walp.] previously inoculated with Bradyrhizobium spp. BR-3256 (CB-756 strain in the presence of 5 mol m-3 NO-3. Twenty-eight-day-old nodulated plants were submitted to water deprivation during 4 consecutive days and afterwards resupplied with nutrient solution during 2 days. The water deprivation caused a rapid increase in the nitrate content in root and a marked reduction in leaf nitrate reductase (NR activity. In contrast nodule NR activity was slightly increased by water deprivation. Concomitantly, in nodules of water stressed plants, leghemoglobin and glutamine synthetase (GS activity declined and a progressive reduction in ureide-N concentration in xylem sap was observed. Leaf-NR activity increased rapidly after rehydration while leaf nitrate content declined. In contrast both GS activity and soluble protein content in the nodule continued to decline in rewatered plants. In addition the concentration of leghemoglobin recovered well, while the xylem ureide-N content experienced a slight increase after rehydration. Despite the nitrate assimilation in leaves and the nodule activity had been both severely affected by water stress, the rapid recovery of nitrate reductase activity suggests that the nitrate assimilation process is less sensitive to drought/rehydration cycle when cowpea plants are nodulated in presence of moderate nitrate level.

  1. A fadD mutant of Sinorhizobium meliloti shows multicellular swarming migration and is impaired in nodulation efficiency on alfalfa roots.

    Science.gov (United States)

    Soto, María José; Fernández-Pascual, Mercedes; Sanjuan, Juan; Olivares, José

    2002-01-01

    Swarming is a form of bacterial translocation that involves cell differentiation and is characterized by a rapid and co-ordinated population migration across solid surfaces. We have isolated a Tn5 mutant of Sinorhizobium meliloti GR4 showing conditional swarming. Swarm cells from the mutant strain QS77 induced on semi-solid minimal medium in response to different signals are hyperflagellated and about twice as long as wild-type cells. Genetic and physiological characterization of the mutant strain indicates that QS77 is altered in a gene encoding a homologue of the FadD protein (long-chain fatty acyl-CoA ligase) of several microorganisms. Interestingly and similar to a less virulent Xanthomonas campestris fadD(rpfB) mutant, QS77 is impaired in establishing an association with its host plant. In trans expression of multicopy fadD restored growth on oleate, control of motility and the symbiotic phenotype of QS77, as well as acyl-CoA synthetase activity of an Escherichia coli fadD mutant. The S. meliloti QS77 strain shows a reduction in nod gene expression as well as a differential regulation of motility genes in response to environmental conditions. These data suggest that, in S. meliloti, fatty acid derivatives may act as intracellular signals controlling motility and symbiotic performance through gene expression.

  2. The Independent Acquisition of Plant Root Nitrogen-Fixing Symbiosis in Fabids Recruited the Same Genetic Pathway for Nodule Organogenesis

    Science.gov (United States)

    Svistoonoff, Sergio; Benabdoun, Faiza Meriem; Nambiar-Veetil, Mathish; Imanishi, Leandro; Vaissayre, Virginie; Cesari, Stella; Diagne, Nathalie; Hocher, Valérie; de Billy, Françoise; Bonneau, Jocelyne; Wall, Luis; Ykhlef, Nadia; Rosenberg, Charles; Bogusz, Didier; Franche, Claudine; Gherbi, Hassen

    2013-01-01

    Only species belonging to the Fabid clade, limited to four classes and ten families of Angiosperms, are able to form nitrogen-fixing root nodule symbioses (RNS) with soil bacteria. This concerns plants of the legume family (Fabaceae) and Parasponia (Cannabaceae) associated with the Gram-negative proteobacteria collectively called rhizobia and actinorhizal plants associated with the Gram-positive actinomycetes of the genus Frankia. Calcium and calmodulin-dependent protein kinase (CCaMK) is a key component of the common signaling pathway leading to both rhizobial and arbuscular mycorrhizal symbioses (AM) and plays a central role in cross-signaling between root nodule organogenesis and infection processes. Here, we show that CCaMK is also needed for successful actinorhiza formation and interaction with AM fungi in the actinorhizal tree Casuarina glauca and is also able to restore both nodulation and AM symbioses in a Medicago truncatula ccamk mutant. Besides, we expressed auto-active CgCCaMK lacking the auto-inhibitory/CaM domain in two actinorhizal species: C. glauca (Casuarinaceae), which develops an intracellular infection pathway, and Discaria trinervis (Rhamnaceae) which is characterized by an ancestral intercellular infection mechanism. In both species, we found induction of nodulation independent of Frankia similar to response to the activation of CCaMK in the rhizobia-legume symbiosis and conclude that the regulation of actinorhiza organogenesis is conserved regardless of the infection mode. It has been suggested that rhizobial and actinorhizal symbioses originated from a common ancestor with several independent evolutionary origins. Our findings are consistent with the recruitment of a similar genetic pathway governing rhizobial and Frankia nodule organogenesis. PMID:23741336

  3. The independent acquisition of plant root nitrogen-fixing symbiosis in Fabids recruited the same genetic pathway for nodule organogenesis.

    Directory of Open Access Journals (Sweden)

    Sergio Svistoonoff

    Full Text Available Only species belonging to the Fabid clade, limited to four classes and ten families of Angiosperms, are able to form nitrogen-fixing root nodule symbioses (RNS with soil bacteria. This concerns plants of the legume family (Fabaceae and Parasponia (Cannabaceae associated with the Gram-negative proteobacteria collectively called rhizobia and actinorhizal plants associated with the Gram-positive actinomycetes of the genus Frankia. Calcium and calmodulin-dependent protein kinase (CCaMK is a key component of the common signaling pathway leading to both rhizobial and arbuscular mycorrhizal symbioses (AM and plays a central role in cross-signaling between root nodule organogenesis and infection processes. Here, we show that CCaMK is also needed for successful actinorhiza formation and interaction with AM fungi in the actinorhizal tree Casuarina glauca and is also able to restore both nodulation and AM symbioses in a Medicago truncatula ccamk mutant. Besides, we expressed auto-active CgCCaMK lacking the auto-inhibitory/CaM domain in two actinorhizal species: C. glauca (Casuarinaceae, which develops an intracellular infection pathway, and Discaria trinervis (Rhamnaceae which is characterized by an ancestral intercellular infection mechanism. In both species, we found induction of nodulation independent of Frankia similar to response to the activation of CCaMK in the rhizobia-legume symbiosis and conclude that the regulation of actinorhiza organogenesis is conserved regardless of the infection mode. It has been suggested that rhizobial and actinorhizal symbioses originated from a common ancestor with several independent evolutionary origins. Our findings are consistent with the recruitment of a similar genetic pathway governing rhizobial and Frankia nodule organogenesis.

  4. Stem formation at alfalfa varieties and correlative dependences with some main parameters

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    Natalya Georgieva

    2015-06-01

    Full Text Available During the period 2006-2009 in IFC-Pleven were studied 9 alfalfa varieties: Europe, Prista 2, Prista 3, Prista 4, Obnova 10, Pleven 6, Dara, Multifoliate and Dama. In the first year of its development alfalfa formed on average 1.42 number of stems per plant. During second and third year their quantity increased to 2.14 and 3.83 number whereafter during fourth year they decreased to 3.28. In all years (except the first year was found a trend of increase in stem number from spring to autumn regrowth with average values 2.11, 2.85 and 3.05 respectively. Average for the four-year period the greatest number of stems (over the average for the nine studied varieties – 2.67 formed the Multifoliate variety followed by Europe, Prista 4, Obnova 10 and Dama. The varieties which are characterized by less density of stand (number of plants per unit area had a greater number of stems per plant (r = -0.530. Correlations with mean and high positive value were found between stem formation and amount and distributions of rainfall during vegetation period (r = 0.989, year of alfalfa development (r = 0.861, nodulation (r = 0.763, weight of root mass (r = 0.411 as well as correlations with mean negative value between number of stems and percentage of damaged stems by Apion seniculus (r = -0.456.

  5. 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.

  6. Short Communication Synergistic effect of rhizobia and plant growth promoting rhizobacteria on the growth and nodulation of lentil seedlings under axenic conditions

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    Muhammad Zafar-ul-Hye

    2013-05-01

    Full Text Available Plant growth promoting rhizobacteria (PGPR containing ACC-deaminase in combination with rhizobia can improve the growth and nodulation in plants by suppressing the endogenous level of ethylene. In the present study, ten strains, each of PGPR and rhizobia from the previously screened cultures were tested for their effect as co-inoculants on growth and nodulation of lentil in growth pouches under axenic conditions. Results showed that most of the combinations improved the lentil growth as compared to the un-inoculated control. Maximum increase in shoot length (1.87 fold, root length (1.97 fold and total biomass (1.98 fold over the un-inoculated control was observed in the treatment where the lentil seedlings were inoculated with the combination Z24P10. Co-inoculation also improved the nodulation in lentil and the maximum number of nodules plant-1 (24 nodules were observed in the combination Z22P10. However, there was no nodulation in few combinations. It is concluded that the co-inoculation with rhizobia and PGPR containing ACC-deaminase has improved the growth and nodulation in lentil under axenic conditions and the selected combinations may be evaluated in pot and field trials

  7. Prohexadione-calcium responsive alfalfa varieties ensure success of corn-interseeded alfalfa production systems

    Science.gov (United States)

    Recent USDA-NASS data indicate alfalfa and corn were planted on about 0.8 and 1.9 million hectares per year, respectively, in the Northeast, Great Lakes, Upper Midwest, and Northern Mountain regions the USA. Because both crops are often grown in rotation, alfalfa could be interseeded at corn plantin...

  8. Induced systemic resistance against Botrytis cinerea by Micromonospora strains isolated from root nodules.

    Science.gov (United States)

    Martínez-Hidalgo, Pilar; García, Juan M; Pozo, María J

    2015-01-01

    Micromonospora is a Gram positive bacterium that can be isolated from nitrogen fixing nodules from healthy leguminous plants, where they could be beneficial to the plant. Their plant growth promoting activity in legume and non-legume plants has been previously demonstrated. The present study explores the ability of Micromonospora strains to control fungal pathogens and to stimulate plant immunity. Micromonospora strains isolated from surface sterilized nodules of alfalfa showed in vitro antifungal activity against several pathogenic fungi. Moreover, root inoculation of tomato plants with these Micromonospora strains effectively reduced leaf infection by the fungal pathogen Botrytis cinerea, despite spatial separation between both microorganisms. This induced systemic resistance, confirmed in different tomato cultivars, is long lasting. Gene expression analyses evidenced that Micromonospora stimulates the plant capacity to activate defense mechanisms upon pathogen attack. The defensive response of tomato plants inoculated with Micromonospora spp. differs from that of non-inoculated plants, showing a stronger induction of jasmonate-regulated defenses when the plant is challenged with a pathogen. The hypothesis of jasmonates playing a key role in this defense priming effect was confirmed using defense-impaired tomato mutants, since the JA-deficient line def1 was unable to display a long term induced resistance upon Micromonospora spp. inoculation. In conclusion, nodule isolated Micromonospora strains should be considered excellent candidates as biocontrol agents as they combine both direct antifungal activity against plant pathogens and the ability to prime plant immunity.

  9. Induced systemic resistance against Botrytis cinerea by Micromonospora strains isolated from root nodules

    Directory of Open Access Journals (Sweden)

    Pilar eMartínez-Hidalgo

    2015-09-01

    Full Text Available Micromonospora is a Gram positive bacterium that can be isolated from nitrogen fixing nodules from healthy leguminous plants, where they could be beneficial to the plant. Their plant growth promoting activity in legume and non-legume plants has been previously demonstrated. The present study explores the ability of Micromonospora strains to control fungal pathogens and to stimulate plant immunity. Micromonospora strains isolated from surface sterilized nodules of alfalfa showed in vitro antifungal activity against several pathogenic fungi. Moreover, root inoculation of tomato plants with these Micromonospora strains effectively reduced leaf infection by the fungal pathogen Botrytis cinerea, despite spatial separation between both microorganisms. This induced systemic resistance, confirmed in different tomato cultivars, is long lasting. Gene expression analyses evidenced that Micromonospora stimulates the plant capacity to activate defense mechanisms upon pathogen attack. The defensive response of tomato plants inoculated with Micromonospora spp. differs from that of non-inoculated plants, showing a stronger induction of jasmonate-regulated defenses when the plant is challenged with a pathogen. The hypothesis of jasmonates playing a key role in this defense priming effect was confirmed using defense-impaired tomato mutants, since the JA-deficient line def1 was unable to display a long term induced resistance upon Micromonospora spp. inoculation.In conclusion, nodule isolated Micromonospora strains should be considered excellent candidates as biocontrol agents as they combine both direct antifungal activity against plant pathogens and the ability to prime plant immunity.

  10. 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.

  11. Changes in microbial contamination levels and prevalence of foodborne pathogens in alfalfa (Medicago sativa) and rapeseed (Brassica napus) during sprout production in manufacturing plants.

    Science.gov (United States)

    Kim, S A; Kim, O M; Rhee, M S

    2013-01-01

    Samples were taken from three sprout processing plants at five different stages of production (a total of 20 investigations). Quantitative analyses comprised aerobic plate counts (APCs) and the measurement of coliforms and Bacillus cereus levels, whereas qualitative analyses involved assessing the levels of Escherichia coli and major foodborne pathogens (E. coli O157:H7, Listeria monocytogenes, Salmonella spp., and Staphylococcus aureus). The APC for alfalfa seeds (3·71-4·61 log CFU g(-1)) and rapeseed (4·25-5·11 log CFU g(-1)) increased by approximately 3 log CFU g(-1) during sprouting, reaching 7·17-7·61 and 7·33-8·28 log CFU g(-1), respectively, by the final stage of production. Similarly, increasing trends were noted in the level of coliforms (0·58-4·03 log CFU g(-1) at the seed stage, increasing to 5·52-6·99 log CFU g(-1) by the sprout stage). Bacillus cereus was detected in eight alfalfa (40%) and 14 rapeseed (70%) sprouts, and L. monocytogenes was isolated from one pregermination soaked alfalfa seed. A slight reduction in the level of bacterial contamination was noted after washing the sprouts with water prior to storage, indicating that improvements to the current washing protocol, or other efficient intervention methods, may be needed. Taken together, these results suggest that improved hygiene control during production and processing and a more sanitary environment are needed. The present study provides comprehensive information regarding the microbiological safety of seeds and sprouts during manufacturing. The present study investigated the levels of microbial contamination present in alfalfa and rapeseed sprouts by examining the samples taken at different stages of the manufacturing process in three actual plants. The results provide detailed information regarding the levels of seed and sprout contamination during production. The results may be useful to those involved in the sprout industry and/or academic research in terms of developing

  12. Involvement of the Sinorhizobium meliloti leuA gene in activation of nodulation genes by NodD1 and luteolin.

    Science.gov (United States)

    Sanjuán-Pinilla, Julio M; Muñoz, Socorro; Nogales, Joaquina; Olivares, José; Sanjuán, Juan

    2002-07-01

    The role of leucine biosynthesis by Sinorhizobium meliloti in the establishment of nitrogen-fixing symbiosis with alfalfa ( Medicago sativa) was investigated. The leuA gene from S. meliloti, encoding alpha-isopropylmalate synthase, which catalyses the first specific step in the leucine biosynthetic pathway, was characterized. S. melilotiLeuA(-) mutants were Leu auxotrophs and lacked alpha-isopropylmalate synthase activity. In addition, leuA auxotrophs were unable to nodulate alfalfa. Alfalfa roots did not seem to secrete enough leucine to support growth of leucine auxotrophs in the rhizosphere. Thus, this growth limitation probably imposes the inability to initiate symbiosis. However, in addition to the leucine auxotrophy, leuA strains were impaired in activation of nodulation genes by the transcriptional activator NodD1 in response to the plant flavone luteolin. By contrast, nod gene activation by NodD3, which does not involve plant-derived inducers, was unaffected. Our results suggest that a leucine-related metabolic intermediate may be involved in activation of nodulation genes by NodD1 and luteolin. This kind of control could be of relevance as a way to link bacterial physiological status to the response to plant signals and initiation of symbiosis.

  13. Stress responses in alfalfa (Medicago sativa L.) 12. Sequence analysis of phenylalanine ammonia-lyase (PAL) cDNA clones and appearance of PAL transcripts in elicitor-treated cell cultures and developing plants.

    Science.gov (United States)

    Gowri, G; Paiva, N L; Dixon, R A

    1991-09-01

    An expression library containing cDNAs derived from transcripts from fungal elicitor-treated alfalfa cell suspension cultures was screened with an antiserum raised against phenylalanine ammonia-lyase (PAL) from alfalfa. A single immunoreactive clone was isolated which encoded a full-length PAL cDNA (APAL1) consisting of a 2175 bp open reading frame, 96 bp 5'-untranslated leader and 128 bp 3'-non-coding region. The deduced amino acid sequence was 86.5% similar to that of the PAL2 gene of bean, and encoded a polypeptide of Mr 78,865. A second PAL cDNA species was isolated, whose 3'-untranslated region was 86% identical to that of APAL1. Southern blot analysis indicated that PAL is encoded by a small multigene family in alfalfa. PAL transcript levels were rapidly and massively induced, and preceded increased PAL extractable activity, on exposure of alfalfa suspension cells to elicitor from baker's yeast. PAL transcripts were most abundant in roots, stems and petioles during growth and development of alfalfa seedlings. These studies provide the basis for an examination of the developmental and environmental control of a key enzyme of phenylpropanoid synthesis in a plant species which is readily amenable to stable genetic transformation.

  14. Alfalfa: bioenergy and more

    Science.gov (United States)

    Alfalfa (Medicago sativa) has the potential to be a significant contributor to America's renewable energy future. In an alfalfa biomass energy production system, alfalfa forage would be separated into stem and leave fractions. The stems would be processed to produce energy, and the leaves would be s...

  15. The sunflower transcription factor HaHB11 confers tolerance to water deficit and salinity to transgenic Arabidopsis and alfalfa plants.

    Science.gov (United States)

    Cabello, Julieta V; Giacomelli, Jorge I; Gómez, María C; Chan, Raquel L

    2017-09-10

    Homeodomain-leucine zipper (HD-Zip) transcription factors are unique to the plant kingdom; members of subfamily I are known to be involved in abiotic stress responses. HaHB11 belongs to this subfamily and it was previously shown that it is able to confer improved yield and tolerance to flooding via a quiescent strategy. Here we show that HaHB11 expression is induced by ABA, NaCl and water deficit in sunflower seedlings and leaves. Arabidopsis transgenic plants expressing HaHB11, controlled either by its own promoter or by the constitutive 35S CaMV, presented rolled leaves and longer roots than WT when grown under standard conditions. In addition, these plants showed wider stems and more vascular bundles. To deal with drought, HaHB11 transgenic plants closed their stomata faster and lost less water than controls, triggering an enhanced tolerance to such stress condition and also to salinity stress. Concomitantly, ABA-synthesis and sensing related genes were differentially regulated in HaHB11 transgenic plants. Either under long-term salinity stress or mild drought stress, HaHB11 transgenic plants did not exhibit yield penalties. Moreover, alfalfa transgenic plants were generated which also showed enhanced drought tolerance. Altogether, the results indicated that HaHB11 was able to confer drought and salinity tolerance via a complex mechanism which involves morphological, physiological and molecular changes. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Distinct patterns of symbiosis-related gene expression in actinorhizal nodules from different plant families

    NARCIS (Netherlands)

    Pawlowski, K.; Swensen, S.; Guan, C.; Hadri, A.E.; Berry, A.M.; Bisseling, T.

    2003-01-01

    Phylogenetic analyses suggest that, among the members of the Eurosid I clade, nitrogen-fixing root nodule symbioses developed multiple times independently, four times with rhizobia and four times with the genus Frankia. In order to understand the degree of similarity between symbiotic systems of dif

  17. 'Ca. Liberibacter asiaticus' proteins orthologous with pSymA-encoded proteins of Sinorhizobium meliloti: hypothetical roles in plant host interation

    Science.gov (United States)

    A nitrogen-fixing alfalfa-nodulating microsymbiont, Sinorhizobium meliloti, has a genome consisting of a 3.5 Mbp circular chromosome and two megaplasmids totaling 3.0 Mbp, one a 1.3 Mbp pSymA carrying nonessential ‘accessory’ genes including nif, nod and others involved in plant interaction. Predict...

  18. Broadening the U.S. alfalfa germplasm base

    Science.gov (United States)

    Over 4000 alfalfa (Medicago sativa L.) plant introductions (PIs) exist in the USDA-ARS National Plant Germplasm System (NPGS). NAAIC has discussed/proposed pre-breeding efforts to utilize this germplasm for creating pre-commercial alfalfa germplasm. Funding constraints have been one impediment to th...

  19. Applied Genetics and Genomics in Alfalfa Breeding

    Directory of Open Access Journals (Sweden)

    E. Charles Brummer

    2012-03-01

    Full Text Available Alfalfa (Medicago sativa L., a perennial and outcrossing species, is a widely planted forage legume for hay, pasture and silage throughout the world. Currently, alfalfa breeding relies on recurrent phenotypic selection, but alternatives incorporating molecular marker assisted breeding could enhance genetic gain per unit time and per unit cost, and accelerate alfalfa improvement. Many major quantitative trait loci (QTL related to agronomic traits have been identified by family-based QTL mapping, but in relatively large genomic regions. Candidate genes elucidated from model species have helped to identify some potential causal loci in alfalfa mapping and breeding population for specific traits. Recently, high throughput sequencing technologies, coupled with advanced bioinformatics tools, have been used to identify large numbers of single nucleotide polymorphisms (SNP in alfalfa, which are being developed into markers. These markers will facilitate fine mapping of quantitative traits and genome wide association mapping of agronomic traits and further advanced breeding strategies for alfalfa, such as marker-assisted selection and genomic selection. Based on ideas from the literature, we suggest several ways to improve selection in alfalfa including (1 diversity selection and paternity testing, (2 introgression of QTL and (3 genomic selection.

  20. Involvement of the smeAB multidrug efflux pump in resistance to plant antimicrobials and contribution to nodulation competitiveness in Sinorhizobium meliloti.

    Science.gov (United States)

    Eda, Shima; Mitsui, Hisayuki; Minamisawa, Kiwamu

    2011-05-01

    The contributions of multicomponent-type multidrug efflux pumps to antimicrobial resistance and nodulation ability in Sinorhizobium meliloti were comprehensively analyzed. Computational searches identified genes in the S. meliloti strain 1021 genome encoding 1 pump from the ATP-binding cassette family, 3 pumps from the major facilitator superfamily, and 10 pumps from the resistance-nodulation-cell division family, and subsequently, these genes were deleted either individually or simultaneously. Antimicrobial susceptibility tests demonstrated that deletion of the smeAB pump genes resulted in increased susceptibility to a range of antibiotics, dyes, detergents, and plant-derived compounds and, further, that specific deletion of the smeCD or smeEF genes in a ΔsmeAB background caused a further increase in susceptibility to certain antibiotics. Competitive nodulation experiments revealed that the smeAB mutant was defective in competing with the wild-type strain for nodulation. The introduction of a plasmid carrying smeAB into the smeAB mutant restored antimicrobial resistance and nodulation competitiveness. These findings suggest that the SmeAB pump, which is a major multidrug efflux system of S. meliloti, plays an important role in nodulation competitiveness by mediating resistance toward antimicrobial compounds produced by the host plant.

  1. Nod Factor-Independent Nodulation in Aeschynomene evenia Required the Common Plant-Microbe Symbiotic Toolkit1

    Science.gov (United States)

    Fabre, Sandrine; Gully, Djamel; Poitout, Arthur; Patrel, Delphine; Arrighi, Jean-François; Cartieaux, Fabienne

    2015-01-01

    Nitrogen fixation in the legume-rhizobium symbiosis is a crucial area of research for more sustainable agriculture. Our knowledge of the plant cascade in response to the perception of bacterial Nod factors has increased in recent years. However, the discovery that Nod factors are not involved in the Aeschynomene-Bradyrhizobium spp. interaction suggests that alternative molecular dialogues may exist in the legume family. We evaluated the conservation of the signaling pathway common to other endosymbioses using three candidate genes: Ca2+/Calmodulin-Dependent Kinase (CCaMK), which plays a central role in cross signaling between nodule organogenesis and infection processes; and Symbiosis Receptor Kinase (SYMRK) and Histidine Kinase1 (HK1), which act upstream and downstream of CCaMK, respectively. We showed that CCaMK, SYMRK, and HK1 are required for efficient nodulation in Aeschynomene evenia. Our results demonstrate that CCaMK and SYMRK are recruited in Nod factor-independent symbiosis and, hence, may be conserved in all vascular plant endosymbioses described so far. PMID:26446590

  2. Photosynthate partitioning and nitrogen fixation of alfalfa and birdsfoot trefoil. [Lotus corniculatus L. ; Medicago sativa L

    Energy Technology Data Exchange (ETDEWEB)

    Shieh, W.J.

    1985-01-01

    Nodule mass and number are usually correlated with rates of nitrogen fixation in legumes. Birdsfoot trefoil (Lotus corniculatus L.) with more than twice the nodule number and mass, however, fixes far less nitrogen than alfalfa (Medicago sativa L.) at the same age. In this research, photosynthesis and photosynthate partitioning and utilization in relation to nitrogen fixation of alfalfa and birdsfoot trefoil were examined in order to determine their relationship to nitrogen fixation potential. Photosynthate to nodules was studied using /sup 14/CO/sub 2/ labeling techniques. Partitioning patterns were altered by shading and dark depletion treatments. Efficiency of photosynthate utilization was examined by determining turnover of /sup 14/C photosynthate in nodule metabolites and by studying rates of cyanide-resistant and cyanide-sensitive O/sub 2/ uptake. Alfalfa nodule activity was greater than trefoil expressed on a hole pot or nodule dry weight basis. Both shading and dark treatments significantly reduced nodule activity as estimated by the acetylene reduction assay. Shoots of both species were found to be the dominant sinks for photosynthate. Percentage /sup 14/C recovered in alfalfa roots was more than twice that of trefoil at 1,2,3,4 and 24 h after labeling. Greater relative specific radioactivity (RSA) in nodules of both species suggests that they were stronger sinks for current photosynthate than roots.

  3. Haemolytic micromethod for rapid estimation of toxic alfalfa saponin

    Directory of Open Access Journals (Sweden)

    Marian Jurzysta

    2015-06-01

    Full Text Available On a plate covered with a thin layer of blood suspension with gelatine 10 microliters of alfalfa juice or alfalfa meal extract is spotted. As a result of saponin diffusion and their reaction with blood a haemolytic ring appears, the diameter of which is proportional to the concentration of toxic saponins. Selected by this method low-saponin individual alfalfa plants proved non toxic for fishes and did not inhibit the growth of Trichoderma viride.

  4. 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.

  5. Thyroid nodule

    Science.gov (United States)

    ... nodules are not always found, but can include: Hashimoto's disease Lack of iodine in the diet Symptoms ... nodules are sometimes found in people who have Hashimoto's disease. This may cause symptoms of an underactive ...

  6. Thyroid Nodules

    Science.gov (United States)

    ... Endocrinologist Search Featured Resource New Mobile App DOWNLOAD Thyroid Nodules March 2010 Download PDFs English Espanol Hindi ... Resources Mayo Clinic American Thyroid Association What are thyroid nodules and who is at risk? A thyroid ...

  7. Thyroid Nodules

    Science.gov (United States)

    ... very accurate for identifying cancerous or “suspicious” nodules. Thyroid ultrasound is used to get an exact picture of ... up may involve a physical exam or a thyroid ultrasound or both. If the nodule gets larger, you ...

  8. Alfalfa microsymbionts from different ITS and nodC lineages of Ensifer meliloti and Ensifer medicae symbiovar meliloti establish efficient symbiosis with alfalfa in Spanish acid soils.

    Science.gov (United States)

    Ramírez-Bahena, Martha-Helena; Vargas, Margarita; Martín, María; Tejedor, Carmen; Velázquez, Encarna; Peix, Álvaro

    2015-06-01

    Alfalfa (Medicago sativa L.) is an important crop worldwide whose cropping in acid soils is hampered by the poor nodulation and yield commonly attributed to the sensitivity of its endosymbionts to acid pH. In this work, we isolated several acid-tolerant strains from alfalfa nodules in three acid soils in northwestern Spain. After grouping by RAPD fingerprinting, most strains were identified as Ensifer meliloti and only two strains as Ensifer medicae according to their 16S-23S intergenic spacer (ITS) sequences that allowed the differentiation of two groups within each one of these species. The two ITS groups of E. meliloti and the ITS group I of E. medicae have been previously found in Medicago nodules; however, the group II of E. medicae has been only found to date in Prosopis alba nodules. The analysis of the nodC gene showed that all strains isolated in this study belong to the symbiovar meliloti, grouping with the type strains of E. meliloti or E. medicae, but some harboured nodC gene alleles different from those found to date in alfalfa nodules. The strains of E. medicae belong to the symbiovar meliloti which should be also recognised in this species, although they harboured a nodC allele phylogenetically divergent to those from E. meliloti strains. Microcosm experiments showed that inoculation of alfalfa with selected acid-tolerant strains significantly increased yields in acid soils representing a suitable agricultural practice for alfalfa cropping in these soils.

  9. Low vapour pressure deficit reduces the beneficial effect of elevated CO{sub 2} on growth of N{sub 2}-fixing alfalfa plants

    Energy Technology Data Exchange (ETDEWEB)

    Luis, I. De; Irigoyen, J.J.; Sanchez-Diaz, M. [Univ. de Navarra, Dept. de Fisioligia Vegetal, Pamplona (Spain)

    2002-11-01

    Plant responses to elevated CO{sub 2} can be modified by many environmental factors, but very little attention has been paid to the interaction between CO{sub 2} and changes in vapour pressure deficit (VPD). Thirty-day-old alfalfa plants (Medicago sativa L. cv. Aragon), which were inoculated with Sinorhizobium meliloti 102F78 strain, were grown for 1 month in controlled environment chambers at 25/15 deg C, 14 h photoperiod, and 600 mol m{sup -2} s{sup -1} photosynthetic photon flux (PPF), using a factorial combination of CO{sub 2} concentration (400 mol mol{sup -1} or 700 mol mol{sup -1}) and vapour pressure deficit (0.48 kPa or 1.74 kPa, which corresponded to relative humidities of 85% and 45% at 25 deg C, respectively). Elevated CO{sub 2} strongly stimulated plant growth under high VPD conditions, but this beneficial effect was not observed under low VPD. Under low VPD, elevated CO{sub 2} also did not enhance plant photosynthesis, and plant water stress was greatest for plants grown at elevated CO{sub 2} and low VPD. Moreover, plants grown under elevated CO{sub 2} and low VPD had a lower leaf soluble protein and photosynthetic activity (photosynthetic rate and carboxylation efficiency) than plants grown under elevated CO{sub 2} and high VPD. Elevated CO{sub 2} significantly increased leaf adaxial and abaxial temperatures. Because the effects of elevated CO{sub 2} were dependent on vapour pressure deficit, VPD needs to be controlled in experiments studying the effect of elevated CO{sub 2} as well as considered in the extrapolations of results to a warmer, high-CO{sub 2} world. (au)

  10. Evaluating effect of biofertilizer on nodulation and soybean (Glycine max L plants growth characteristics under water deficit stress of seed

    Directory of Open Access Journals (Sweden)

    M. Tajik Khaveh

    2016-05-01

    Full Text Available In order to evaluate the effects of biofertilizer on soybean (Glycine max L. seed vigor that produced under water deficit condition and related traits, an experiment was conducted in a factorial layout based of complete randomized block design with four replications at the research greenhouse of Aboureihan campus- Tehran University, Iran. Experimental treatments were include biofertilizer (seed inoculation with Bradyrhizobium japonicum, co-inoculation with Bradyrhizobium japonicum and Pseudomonas fluorescens, co-inoculation with Bradyrhizobium japonicum and Glomus mosseae, Cultivar (Zalta Zalha and Clark×Hobbit line and water deficit stress [irrigation plants after 50 (normal irrigation, 100 (medium stress, 150 (sever stress mm evaporation from pan class A, in parents field]. Results showed that the water deficit stress had negative effects on seed quality and seedling emergence percentage, mean daily seedling emergence, root, leaf and shoot dry weight, number of nodule were decreased. ZaltaZalha cultivar had higher shoot dry weight and number of leaf compared with other cultivars. Applications of biofertilzer was effective on stem diameter, root, leaf and shoot dry weight, number of leaf and nodule and those attributes increased by co-inoculation of Bradyrhizobium japonicum and Glomus mosseae. Also, use of biofertilizer in stress levels was effective on stem dry weight. Stem dry weight was increased by Co-inoculation of cultivar seeds with Bradyrhizobium japonicum and Glomus mosseae.

  11. Root Exudates of Various Host Plants of Rhizobium leguminosarum Contain Different Sets of Inducers of Rhizobium Nodulation Genes.

    Science.gov (United States)

    Zaat, S A; Wijffelman, C A; Mulders, I H; van Brussel, A A; Lugtenberg, B J

    1988-04-01

    Rhizobium promoters involved in the formation of root nodules on leguminous plants are activated by flavonoids in plant root exudate. A series of Rhizobium strains which all contain the inducible Rhizobium leguminosarum nodA promoter fused to the Escherichia coli lacZ gene, and which differ only in the source of the regulatory nodD gene, were recently used to show that the regulatory nodD gene determines which flavonoids are able to activate the nodA promoter (HP Spaink, CA Wijffelman, E Pees, RJH Okker, BJJ Lugtenberg 1987 Nature 328: 337-340). Since these strains therefore are able to discriminate between various flavonoids, they were used to determine whether or not plants that are nodulated by R. leguminosarum produce different inducers. After chromatographic separation of root exudate constituents from Vicia sativa L. subsp. nigra (L.), V. hirsuta (L.) S.F. Gray, Pisum sativum L. cv Rondo, and Trifolium subterraneum L., the fractions were tested with a set of strains containing a nodD gene of R. leguminosarum, R. trifolii, or Rhizobium meliloti, respectively. It appeared that the source of nodD determined whether, and to what extent, the R. leguminosarum nodA promoter was induced. Lack of induction could not be attributed to the presence of inhibitors. Most of the inducers were able to activate the nodA promoter in the presence of one particular nodD gene only. The inducers that were active in the presence of the R. leguminosarum nodD gene were different in each root exudate.

  12. 76 FR 5780 - Determination of Regulated Status of Alfalfa Genetically Engineered for Tolerance to the...

    Science.gov (United States)

    2011-02-02

    ... Animal and Plant Health Inspection Service Determination of Regulated Status of Alfalfa Genetically... regulated status of alfalfa genetically engineered for tolerance to the herbicide glyphosate based on APHIS... decision and determination on the petition regarding the regulated status of alfalfa genetically engineered...

  13. Alfalfa nitrogen credit to first-year corn: potassium, regrowth, and tillage timing effects

    Science.gov (United States)

    Compared to corn (Zea mays L.) following corn, N guidelines for corn following alfalfa (Medicago sativa L.) in the U.S. Corn Belt suggest that N rates for first-year corn after alfalfa be reduced by about 168 kg N/ha when 43 to 53 alfalfa plants per square meter are present at termination; however, ...

  14. Modeling feral alfalfa (Medicago sativa subsp. sativa L.) occurrence using topographical and environmental variables

    Science.gov (United States)

    Because alfalfa is a perennial species cross pollinated by bees and can establish along roadsides and ruderal areas, there is concern that feral plants can serve as reservoirs and conduits for transgenic genes. The objective of this study was to survey feral alfalfa in alfalfa seed production areas ...

  15. Deletion of the SACPD-C locus alters the symbiotic relationship between Bradyrhizobium japonicum USDA110 and soybean, resulting in elicitation of plant defense response and nodulation defects

    Science.gov (United States)

    Legumes form symbiotic association with soil-dwelling bacteria collectively called rhizobia. This association results in the formation of nodules, unique plant-derived organs, within which the rhizobia are housed. Rhizobia encoded-nitrogenase facilitates the conversation of atmospheric nitrogen int...

  16. Draft Genome Sequence of Frankia Strain G2, a Nitrogen-Fixing Actinobacterium Isolated from Casuarina equisetifolia and Able To Nodulate Actinorhizal Plants of the Order Rhamnales.

    Science.gov (United States)

    Nouioui, Imen; Gtari, Maher; Göker, Markus; Ghodhbane-Gtari, Faten; Tisa, Louis S; Fernandez, Maria P; Normand, Philippe; Huntemann, Marcel; Clum, Alicia; Pillay, Manoj; Varghese, Neha; Reddy, T B K; Ivanova, Natalia; Woyke, Tanja; Kyrpides, Nikos C; Klenk, Hans-Peter

    2016-05-26

    Frankia sp. strain G2 was originally isolated from Casuarina equisetifolia and is characterized by its ability to nodulate actinorhizal plants of the Rhamnales order, but not its original host. It represents one of the largest Frankia genomes so far sequenced (9.5 Mbp).

  17. Complete Genome Sequence of Micromonospora Strain L5, a Potential Plant-Growth-Regulating Actinomycete, Originally Isolated from Casuarina equisetifolia Root Nodules

    Energy Technology Data Exchange (ETDEWEB)

    Hirsch, A. M.; Alvarado, J.; Bruce, D.; Chertkov, O.; De Hoff, P. L.; Detter, J. C.; Fujishige, N. A.; Goodwin, L. A.; Han, J.; Han, S.; Ivanova, N.; Land, M. L.; Lum, M. R.; Milani-Nejad, N.; Nolan, M.; Pati, A.; Pitluck, S.; Tran, S. S.; Woyke, T.; Valdes, M.

    2013-08-29

    Micromonospora species live in diverse environments and exhibit a broad range of functions including antibiotic production, biocontrol, and ability to degrade complex polysaccharides. To learn more about these versatile actinomycetes, we sequenced the genome of strain L5, originally isolated from root nodules of an actinorhizal plant growing in Mexico.

  18. Investigation of Antileishmanial Effect of Alcoholic Extract and Essential Oil of Medicinal Plant Leaf Black Alfalfa (Medicago Lupulina, on The Number of Clinical Isolates of Leishmania Major Promastigotes in Vitro

    Directory of Open Access Journals (Sweden)

    E Gharirvand Eskandari

    2016-05-01

    Full Text Available Introduction: Leishmaniasis has created enormous global health problems. Side effects, drug resistance and the lack of effective vaccines had led to the new effective compounds effective of plants. The aim of this study was to introduce a traditional medicinal plant called Black alfalfa (Medicago Lupulina that can be used as a valuable resource against cutaneous leishmaniasis. Methods: In this experimental study, alcoholic extract was prepared by maceration and essential oil by distillation water method. Leishmania major promastigotes were cultured at 25 ± 2° C in N.N.N culturemedium, then in Schneider and next were cultured in RPMI- 1640. afterward, using MTT (Methyl Thiazole Tetrazolium, the IC50 (Inhibitory Concentrations 50% for extracts, essence and Glucantime were determined. The results using Tukey and t-test were analyzed and were presented by software SPSS16. MTT assay were repeated. 3 times for each sample. Results: IC50 for alfalfa leaf extract and essential oil of black alfalfa and Glucantime against L. major promastigotes was determined after 24, 48 and 72 hours, 240, 130 and 69 micrograms per ml, and 801, 340 and 190 micrograms per ml, also 26, 19 and 11 micrograms per ml , respectively. There was a significant differences between the IC50 plant extract and essential oil and Glucantime after 24, 48 and 72 hours. Conclusion: Alcoholic extracts and essential oil the plant had significant anti leishmaniasis effects in vitro. In this way, it can be considered as an anti-leishmaniasis among the herbs.

  19. Colonization and plant growth promoting characterization of endophytic Pseudomonas chlororaphis strain Zong1 isolated from Sophora alopecuroides root nodules.

    Science.gov (United States)

    Zhao, Long Fei; Xu, Ya Jun; Ma, Zhan Qiang; Deng, Zhen Shan; Shan, Chang Juan; Wei, Ge Hong

    2013-01-01

    The endophytic strain Zong1 isolated from root nodules of the legume Sophora alopecuroides was characterized by conducting physiological and biochemical tests employing gfp-marking, observing their plant growth promoting characteristics (PGPC) and detecting plant growth parameters of inoculation assays under greenhouse conditions. Results showed that strain Zong1 had an effective growth at 28 ºC after placed at 4-60 ºC for 15 min, had a wide range pH tolerance of 6.0-11.0 and salt tolerance up to 5% of NaCl. Zong1 was resistant to the following antibiotics (μg/mL): Phosphonomycin (100), Penicillin (100) and Ampicillin (100). It could grow in the medium supplemented with 1.2 mmol/L Cu, 0.1% (w/v) methylene blue and 0.1-0.2% (w/v) methyl red, respectively. Zong1 is closely related to Pseudomonas chlororaphis based on analysis the sequence of 16S rRNA gene. Its expression of the gfp gene indicated that strain Zong1 may colonize in root or root nodules and verified by microscopic observation. Furthermore, co-inoculation with Zong1 and SQ1 (Mesorhizobium sp.) showed significant effects compared to single inoculation for the following PGPC parameters: siderophore production, phosphate solubilization, organic acid production, IAA production and antifungal activity in vitro. These results suggest strains P. chlororaphi Zong1 and Mesorhizobium sp. SQ1 have better synergistic or addictive effect. It was noteworthy that each growth index of co-inoculated Zong1+SQ1 in growth assays under greenhouse conditions is higher than those of single inoculation, and showed a significant difference (p growth promoting agent.

  20. Colonization and plant growth promoting characterization of endophytic Pseudomonas chlororaphis strain Zong1 isolated from Sophora alopecuroides root nodules

    Directory of Open Access Journals (Sweden)

    Long Fei Zhao

    2013-01-01

    Full Text Available The endophytic strain Zong1 isolated from root nodules of the legume Sophora alopecuroides was characterized by conducting physiological and biochemical tests employing gfp-marking, observing their plant growth promoting characteristics (PGPC and detecting plant growth parameters of inoculation assays under greenhouse conditions. Results showed that strain Zong1 had an effective growth at 28 ºC after placed at 4-60 ºC for 15 min, had a wide range pH tolerance of 6.0-11.0 and salt tolerance up to 5% of NaCl. Zong1 was resistant to the following antibiotics (µg/mL: Phosphonomycin (100, Penicillin (100 and Ampicillin (100. It could grow in the medium supplemented with 1.2 mmol/L Cu, 0.1% (w/v methylene blue and 0.1-0.2% (w/v methyl red, respectively. Zong1 is closely related to Pseudomonas chlororaphis based on analysis the sequence of 16S rRNA gene. Its expression of the gfp gene indicated that strain Zong1 may colonize in root or root nodules and verified by microscopic observation. Furthermore, co-inoculation with Zong1 and SQ1 (Mesorhizobium sp. showed significant effects compared to single inoculation for the following PGPC parameters: siderophore production, phosphate solubilization, organic acid production, IAA production and antifungal activity in vitro. These results suggest strains P. chlororaphi Zong1 and Mesorhizobium sp. SQ1 have better synergistic or addictive effect. It was noteworthy that each growth index of co-inoculated Zong1+SQ1 in growth assays under greenhouse conditions is higher than those of single inoculation, and showed a significant difference (p < 0.05 when compared to a negative control. Therefore, as an endophyte P. chlororaphis Zong1 may play important roles as a potential plantgrowth promoting agent.

  1. Intended release and actual retention of alfalfa leafcutting bees (hymenoptera: megachilidae) for pollination in commercial alfalfa seed fields

    Science.gov (United States)

    Low, medium, and high stocking densities of Megachile rotundata, the alfalfa leafcutting bee, were released over four years in three research plots of Utah alfalfa planted at seed-production rates. A low number of bees (46-79% of released) survived the incubation and field emergence processes, and ...

  2. 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.

  3. Effects of N(6)-benzylaminopurine and Indole Acetic Acid on In Vitro Shoot Multiplication, Nodule-like Meristem Proliferation and Plant Regeneration of Malaysian Bananas (Musa spp.).

    Science.gov (United States)

    Sipen, Philip; Davey, Michael R

    2012-12-01

    Different concentrations of N(6)-benzylaminopurine (BAP) and indole acetic acid (IAA) in Murashige and Skoog based medium were assessed for their effects on shoot multiplication, nodule-like meristem proliferation and plant regeneration of the Malaysian banana cultivars Pisang Mas, Pisang Nangka, Pisang Berangan and Pisang Awak. BAP at 1-14 mg L(-1) with or without 0.2 mg L(-1) IAA, or BAP at 7-14 mg L(-1) with the same concentration of IAA, was evaluated for shoot multiplication from shoot tips and the proliferation of nodule-like meristems from scalps, respectively. Plant regeneration from scalps was assessed using 1 mg L(-1) BAP and 0.2 mg L(-1) IAA separately, or a combination of these two growth regulators. Data on shoot multiplication, the proliferation of nodule-like meristems with associated plant regeneration were recorded after 30 days of culture. A maximum of 5 shoots per original shoot tip was achieved on medium supplemented with BAP at 5 mg L(-1) (Pisang Nangka), 6 mg L(-1) (Pisang Mas and Pisang Berangan), or 7 mg L(-1) (Pisang Awak), with 0.2 mg L(-1) IAA. BAP at 11 mg L(-1) with 0.2 mg L(-1) IAA induced the most highly proliferating nodule-like meristems in the four banana cultivars. Plant regeneration from scalps was optimum in all cases on medium containing 1 mg L(-1) BAP and 0.2 mg L(-1) IAA. This is the first report on the successful induction of highly proliferating nodule-like meristems and plant regeneration from scalps of the Malaysian banana cultivars Pisang Mas, Pisang Nangka, Pisang Berangan and Pisang Awak.

  4. Position and quantity of endogensis rhizobia in alfalfa plant%苜蓿内生根瘤菌分布部位与数量变化动态

    Institute of Scientific and Technical Information of China (English)

    李剑峰; 张淑卿; 师尚礼; 霍平慧

    2009-01-01

    为探明根瘤菌在苜蓿植株及其种子内的分布规律,对苜蓿植株及种子各部位在不同生育期内生根瘤菌的分布和数量变化进行了研究.结果表明:苜蓿植株内根瘤菌的分布和数量在空间和时间上都具有很大异质性.空间分布上,不同部位组织的内生根瘤菌数量随植株光合产物源-库的运输方向呈逐渐增大的趋势,绝大部分内生根瘤菌分布于植株的根系,并主要分布于毛根,在主根内主要存在于表皮和皮层,中柱分布较少;在植株地上部分,内生根瘤菌主要在营养期末分布于花芽[8.6~9.6×10~3 cfu·g~(-1) (FW)],在蕾期和花期分布于雌蕊子房的子房壁,在结英期分布干荚果果皮(1.07×10~3 cfu·pod~(-1)),在种子成熟期则主要存在于新生的种子中;茎内的根瘤菌数量在营养期和结荚期不足2×10~2 cfu·g~(-1) (FW),而在蕾期和花期完全绝迹;叶片内则始终不存在根瘤菌.在时间上,结荚期根、荚果皮内的根瘤菌数量明显高于其他时期;花内各器官(不包括花梗)在授粉后根瘤菌数量迅速增加;由子房向荚果发育的过程中,子房壁和胚珠内的根瘤菌数量随时间呈对数增长;胚珠在受精后即存在有内生根瘤菌.并且幼嫩种子内生根瘤菌数量远高于受精胚珠,证明内生根瘤菌能被转运并定殖在发育早期的种子中.两个苜蓿品种成熟种子在收获120 d后,根瘤菌数量比刚收获时分别增加131.46倍("陇东")和11.76倍("游客"),说明根瘤菌进入种子后,仍然有一个继续繁殖增长的过程.%To determine the distribution characteristics of rhizobium in alfalfa plants and seeds, the distribution and abundance of endogenous rhizobium in different parts and seed of alfalfa at different growth stages were analyzed. The result indicates significant spatial and temporal heterogeneity in endogenous rhizobium abundance in alfalfa plant. For the spatial distribution, rhizobium abundance

  5. Development and characterization of the first infectious clone of alfalfa latent virus, a strain of Pea streak virus

    Science.gov (United States)

    Alfalfa (Medicago sativa) is a natural host plant for many plant pathogens including fungi, bacteria, nematodes and viruses. Alfalfa latent virus (ALV) is a member of the carlavirus group and occurs symptomlessly in alfalfa. The first complete genomic sequence of the ALV that was recently obtained i...

  6. Effect of plant growth promoting rhizobacteria containing ACC-deaminase on maize (Zea mays L.) growth under axenic conditions and on nodulation in mung bean (Vigna radiata L.).

    Science.gov (United States)

    Shaharoona, B; Arshad, M; Zahir, Z A

    2006-02-01

    This study was conducted to test the hypothesis that the bacterial strains possessing 1-aminocyclopropane-1-carboxylic acid (ACC)-deaminase activity may also promote growth of inoculated plants and could increase nodulation in legumes upon co-inoculation with rhizobia. Several rhizobacteria were isolated from maize rhizosphere through enrichment on ACC as a sole N source. Purified isolates were screened for growth promotion in maize under axenic conditions and for in vitro ACC-deaminase activity. A significant positive correlation was observed between in vitro ACC-deaminase activity of bacterial cells and root elongation. None of the isolates produced auxins. Bradyrhizobium japonicum produced less amount of auxins but did not carry ACC-deaminase activity. Results of pot experiment revealed that co-inoculation with Bradyrhizobium and plant growth promoting rhizobacteria (PGPR) isolates enhanced the nodulation in mung bean compared with inoculation with Bradyrhizobium alone. It is highly expected that inoculation with rhizobacteria containing ACC-deaminase hydrolysed endogenous ACC into ammonia and alpha-ketobutyrate instead of ethylene. Consequently, root and shoot growth as well as nodulation were promoted. The ACC-deaminase trait could be employed as an efficient tool to screen effective PGPR, which could be successfully used as biofertilizers to increase the growth of inoculated plants as well as nodulation in legumes.

  7. 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.

  8. Manganese nodules

    Science.gov (United States)

    Hein, James R.; Harff, Jan; Petersen, Sven; Thiede, Jorn

    2016-01-01

    The existence of manganese (Mn) nodules (Fig. 1) has been known since the late 1800s when they were collected during the Challenger expedition of 1873–1876. However, it was not until after WWII that nodules were further studied in detail for their ability to adsorb metals from seawater. Many of the early studies did not distinguish Mn nodules from Mn crusts. Economic interest in Mn nodules began in the late 1950s and early 1960s when John Mero finished his Ph.D. thesis on this subject, which was published...

  9. Cultivation of alfalfa (medicago sativa L).

    Science.gov (United States)

    Rashmi, R; Sarkar, M; Vikramaditya

    1997-10-01

    Madicago sativa Linn, commonly known as 'Alfalfa', is a tonic plant rich in proteins, minerals, enzymes and vitamins, Bulk quantity of the whole plant is required in the pharmaceutical industries especially in homoecopathic pharmacies, Hence, there is a great need to cultivate this plant for sustained supply of the drug. Use of good and adequate phosphatecontaining farm yard manure, timely irrigation and appropriate spacing between plants results in good yield.

  10. Alfalfa non-feed uses

    Science.gov (United States)

    Non-feed uses for alfalfa such as biomass energy and phytoremediation could increase alfalfa acreage and improve farm profitability. The new bio-energy alfalfa and production system increased forage yield and ethanol production. New alfalfas with enhanced nitrogen cycling capacities would protect wa...

  11. Metal uptake of tomato and alfalfa plants as affected by water source, salinity, and Cd and Zn levels under greenhouse conditions.

    Science.gov (United States)

    Gharaibeh, Mamoun A; Marschner, Bernd; Heinze, Stefanie

    2015-12-01

    Irrigation with wastewater is a promising option to improve crop yields and to reduce pressure on freshwater sources. However, heavy metal concentrations in wastewater may cause health concerns. A greenhouse pot experiment was conducted in order to determine cadmium (Cd) and zinc (Zn) concentrations in sandy soil and plant tissues of tomato (Lycopersicon esculentum L.) and alfalfa (Medicago sativa L.). A 2 × 2 × 4 × 2 factorial treatment arrangement was utilized. Two water sources, fresh (FW) or treated wastewater (TWW), at two salinity levels (1 and 3 dS m(-1)) containing different levels of Cd and Zn were used. Samples were collected after a 90-day growth period. It was observed that the growth of both plants was depressed at the highest metal level (L3). Metal accumulation in plant parts increased with the increase of metal concentration and salinity in irrigation water. At low salinity, water source was the main factor which controlled metal accumulation, whereas, at high salinity, chloride appeared to be the principal factor controlling metal uptake regardless of water source. Metal translocation from roots to shoots increased in TWW-irrigated plants, even in the controls. Tomatoes accumulated Cd up to and above critical levels safe for human consumption, even though Cd concentration in irrigation water did not exceed the current recommended values. Therefore, food production in sandy soils may well pose a health hazard when irrigated with TWW containing heavy metals. Complexation with dissolved organic compounds (DOC) in TWW may be to be the principal factor responsible for increased metal uptake and transfer at low salinity, thereby increasing the risk of heavy metal contamination of food and forage crops.

  12. Photosynthesis, N(2) fixation and taproot reserves during the cutting regrowth cycle of alfalfa under elevated CO(2) and temperature.

    Science.gov (United States)

    Erice, G; Sanz-Sáez, A; Aranjuelo, I; Irigoyen, J J; Aguirreolea, J; Avice, J-C; Sánchez-Díaz, M

    2011-11-15

    Future climatic conditions, including rising atmospheric CO(2) and temperature may increase photosynthesis and, consequently, plant production. A larger knowledge of legume performance under the predicted growth conditions will be crucial for safeguarding crop management and extending the area under cultivation with these plants in the near future. N(2) fixation is a key process conditioning plant responsiveness to varying growth conditions. Moreover, it is likely to increase under future environments, due to the higher photosynthate availability, as a consequence of the higher growth rate under elevated CO(2). However, as described in the literature, photosynthesis performance is frequently down-regulated (acclimated) under long-term exposure to CO(2), especially when affected by stressful temperature and water availability conditions. As growth responses to elevated CO(2) are dependent on sink-source status, it is generally accepted that down-regulation occurs in situations with insufficient plant C sink capacity. Alfalfa management involves the cutting of shoots, which alters the source-sink relationship and thus the photosynthetic behaviour. As the growth rate decreases at the end of the pre-cut vegetative growth period, nodulated alfalfa plants show photosynthetic down-regulation, but during regrowth following defoliation, acclimation to elevated CO(2) disappears. The shoot harvest also leads to a drop in mineral N uptake and C translocation to the roots, resulting in a reduction in N(2) fixation due to the dependence on photosynthate supply to support nodule function. Therefore, the production of new shoots during the first days following cutting requires the utilization of reduced C and N compounds that have been stored previously in reserve organs. The stored reserves are mediated by phytohormones such as methyl jasmonate and abscisic acid and in situations where water stress reduces shoot production this potentially enables the enhancement of taproot

  13. Alfalfa benefits from Medicago truncatula: the RCT1 gene from M. truncatula confers broad-spectrum resistance to anthracnose in alfalfa.

    Science.gov (United States)

    Yang, Shengming; Gao, Muqiang; Xu, Chenwu; Gao, Jianchang; Deshpande, Shweta; Lin, Shaoping; Roe, Bruce A; Zhu, Hongyan

    2008-08-26

    Alfalfa is economically the most important forage legume worldwide. A recurrent challenge to alfalfa production is the significant yield loss caused by disease. Although knowledge of molecular mechanisms underlying host resistance should facilitate the genetic improvement of alfalfa, the acquisition of such knowledge is hampered by alfalfa's tetrasomic inheritance and outcrossing nature. However, alfalfa is congeneric with the reference legume Medicago truncatula, providing an opportunity to use M. truncatula as a surrogate to clone the counterparts of many agronomically important genes in alfalfa. In particular, the high degree of sequence identity and remarkably conserved genome structure and function between the two species enables M. truncatula genes to be used directly in alfalfa improvement. Here we report the map-based cloning of RCT1, a host resistance (R) gene in M. truncatula that confers resistance to multiple races of Colletotrichum trifolii, a hemibiotrophic fungal pathogen that causes anthracnose disease of alfalfa. RCT1 is a member of the Toll-interleukin-1 receptor/nucleotide-binding site/leucine-rich repeat (TIR-NBS-LRR) class of plant R genes and confers broad-spectrum anthracnose resistance when transferred into susceptible alfalfa plants. Thus, RCT1 provides a novel resource to develop anthracnose-resistant alfalfa cultivars and contributes to our understanding of host resistance against the fungal genus Colletotrichum. This work demonstrates the potential of using M. truncatula genes for genetic improvement of alfalfa.

  14. Effect of plant growth promoting micro organisms on increasing water use efficiency of alfalfa in water-stress conditions

    Directory of Open Access Journals (Sweden)

    M. Zafari

    2015-11-01

    Full Text Available In order to study the effect of bacterial growth on water use efficiency of alfalfa, a greenhouse experiment, as factorial based on completely randomized blocks design with three replications, was conducted at Faculty of Agriculture, University of Mohaghegh Ardabili, Ardabil, Iran, in 2012. Treatments consisted of 3 levels of water stress (75, 55 and 35% of field capacity and seed inoculation at 4 levels (no inoculation (control, inoculation with mycorhhiza G. mosseae, inoculation with rhyzobium S. meliloti, and inoculation with combination of mycorhhiza and rhyzobium. Results showed that water stress and seed inoculation have significant effect (P&le0.01 on leaf nutrients content. Water stress reduced absorption of phosphorus (23%, potassium (8%, iron (4% and increased sodium absorption (14% in non-inoculated seeds. Inoculation of seeds reduced stress effects and combined inoculation had the highest effect. Stomatal conductance and water use efficiency were affected (P&le0.01 by inoculation and water stress. Stomatal conductance was decreased during the stress period and seed inoculation with mycorhhiza G. mosseae was most effective on increasing stomatal conductance (47% at the highest level of stress. Water use efficiency increased as a result of water stress and inoculation. The highest value of water use efficiency (0.166 mg/kg was obtained in the combined inoculation with 35% field capacity treatment. Results of regression equations showed that during the inoculation, contribution of phosphorus and potassium in regulation of stomatal conductance was increased and contribution of sodium was decreased.  However, during the stress period, the share of potassium and sodium was increased in stomatal conductance and the share of phosphorus was reduced. Also, stress increased the role of stomatal conductance in water use efficiency. However, inoculation reduced the role of stomatal conductance in water use efficiency.

  15. Enhanced nodulation and nodule development by nolR mutants of Sinorhizobium medicae on specific Medicago host genotypes.

    Science.gov (United States)

    Sugawara, Masayuki; Sadowsky, Michael J

    2014-04-01

    The nolR gene encodes a negatively acting, transcriptional regulatory protein of core Nod-factor biosynthetic genes in the sinorhizobia. Although previous reports showed that nolR modulates Nod-factor production and enhances nodulation speed of Sinorhizobium meliloti on alfalfa, there have been no reports for the symbiotic function of this gene in the S. medicae-Medicago truncatula symbiosis. Here, we constructed an nolR mutant of S. medicae WSM419 and evaluated mutant and wild-type strains for their nodulation ability, competitiveness, host specificity, and density-dependent nodulation phenotypes. When the mutant was inoculated at low and medium population densities, it showed enhanced nodule formation during the initial stages of nodulation. Results of quantitative competitive nodulation assays indicated that an nolR mutant had 2.3-fold greater competitiveness for nodulation on M. truncatula 'A17' than did the wild-type strain. Moreover, the nodulation phenotype of the nolR mutant differed among Medicago genotypes and showed significantly enhanced nodule development on M. tricycla. Taken together, these results indicated that mutation of nolR in S. medicae positively influenced nodule initiation, competitive nodulation, and nodule development at later nodulation stages. This may allow nolR mutants of S. medicae to have a selective advantage under field conditions.

  16. Rhizobia from Lanzarote, the Canary Islands, that nodulate Phaseolus vulgaris have characteristics in common with LMW RNA group II Sinorhizobium meliloti of Medicago, Melilotus and Trigonella from soils of mainland Spain

    Science.gov (United States)

    Several isolates from nodules of Phaseolus vulgaris grown in soil of Lanzarote, an island of the Canaries, had electrophoretic LMW RNA patterns identical with a less common pattern within S. meliloti (assigned as group II) obtained from nodules of alfalfa and alfalfa-related legumes grown in northe...

  17. Enhanced tolerance and remediation to mixed contaminates of PCBs and 2,4-DCP by transgenic alfalfa plants expressing the 2,3-dihydroxybiphenyl-1,2-dioxygenase.

    Science.gov (United States)

    Wang, Yan; Ren, Hejun; Pan, Hongyu; Liu, Jinliang; Zhang, Lanying

    2015-04-01

    Polychlorinated biphenyls (PCBs) and 2,4-dichlorophenol (2,4-DCP) generally led to mixed contamination of soils as a result of commercial and agricultural activities. Their accumulation in the environment poses great risks to human and animal health. Therefore, the effective strategies for disposal of these pollutants are urgently needed. In this study, genetic engineering to enhance PCBs/2,4-DCP phytoremediation is a focus. We cloned the 2,3-dihydroxybiphenyl-1,2-dioxygenase (BphC.B) from a soil metagenomic library, which is the key enzyme of aerobic catabolism of a variety of aromatic compounds, and then it was expressed in alfalfa driven by CaMV 35S promoter using Agrobacterium-mediated transformation. Transgenic line BB11 was selected out through PCR, Western blot analysis and enzyme activity assays. Its disposal and tolerance to both PCBs and 2,4-DCP were examined. The tolerance capability of transgenic line BB11 towards complex contaminants of PCBs/2,4-DCP significantly increased compared with non-transgenic plants. Strong dissipation of PCBs and high removal efficiency of 2,4-DCP were exhibited in a short time. It was confirmed expressing BphC.B would be a feasible strategy to help achieving phytoremediation in mixed contaminated soils with PCBs and 2,4-DCP.

  18. Manganese nodules

    Science.gov (United States)

    Hein, James R.; Harff, Jan; Petersen, Sven; Thiede, Jorn

    2016-01-01

    The existence of manganese (Mn) nodules (Figure 1) has been known since the late 1800s when they were collected during the Challenger expedition of 1873–1876. However, it was not until after WWII that nodules were further studied in detail for their ability to adsorb metals from seawater. Many of the early studies did not distinguish Mn nodules from Mn crusts. Economic interest in Mn nodules began in the late 1950s and early 1960s when John Mero finished his Ph.D. thesis on this subject, which was published in the journal Economic Geology (Mero, 1962) and later as a book (Mero, 1965). By the mid-1970s, large consortia had formed to search for and mine Mn nodules that occur between the Clarion and Clipperton fracture zones (CCZ) in the NE Pacific (Figure 2). This is still the area considered of greatest economic potential in the global ocean because of high nickel (Ni), copper (Cu), and Mn contents and the dense distribution of nodules in the area. While the mining of nodules was fully expected to begin in the late 1970s or early 1980s, this never occurred due to a downturn in the price of metals on the global market. Since then, many research cruises have been undertaken to study the CCZ nodules, and now 15 contracts for exploration sites have been given or are pending by the International Seabed Authority (ISA). Many books and science journal articles have been published summarizing the early work (e.g., Baturin, 1988; Halbach et al., 1988), and research has continued to the present day (e.g., ISA, 1999; ISA, 2010). Although the initial attraction for nodules was their high Ni, Cu, and Mn contents, subsequent work has shown that nodules host large quantities of other critical metals needed for high-tech, green-tech, and energy applications (Hein et al., 2013; Hein and Koschinsky, 2014).

  19. Experimental Study on Characteristics of High Water Plant Alfalfa%高水分植物苜蓿强化干燥特性的试验研究

    Institute of Scientific and Technical Information of China (English)

    车刚; 王鑫; 万霖

    2015-01-01

    针对苜蓿为非均一性物质的特性,亟待提高各组分干燥程度的一致性。通过紫花苜蓿的生理和物理特性的试验分析,建立叶片、茎秆和草段的干燥特性曲线。利用基本的物理方法,确定最长干燥存留时间与苜蓿含水率的关系,机械处理次数和草段长度对苜蓿干燥速率的影响规律。试验研究表明:用机械压裂茎秆的方式处理的苜蓿与未处理的相比,干燥速度提高1.5~2倍。选择光滑槽压扁一次和纵向槽重复压扁三次处理的干燥效果最好。%According to the characteristics of uniformity material on alfalfa,the drying consistency of alfalfa components would be improved. Drying characteristic curve of leaf,stem and grass were established by physiological and physical properties of alfalfa. Using the basic physical methods,the relationship of the longest drying time and moisture content of alfalfa was determined,and the effect law of mechanical processing times and grass length to alfalfa drying rate were studied. The experimental study showed the mechanical way of fracturing alfalfa was compared with untreated alfalfa,the drying rate was increased by 1.5-2 times. The best drying effect of squashed alfalfa stem was to use smooth slot one time and longitudinal slot three times.

  20. Rapid phenotyping of alfalfa root system architecture

    Science.gov (United States)

    Root system architecture (RSA) influences the capacity of an alfalfa plant for symbiotic nitrogen fixation, nutrient uptake and water use efficiency, resistance to frost heaving, winterhardiness, and some pest and pathogen resistance. However, we currently lack a basic understanding of root system d...

  1. Virulence of Fusarium species to alfalfa seedlings

    Directory of Open Access Journals (Sweden)

    Krnjaja Vesna

    2005-01-01

    Full Text Available In in vitro conditions, virulence of 91 isolates of species Fusarium genus (F. oxysporum, F. solani, F. acuminatum, F. equiseti, F. arthrosporioides, F. prolifera- tum, F. avenaceum, F. semitectum, F. tricinctum, F. sporotrichioides and F. graminearum towards alfalfa seedlings was investigated. Isolates of investigated species originated from diseased alfalfa plants collected at four locations in Serbia based on symptoms of wilting caused by Fusarium and root rotting. Pathogenicity and virulence of investigated isolates of Fusarium spp. were determined by visual evaluation of inoculated seedlings of cultivar K28 in laboratory conditions. All isolated of investigated species had pathogenic effect on alfalfa seedlings which expressed symptoms such as necrosis of root, moist rotting and "melting of seedlings". Colour of necrotic root tissue varied from light brown, brown lipstick red to explicit black, depending on the Fusarium species. Strong virulence was established in 48 isolates, medium virulence in 31 and weak virulence in 12 isolates.

  2. Virulence of Fusarium species to alfalfa seedlings

    Directory of Open Access Journals (Sweden)

    Krnjaja Vesna

    2005-01-01

    Full Text Available In in vitro conditions, virulence of 91 isolates of species Fusarium genus (F. oxysporum, F. solani, F. acuminatum, F. equiseti, F. arthrosporioides, F. proliferatum, F. avenaceum, F. semitectum, F. tricinctum, F. sporotrichioides and F. graminearum towards alfalfa seedlings was investigated. Isolates of investigated species originated from diseased alfalfa plants collected on four locations in Serbia based on symptoms of wilting caused by fusarium and root rotting. Pathogenicity and virulence of investigated isolates of Fusarium spp. were determined by visual evaluation of inoculated seedlings of cultivars K28 in laboratory conditions. All isolated of investigated species had pathogenic effect on alfalfa seedlings, which expressed symptoms such as necrosis of root, moist rotting and "melting of seedlings". Colour of necrotic root tissue varied from light brown, brown, lipstick red to explicit black, depending on the Fusarium species. Strong virulence was established in 48 isolates, medium virulence in 31 and weak virulence in 12 isolates.

  3. Knockdown of LjALD1, AGD2-like defense response protein 1, influences plant growth and nodulation in Lotus japonicus.

    Science.gov (United States)

    Chen, Wei; Li, Xueliu; Tian, Lu; Wu, Pingzhi; Li, Meiru; Jiang, Huawu; Chen, Yaping; Wu, Guojiang

    2014-11-01

    The discovery of the enzyme L,L-diaminopimelate aminotransferase (LL-DAP-AT, EC 2.6.1.83) uncovered a unique step in the L-lysine biosynthesis pathway in plants. In Arabidopsis thaliana, LL-DAP-AT has been shown to play a key role in plant-pathogen interactions by regulation of the salicylic acid (SA) signaling pathway. Here, a full-length cDNA of LL-DAP-AT named as LjALD1 from Lotus japonicus (Regel) Larsen was isolated. The deduced amino acid sequence shares 67% identity with the Arabidopsis aminotransferase AGD2-LIKE DEFENSE RESPONSE PROTEIN1 (AtALD1) and is predicted to contain the same key elements: a conserved aminotransferase domain and a pyridoxal-5'-phosphate cofactor binding site. Quantitative real-time PCR analysis showed that LjALD1 was expressed in all L. japonicus tissues tested, being strongest in nodules. Expression was induced in roots that had been infected with the symbiotic rhizobium Mesorhizobium loti or treated with SA agonist benzo-(1, 2, 3)-thiadiazole-7-carbothioic acid. LjALD1 Knockdown exhibited a lower SA content, an increased number of infection threads and nodules, and a slight reduction in nodule size. In addition, compared with wild-type, root growth was increased and shoot growth was suppressed in LjALD1 RNAi plant lines. These results indicate that LjALD1 may play important roles in plant development and nodulation via SA signaling in L. japonicus. © 2014 Institute of Botany, Chinese Academy of Sciences.

  4. The evolution of nodulation

    NARCIS (Netherlands)

    Gualtieri, G.; Bisseling, T.

    2000-01-01

    In this review we will first describe the different steps leading to nodule formation, and these will be compared with processes of non-symbiotic plant development and growth. In general, aspects of both actinorhizal as well as rhizobial symbiosis are described, but in several cases, the emphasis wi

  5. The Sinorhizobium meliloti RNA chaperone Hfq influences central carbon metabolism and the symbiotic interaction with alfalfa

    Directory of Open Access Journals (Sweden)

    Jiménez-Zurdo José I

    2010-03-01

    Full Text Available Abstract Background The bacterial Hfq protein is able to interact with diverse RNA molecules, including regulatory small non-coding RNAs (sRNAs, and thus it is recognized as a global post-transcriptional regulator of gene expression. Loss of Hfq has an extensive impact in bacterial physiology which in several animal pathogens influences virulence. Sinorhizobium meliloti is a model soil bacterium known for its ability to establish a beneficial nitrogen-fixing intracellular symbiosis with alfalfa. Despite the predicted general involvement of Hfq in the establishment of successful bacteria-eukaryote interactions, its function in S. meliloti has remained unexplored. Results Two independent S. meliloti mutants, 2011-3.4 and 1021Δhfq, were obtained by disruption and deletion of the hfq gene in the wild-type strains 2011 and 1021, respectively, both exhibiting similar growth defects as free-living bacteria. Transcriptomic profiling of 1021Δhfq revealed a general down-regulation of genes of sugar transporters and some enzymes of the central carbon metabolism, whereas transcripts specifying the uptake and metabolism of nitrogen sources (mainly amino acids were more abundant than in the wild-type strain. Proteomic analysis of the 2011-3.4 mutant independently confirmed these observations. Symbiotic tests showed that lack of Hfq led to a delayed nodulation, severely compromised bacterial competitiveness on alfalfa roots and impaired normal plant growth. Furthermore, a large proportion of nodules (55%-64% elicited by the 1021Δhfq mutant were non-fixing, with scarce content in bacteroids and signs of premature senescence of endosymbiotic bacteria. RT-PCR experiments on RNA from bacteria grown under aerobic and microoxic conditions revealed that Hfq contributes to regulation of nifA and fixK1/K2, the genes controlling nitrogen fixation, although the Hfq-mediated regulation of fixK is only aerobiosis dependent. Finally, we found that some of the recently

  6. Animal feed compositions containing phytase derived from transgenic alfalfa and methods of use thereof

    Science.gov (United States)

    Austin-Phillips, Sandra; Koegel, Richard G.; Straub, Richard J.; Cook, Mark

    1999-01-01

    A value-added composition of matter containing plant matter from transgenic alfalfa which expresses exogenous phytase activity is disclosed. The phytase activity is a gene product of an exogenous gene encoding for phytase which has been stably incorporated into the genome of alfalfa plants. The transgenic alfalfa expresses phytase activity in nutritionally-significant amounts, thereby enabling its use in animal feeds to eliminate the need for phosphorous supplementation of livestock, poultry, and fish feed rations.

  7. Animal feed compositions containing phytase derived from transgenic alfalfa and methods of use thereof

    Energy Technology Data Exchange (ETDEWEB)

    Austin-Phillips, Sandra (Madison, WI); Koegel, Richard G. (Madison, WI); Straub, Richard J. (Brooklyn, WI); Cook, Mark (Madison, WI)

    2001-01-01

    A value-added composition of matter containing plant matter from transgenic alfalfa which expresses exogenous phytase activity is disclosed. The phytase activity is a gene product of an exogenous gene encoding for phytase which has been stably incorporated into the genome of alfalfa plants. The transgenic alfalfa expresses phytase activity in nutritionally-significant amounts, thereby enabling its use in animal feeds to eliminate the need for phosphorous supplementation of livestock, poultry, and fish feed rations.

  8. Knockdown of LjALD1, AGD2-like defense response protein 1, influences plant growth and nodulation in Lotus japonicus

    Institute of Scientific and Technical Information of China (English)

    Wei Chen; Xueliu Li; Lu Tian; Pingzhi Wu; Meiru Li; Huawu Jiang; Yaping Chen; and Guojiang Wu

    2014-01-01

    The discovery of the enzyme L,L‐diaminopimelate aminotransferase (LL‐DAP‐AT, EC 2.6.1.83) uncovered a unique step in the L‐lysine biosynthesis pathway in plants. In Arabidopsis thaliana, LL‐DAP‐AT has been shown to play a key role in plant‐pathogen interactions by regulation of the salicylic acid (SA) signaling pathway. Here, a ful‐length cDNA of LL‐DAP‐AT named as LjALD1 from Lotus japonicus (Regel) Larsen was isolated. The deduced amino acid sequence shares 67%identity with the Arabidopsis aminotransferase AGD2‐LIKE DEFENSE RESPONSE PROTEIN1 (AtALD1) and is predicted to contain the same key elements:a conserved aminotransferase domain and a pyridoxal‐5’‐phosphate cofactor binding site. Quantitative real‐time PCR analysis showed that LjALD1 was expressed in al L. japonicus tissues tested, being strongest in nodules. Expression was induced in roots that had been infected with the symbiotic rhizobium Mesorhizobium loti or treated with SA agonist benzo‐(1, 2, 3)‐thiadiazole‐7‐carbothioic acid. LjALD1 Knockdown exhibited a lower SA content, an increased number of infection threads and nodules, and a slight reduction in nodule size. In addition, compared with wild‐type, root growth was increased and shoot growth was suppressed in LjALD1 RNAi plant lines. These results indicate that LjALD1 may play important roles in plant development and nodulation via SA signaling in L. japonicus.

  9. Ectopic expression of miR156 represses nodulation and causes morphological and developmental changes in Lotus japonicus.

    Science.gov (United States)

    Wang, Ying; Wang, Zhishuo; Amyot, Lisa; Tian, Lining; Xu, Ziqin; Gruber, Margaret Y; Hannoufa, Abdelali

    2015-04-01

    The effects of microRNA156 overexpression on general plant architecture, branching, flowering time and nodulation were investigated in the model legume, Lotus japonicus. We cloned an miR156 homolog, LjmiR156a, from L. japonicus, and investigated its SQUAMOSA PROMOTER BINDING PROTEIN LIKE (SPL) genes and its biological function at enhancing vegetative biomass yield, extending flowering time, and its impact on nodulation. Thirteen potential targets for LjmiR156 were identified in vitro and their expression profiles were determined in aerial and underground parts of mature plants, including genes coding for eight SPLs, one WD-40, one RNA-directed DNA polymerase, two transport proteins, and one histidine-phosphotransfer protein. Two SPL and one WD-40 cleavage targets for LjmiR156-TC70253, AU089191, and TC57859-were identified. Transgenic plants with ectopic expression of LjmiR156a showed enhanced branching, dramatically delayed flowering, underdeveloped roots, and reduced nodulation. We also examined the transcript levels of key genes involved in nodule organogenesis and infection thread formation to determine the role of miR156 in regulating symbiosis. Overexpression of LjmiR156a led to repression of several nodulation genes during the early stages of root development such as three ENOD genes, SymPK, POLLUX, CYCLOPS, Cerberus, and Nsp1, and the stimulation of NFR1. Our results show that miR156 regulates vegetative biomass yield, flowering time and nodulation by silencing downstream target SPLs and other genes, suggesting that the miR156 regulatory network could be modified in forage legumes (such as alfalfa and trefoils) and in leafy vegetables (like lettuce and spinach) to positively impact economically valuable crop species.

  10. Effect of Planting Methods and Seeding Rates on Yield of Alfalfa (Medicago sativa L. CV. Hamedani in Bajgah, Fars Province

    Directory of Open Access Journals (Sweden)

    A. Yazdani

    2015-06-01

    ha-1, respectively. Seeding rates also had a significant effect on number of weeds so that maximum and minimum weed numbers were obtained in 20 kg and 5 kg seed ha-1. Our results showed that 20 kg seed ha-1 and furrow planting method was the best treatment to gain maximum forage yield and minimum weed's detrimental impact.

  11. 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 alfalfa silage (P alfalfa silage treated with pesticides (P alfalfa silage (P 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.

  12. Aluminium-induced reduction of plant growth in alfalfa (Medicago sativa) is mediated by interrupting auxin transport and accumulation in roots

    OpenAIRE

    Shengyin Wang; Xiaoyan Ren; Bingru Huang; Ge Wang; Peng Zhou; Yuan An

    2016-01-01

    The objective of this study was to investigate Al3+-induced IAA transport, distribution, and the relation of these two processes to Al3+-inhibition of root growth in alfalfa. Alfalfa seedlings with or without apical buds were exposed to 0 or 100 μM AlCl3 and were foliar sprayed with water or 6 mg L−1 IAA. Aluminium stress resulted in disordered arrangement of cells, deformed cell shapes, altered cell structure, and a shorter length of the meristematic zone in root tips. Aluminium stress signi...

  13. Lung Nodules: Overview

    Science.gov (United States)

    ... Research & Science Education & Training Home Conditions Lung Nodules Lung Nodules Make an Appointment Find a Doctor Ask ... Kern, MD (June 01, 2016) What is a lung nodule? A lung nodule is also called a ...

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

    Science.gov (United States)

    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.

  15. Roadside alfalfa: Innocent bystanders or conveyers of genetically-engineered traits?

    Science.gov (United States)

    Clumps of alfalfa are a common sight along roads and vacant lots in areas that grow alfalfa for hay or seed. So what role do feral roadside plants play in dispersing transgenes? Is there a risk that transgenic feral plants serve as reservoirs or conduits that might facilitate the movement of transg...

  16. Multiwalled carbon nanotubes in alfalfa and wheat: toxicology and uptake

    OpenAIRE

    Miralles, Pola; Johnson, Errin; Tamara L. Church; Harris, Andrew T.

    2012-01-01

    Data on the bioavailability and toxicity of carbon nanotubes (CNTs) in the environment, and, in particular, on their interactions with vascular plants, are limited. We investigated the effects of industrial-grade multiwalled CNTs (75 wt% CNTs) and their impurities on alfalfa and wheat. Phytotoxicity assays were performed during both seed germination and seedling growth. The germinations of both species were tolerant of up to 2560 mg l−1 CNTs, and root elongation was enhanced in alfalfa and wh...

  17. Growth With of Alfalfa Mutant in Different Nitrogen Fertilizer and Defoliation Intensity

    Directory of Open Access Journals (Sweden)

    Widyati Slamet

    2012-10-01

    Full Text Available The research was conducted to evaluate growth  of alfalfa mutan (plant height increment, number of leaves and dry matter production in different  Nitrogen Fertilizer and defoliation intensity. The design used was randomized block design 4x2 factorial with 3 replications. The first was dosage of Nitrogen fertilizerNitrogen   (0, 30, 60 and 90 kg N / ha,  the second factor was defoliation intensity (5 and 10 cm. Variables observed alfalfa growth (plant height increment, number of leaves, the production of dry matter (DM forage. The results showed that different N fertilization did not affect the growth of alfalfa mutants. Defoliation intensity  affectedmnumber of leaves and   DM  production of alfalfa mutant. Fertilization to 90 kg N / ha has not affected the growth and   defoliation  intensity 10 cm gave better growth on alfalfa mutant.   [Keywords: alfalfa mutant; Nitrogen fertilizer; defoliation; growth

  18. Differential Pathogenicity of Four Pratylenchus neglectus Populations on Alfalfa

    OpenAIRE

    1991-01-01

    A Pratylenchus neglectus population from lltah (UT3) was more virulent to Lahontan alfalfa than other P. neglectus populations from Utah (UT1, UT2) and Wyoming (WY). All alfalfa plants survived at 24 ± 3 C when inoculated with WY, UT1, or UT2 at initial populations (Pi) of 500, 1,000, and 5,000 nematodes per plant. At Pi 10,000 with WY, UT1, or UT2, plant mortality was 15, 15, and 20%, respectively; at Pi 5,000 and 10,000 with UT3, plant mortality was 10 and 40%. The WY, UT1, and UT2 populati...

  19. Transcriptome Analysis of Resistant and Susceptible Alfalfa Cultivars Infected With Root-Knot Nematode Meloidogyne incognita

    OpenAIRE

    Postnikova, Olga A.; Maria Hult; Jonathan Shao; Andrea Skantar; Nemchinov, Lev G.

    2015-01-01

    Nematodes are one of the major limiting factors in alfalfa production. Root-knot nematodes (RKN, Meloidogyne spp.) are widely distributed and economically important sedentary endoparasites of agricultural crops and they may inflict significant damage to alfalfa fields. As of today, no studies have been published on global gene expression profiling in alfalfa infected with RKN or any other plant parasitic nematode. Very little information is available about molecular mechanisms that contribute...

  20. 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.

  1. 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.

  2. Nodules Initiated by Rhizobium meliloti Exopolysaccharide Mutants Lack a Discrete, Persistent Nodule Meristem 1

    Science.gov (United States)

    Yang, Cheng; Signer, Ethan R.; Hirsch, Ann M.

    1992-01-01

    Infection of alfalfa with Rhizobium meliloti exo mutants deficient in exopolysaccharide results in abnormal root nodules that are devoid of bacteria and fail to fix nitrogen. Here we report further characterization of these abnormal nodules. Tightly curled root hairs or shepherd's crooks were found after inoculation with Rm 1021-derived exo mutants, but curling was delayed compared with wild-type Rm 1021. Infection threads were initiated in curled root hairs by mutants as well as by wild-type R. meliloti, but the exo mutant-induced threads aborted within the peripheral cells of the developing nodule. Also, nodules elicited by Rm 1021-derived exo mutants were more likely to develop on secondary roots than on the primary root. In contrast with wild-type R. meliloti-induced nodules, the exo mutant-induced nodules lacked a well defined apical meristem, presumably due to the abortion of the infection threads. The relationship of these findings to the physiology of nodule development is discussed. ImagesFigure 3Figure 1Figure 2Figure 4 PMID:16668605

  3. Studies on the Alfalfa Growing Effects of Alfalfa Rhizobium%苜蓿根瘤菌剂对紫花苜蓿生长影响研究

    Institute of Scientific and Technical Information of China (English)

    樊川; 杨旭升; 曹亚斌

    2015-01-01

    通过不同剂量苜蓿根瘤菌接种两种紫花苜蓿试验,比较了接种根瘤菌剂对紫花苜蓿生长的影响。试验结果表明:接种苜蓿根瘤菌剂能够提高紫花苜蓿结瘤数量和根瘤重,促进紫花苜蓿的生长发育,增加苜蓿草的产草量。在所试验的接种剂量中,以每 kg 苜蓿种子接种12mL 根瘤菌剂为最佳接种量。%This paper studied the alfalfa growing effects of alfalfa rhizobium by inoculated different amounts.The experimental results showed that inoculating rhizobium can raise the nodulating amount of alfalfa and the weight of nodulations,promoting the growth of alfalfas and augment the productions of alfalfas.Among the different inoculated amounts,the amount of 12ml is the best inoculated amount.

  4. Diversity and numbers of root-nodule bacteria (rhizobia in Polish soils

    Directory of Open Access Journals (Sweden)

    Stefan Martyniuk

    2011-01-01

    Full Text Available Using a sand pouch-plant infection method, populations of several species of root-nodule bacteria (rhizobia were enumerated in eighty soils collected throughout Poland. Rhizobium leguminosarum bv. viciae (symbionts of pea, faba bean, vetch and R. leguminosarum bv. trifolii (symbionts of clover were detected in 77 and 76 soils, respectively. Most of these soils contained moderate and high numbers of these species of the rhizobia. Symbionts of beans, R. leguminosarum bv. phaseoli, were assessed in 76 soils; of this number 15 soils had no detectable populations of bean rhizobia and in 40 soils high or moderate numbers of these bacteria were found. Bradyrhizobium sp. (Lupinus, root-nodule bacteria of lupine and serradella, were absent in 19 soils, out of 80 tested, and 34 soils were colonised by high or moderate populations of bradyrhizobia. Sinorhizobium meliloti, rhizobia nodulating alfalfa, were sparse in the examined soils; with 56 soil containing no detectable numbers of S. meliloti and only 6 soils harbouring high or moderate populations of this species. The estimated numbers of the rhizobia in the studied soils were also related to some physical and chemical properties of these soils.

  5. Transcriptional networks leading to symbiotic nodule organogenesis.

    Science.gov (United States)

    Soyano, Takashi; Hayashi, Makoto

    2014-08-01

    The symbiosis with nitrogen-fixing bacteria leading to root nodules is a relatively recent evolutionary innovation and limited to a distinct order of land plants. It has long been a mystery how plants have invented this complex trait. However, recent advances in molecular genetics of model legumes has elucidated genes involved in the development of root nodules, providing insights into this process. Here we discuss how the de novo assembly of transcriptional networks may account for the predisposition to nodulate. Transcriptional networks and modes of gene regulation from the arbuscular mycorrhizal symbiosis, nitrate responses and aspects of lateral root development have likely all contributed to the emergence and development of root nodules.

  6. Proline auxotrophy in Sinorhizobium meliloti results in a plant-specific symbiotic phenotype.

    Science.gov (United States)

    diCenzo, George C; Zamani, Maryam; Cowie, Alison; Finan, Turlough M

    2015-12-01

    In order to effectively manipulate rhizobium-legume symbioses for our benefit, it is crucial to first gain a complete understanding of the underlying genetics and metabolism. Studies with rhizobium auxotrophs have provided insight into the requirement for amino acid biosynthesis during the symbiosis; however, a paucity of available L-proline auxotrophs has limited our understanding of the role of L-proline biosynthesis. Here, we examined the symbiotic phenotypes of a recently described Sinorhizobium meliloti L-proline auxotroph. Proline auxotrophy was observed to result in a host-plant-specific phenotype. The S. meliloti auxotroph displayed reduced symbiotic capability with alfalfa (Medicago sativa) due to a decrease in nodule mass formed and therefore a reduction in nitrogen fixed per plant. However, the proline auxotroph formed nodules on white sweet clover (Melilotus alba) that failed to fix nitrogen. The rate of white sweet clover nodulation by the auxotroph was slightly delayed, but the final number of nodules per plant was not impacted. Examination of white sweet clover nodules by confocal microscopy and transmission electron microscopy revealed the presence of the S. meliloti proline auxotroph cells within the host legume cells, but few differentiated bacteroids were identified compared with the bacteroid-filled plant cells of WT nodules. Overall, these results indicated that L-proline biosynthesis is a general requirement for a fully effective nitrogen-fixing symbiosis, likely due to a transient requirement during bacteroid differentiation.

  7. The effects of Pantoea sp. strain Y4-4 on alfalfa in the remediation of heavy-metal-contaminated soil, and auxiliary impacts of plant residues on the remediation of saline-alkali soils.

    Science.gov (United States)

    Li, Shuhuan; Wang, Jie; Gao, Nanxiong; Liu, Lizhu; Chen, Yahua

    2017-04-01

    The plant-growth-promoting rhizobacterium (PGPR) Y4-4 was isolated from plant rhizosphere soil and identified as Pantoea sp. by 16S rRNA sequence analysis. The effects of strain Y4-4 on alfalfa grown in heavy-metals-contaminated soil was investigated using a pot experiment. In a Cu-rich environment, the shoot dry mass and total dry mass of plants inoculated with strain Y4-4 increased by 22.6% and 21%, and Cu accumulation increased by 15%. In a Pb-Zn-rich environment, the shoot dry mass and total dry mass of plants inoculated with strain Y4-4 increased by 23.4% and 22%, and Zn accumulation increased by 30.3%. In addition, the salt tolerance and biomass of wheat seedlings could be improved by applying strain Y4-4 mixed with plant residue as a result of the Cu-rich plant residues providing copper nutrition to wheat. This study offers an efficient PGPR with strong salt tolerance and a safe strategy for the post-treatment of plant residue.

  8. The Productivity and Quality of Alfalfa (Medicago sativa L. in Romanian Forest Steppe

    Directory of Open Access Journals (Sweden)

    Mihai STAVARACHE

    2015-04-01

    Full Text Available Alternative use of alfalfa, for various purposes, including the production of biofuels or food supplement for human alimentation, is a study topic still in its early stages of research. Studying and understanding the biology of alfalfa and the factors with a major influence on it are very important activities. The productivity and quality of alfalfa are two indicators that help determine, in addition to economic value, the way in which alfalfa can be used. Evolution of alfalfa yield and quality depends on many factors, such as the growth stage of alfalfa plants at harvesting. It was observed over three years of vegetation the influence of alfalfa plant growth stage at harvest on plant height, leaves/stems ratio, production of leaves, stems and whole plant (DM - dry matter per hectare and on quality indicators (CP - crude protein, NDF - neutral detergent fiber and ADF - acid detergent fiber. The results showed that, with the advancement of phenological phases, from early bud stage to complete flowering, the total biomass output raised from 2.79 Mg·ha-1 to 4.60 Mg·ha-1, the neutral detergent fiber raised from 48.4-50.6% to 62.0-67.7%, while crude protein content decreased from 21.2-24.0% to 13.3-16.5%. The parameter values were correlated with alfalfa growth stage during the harvesting (significant at the 0.05 and 0.01 probability levels.

  9. Economics of growth regulator treatment of alfalfa seed for interseeding into silage corn

    Science.gov (United States)

    Previous studies have focused on interseeding of alfalfa into corn for use as a temporary cover crop rather than as a means of jump-starting alfalfa production after corn. In ongoing field studies, we are evaluating whether plant growth regulators (PGR) may be used to aid the establishment of inters...

  10. 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...

  11. Relationship of red and photographic infrared spectral radiances to alfalfa biomass, forage water content, percentage canopy cover, and severity of drought stress

    Science.gov (United States)

    Tucker, C. J.; Elgin, J. H., Jr.; Mcmurtrey, J. E., III

    1979-01-01

    Red and photographic infrared spectral data were collected using a handheld radiometer for two cuttings of alfalfa. Significant linear and non-linear correlation coefficients were found between the spectral variables and plant height, biomass, forage water content, and estimated canopy cover for the earlier alfalfa cutting. The alfalfa of later cutting experienced a period of severe drought stress which limited growth. The spectral variables were found to be highly correlated with the estimated drought scores for this alfalfa cutting.

  12. Oxidative burst in alfalfa-Sinorhizobium meliloti symbiotic interaction.

    Science.gov (United States)

    Santos, R; Hérouart, D; Sigaud, S; Touati, D; Puppo, A

    2001-01-01

    Reactive oxygen species are produced as an early event in plant defense response against avirulent pathogens. We show here that alfalfa responds to infection with Sinorhizobium meliloti by production of superoxide and hydrogen peroxide. This similarity in the early response to infection by pathogenic and symbiotic bacteria addresses the question of which mechanism rhizobia use to counteract the plant defense response.

  13. Overexpression of BetS, a Sinorhizobium meliloti high-affinity betaine transporter, in bacteroids from Medicago sativa nodules sustains nitrogen fixation during early salt stress adaptation.

    Science.gov (United States)

    Boscari, Alexandre; Van de Sype, Ghislaine; Le Rudulier, Daniel; Mandon, Karine

    2006-08-01

    Sinorhizobium meliloti possesses several betaine transporters to cope with salt stress, and BetS represents a crucial high-affinity glycine and proline betaine uptake system involved in the rapid acquisition of betaines by cells subjected to osmotic upshock. Using a transcriptional lacZ (beta-galactosidase) fusion, we showed that betS is expressed during the establishment of the symbiosis and in mature nitrogen-fixing nodules. However, neither Nod nor Fix phenotypes were impaired in a betS mutant. BetS is functional in isolated bacteroids, and its activity is strongly activated by high osmolarity. In bacteroids from a betS mutant, glycine betaine and proline betaine uptake was reduced by 85 to 65%, indicating that BetS is a major component of the overall betaine uptake activity in bacteroids in response to osmotic stress. Upon betS overexpression (strain UNA349) in free-living cells, glycine betaine transport was 2.3-fold higher than in the wild-type strain. Interestingly, the accumulation of proline betaine, the endogenous betaine synthesized by alfalfa plants, was 41% higher in UNA349 bacteroids from alfalfa plants subjected to 1 week of salinization (0.3 M NaCl) than in wild-type bacteroids. In parallel, a much better maintenance of nitrogen fixation activity was observed in 7-day-salinized plants nodulated with the overexpressing strain than in wild-type nodulated plants. Taken altogether, these results are consistent with the major role of BetS as an emergency system involved in the rapid uptake of betaines in isolated and in planta osmotically stressed bacteroids of S. meliloti.

  14. Avaliação da eficiência de herbicidas no controle de plantas daninhas em alfafa Decreasing of weed plants using herbicides and herbicides mix in alfalfa crop

    Directory of Open Access Journals (Sweden)

    Wilson da Silva

    2004-08-01

    Full Text Available Objetivou-se neste trabalho avaliar a seletividade e a eficiência de herbicidas no controle de plantas daninhas na cultura da alfafa. Foram desenvolvidos dois experimentos em Anápolis, GO. No primeiro, avaliaram-se oito herbicidas (imazethapyr, MSMA, fomesafen, bentazon, chlorimuron-ethyl, imazamox aplicados em pós-emergência da alfafa, e no segundo, seis misturas de herbicidas [(diuron + MSMA, (diuron + MSMA, (diuron + paraquat, (diuron + paraquat] aplicados um dia após o corte da alfafa. Em ambos os experimentos, foram incluídas duas testemunhas (capinada e sem capina, sendo os tratamentos distribuídos em blocos ao acaso com quatro repetições. Os herbicidas como imazethapyr, MSMA, fomesafen, bentazon, chlorimuron-ethyl e imazamox foram seletivos à cultura da alfafa e eficientes no controle de plantas daninhas. As misturas dos herbicidas (diuron + MSMA e (diuron + paraquat aplicados logo após o corte da alfafa causaram danos às folhas e aos caules remanescentes. As brotações, todavia, surgiram isentas de toxicidade, apresentando crescimento e desenvolvimento normais. Essas misturas promoveram eficiente controle de plantas daninhas, e não afetaram o acúmulo de biomassa seca e altura de plantas em relação à testemunha capinada. O não-controle das plantas daninhas reduziu a produção de biomassa seca e a altura das plantas de alfafa em 74,5% e 42,8%, respectivamente.The objectives of this study were to evaluate the efficiency and selectivity of herbicides on the weed plants control at the alfalfa crop. Two experiments were carried out at the Experimental Station of Agenciarural, located in Anápolis, GO. In the first experiment it was evaluated eight herbicides (imazethapyr, MSMA, fomesafen, bentazon, chlorimuron-ethyl and imazamox post emergency and in the second experiment six different mixed of herbicides [(diuron + MSMA, (diuron + MSMA, (diuron + paraquat, (diuron + paraquat] applied one day after the alfalfa harvesting

  15. Multiwalled carbon nanotubes in alfalfa and wheat: toxicology and uptake.

    Science.gov (United States)

    Miralles, Pola; Johnson, Errin; Church, Tamara L; Harris, Andrew T

    2012-12-07

    Data on the bioavailability and toxicity of carbon nanotubes (CNTs) in the environment, and, in particular, on their interactions with vascular plants, are limited. We investigated the effects of industrial-grade multiwalled CNTs (75 wt% CNTs) and their impurities on alfalfa and wheat. Phytotoxicity assays were performed during both seed germination and seedling growth. The germinations of both species were tolerant of up to 2560 mg l(-1) CNTs, and root elongation was enhanced in alfalfa and wheat seedlings exposed to CNTs. Remarkably, catalyst impurities also enhanced root elongation in alfalfa seedlings as well as wheat germination. Thus the impurities, not solely the CNTs, impacted the plants. CNT internalization by plants was investigated using electron microscopy and two-dimensional Raman mapping. The latter showed that CNTs were adsorbed onto the root surfaces of alfalfa and wheat without significant uptake or translocation. Electron microscopy investigations of internalization were inconclusive owing to poor contrast, so Fe(3)O(4)-functionalized CNTs were prepared and studied using energy-filter mapping of Fe(3)O(4). CNTs bearing Fe(3)O(4) nanoparticles were detected in the epidermis of one wheat root tip only, suggesting that internalization was possible but unusual. Thus, alfalfa and wheat tolerated high concentrations of industrial-grade multiwalled CNTs, which adsorbed onto their roots but were rarely taken up.

  16. Analysis of nodule meristem persistence and ENOD40 functioning in Medicago truncatula nodule formation

    NARCIS (Netherlands)

    Wan Xi,

    2007-01-01

    Medicago root nodules are formed as a result of the interaction of the plant with the soil-borne bacterium Sinorhizobium meliloti. Several plant genes are induced during nodule formation and MtENOD40 is one of the earliest genes activated. The precise function as well as the molecule harboring the b

  17. Gene-Transformation-Induced Changes in Chemical Functional Group Features and Molecular Structure Conformation in Alfalfa Plants Co-Expressing Lc-bHLH and C1-MYB Transcriptive Flavanoid Regulatory Genes: Effects of Single-Gene and Two-Gene Insertion

    Directory of Open Access Journals (Sweden)

    Ravindra G. Heendeniya

    2017-03-01

    Full Text Available Alfalfa (Medicago sativa L. genotypes transformed with Lc-bHLH and Lc transcription genes were developed with the intention of stimulating proanthocyanidin synthesis in the aerial parts of the plant. To our knowledge, there are no studies on the effect of single-gene and two-gene transformation on chemical functional groups and molecular structure changes in these plants. The objective of this study was to use advanced molecular spectroscopy with multivariate chemometrics to determine chemical functional group intensity and molecular structure changes in alfalfa plants when co-expressing Lc-bHLH and C1-MYB transcriptive flavanoid regulatory genes in comparison with non-transgenic (NT and AC Grazeland (ACGL genotypes. The results showed that compared to NT genotype, the presence of double genes (Lc and C1 increased ratios of both the area and peak height of protein structural Amide I/II and the height ratio of α-helix to β-sheet. In carbohydrate-related spectral analysis, the double gene-transformed alfalfa genotypes exhibited lower peak heights at 1370, 1240, 1153, and 1020 cm−1 compared to the NT genotype. Furthermore, the effect of double gene transformation on carbohydrate molecular structure was clearly revealed in the principal component analysis of the spectra. In conclusion, single or double transformation of Lc and C1 genes resulted in changing functional groups and molecular structure related to proteins and carbohydrates compared to the NT alfalfa genotype. The current study provided molecular structural information on the transgenic alfalfa plants and provided an insight into the impact of transgenes on protein and carbohydrate properties and their molecular structure’s changes.

  18. Standardized mapping of nodulation patterns in legume roots.

    Science.gov (United States)

    Remmler, Lauren; Clairmont, Lindsey; Rolland-Lagan, Anne-Gaëlle; Guinel, Frédérique Catherine

    2014-05-01

    Optimizing nodulation in legumes is a target for crop improvement, and the spatial control of nodulation is just beginning to be unravelled. However, there is currently no method for standard phenotyping of nodulation patterns. Here we present a method and software for the quantitative analysis of nodulation phenotypes. Roots of nodulated peas (Pisum sativum), wild-type and two mutants, were photographed. Data from the photographs were extracted using custom image and data analysis software. The software makes it possible to extract each nodule's position along primary and lateral roots, and to represent the nodulated root system in a standardized way independent of the way roots are arranged in the soil. A wide variety of nodulation and root variables are calculated, and average spatial nodulation patterns can be computed from multiple samples. Standardized spatial analysis of nodulation patterns opens the way for comparative analyses among genotypes of a single legume species, as here in pea. This approach could also be used to compare nodulation patterns among crops, among plants grown under different environmental conditions, or among plants exposed to different pharmacological treatments. The proposed method should therefore prove useful for studies on nodule organogenesis and nodule physiology and for optimizing nodulation in crops. © 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.

  19. Vigna mungo, V. radiata and V. unguiculata plants sampled in different agronomical-ecological-climatic regions of India are nodulated by Bradyrhizobium yuanmingense.

    Science.gov (United States)

    Appunu, Chinnaswamy; N'Zoue, Angèle; Moulin, Lionel; Depret, Géraldine; Laguerre, Gisèle

    2009-10-01

    Vigna mungo, Vigna radiata and Vigna unguiculata are important legume crops cultivated in India, but little is known about the genetic resources in native rhizobia that nodulate these species. To identify these bacteria, a core collection of 76 slow-growing isolates was built from root nodules of V. mungo, V. radiata and V. unguiculata plants grown at different sites within three agro-ecological-climatic regions of India. The genetic diversity of the bacterial collection was assessed by restriction fragment length polymorphism (RFLP) analysis of PCR-amplified DNA fragments of the 16S-23S rDNA intergenic spacer (IGS) region, and the symbiotic genes nifH and nodC. One rDNA IGS type grouped 91% of isolates, but more diversity was found at the symbiotic loci (17 symbiotic genotypes). Overall, no host plant specificity was shown, the three host plant species sharing common bradyrhizobial genotypes that represented 62% of the collection. Similarly, the predominant genotypes were found at most sampling sites and in all agro-ecological-climatic regions. Phylogenies inferred from IGS sequencing and multi-locus sequence analysis of the dnaK, glnII and recA genes indicated that all isolates but one were clustered with the Bradyrhizobium yuanmingense species. The nifH phylogeny also grouped the different nif haplotypes within a cluster including B. yuanmingense, except for one infrequent nif haplotype which formed a new lineage within the Bradyrhizobium genus. These results may reflect a long history of co-evolution between B. yuanmingense and Vigna spp. in India, while intra-species polymorphism detected in the symbiotic loci may be linked with the long history of diversification of B. yuanmingense coinciding with that of its host legumes.

  20. Why is my alfalfa yellow?

    Science.gov (United States)

    In 2016, many parts of the Midwest experienced far wetter than normal summer weather and by August or September, many growers were asking, “Why is my alfalfa yellow?” When all or part of an alfalfa field is yellow, it is a certain sign that something has gone wrong. In this case the problem in most ...

  1. Growth and nodulation of soybean plants fertilized with poultry litter Desenvolvimento e nodulação de plantas de soja adubadas com cama de aves

    Directory of Open Access Journals (Sweden)

    Vilmar Antonio Ragagnin

    2013-02-01

    Full Text Available Nowadays much of the chemical fertilizers used in Brazil come from imports, what creates the necessity to seek for alternative fertilizers. One possibility is organic fertilizers, including poultry litter, which presents increasing availability in various regions of the country, due to the expansion of the poultry industry. Despite its availability, there are no enough studies that show the benefits of using poultry litter on soybeans in substitution of mineral fertilizer. The present study aimed to evaluate the effect of different doses of poultry litter on leaf chlorophyll content, nodulation, and development of soybean plants. It was used a completely randomized design with five doses of poultry litter (0, 1, 2, 4 and 8 t ha-1 and a control with mineral fertilizer with four repetitions. On stages R1 and R5, chlorophyll readings were done using a portable meter and also the measurement of plant height. Plants were cut near the surface, roots were washed and the nodules were detached and counted. After that, it was determined the dry weight of plant roots, nodules, and shoots. Soybean fertilization with poultry litter increases the chlorophyll content of leaves, plant height, shoot dry matter, root dry matter, and nodulation. The use of poultry litter on soybean favors its growth with decreasing increments, except for root dry matter, and with an optimum dose for each variable. The supply of poultry litter can replace mineral fertilizers in terms of soybean growth.Atualmente, boa parte dos fertilizantes utilizados no Brasil é importada, o que cria a necessidade de busca por fertilizantes alternativos. Uma possibilidade são os fertilizantes orgânicos, incluindo a cama de aves, que apresenta crescente disponibilidade em várias regiões do país, em razão da expansão da indústria avícola. Apesar da disponibilidade e potencial, não há estudos suficientes que mostrem os benefícios do uso da cama de aves na cultura da soja em substitui

  2. Mechanistic action of gibberellins in legume nodulation.

    Science.gov (United States)

    Hayashi, Satomi; Gresshoff, Peter M; Ferguson, Brett J

    2014-10-01

    Legume plants are capable of entering into a symbiotic relationship with rhizobia bacteria. This results in the formation of novel organs on their roots, called nodules, in which the bacteria capture atmospheric nitrogen and provide it as ammonium to the host plant. Complex molecular and physiological changes are involved in the formation and establishment of such nodules. Several phytohormones are known to play key roles in this process. Gibberellins (gibberellic acids; GAs), a class of phytohormones known to be involved in a wide range of biological processes (i.e., cell elongation, germination) are reported to be involved in the formation and maturation of legume nodules, highlighted by recent transcriptional analyses of early soybean symbiotic steps. Here, we summarize what is currently known about GAs in legume nodulation and propose a model of GA action during nodule development. Results from a wide range of studies, including GA application, mutant phenotyping, and gene expression studies, indicate that GAs are required at different stages, with an optimum, tightly regulated level being key to achieve successful nodulation. Gibberellic acids appear to be required at two distinct stages of nodulation: (i) early stages of rhizobia infection and nodule primordium establishment; and (ii) later stages of nodule maturation.

  3. Mechanistic action of gibberellins in legume nodulation

    Institute of Scientific and Technical Information of China (English)

    Satomi Hayashi; Peter MGresshoff; Brett J.Ferguson

    2014-01-01

    Legume plants are capable of entering into a symbiotic relationship with rhizobia bacteria. This results in the formation of novel organs on their roots, cal ed nodules, in which the bacteria capture atmospheric nitrogen and provide it as ammonium to the host plant. Complex molecular and physiological changes are involved in the formation and establishment of such nodules. Several phytohormones are known to play key roles in this process. Gibberel ins (gibberel ic acids;GAs), a class of phytohormones known to be involved in a wide range of biological processes (i.e., cellelongation, germination) are reported to be involved in the formation and maturation of legume nodules, highlighted by recent tran-scriptional analyses of early soybean symbiotic steps. Here, we summarize what is currently known about GAs in legume nodulation and propose a model of GA action during nodule development. Results from a wide range of studies, including GA application, mutant phenotyping, and gene expression studies, indicate that GAs are required at different stages, with an optimum, tightly regulated level being key to achieve successful nodulation. Gibberel ic acids appear to be required at two distinct stages of nodulation:(i) early stages of rhizobia infection and nodule primordium establishment;and (i ) later stages of nodule maturation.

  4. Aluminium-induced reduction of plant growth in alfalfa (Medicago sativa) is mediated by interrupting auxin transport and accumulation in roots.

    Science.gov (United States)

    Wang, Shengyin; Ren, Xiaoyan; Huang, Bingru; Wang, Ge; Zhou, Peng; An, Yuan

    2016-07-20

    The objective of this study was to investigate Al(3+)-induced IAA transport, distribution, and the relation of these two processes to Al(3+)-inhibition of root growth in alfalfa. Alfalfa seedlings with or without apical buds were exposed to 0 or 100 μM AlCl3 and were foliar sprayed with water or 6 mg L(-1) IAA. Aluminium stress resulted in disordered arrangement of cells, deformed cell shapes, altered cell structure, and a shorter length of the meristematic zone in root tips. Aluminium stress significantly decreased the IAA concentration in apical buds and root tips. The distribution of IAA fluorescence signals in root tips was disturbed, and the IAA transportation from shoot base to root tip was inhibited. The highest intensity of fluorescence signals was detected in the apical meristematic zone. Exogenous application of IAA markedly alleviated the Al(3+)-induced inhibition of root growth by increasing IAA accumulation and recovering the damaged cell structure in root tips. In addition, Al(3+) stress up-regulated expression of AUX1 and PIN2 genes. These results indicate that Al(3+)-induced reduction of root growth could be associated with the inhibitions of IAA synthesis in apical buds and IAA transportation in roots, as well as the imbalance of IAA distribution in root tips.

  5. Effects of two alfalfa preparations with different particle sizes on the gastric mucosa in weanlings: alfalfa chaff versus alfalfa pellets

    OpenAIRE

    Vondran, Sarah; Venner, Monica; Vervuert, Ingrid

    2016-01-01

    Background Feeding alfalfa hay is often recommended for its buffering components, like protein and calcium, to prevent lesions of the gastric mucosa in horses. Until now, there has been no information regarding the influence of alfalfa particle size on the gastric mucosa. The aim of this study was to investigate the effects of feeding two alfalfa preparations with different particle sizes (alfalfa chaff vs alfalfa pellets) in comparison with grass hay on the gastric mucosa in weanling horses....

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

    Science.gov (United States)

    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.

  7. Occurrence of alfalfa (Medicago sativa L.) populations along roadsides in southern Manitoba, Canada and their potential role in intraspecific gene flow.

    Science.gov (United States)

    Bagavathiannan, Muthukumar V; Gulden, Robert H; Van Acker, Rene C

    2011-04-01

    Alfalfa is a highly outcrossing perennial species that can be noticed in roadsides as feral populations. There remains little information available on the extent of feral alfalfa populations in western Canadian prairies and their role in gene flow. The main objectives of this study were (a) to document the occurrence of feral alfalfa populations, and (b) to estimate the levels of outcrossing facilitated by feral populations. A roadside survey confirmed widespread occurrence of feral alfalfa populations, particularly in alfalfa growing regions. The feral populations were dynamic and their frequency ranged from 0.2 to 1.7 populations km(-1). In many cases, the nearest feral alfalfa population from alfalfa production field was located within a distance sufficient for outcrossing in alfalfa. The gene flow study confirmed that genes can move back and forth between feral and cultivated alfalfa populations. In this study, the estimated outcrossing levels were 62% (seed fields to feral), 78% (feral to seed fields), 82% (hay fields to feral) and 85% (feral to feral). Overall, the results show that feral alfalfa plants are prevalent in alfalfa producing regions in western Canada and they can serve as bridges for gene flow at landscape level. Management of feral populations should be considered, if gene flow is a concern. Emphasis on preventing seed spill/escapes and intentional roadside planting of alfalfa cultivars will be particularly helpful. Further, realistic and pragmatic threshold levels should be established for markets sensitive to the presence of GE traits.

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

  9. Root developmental programs shape the Medicago truncatula nodule meristem

    NARCIS (Netherlands)

    Franssen, H.; Xiao, T.T.; Kulikova, O.; Wan, X.; Bisseling, T.; Scheres, B.; Heidstra, R.

    2015-01-01

    Nodules on the roots of legume plants host nitrogen-fixing Rhizobium bacteria. Several lines of evidence indicate that nodules are evolutionarily related to roots. We determined whether developmental control of the Medicago truncatula nodule meristem bears resemblance to that in root meristems

  10. The mouthparts enriched odorant binding protein 11 of the alfalfa plant bug Adelphocoris lineolatus displays a preferential binding behaviour to host plant secondary metabolites

    Directory of Open Access Journals (Sweden)

    Liang eSun

    2016-06-01

    Full Text Available Odorant binding proteins (OBPs are proposed to be directly required for odorant discrimination and represent potential interesting targets for pest control. In the notoriously agricultural pest Adelphocoris lineolatus, our previous functional investigation of highly expressed antennal OBPs clearly supported this viewpoint, whereas the findings of the current study by characterizing of AlinOBP11 rather indicated that OBP in hemipterous plant bugs might fulfill a different and tantalizing physiological role. The phylogenetic analysis uncovered that AlinOBP11 together with several homologous bug OBP proteins are potential orthologs, implying they could exhibit a conserved function. Next, the results of expression profiles solidly showed that AlinOBP11 was predominantly expressed at adult mouthparts, the most important gustatory organ of Hemiptera mirid bug. Finally, a rigorously selective binding profile was observed in the fluorescence competitive binding assay, in which recombinant AlinOBP11 displayed much stronger binding abilities to non-volatile secondary metabolite compounds than the volatile odorants. These results reflect that AlinOBP11, even its orthologous proteins across bug species, could be associated with a distinctively conserved physiological role such as a crucial carrier for non-volatiles host secondary metabolites in gustatory system.

  11. Biological and Molecular Variability of Alfalfa mosaic virus Affecting Alfalfa Crop in Riyadh Region

    Directory of Open Access Journals (Sweden)

    Mohammed A. AL-Saleh

    2013-12-01

    Full Text Available In 2011–2012, sixty nine samples were collected from alfalfa plants showing viral infection symptoms in Riyadh region. Mechanical inoculation with sap prepared from two collected samples out of twenty five possitive for Alfalfa mosaic virus (AMV by ELISA were produced systemic mosaic on Vigna unguiculata and Nicotiana tabacum, local lesion on Chenopodium amaranticolor and C. quinoa. Vicia faba indicator plants that induce mosaic and mottle with AMV-Sagir isolate and no infection with AMV-Wadi aldawasser isolate. Approximately 700-bp was formed by RT-PCR using AMV coat protein specific primer. Samples from infected alfalfa gave positive results, while healthy plant gave negative result using dot blot hybridization assay. The nucleotide sequences of the Saudi isolates were compared with corresponding viral nucleotide sequences reported in GenBank. The obtained results showed that the AMV from Australia, Brazil, Puglia and China had the highest similarity with AMV-Sajer isolate. While, the AMV from Spain and New Zealaland had the lowest similarity with AMV-Sajer and Wadi aldawasser isolates. The data obtained in this study has been deposited in the GenBank under the accession numbers KC434083 and KC434084 for AMV-Sajer and AMV- Wadialdawasser respectively. This is the first report regarding the gnetic make up of AMV in Saudi Arabia.

  12. Interação entre cultivares, estirpes comerciais de Rhizobium meliloti e fungicidas no incremento da produção de alfafa Interactions of cultivars, commercial strains of Rhizobium meliloti and fungicides on the production of alfalfa

    Directory of Open Access Journals (Sweden)

    Patrícia Perondi Anchão Oliveira

    1999-03-01

    Full Text Available A cultura da alfafa (Medicago sativa é importante para a produção animal, por sua alta qualidade nutritiva, alto potencial de produção e sucesso na fixação de nitrogênio, através da simbiose com o Rhizobium meliloti, dispensando o uso de adubação nitrogenada. Neste trabalho objetivou-se selecionar estirpes comerciais de R. meliloti (SEMIA-116, SEMIA-134 e SEMIA-135 em relação às cultivares de alfafa Flórida 77, Pioneer 5929, CUF 101 e Crioula, e avaliar os fungicidas Iprodione e Thiram em tratamento de semente e solo na cultivar Crioula. Não houve interação entre as estirpes de R. meliloti e as cultivares de alfafa quanto à produção de matéria seca, e as plantas infestadas com a SEMIA-116 apresentaram maior número de nódulos. Houve interações entre os diferentes tipos e doses de fungicidas e as estirpes de R. meliloti testadas. A estirpe SEMIA-116 apresentou superioridade, representada pela maior produção de biomassa e número de nódulos. Pelos resultados obtidos, não se recomenda a inoculação da estirpe SEMIA-135, quando em associação com o fungicida Iprodione, em solo ou semente.The cultivation of alfalfa (Medicago sativa is not only important because of its nutrient, content, but also for its high productivity and its ability to fix nitrogen through its association with Rhizobium meliloti. To better understand this symbiotic association, the interaction between R. meliloti strains SEMIA-116, SEMIA-134 and SEMIA-135 with different cultivars of alfalfa Florida 77, Pioneer 5929, CUF 101 and Crioula was evaluated. In addition to the above treatments, the fungicides Iprodione and Thiram were used with the cultivar Crioula to study their effects on the alfalfa/Rhizobium interaction. Plants inoculated with SEMIA-116 had a greater number of nodules. However, no significant difference in the production of plant dry matter was observed between the various Rhizobium strains and the alfalfa cultivars tested. In the

  13. Evaluation of somatic embryos of alfalfa for recombinant protein expression.

    Science.gov (United States)

    Fu, Guohua; Grbic, Vojislava; Ma, Shengwu; Tian, Lining

    2015-02-01

    Somatic embryos of alfalfa can accumulate higher levels of recombinant proteins comparing to vegetative organs. Somatic embryos may be explored as a new system for new protein production for plants. Plants have been explored via genetic engineering as an inexpensive system for recombinant protein production. However, protein expression levels in vegetative tissues have been low, which limits the commercial utilization of plant expression systems. Somatic embryos resemble zygotic embryos in many aspects and may accumulate higher levels of proteins as true seed. In this study, somatic embryo of alfalfa (Medicago sativa L.) was investigated for the expression of recombinant proteins. Three heterologous genes, including the standard scientific reporter uid that codes for β-glucuronidase and two genes of interest: ctb coding for cholera toxin B subunit (CTB), and hIL-13 coding for human interleukin 13, were independently introduced into alfalfa via Agrobacterium-mediated transformation. Somatic embryos were subsequently induced from transgenic plants carrying these genes. Somatic embryos accumulated approximately twofold more recombinant proteins than vegetative organs including roots, stems, and leaves. The recombinant proteins of CTB and hIL-13 accumulated up to 0.15 and 0.18 % of total soluble protein in alfalfa somatic embryos, respectively. The recombinant proteins expressed in somatic embryos also exhibited biological activities. As somatic embryos can be induced in many plant species and their production can be scaled up via different avenues, somatic embryos may be developed as an efficient expression system for recombinant protein production.

  14. Analysis of nodulation kinetics in Frankia-Discaria trinervis symbiosis reveals different factors involved in the nodulation process.

    Science.gov (United States)

    Gabbarini, Luciano Andrés; Wall, Luis Gabriel

    2008-08-01

    The induction of root nodule development in actinorhizal symbiosis would depend on the concentration of factors produced by the bacteria and the plant. A detailed analysis of nodulation description parameters revealed different factors related to the nodulation process. The initial time for nodulation (t(0)), the initial nodulation rate (v(0)) and the total time of nodule development (t(NOD)) were defined and consequently quantified in different experimental conditions: co-inoculation of Discaria trinervis with increasing concentrations of different non-infective bacteria together with the full compatible infective Frankia strain (the indicator strain) used at a limiting concentration or by changing plant factor(s) concentration. All the above nodulation parameters were modified by changing doses of full compatibility infective strain Frankia BCU110501; v(0) appears to be an expression of symbiotic recognition between partners as only fully symbiotic indicator Frankia BCU110501 was able to change it; t(0) seems not to reflect symbiotic recognition because it can also be modified by non-infective Frankia but suggest the existence of a basic level of plant microbe recognition. The initial time for nodulation t(0), reflecting the time required for the early interactions toward nodulation, is an inverse measure of the ability to establish early interactions toward nodulation. The increase in plant factors concentration also reduces t(0) values, suggesting that a plant factor is involved and favors very early interactions. Increases in plant factors concentration also modify the final number of nodules per plant and the nodule cluster profile along the taproot as an expression of the autoregulation phenomenon. Meanwhile, Frankia inoculums' concentration, either infective or not, modified t(NOD) in an opposite way plant factors did. In conclusion, the analysis of nodulation kinetics appears to be an appropriate tool to investigate factors involved in the symbiotic

  15. NUTRITIONAL ENHANCEMENT OF ALFALFA THROUGH GENETIC ENGINEERING

    Directory of Open Access Journals (Sweden)

    J. Faragó

    2008-09-01

    Full Text Available Alfalfa (Medicago sativa L. is a pasture legume crop of primary importance to animal production throughout the world. The nutritional quality of alfalfa, as of other leguminous forage crops, is mainly determined by their content in selected essential amino acids (EAAs, such as methionine (Met and cysteine (Cys. In alfalfa, however, these S-containing amino acids constitute only about 1% or less of crude proteins (Frame et al., 1998. This is significantly less than the 3.5% Met+Cys content in the recommended FAO reference protein (FAO, 1973. Recent advances in genetic engineering allow to use the transgenic approach to increase the content of specific essential amino acids in target plant species. A number of different molecular approaches have been developed to address this issue, such as over-expression of a heterologous or homologous Met-rich protein, expression of a synthetic protein, modification of protein sequence, and metabolic engineering of the free amino acid pool and protein sink. To study the possibility of transgenic enhancement of nutritional quality of alfalfa, we used the approach of expression of a heterologous protein rich in Met+Cys in cells of alfalfa. The T-DNA introduced into the genome of alfalfa, using Agrobacterium tumefaciens-mediated genetic transformation, contained the selectable merker gene nptII for kanamycin (Kn resistance, and a cDNA of Ov gene from Japanese quail (Coturnix coturnix coding for a high Met+Cys containing ovalbumine (Mucha et al., 1991, both under constitutive promoters. After cocultivation of petiole segment- and leaf blade-explants of two highly embryogenic alfalfa genotypes Rg9/I-14-22 and Rg11/I-10-68 (Faragó et al., 1997 with cells of A. tumefaciens strain AGL1 carrying the nptII and Ov genes, and selection of transgenic cells on Kn containing selective media, more than one hundred putatively transgenic regenerants were obtained through somatic embryogenesis. Biological (Kn rooting assay

  16. Direct detection of radicals in intact soybean nodules

    DEFF Research Database (Denmark)

    Mathieu, C; Moreau, S; Frendo, P

    1998-01-01

    Electron paramagnetic resonance spectroscopy has been employed to examine the nature of the metal ions and radicals present in intact root nodules of soybean plants grown in the absence of nitrate. The spectra obtained from nodules of different ages using this non-invasive technique show dramatic...... of the soybean plants, in a manner analogous to that recently described for Lupinus albus. This Lb-NO complex is present at lower concentrations in older nodules, and is almost completely absent from senescent nodules. Exposure of young and mature nodules to oxidant stress, in the form of hydrogen peroxide...

  17. Auxin and nitric oxide control indeterminate nodule formation

    Directory of Open Access Journals (Sweden)

    Spena Angelo

    2007-05-01

    Full Text Available Abstract Background Rhizobia symbionts elicit root nodule formation in leguminous plants. Nodule development requires local accumulation of auxin. Both plants and rhizobia synthesise auxin. We have addressed the effects of bacterial auxin (IAA on nodulation by using Sinorhizobium meliloti and Rhizobium leguminosarum bacteria genetically engineered for increased auxin synthesis. Results IAA-overproducing S. meliloti increased nodulation in Medicago species, whilst the increased auxin synthesis of R. leguminosarum had no effect on nodulation in Phaseolus vulgaris, a legume bearing determinate nodules. Indeterminate legumes (Medicago species bearing IAA-overproducing nodules showed an enhanced lateral root development, a process known to be regulated by both IAA and nitric oxide (NO. Higher NO levels were detected in indeterminate nodules of Medicago plants formed by the IAA-overproducing rhizobia. The specific NO scavenger cPTIO markedly reduced nodulation induced by wild type and IAA-overproducing strains. Conclusion The data hereby presented demonstrate that auxin synthesised by rhizobia and nitric oxide positively affect indeterminate nodule formation and, together with the observation of increased expression of an auxin efflux carrier in roots bearing nodules with higher IAA and NO content, support a model of nodule formation that involves auxin transport regulation and NO synthesis.

  18. Molecular mechanisms controlling legume autoregulation of nodulation

    National Research Council Canada - National Science Library

    Dugald E. Reid; Brett J. Ferguson; Satomi Hayashi; Yu-Hsiang Lin; Peter M. Gresshoff

    2011-01-01

    .... • Scope Understanding how legume plants maintain control of nodulation to balance the nitrogen gains with their energy needs and developmental costs will assist in increasing their productivity and relative advantage...

  19. The demography of feral alfalfa (Medicago sativa L.) populations occurring in roadside habitats in Southern Manitoba, Canada: implications for novel trait confinement.

    Science.gov (United States)

    Bagavathiannan, Muthukumar V; Gulden, Robert H; Begg, Graham S; Van Acker, Rene C

    2010-09-01

    Feral populations of cultivated crops can act as reservoirs for novel genetically engineered (GE) traits and aid in trait movement at the landscape level. However, little information is available on the potential of cultivated crops to become feral. In this study, we investigated the ferality of alfalfa populations (non-GE version) occurring in roadside habitats. Knowledge on the nature of roadside alfalfa populations would be useful for designing efficient trait confinement protocols and coexistence strategies in alfalfa. We investigated roadside alfalfa populations from 2006 to 2009 in three rural municipalities (Hanover, MacDonald, and Springfield) in Southern Manitoba, Canada. We studied the demography of these populations including seedbank, seedling recruitment, and fecundity and examined the impact of road verge mowing on key life stages of these populations. We also compared the growth and reproductive attributes of roadside and cultivated alfalfa populations. Alfalfa is reproductively successful in roadside habitats and capable of establishing self-perpetuating populations. A substantial portion of the alfalfa seeds we extracted from seedbank samples were viable but not germinable, suggesting some degree of seedbank persistence in roadside habitats. In the roadside habitat, alfalfa seedlings recruited successfully, however, seedling mortality was high when seedlings were in close proximity to well-established alfalfa plants. Mowing dramatically reduced the reproductive success of roadside alfalfa. Generally, the growth and reproduction of roadside alfalfa was comparable to cultivated alfalfa except for total fecundity. Considering the long lifespan (>10 years) of alfalfa and the levels of fecundity, seedbank, and seedling survival we observed, long-term persistence of roadside alfalfa populations seems reasonable. In the context of novel trait confinement, our results suggest that feral alfalfa populations required to be managed if there is a desire

  20. Characterization of alfalfa populations contrasting for root system architecture (RSA)

    Science.gov (United States)

    The root system architecture affects the capacity for nutrient and water uptake thus impacting biomass yield production and may contribute to the persistence of perennial plants. The objectives of this study were to phenotype the roots of three alfalfa populations and identify differences between di...

  1. Activation of cell divisions in legume nodulation

    DEFF Research Database (Denmark)

    Nadzieja, Marcin

    Leguminous plants engage into symbiotic relationships with soil bacteria, rhizobia, and develop root nodules. This process initiates with recognition of bacteria derived signalling molecules called nod factors. The subsequent events lead to symbiotic infection and, occurring in parallel, de novo......) vasculature derived auxin contributes to auxin maxima in root cortex, vi) cytokinin and auxin cooperate with symbiotic transcription factors to determine position of the nodule founder cell and maintain cell divisions in nodule primordia, vii) deprivation of the vasculature derived auxin at the site...... regulation of symbiotic infection and organogenesis. Importantly, the developed tools and obtained results open new opportunities for future investigation of plant-rhizobia interaction....

  2. Adaptability of Alfalfa and Ryegrass to New Reclamation Materials

    Institute of Scientific and Technical Information of China (English)

    WANG ZHAO-FENG; ZHANG Lei-na; FENG Yong-jun; KANG Jing-tao

    2006-01-01

    This paper discussed the adaptability of the alfalfa and ryegrass to the new artificial soil that was composed of fly ash and furfural residue. The potted experiments were taken to compare the growth conditions, yields, and qualities of both grasses. Some pots were covered with wheat stalks and others without. The medium Fluvo-aquic soil was taken as control. The results indicated that the growth of two grasses in the mixture were inferior to their growth in the control in initial period. With the leaching of rain and irrigation, the two grasses improved gradually. The alfalfa and ryegrass both could grow in the new mixture, and the ryegrass would be better to plant in no covered material and the alfalfa in covered. This will be a new way to reclaim subsided land and to improve the environment in mining area.

  3. Biochemical and Molecular Phylogenetic Study of Agriculturally Useful Association of a Nitrogen-Fixing Cyanobacterium and Nodule Sinorhizobium with Medicago sativa L.

    Directory of Open Access Journals (Sweden)

    E. V. Karaushu

    2015-01-01

    Full Text Available Seed inoculation with bacterial consortium was found to increase legume yield, providing a higher growth than the standard nitrogen treatment methods. Alfalfa plants were inoculated by mono- and binary compositions of nitrogen-fixing microorganisms. Their physiological and biochemical properties were estimated. Inoculation by microbial consortium of Sinorhizobium meliloti T17 together with a new cyanobacterial isolate Nostoc PTV was more efficient than the single-rhizobium strain inoculation. This treatment provides an intensification of the processes of biological nitrogen fixation by rhizobia bacteria in the root nodules and an intensification of plant photosynthesis. Inoculation by bacterial consortium stimulates growth of plant mass and rhizogenesis and leads to increased productivity of alfalfa and to improving the amino acid composition of plant leaves. The full nucleotide sequence of the rRNA gene cluster and partial sequence of the dinitrogenase reductase (nifH gene of Nostoc PTV were deposited to GenBank (JQ259185.1, JQ259186.1. Comparison of these gene sequences of Nostoc PTV with all sequences present at the GenBank shows that this cyanobacterial strain does not have 100% identity with any organisms investigated previously. Phylogenetic analysis showed that this cyanobacterium clustered with high credibility values with Nostoc muscorum.

  4. Biochemical and Molecular Phylogenetic Study of Agriculturally Useful Association of a Nitrogen-Fixing Cyanobacterium and Nodule Sinorhizobium with Medicago sativa L.

    Science.gov (United States)

    Karaushu, E V; Lazebnaya, I V; Kravzova, T R; Vorobey, N A; Lazebny, O E; Kiriziy, D A; Olkhovich, O P; Taran, N Yu; Kots, S Ya; Popova, A A; Omarova, E; Koksharova, O A

    2015-01-01

    Seed inoculation with bacterial consortium was found to increase legume yield, providing a higher growth than the standard nitrogen treatment methods. Alfalfa plants were inoculated by mono- and binary compositions of nitrogen-fixing microorganisms. Their physiological and biochemical properties were estimated. Inoculation by microbial consortium of Sinorhizobium meliloti T17 together with a new cyanobacterial isolate Nostoc PTV was more efficient than the single-rhizobium strain inoculation. This treatment provides an intensification of the processes of biological nitrogen fixation by rhizobia bacteria in the root nodules and an intensification of plant photosynthesis. Inoculation by bacterial consortium stimulates growth of plant mass and rhizogenesis and leads to increased productivity of alfalfa and to improving the amino acid composition of plant leaves. The full nucleotide sequence of the rRNA gene cluster and partial sequence of the dinitrogenase reductase (nifH) gene of Nostoc PTV were deposited to GenBank (JQ259185.1, JQ259186.1). Comparison of these gene sequences of Nostoc PTV with all sequences present at the GenBank shows that this cyanobacterial strain does not have 100% identity with any organisms investigated previously. Phylogenetic analysis showed that this cyanobacterium clustered with high credibility values with Nostoc muscorum.

  5. Evaluation of abundance of aerobic bacteria in the rhizosphere of transgenic and non-transgenic alfalfa lines.

    Science.gov (United States)

    Faragová, N; Faragó, J; Drábeková, J

    2005-01-01

    Fourteen genetically modified lines of alfalfa (Medicago sativa) containing the gene Ov from Japanese quail, coding for a methionine-rich protein ovalbumin, were evaluated for nodulation ability and concentration of aerobic bacteria in the rhizosphere. The transgenic lines were derived from a highly regenerable genotype Rg9/I-14-22, selected from cv. Lucia. On selective media, a higher concentration of ammonifying bacteria, bacterial spores, denitrifying and nitrifying bacteria were observed in the rhizosphere of transgenic clonesand, on the other hand, lower concentration of cellulolytic bacteria and Azotobacter spp. compared with the rhizosphere of non-transgenic clone SE/22-GT2. A statistically significant difference in the concentration of all the bacterial types was found between samples taken from two types of substrates (i.e. sterile vs. nonsterile). Higher bacterial concentration (measured as colony forming units per g soil dry mass) were observed for all tested groups of culturable bacteria in the non-sterile substrate. The presence of Azotobacter spp. was found only in the rhizosphere of plants grown in non-sterile soil in which the highest number of fertile soil particles (97 %) was observed in transgenic clones SE/22-9-1-12 and SE/22-11-1-1S.1. Concentration of bacteria involved in the N cycle in the soil was increased in the rhizosphere of transgenic clones and decreased in the rhizosphere of non-transgenic plants compared with the average value. In spite of some differences in colony numbers in samples isolated from the root rhizosphere of transgenic and nontransgenic alfalfa plants, we could not detect any statistically significant difference between individual lines.

  6. Root developmental programs shape the Medicago truncatula nodule meristem.

    Science.gov (United States)

    Franssen, Henk J; Xiao, Ting Ting; Kulikova, Olga; Wan, Xi; Bisseling, Ton; Scheres, Ben; Heidstra, Renze

    2015-09-01

    Nodules on the roots of legume plants host nitrogen-fixing Rhizobium bacteria. Several lines of evidence indicate that nodules are evolutionarily related to roots. We determined whether developmental control of the Medicago truncatula nodule meristem bears resemblance to that in root meristems through analyses of root meristem-expressed PLETHORA genes. In nodules, MtPLETHORA 1 and 2 are preferentially expressed in cells positioned at the periphery of the meristem abutting nodule vascular bundles. Their expression overlaps with an auxin response maximum and MtWOX5, which is a marker for the root quiescent center. Strikingly, the cells in the central part of the nodule meristem have a high level of cytokinin and display MtPLETHORA 3 and 4 gene expression. Nodule-specific knockdown of MtPLETHORA genes results in a reduced number of nodules and/or in nodules in which meristem activity has ceased. Our nodule gene expression map indicates that the nodule meristem is composed of two distinct domains in which different MtPLETHORA gene subsets are expressed. Our mutant studies show that MtPLETHORA genes function redundantly in nodule meristem maintenance. This indicates that Rhizobium has recruited root developmental programs for nodule formation. © 2015. Published by The Company of Biologists Ltd.

  7. Exopolysaccharide production by nitrogen-fixing bacteria within nodules of Medicago plants exposed to chronic radiation in the Chernobyl exclusion zone.

    Science.gov (United States)

    Pawlicki-Jullian, Nathalie; Courtois, Bernard; Pillon, Michelle; Lesur, David; Le Flèche-Mateos, Anne; Laberche, Jean-Claude; Goncharova, Nadia; Courtois, Josiane

    2010-03-01

    Nitrogen-fixing bacteria isolated from root nodules of Medicago plants growing in the 10 km zone around the Chernobyl nuclear power plant were screened for the production of new water-soluble acidic exopolysaccharides (EPSs). The different strains belonged to the Enteriobacteriaceae family (Enterobacter ludwigii, Raoultella terrigena, Klebsiella oxytoca), except for one which belonged to the Rhizobiaceae family (Sinorhizobium meliloti). All of the bacteria produced highly viscous EPS with an average molecular weight comprised between 1 x 10(6) and 3 x 10(6) Da. Five different compositions of EPS were characterized by physico-chemical analyses and (1)H NMR spectroscopy: galactose/mannose (2/1), galactose/glucose (1/1), galactose/glucose/mannose (1/2/1), fucose/galactose/glucose (2/1/1) and fucose/galactose/glucose/mannose (2/2/1/1 or 1/1/2/4). Glucuronic acid, a charged monosaccharide, was also recovered in most of the different EPSs. Copyright 2010 Elsevier Masson SAS. All rights reserved.

  8. Non-nodulated bacterial leaf symbiosis promotes the evolutionary success of its host plants in the coffee family (Rubiaceae)

    DEFF Research Database (Denmark)

    Verstraete, Brecht; Janssens, Steven; Rønsted, Nina

    2017-01-01

    Every plant species on Earth interacts in some way or another with microorganisms and it is well known that certain forms of symbiosis between different organisms can drive evolution. Within some clades of Rubiaceae (coffee family), a specific plant-bacteria interaction exists in which non...

  9. CAROTENOID CLEAVAGE DIOXYGENASE 7 modulates plant growth, reproduction, senescence, and determinate nodulation in the model legume Lotus japonicas

    NARCIS (Netherlands)

    Liu, J.; Novero, M.; Charnikhova, T.; Ferrandino, A.; Schubert, A.; Ruyter-Spira, C.P.; Biofante, P.; Lovisolo, C.; Bouwmeester, H.J.

    2013-01-01

    Strigolactones (SLs) are newly identified hormones that regulate multiple aspects of plant development, infection by parasitic weeds, and mutualistic symbiosis in the roots. In this study, the role of SLs was studied for the first time in the model plant Lotus japonicus using transgenic lines silenc

  10. Nodulação, fixação de nitrogênio e produção de matéria seca de alfafa em resposta a doses de calcário, com diferentes relações cálcio:magnésio Nodulation, nitrogen fixation and alfalfa dry matter production as affected by rates of limestone with different calcium:magnesium ratios applyed

    Directory of Open Access Journals (Sweden)

    Fernando Teixeira Gomes

    2002-12-01

    Full Text Available Altas concentrações de alumínio reduzem a eficência da nodulação e da fixação biológica do nitrogênio atmosférico em leguminosas. O presente ensaio foi realizado em casa de vegetação com o objetivo de avaliar a necessidade de calagem para o cultivo da alfafa em LATOSSOLO VERMELHO-AMARELO álico, textura argilosa. Também foi avaliado o efeito da relação Ca:Mg do calcário aplicado ao solo, sobre a nodulação, os teores de nitrogênio e a produção de matéria seca da parte aérea. Os tratamentos consistiram de duas cultivares de alfafa (Flórida 77 e Crioula, quatro doses de calcário (1,2; 2,4; 4,8; 7,2 t ha-1 e cinco relações percentuais estequiométricas entre Ca e Mg (100:0; 75:25; 50:50; 25:75; 0:100, em delineamento fatorial inteiramente casualizado, com quatro repetições. A nodulação e os teores de nitrogênio na parte aérea aumentaram com a adição de doses crescentes de calcário ao solo. A produção de matéria seca da parte aérea das duas cultivares aumentou em função do aumento das doses de calcário, sendo que somente o cultivar Flórida 77 apresentou diferenças significativas entre as relações Ca:Mg. A relação 100:0 foi a que rendeu maior produção de matéria seca da parte aérea.High aluminum concentrations decrease the nodulation potential, hence the biological atmosferic nitrogen fixation of legume. This greenhouse experiment was aimed to evaluate the effects of limestone applyet on alfalfa cultivation in alic YELLOWISH RED LATOSOL SOIL, clay texture. It evaluated the ideal Ca2+ and Mg2+ ratios in neutralizing soil acidity. Treatments were two alfalfa cultivars (Florida 77 and Creoula, four limestone rates (1.2; 2.4; 4.8; 7.2t.ha-1 and five stoichiometric percentual ratios Ca:Mg between (100:0; 75:25; 50:50; 25:75; 0:100, in a factorial set up into a completely randomized design with four replications. Nodulation and nitrogen contents in the shoots increased with increased limestone rates

  11. Overexpression of a Cytosolic Ascorbate Peroxidase Gene, OsAPX2, Increases Salt Tolerance in Transgenic Alfalfa

    Institute of Scientific and Technical Information of China (English)

    ZHANG Qian; MA Cui; XUE Xin; XU Ming; LI Jing; WU Jin-xia

    2014-01-01

    Alfalfa (Medicago sativa L.) is an important forage crop in the world and it is of great signiifcance for the improvement of its salt tolerance. To improve salt tolerance in alfalfa, a rice ascorbate peroxidase gene (OsAPX2) was introduced into alfalfa using Agrobacterium tumefaciens-mediated transformation with marker gene bar. The different T-DNA insertions in T1 transgenic alfalfa were identiifed by Southern hybridization. Three independent T2 transgenic lines were selected for stress analysis and the results showed that all of them were salt tolerant compared with wild-type plants. The transgenic plants had low levels of H2O2, malondialdehyde and relative electrical conductivity under salt and drought stresses. Moreover, the contents of chlorophyll and proline, and APX activity were high in transgenic plants under salt and drought stresses. Taken together, the overexpression of OsAPX2 enhances salt tolerance in alfalfa through scavenging reactive oxygen species.

  12. Dynamic Transportation of Endophytic Rhizobia in Plants during Growth Development of Alfalfa%苜蓿生育过程中植株体内内生根瘤菌的运移动态

    Institute of Scientific and Technical Information of China (English)

    张淑卿; 李剑峰; 师尚礼

    2009-01-01

    [Objective] The aim of this study was to investigate the dynamic distribution of endophytic rhizobia in different plant parts during seed development of alfalfa and the carrying mechanism of endophytic rhizobia from alfalfa seeds. [Method] Root, stem, leaf, flower, pod and seed from two varieties of alfalfa in different development stages were used for surface disinfection, and then the present site and quantity change of rhizobia in alfalfa were studied by the dilution-plate method. [Result] The number of rhizobia from underground plants in the same stage was as follows: main root<lateral root<hair root. The number of rhizobia from main root and hair root in different stages was as follows: bearing pod stage>branching stage>florescence, while that from lateral root was as follows: branching stage>florescence>bearing pod stage. The distribution of endophytic rhizobia from aerial plants was as follows: few rhizobia was found in stem during branching stage and bearing pod stage, which accounted for 155 cfu/g and 125 cfu/g respectively. Rhizobia mainly distributed in flower bud during branching stage accounting for 9 400 cfu/g, while only in ovary wall during squaring stage accounting for 18 cfu/5 flowers, and few in pollen (26 cfu/5 flowers), fertilized ovules (26 cfu/50 grains), receptacle (15 cfu/5 flowers) and ovary wall (95 cfu/5 flowers) during florescence. Rhizobia existed in pod peel (3 550 cfu/5 pods) and immature seed (1 135 cfu/50 grains) during bearing pod stage, and greatly distributed in seed coat of mature seeds (1 435 cfu/50 grains), but only few in embryo (32 cfu/50 grains) and cotyledon (22 cfu/50 grains). [Conclusion] There is a significant difference in the distribution of endophytic rhizobia from different parts of aerial alfalfa, which only exists in the parts directly related to seed development.%[目的]了解苜蓿种子形成过程中根瘤菌在植株各部位的动态分布,探明苜蓿种子内生根瘤菌的携带机理.[方

  13. Alfalfa varieties differ markedly in seedling survival when interseeded into corn and treated with prohexadione-calcium

    Science.gov (United States)

    Interseeded alfalfa could serve as a dual purpose crop for providing groundcover during silage corn production and forage during subsequent years of production, but this system has been unworkable because competition between the co-planted crops often leads to stand failure of interseeded alfalfa. R...

  14. Non-nodulated bacterial leaf symbiosis promotes the evolutionary success of its host plants in the coffee family (Rubiaceae)

    DEFF Research Database (Denmark)

    Verstraete, Brecht; Janssens, Steven; Rønsted, Nina

    2017-01-01

    Every plant species on Earth interacts in some way or another with microorganisms and it is well known that certain forms of symbiosis between different organisms can drive evolution. Within some clades of Rubiaceae (coffee family), a specific plant-bacteria interaction exists in which non...... in the Late Miocene, a period when the savannah habi- tat is believed to have expanded on the African continent and herbivore pressure increased. The presence of bacterial leaf endophytes appears to be restricted to Old World lineages so far. Plant lineages with leaf endophytes show a significantly higher...

  15. Mezclas de Alfalfa y Gramíneas en la Serie Sabana de Bogotá Tipo Franco Arcilloso.

    Directory of Open Access Journals (Sweden)

    Chaverra Gil. Hernán

    1960-06-01

    36 y 52 %, en comparación con la alfalfa su producción fue inferior en un 4 a 10 %. La producción promedia de forraje de las mezclas alfalfa ryegrass anual, en la siembra de gramíneas en alfalfa establecida fue de 2345 Kg./hect, de forraje seco por corte en comparación con 2340 de la alfalfa sola. El 55% de la producción total de forraje la proporcionó la gramínea. El agua recibida entre los cortes tuvo una influencia bastante marcada en las fluctuaciones de la composición botánica. De acuerdo con los resultados de este estudio y como práctica agrícola, la siembra simultánea de alfalfa en mezcla con ryegrass inglés, ryegrass anual y orchoro, en este tipo de suelo, sería aconsejable hacerla mediante los siguientes métodos: aAlfalfa-orchoro. Alfalfa en surcos separados 25 cm. A razón de 15 Kg./hect, de semilla y orchoro al voleo con una densidad de siembra de 6Kg./hect. Se puede utilizar también la siembra en el mismo surco para facilitar la siembra total a máquina. bAlfalfa-ryegrass inglés, alfalfa-ryegrass annual. Leguminosas y gramíneas en surcos alternos, 7 Kg./hect. de alfalfa por 4kg./hect. de ryegrass inglés o anual. cEn la alfalfa catablecida puede sembrarse al voleo cualquier gramínea sin que se presente mucha competencia para la alfalfa./Abstract Two experiments were conducted with mixtures of grasses and alfalfa on the soil series. Sabana de Bogotá, a clay loam, at the National Center of Agricultural Investigation, Tibaitatá. In the first experiments alfalfa was seeded at the same time as English rygrass, Italian rygrass, orchard grass and white clover. The mixtures were seeded in the following manner: aAlfalfa and grasses broadcast, b in the same row, c in the alternate rows, d alfalfa in rows and the grasses broadcast, and e alfalfa in rows and white clover broadcast. In the second experiment the alfalfa was planted in rows 25 cm. apart. After the fourth uniform hervest the following grasses were broadcast between the

  16. Modelling the dynamics of feral alfalfa populations and its management implications.

    Directory of Open Access Journals (Sweden)

    Muthukumar V Bagavathiannan

    Full Text Available BACKGROUND: Feral populations of cultivated crops can pose challenges to novel trait confinement within agricultural landscapes. Simulation models can be helpful in investigating the underlying dynamics of feral populations and determining suitable management options. METHODOLOGY/PRINCIPAL FINDINGS: We developed a stage-structured matrix population model for roadside feral alfalfa populations occurring in southern Manitoba, Canada. The model accounted for the existence of density-dependence and recruitment subsidy in feral populations. We used the model to investigate the long-term dynamics of feral alfalfa populations, and to evaluate the effectiveness of simulated management strategies such as herbicide application and mowing in controlling feral alfalfa. Results suggest that alfalfa populations occurring in roadside habitats can be persistent and less likely to go extinct under current roadverge management scenarios. Management attempts focused on controlling adult plants alone can be counterproductive due to the presence of density-dependent effects. Targeted herbicide application, which can achieve complete control of seedlings, rosettes and established plants, will be an effective strategy, but the seedbank population may contribute to new recruits. In regions where roadside mowing is regularly practiced, devising a timely mowing strategy (early- to mid-August for southern Manitoba, one that can totally prevent seed production, will be a feasible option for managing feral alfalfa populations. CONCLUSIONS/SIGNIFICANCE: Feral alfalfa populations can be persistent in roadside habitats. Timely mowing or regular targeted herbicide application will be effective in managing feral alfalfa populations and limit feral-population-mediated gene flow in alfalfa. However, in the context of novel trait confinement, the extent to which feral alfalfa populations need to be managed will be dictated by the tolerance levels established by specific production

  17. Membrane lipids analysis of a bacterial strain 17560 isolated from nodule of Alfalfa and cloning and expression of the genes for diacylglyceryl trimethylhomoserine (DGTS) biosynthesis%苜蓿根瘤菌17560细胞膜膜脂组成分析及DGTS合成基因克隆与表达

    Institute of Scientific and Technical Information of China (English)

    刘华伟; 马晓彤; 王旭明; 李秀爱; 邹小琳; 孙建光; 高俊莲

    2013-01-01

    pLH01可以在IPTG诱导后合成无磷脂DGTS,而转入空载体pET-30a(+)的阴性对照菌株E.coli BL21 (DE3)·pET-30a(+)则不能合成.[结论]系统发育地位相同的苜蓿根瘤菌株的细胞膜脂组成明显不同;苜蓿根瘤菌的细胞膜组成随培养基中的磷含量不同而变化,低磷胁迫条件下其细胞膜脂主要由OL和DGTS等无磷脂组成;在Sinorhizobium膜脂中首次发现一种未知的氨基磷脂及3种不同类型的鸟氨酸脂(OLs);从菌株17560中克隆获得2个DGTS合成基因btaA和btaB,在大肠杆菌中成功表达,并证实了所表达基因的功能.%[Objective] The aim of this study is to analyse membrane lipids composition of a rhizobial strain isolated from nodule of Alfalfa in Heilongjiang province under phosphorus-limited condition and to clone and identify the genes required for diacylglyceryl trimethylhomoserine (DGTS) biosynthesis.[Methods] The rhizobial strain was cultured in Sherwood minimal medium containing either normal or low concentrations of inorganic phosphate.The membrane iipids were extracted by Bligh-Dyer method and were analysed by thin-layer chromatography (TLC) using both the lipids of Sinorhizobium meliloti 1021 and standard samples of some phospholipid as a reference.PCR primers was designed according to the sequences of btaA and btaB in the GenBank.The PCR-amplified btaA and btaB genes were expressed in Escherichia coli BL21(DE3).The gene function was verified by testing DGTS production.16S rRNA gene sequences of the strain 17560 was analysed.[Results] The strain 17560 isolated from Alfalfa in Heilongjiang province shares 99.8% 16S rRNA gene sequence identity with Sinorhizobium meliloti.But its lipid compositions were quite different from that of the reference strain S.meliloti 1021.Under phosphorus-limited conditions,the main membrane lipids of the strain17560 were phosphorus-free lipids,such as ornithine lipid (OL) and DGTS.Strain 17560 contain three different types of OLs meanwhile

  18. Managing the rotation from alfalfa to corn

    Science.gov (United States)

    Alfalfa provides many benefits to cropping systems. These benefits occur both during alfalfa production and during the subsequent crops that follow. Some of the common benefits during alfalfa production are increased soil organic matter, decreased soil erosion, and decreased soil nitrate leaching lo...

  19. Molecular Characterization of the 16S rRNA Gene of Phytoplasmas Detected in Two Leafhopper Species Associated with Alfalfa Plants Infected with Witches' Broom in Oman

    Directory of Open Access Journals (Sweden)

    A.J. Khan

    2003-12-01

    Full Text Available Two leafhopper species, Austroagallia avicula and Empoasca sp., were consistently found in alfalfa fields infected with witches’ broom phytoplasma (OmanAlfWB in the Al-Batinah, Dakhliya, North and South Sharqiya, Muscat, and Al-Bureimi regions of the Sultanate of Oman. Phytoplasmas from both leafhoppers were detected by specific polymerase chain reaction (PCR amplification of the 16S rRNA gene and the spacer region in direct PCR using P1/P7 primer pairs. Comparative RFLP profiles of the amplified rRNA gene and the spacer region from leafhopper phytoplasmas and from 20 phytoplasma controls yielded patterns referable to phytoplasmas belonging to the peanut witches’ broom group (16SrII group. In particular, extensive RFLP analyses with the endonucleases HpaII, Tru9I, Tsp509I, and RsaI indicated that the phytoplasmas in A. avicula and Empoasca sp. were identical but showed some differences from OmanAlfWB; however, RFLP patterns obtained with TaqI showed the OmanAlfWB and the phytoplasmas from the two leafhoppers to be identical. Direct PCR products amplified from phytoplasma leafhopper DNA using the P1/P7 primer pair were cloned and sequenced yielding 1758 bp and 1755 bp products from A. avicula and Empoasca sp. respectively; the homology of these sequences with OmanAlfWB and papaya yellow crinkle phytoplasmas was more than 98%. A phylogenetic tree based on the 16S rRNA gene and spacer region sequences from 44 phytoplasmas revealed that the phytoplasmas from the leafhoppers clustered with OmanAlfWB, papaya yellow crinkle, and gerbera phyllody phytoplasmas, all belonging to 16SrII group, but were distinct from lime witches’ broom phytoplasma, the most commonly found phytoplasma in the Sultanate of Oman.

  20. The Distribution of ALFALFA Galaxies

    CERN Document Server

    Martin, Ann M

    2007-01-01

    The ALFALFA blind HI survey will enable a census of the distribution of gas-rich galaxies in the local Universe. Sensitive to an HI mass of 10**7 solar masses at the distance of the Virgo cluster, ALFALFA will probe the smallest objects locally and provide a new consideration of near-field cosmology. Additionally, with a larger, cosmologically significant sample volume and wider bandwidth than previous blind surveys, a much larger number of detections in each mass bin is possible, with adequate angular resolution to eliminate the need for extensive follow-up observations. This increased sensitivity will greatly enhance the utility of cosmological probles in HI. ALFALFA will eventually measure the correlation function of HI selected galaxies in a large local volume. The larger sample and volume size of the ALFALFA dataset will also robustly measure the HI mass function (HIMF). Here, we present the preliminary results on the distribution of local gas-rich galaxies from a first ALFALFA catalog covering 540 deg**...

  1. Alfalfa snakin-1 prevents fungal colonization and probably coevolved with rhizobia.

    Science.gov (United States)

    García, Araceli Nora; Ayub, Nicolás Daniel; Fox, Ana Romina; Gómez, María Cristina; Diéguez, María José; Pagano, Elba María; Berini, Carolina Andrea; Muschietti, Jorge Prometeo; Soto, Gabriela

    2014-09-17

    The production of antimicrobial peptides is a common defense strategy of living cells against a wide range of pathogens. Plant snakin peptides inhibit bacterial and fungal growth at extremely low concentrations. However, little is known of their molecular and ecological characteristics, including origin, evolutionary equivalence, specific functions and activity against beneficial microbes. The aim of this study was to identify and characterize snakin-1 from alfalfa (MsSN1). Phylogenetic analysis showed complete congruence between snakin-1 and plant trees. The antimicrobial activity of MsSN1 against bacterial and fungal pathogens of alfalfa was demonstrated in vitro and in vivo. Transgenic alfalfa overexpressing MsSN1 showed increased antimicrobial activity against virulent fungal strains. However, MsSN1 did not affect nitrogen-fixing bacterial strains only when these had an alfalfa origin. The results reported here suggest that snakin peptides have important and ancestral roles in land plant innate immunity. Our data indicate a coevolutionary process, in which alfalfa exerts a selection pressure for resistance to MsSN1 on rhizobial bacteria. The increased antimicrobial activity against virulent fungal strains without altering the nitrogen-fixing symbiosis observed in MsSN1-overexpressing alfalfa transgenic plants opens the way to the production of effective legume transgenic cultivars for biotic stress resistance.

  2. [Nitrogenase, hydrogenase and nitrate reductase activities, oxygen consumption, and ATP content in nodules formed by strains of Rhizobium leguminosarum 128C53 and 300 in symbiosis with pea plants].

    Science.gov (United States)

    Bedmar, E J; Olivares, J

    1986-10-01

    The nitrogenase activity, nitrate reductase activity and oxygen uptake as well as the hydrogen incorporation and ATP content were examined in the root nodules and bacteroids, respectively, formed by Rhizobium leguminosarum strains 128C53 (hydrogenase positive) and 300 (hydrogenase negative) in symbiosis with Pisum sativum plants grown in the presence of 2 mM KNO3. The strain 128C53 showed the greatest values for all parameters analyzed, except for the nitrate reductase activity, which was higher for the strain 300. Similarly, nodule nitrate reductase activity in strain 300 was greater than that in strain 128C53 when plants grew in the absence of combined nitrogen. In general, the highest values were obtained when determinations were made after 7 hours of plant illumination. However, the hydrogenase activity of strain 128C53 and the nitrate reductase activities of both strains increased with the light period, reaching a maximum after 14 hours of illumination. These results suggest that the benefits derived from the superior symbiotic properties and from the presence of hydrogenase activity in strain 128C53 could be counteracted by the higher rates of the nodule nitrate reductase activity in strain 300.

  3. Role of silicon counteracting cadmium toxicity in Alfalfa (Medicago sativa L.

    Directory of Open Access Journals (Sweden)

    Ahmad H. Kabir

    2016-07-01

    Full Text Available 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 (CAT, ascorbate peroxidase (APX and superoxide dismutase (SOD 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.

  4. 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. PMID:27512401

  5. 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.

  6. Effects of additive application upon ad libitum intake, in vivo digestibility and nitrogen balance of alfalfa haylage

    Directory of Open Access Journals (Sweden)

    Mladen Knežević

    2009-09-01

    Full Text Available The research objective was to determine the effect of the additive Sill-All application on ad libitum intake, in vivo digestibility and nitrogen balance of alfalfa haylage. About 40 % alfalfa plants was ensiled at the phonological flowering stage into plastic foil-wrapped bales without or with additive (2 L t-1 plant mass. No statistically significant differences were found between alfalfa ensiled without additive and alfalfa ensiled with additive for the content of dry matter (DM (632 g and 631 g DM kg-1 fresh sample, respectively. Alfalfa ensiled without additive contained 921 g organic matter (OM kg-1 DM, which was significantly higher (P<0.001 compared to alfalfa ensiled with additive (902 g OM kg-1 DM. Alfalfa ensiled without additive contained 141 g crude proteins (CP kg-1 DM, which was significantly higher (P<0.001 compared to alfalfa ensiled with additive (139 g CP kg-1 DM. Alfalfa ensiled with additive contained significantly less acid detergent fibers (ADF (P<0.001 compared to alfalfa ensiled without additive (445 g kg-1 DM and 456 g kg-1 DM, respectively and had a lower pH value (P<0.001 (5.29 and 5.56, respectively. No statistically significant differences were found between the studied feeding treatments for ad libitum intake of fresh ration and DM ration, for the measured parameters of digestibility and N balance. It was concluded that addition of the additive to alfalfa haylage led to significant changes in chemical composition; however, changes in chemical composition had no impact on measured biological parameters (ad libitum intake, in vivo digestibility and nitrogen balance.

  7. Medicago truncatula Rop GTPases expression in young nodules

    Directory of Open Access Journals (Sweden)

    Lucia Kusu

    2012-02-01

    Full Text Available Small GTPases are monomeric guanine nucleotide binding proteins with molecular weight of 21 to 30 kD. Plants have plantspecific small GTPases, termed Rop (Rho GTPases from plants which serve as signaling proteins in plant signal transduction. To have better understanding on nodulation process in legume, which Rop proteins that play a dominant role in nodule formation would be determined. Therefore, the expression of five Medicago truncatula Rop GTPase genes (MtRops in young nodules were studied in this experiment. RT-PCR method was used to examine the expression of MtRops. The MtRops expression in inoculated M. truncatula plants were compared with those in uninoculated plants. Results of this studies showed that the expression of MtRop5 and MtRop6 were significantly induced in young nodules, it was suggesting that these Rops may have important role during nodule formation.

  8. 植物激素在豆科植物根瘤形成和发育过程中的调控作用%Regulation of Plant Hormones on the Formation and Development of Legumes Root Nodules

    Institute of Scientific and Technical Information of China (English)

    刘薇; 赵振芳; 冯永君; 吴存祥

    2013-01-01

    豆科植物能够利用与根瘤菌形成的共生系统高效地固定氮素养分.根瘤形成过程包括表皮细胞中侵染线形成和皮层细胞根瘤原基形成2个阶段.豆科植物根瘤的形成和发育存在严格的调控机制,综述了多种激素正向(如细胞分裂素、生长素、赤霉素等)或负向(乙烯、逆境激素等)或协同调控根瘤的形成和发育的研究进展.并提出对激素调控结瘤机制的深入研究,将有助于提高豆科作物的产量,减缓氮肥过度使用造成的环境问题.%Legumes have evolved symbiotic interactions with rhizobial bacteria to efficiently fix nitrogen nutrient. The formation of a nitrogen-fixing nodule involves two developmental processes: infection thread initiation in epidermal cells and nodule pri-mordia formation in the cortex. Legumes develop a strict mechanism to regulate nodulation. The researches of several plant hormones have been reported positively(cytokinins,auxin,gibberellins,etc. )or negatively(ethylene,stress hormones,etc. )or coordinated regulate the formation and development of nodulation were summarized. It put forward that profounder researches on the mechanism of nodule regulation would contribute to the improvement of legumes production and the reducement of environmental problems caused by overusing of nitrogen fertilizers.

  9. Root nodule organogenesis. Molecular characterization of the zonation of the central tissue.

    NARCIS (Netherlands)

    Yang, W.C.

    1994-01-01

    Legume plants form root nodules by interacting with the soil bacterium, Rhizobium. In these nodules bacteria are able to convert atmospheric nitrogen into ammonia which is used by the host plants as nitrogen source. Therefore symbiotic nitrogen fixation in root nodules is of great importance for agr

  10. Molecular mechanisms controlling legume autoregulation of nodulation.

    Science.gov (United States)

    Reid, Dugald E; Ferguson, Brett J; Hayashi, Satomi; Lin, Yu-Hsiang; Gresshoff, Peter M

    2011-10-01

    High input costs and environmental pressures to reduce nitrogen use in agriculture have increased the competitive advantage of legume crops. The symbiotic relationship that legumes form with nitrogen-fixing soil bacteria in root nodules is central to this advantage. Understanding how legume plants maintain control of nodulation to balance the nitrogen gains with their energy needs and developmental costs will assist in increasing their productivity and relative advantage. For this reason, the regulation of nodulation has been extensively studied since the first mutants exhibiting increased nodulation were isolated almost three decades ago. Nodulation is regulated primarily via a systemic mechanism known as the autoregulation of nodulation (AON), which is controlled by a CLAVATA1-like receptor kinase. Multiple components sharing homology with the CLAVATA signalling pathway that maintains control of the shoot apical meristem in arabidopsis have now been identified in AON. This includes the recent identification of several CLE peptides capable of activating nodule inhibition responses, a low molecular weight shoot signal and a role for CLAVATA2 in AON. Efforts are now being focused on directly identifying the interactions of these components and to identify the form that long-distance transport molecules take.

  11. Physiological distinctions of pea symbiotic mutants with various nodulation level under inoculation by Rhizobium leguminosarum

    Directory of Open Access Journals (Sweden)

    Akimova G. P.

    2012-11-01

    Full Text Available The study was focused on morphological and physiological peculiarities of pea plants with various nodulation ability under the infection by Rhizobium leguminosarum. Microscopic studies identified distinct features in formation of root nodules in different pea mutants. We revealed differences in the content of indole-acetic acid (IAA in mutants in the root zones differing by sensibility to R. leguminosarum. It was concluded that hormonal system of plants plays an important role in control of nodulation process, and emergence of mutations inducing disturbance of nodule-forming ability apparently depends on root hormonal status, in particular, IAA concentration ensuring both formation of nodule meristem and further organogenesis of the nodule.

  12. First report of Alfalfa mosaic virus infecting basil (Ocimum basilicum L.) in California.

    Science.gov (United States)

    Basil (Ocimum basilicum L.) plants collected from a field in Imperial County, CA in May, 2011 were found to exhibit yellowing, chlorotic sectors and spots on leaves, resulting in plants being unmarketable. Total nucleic acid was extracted from plants and tested by RT-PCR for the presence of Alfalfa...

  13. 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.

  14. Contrasting Strategies of Alfalfa Stem Elongation in Response to Fall Dormancy in Early Growth Stage: The Tradeoff between Internode Length and Internode Number

    Science.gov (United States)

    Wang, Zongli; Sun, Qizhong

    2015-01-01

    Fall dormancy (FD) in alfalfa (Medicago sativa L.) can be described using 11 FD ratings, is widely used as an important indicator of stress resistance, productive performance and spring growth. However, the contrasting growth strategies in internode length and internode number in alfalfa cultivars with different FD rating are poorly understood. Here, a growth chamber study was conducted to investigate the effect of FD on plant height, aboveground biomass, internode length, and internode number in alfalfa individuals in the early growth stages. In order to simulate the alfalfa growth environment in the early stage, 11 alfalfa cultivars with FD ratings from one to 11 were chosen and seeded at the greenhouse, and then were transplanted into an artificial growth chamber. The experimental design was a randomized complete block in a split-plot arrangement with three replicates. Plant height, above-ground biomass, internode length, and internode number were measured in early growth stage in all individuals. Our findings showed that plant height and the aboveground biomass of alfalfa did not significantly differ among 11 different FD rated cultivars. Also, internode length and internode number positively affected plant height and the aboveground biomass of alfalfa individuals and the average internode length significantly increased with increasing FD rating. However, internode number tended to sharply decline when the FD rating increased. Moreover, there were no correlations, slightly negative correlations, and strongly negative correlations between internode length and internode number in alfalfa individuals among the three scales, including within-FD ratings, within-FD categories and inter-FD ratings, respectively. Therefore, our results highlighted that contrasting growth strategies in stem elongation were adopted by alfalfa with different FD ratings in the early growth stage. Alfalfa cultivars with a high FD rating have longer internodes, whereas more dormant alfalfa

  15. Search for Nodulation and Nodule Development-Related Cystatin Genes in the Genome of Soybean (Glycine max).

    Science.gov (United States)

    Yuan, Songli; Li, Rong; Wang, Lei; Chen, Haifeng; Zhang, Chanjuan; Chen, Limiao; Hao, Qingnan; Shan, Zhihui; Zhang, Xiaojuan; Chen, Shuilian; Yang, Zhonglu; Qiu, Dezhen; Zhou, Xinan

    2016-01-01

    Nodulation, nodule development and senescence directly affects nitrogen fixation efficiency, and previous studies have shown that inhibition of some cysteine proteases delay nodule senescence, so their nature inhibitors, cystatin genes, are very important in nodulation, nodule development, and senescence. Although several cystatins are actively transcribed in soybean nodules, their exact roles and functional diversities in legume have not been well explored in genome-wide survey studies. In this report, we performed a genome-wide survey of cystatin family genes to explore their relationship to nodulation and nodule development in soybean and identified 20 cystatin genes that encode peptides with 97-245 amino acid residues, different isoelectric points (pI) and structure characteristics, and various putative plant regulatory elements in 3000 bp putative promoter fragments upstream of the 20 soybean cystatins in response to different abiotic/biotic stresses, hormone signals, and symbiosis signals. The expression profiles of these cystatin genes in soybean symbiosis with rhizobium strain Bradyrhizobium japonicum strain 113-2 revealed that 7 cystatin family genes play different roles in nodulation as well as nodule development and senescence. However, these genes were not root nodule symbiosis (RNS)-specific and did not encode special clade cystatin protein with structures related to nodulation and nodule development. Besides, only two of these soybean cystatins were not upregulated in symbiosis after ABA treatment. The functional analysis showed that a candidate gene Glyma.15G227500 (GmCYS16) was likely to play a positive role in soybean nodulation. Besides, evolutionary relationships analysis divided the cystatin genes from Arabidopsis thaliana, Nicotiana tabacum, rice, barley and four legume plants into three groups. Interestingly, Group A cystatins are special in legume plants, but only include one of the above-mentioned 7 cystatin genes related to nodulation and

  16. A reduced fraction of plant N derived from atmospheric N (%Ndfa) and reduced rhizobial nifH gene numbers indicate a lower capacity for nitrogen fixation in nodules of white clover exposed to long-term CO2 enrichment

    Science.gov (United States)

    Watanabe, T.; Bowatte, S.; Newton, P. C. D.

    2013-12-01

    Using the δ15N natural abundance method, we found that the fraction of nitrogen derived from atmospheric N (%Ndfa) in field-grown white clover (Trifolium repens L.) plants was significantly lower (72.0% vs. 89.8%, p = 0.047 in a grassland exposed to elevated CO2 for 13 yr using free air carbon dioxide enrichment (FACE). Twelve months later we conducted an experiment to investigate the reasons behind the reduced N fixation. We took cuttings from white clover plants growing in the FACE and established individual plants in a glasshouse using soil from the appropriate ambient or elevated CO2 treatments. The established plants were then transplanted back into their "rings of origin" and sampled over a 6-week period. We used molecular ecological analyses targeting nifH genes and transcripts of rhizobia in symbiosis with white clover (Trifolium repens L.) to understand the potential mechanisms. Shoot biomass was significantly lower in eCO2, but there was no difference in nodule number or mass per plant. The numbers of nifH genes and gene transcripts per nodule were significantly reduced under eCO2, but the ratio of gene to transcript number and the strains of rhizobia present were the same in both treatments. We conclude that the capacity for biological nitrogen fixation was reduced by eCO2 in white clover and was related to the reduced rhizobia numbers in nodules. We discuss the finding of reduced gene number in relation to factors controlling bacteroid DNA amount, which may imply an influence of nitrogen as well as phosphorus.

  17. Phytosulfokine Is Involved in Positive Regulation of Lotus japonicus Nodulation.

    Science.gov (United States)

    Wang, Chao; Yu, Haixiang; Zhang, Zhongming; Yu, Liangliang; Xu, Xiaoshu; Hong, Zonglie; Luo, Li

    2015-08-01

    Phytosulfokine (PSK) is a tyrosine-sulfated peptide that is widely distributed in plants, participating in cell proliferation, differentiation, and innate immunity. The potential role of PSK in nodulation in legumes has not been reported. In this work, five PSK precursor genes were identified in Lotus japonicas, designated as LjPSK1 to LjPSK5. Three of them (LjPSK1, LjPSK4, and LjPSK5) were found to be expressed in nitrogen-fixing root nodules. LjPSK1 and LjPSK4 were not induced at the early stage of nodulation. Interestingly, while the expression of LjPSK4 was also found in spontaneous nodules without rhizobial colonization, LjPSK1 was not induced in these pseudo nodules. Promoter-β-glucuronidase analysis revealed that LjPSK1 was highly expressed in enlarged symbiotic cells of nodules. Exogenous addition of 1 1M synthetic PSK peptide resulted in increased nodule numbers per plant. Consistently, the number of mature nodules but not the events of rhizobial infection and nodule initiation was increased by overexpressing LjPSK1 in transgenic hairy roots, in which the expression of jasmonate-responsive genes was found to be repressed. These results suggest that PSK is a new peptide signal that regulates nodulation in legumes, probably through cross-talking with other phytohormones.

  18. 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.

  19. Novel expression pattern of cytosolic gln synthetase in nitrogen-fixing root nodules of the actinorhizal host, Datisca glomerata

    NARCIS (Netherlands)

    Berry, A.M.; Murphy, T.M.; Okubara, P.A.; Jacobsen, K.R.; Swensen, S.M.; Pawlowski, K.

    2004-01-01

    Gln synthetase (GS) is the key enzyme of primary ammonia assimilation in nitrogen-fixing root nodules of legumes and actinorhizal (Frankia-nodulated) plants. In root nodules of Datisca glomerata (Datiscaceae), transcripts hybridizing to a conserved coding region of the abundant nodule isoform, DgGS1

  20. Novel expression pattern of cytosolic gln synthetase in nitrogen-fixing root nodules of the actinorhizal host, Datisca glomerata

    NARCIS (Netherlands)

    Berry, A.M.; Murphy, T.M.; Okubara, P.A.; Jacobsen, K.R.; Swensen, S.M.; Pawlowski, K.

    2004-01-01

    Gln synthetase (GS) is the key enzyme of primary ammonia assimilation in nitrogen-fixing root nodules of legumes and actinorhizal (Frankia-nodulated) plants. In root nodules of Datisca glomerata (Datiscaceae), transcripts hybridizing to a conserved coding region of the abundant nodule isoform,

  1. Alfalfa Responses to Gypsum Application Measured Using Undisturbed Soil Columns

    Directory of Open Access Journals (Sweden)

    Rebecca Tirado-Corbalá

    2017-07-01

    Full Text Available Gypsum is an excellent source of Ca and S, both of which are required for crop growth. Large amounts of by-product gypsum [Flue gas desulfurization gypsum-(FGDG] are produced from coal combustion in the United States, but only 4% is used for agricultural purposes. The objective of this study was to evaluate the effects of (1 untreated, (2 short-term (4-year annual applications of gypsum totaling 6720 kg ha−1, and (3 long-term (12-year annual applications of gypsum totaling 20,200 kg ha−1 on alfalfa (Medicago sativa L. growth and nutrient uptake, and gypsum movement through soil. The study was conducted in a greenhouse using undisturbed soil columns of two non-sodic soils (Celina silt loam and Brookston loam. Aboveground growth of alfalfa was not affected by gypsum treatments when compared with untreated (p > 0.05. Total root biomass (0–75 cm for both soils series was significantly increased by gypsum application (p = 0.04, however, increased root growth was restricted to 0–10 cm depth. Soil and plant analyses indicated no unfavorable environmental impact from of the 4-year and 12-year annual application of FGDG. We concluded that under sufficient water supply, by-product gypsum is a viable source of Ca and S for land application that might benefit alfalfa root growth, but has less effect on aboveground alfalfa biomass production. Undisturbed soil columns were a useful adaptation of the lysimeter method that allowed detailed measurements of alfalfa nutrient uptake, root biomass, and yield and nutrient movement in soil.

  2. Molecular cloning of a bifunctional beta-xylosidase/alpha-L-arabinosidase from alfalfa roots: heterologous expression in Medicago truncatula and substrate specificity of the purified enzyme.

    Science.gov (United States)

    Xiong, Jin-Song; Balland-Vanney, Maud; Xie, Zhi-Ping; Schultze, Michael; Kondorosi, Adam; Kondorosi, Eva; Staehelin, Christian

    2007-01-01

    Glycoside hydrolases are often members of a multigene family, suggesting individual roles for each isoenzyme. Various extracellular glycoside hydrolases have an important but poorly understood function in remodelling the cell wall during plant growth. Here, MsXyl1, a concanavalin A-binding protein from alfalfa (Medicago sativa L.) belonging to the glycoside hydrolase family 3 (beta-D-xylosidase branch) is characterized. Transcripts of MsXyl1 were detected in roots (particularly root tips), root nodules, and flowers. MsXyl1 under the control of the CaMV 35S promoter was expressed in the model legume Medicago truncatula (Gaertner). Concanavalin A-binding proteins from the transgenic plants exhibited 5-8-fold increased activities towards three p-nitrophenyl (PNP) glycosides, namely PNP-beta-D-xyloside, PNP-alpha-L-arabinofuranoside, and PNP-alpha-L-arabinopyranoside. An antiserum raised against a synthetic peptide recognized MsXyl1, which was processed to a 65 kDa form. To characterize the substrate specificity of MsXyl1, the recombinant protein was purified from transgenic M. truncatula leaves by concanavalin A and anion chromatography. MsXyl1cleaved beta-1,4-linked D-xylo-oligosaccharides and alpha-1,5-linked L-arabino-oligosaccharides. Arabinoxylan (from wheat) and arabinan (from sugar beet) were substrates for MsXyl1, whereas xylan (from oat spelts) was resistant to degradation. Furthermore, MsXyl1 released xylose and arabinose from cell wall polysaccharides isolated from alfalfa roots. These data suggest that MsXyl1 is a multifunctional beta-xylosidase/alpha-L-arabinofuranosidase/alpha-L-arabinopyranosidase implicated in cell wall turnover of arabinose and xylose, particularly in rapidly growing root tips. Moreover, the findings of this study demonstrate that stable transgenic M. truncatula plants serve as an excellent expression system for purification and characterization of proteins.

  3. Alfalfa stem tissues: Cell wall deposition, composition, and degradability

    NARCIS (Netherlands)

    Jung, H.G.; Engels, F.M.

    2002-01-01

    Declining cell wall degradability of alfalfa (Medicago sativa L.) stems with maturation limits the nutritional value of alfalfa for ruminants. This study characterized changes in cell wall concentration, composition, and degradability by rumen microbes resulting from alfalfa stem tissue proliferatio

  4. Alfalfa stem tissues: Cell wall deposition, composition, and degradability

    NARCIS (Netherlands)

    Jung, H.G.; Engels, F.M.

    2002-01-01

    Declining cell wall degradability of alfalfa (Medicago sativa L.) stems with maturation limits the nutritional value of alfalfa for ruminants. This study characterized changes in cell wall concentration, composition, and degradability by rumen microbes resulting from alfalfa stem tissue

  5. Alfalfa stem tissues: Cell wall deposition, composition, and degradability

    NARCIS (Netherlands)

    Jung, H.G.; Engels, F.M.

    2002-01-01

    Declining cell wall degradability of alfalfa (Medicago sativa L.) stems with maturation limits the nutritional value of alfalfa for ruminants. This study characterized changes in cell wall concentration, composition, and degradability by rumen microbes resulting from alfalfa stem tissue proliferatio

  6. Nodule activity and allocation of photosynthate of soybean during recovery from water stress

    Science.gov (United States)

    Fellows, R. J.; Patterson, R. P.; Raper, C. D. Jr; Harris, D.; Raper CD, J. r. (Principal Investigator)

    1987-01-01

    Nodulated soybean plants (Glycine max [L.] Merr. cv Ransom) in a growth-chamber study were subjected to a leaf water potential (psi w) of -2.0 megapascal during vegetative growth. Changes in nonstructural carbohydrate contents of leaves, stems, roots, and nodules, allocation of dry matter among plant parts, in situ specific nodule activity, and in situ canopy apparent photosynthetic rate were measured in stressed and nonstressed plants during a 7-day period following rewatering. Leaf and nodule psi w also were determined. At the time of maximum stress, concentration of nonstructural carbohydrates had declined in leaves of stressed, relative to nonstressed, plants, and the concentration of nonstructural carbohydrates had increased in stems, roots, and nodules. Sucrose concentrations in roots and nodules of stressed plants were 1.5 and 3 times greater, respectively, than those of nonstressed plants. Within 12 hours after rewatering, leaf and nodule psi w of stressed plants had returned to values of nonstressed plants. Canopy apparent photosynthesis and specific nodule activity of stressed plants recovered to levels for nonstressed plants within 2 days after rewatering. The elevated sucrose concentrations in roots and nodules of stressed plants also declined rapidly upon rehydration. The increase in sucrose concentration in nodules, as well as the increase of carbohydrates in roots and stems, during water stress and the rapid disappearance upon rewatering indicates that inhibition of carbohydrate utilization within the nodule may be associated with loss of nodule activity. Availability of carbohydrates within the nodules and from photosynthetic activity following rehydration of nodules may mediate the rate of recovery of N2-fixation activity.

  7. Nodule activity and allocation of photosynthate of soybean during recovery from water stress

    Science.gov (United States)

    Fellows, R. J.; Patterson, R. P.; Raper, C. D. Jr; Harris, D.; Raper CD, J. r. (Principal Investigator)

    1987-01-01

    Nodulated soybean plants (Glycine max [L.] Merr. cv Ransom) in a growth-chamber study were subjected to a leaf water potential (psi w) of -2.0 megapascal during vegetative growth. Changes in nonstructural carbohydrate contents of leaves, stems, roots, and nodules, allocation of dry matter among plant parts, in situ specific nodule activity, and in situ canopy apparent photosynthetic rate were measured in stressed and nonstressed plants during a 7-day period following rewatering. Leaf and nodule psi w also were determined. At the time of maximum stress, concentration of nonstructural carbohydrates had declined in leaves of stressed, relative to nonstressed, plants, and the concentration of nonstructural carbohydrates had increased in stems, roots, and nodules. Sucrose concentrations in roots and nodules of stressed plants were 1.5 and 3 times greater, respectively, than those of nonstressed plants. Within 12 hours after rewatering, leaf and nodule psi w of stressed plants had returned to values of nonstressed plants. Canopy apparent photosynthesis and specific nodule activity of stressed plants recovered to levels for nonstressed plants within 2 days after rewatering. The elevated sucrose concentrations in roots and nodules of stressed plants also declined rapidly upon rehydration. The increase in sucrose concentration in nodules, as well as the increase of carbohydrates in roots and stems, during water stress and the rapid disappearance upon rewatering indicates that inhibition of carbohydrate utilization within the nodule may be associated with loss of nodule activity. Availability of carbohydrates within the nodules and from photosynthetic activity following rehydration of nodules may mediate the rate of recovery of N2-fixation activity.

  8. Characterization of green-tissue protein extract from alfalfa (Medicago sativa) exploiting a 3-D technique.

    Science.gov (United States)

    Aguilar, Oscar; Glatz, Charles E; Rito-Palomares, Marco

    2009-09-01

    There is a growing interest of pharmaceutical companies for plant-based production systems. To facilitate the general acceptance of plants as bioreactors, the establishment of efficient downstream operations is critical. It has been proposed that a better understanding of the properties of the contaminant proteins can benefit downstream processing design and operation. The coupled application of 2-DE with aqueous two-phase partitioning has been suggested as a practical 3-D method to characterize potential contaminant proteins from plant extracts. The application of this novel 3-D approach to a complex protein extract from alfalfa (Medicago sativa) containing a model recombinant protein (human granulocyte colony stimulating factor (hG-CSF)) resulted in the quantification of 55 protein spots. The 3-D properties (M(r), pI, and K(p)) obtained for 17 proteins comprising 69% of the alfalfa proteins, allowed the proposal of a prefractionation step as well as the identification of the target molecule (rG-CSF) from bulk of alfalfa proteins. The information obtained from this experimental approach was useful for the identification of the potential contaminant proteins that will occur in alfalfa when this plant is used as a host for recombinant proteins. Additionally, this method will assist in the design of adequate purification strategies for recombinant proteins expressed in alfalfa green tissue.

  9. Enzymatic role for soybean ecto-apyrase in nodulation.

    Science.gov (United States)

    Tanaka, Kiwamu; Nguyen, Tran H N; Stacey, Gary

    2011-07-01

    Root nodulation is regulated by a variety of mechanisms. Ecto-apyrase is an enzyme proposed to control the concentration of extracellular nucleotides. Transgenic expression of the soybean GS52 ecto-apyrase was shown to stimulate nodulation. However, mutation of the enzyme to disrupt enzymatic activity prevented this effect. Therefore, the data suggest that the enzymatic activity of the ecto-apyrase is critical for nodulation enhancement, suggesting a direct effect on extracellular nucleotide hydrolysis. In this article, we propose a hypothetical mechanism for plant ecto-apyrase function during nodulation.

  10. Transcription profiling of soybean nodulation by Bradyrhizobium japonicum

    Science.gov (United States)

    Legumes interact with nodulating bacteria to convert atmospheric nitrogen into ammonia for plant use. This nitrogen fixation takes place within root nodules that form after infection of root hairs by compatible rhizobium. Using cDNA microarrays, we monitored gene expression in soybean (Glycine max) ...

  11. Potato leafhopper (Hemiptera: Cicadellidae) ecology and integrated pest management focused on alfalfa

    Science.gov (United States)

    Knowledge to date on biology of the potato leafhopper, Empoasca fabae (Harris), is summarized, including geographic distribution, development, migration, agricultural host plants, and the mechanism of injury to host plants. Damage to alfalfa, potato, soybean and snap bean, as well as treatment guide...

  12. [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.

  13. 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

  14. 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

  15. 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

  16. CLE peptides control Medicago truncatula nodulation locally and systemically.

    Science.gov (United States)

    Mortier, Virginie; Den Herder, Griet; Whitford, Ryan; Van de Velde, Willem; Rombauts, Stephane; D'Haeseleer, Katrien; Holsters, Marcelle; Goormachtig, Sofie

    2010-05-01

    The CLAVATA3/embryo-surrounding region (CLE) peptides control the fine balance between proliferation and differentiation in plant development. We studied the role of CLE peptides during indeterminate nodule development and identified 25 MtCLE peptide genes in the Medicago truncatula genome, of which two genes, MtCLE12 and MtCLE13, had nodulation-related expression patterns that were linked to proliferation and differentiation. MtCLE13 expression was up-regulated early in nodule development. A high-to-low expression gradient radiated from the inner toward the outer cortical cell layers in a region defining the incipient nodule. At later stages, MtCLE12 and MtCLE13 were expressed in differentiating nodules and in the apical part of mature, elongated nodules. Functional analysis revealed a putative role for MtCLE12 and MtCLE13 in autoregulation of nodulation, a mechanism that controls the number of nodules and involves systemic signals mediated by a leucine-rich repeat receptor-like kinase, SUNN, which is active in the shoot. When MtCLE12 and MtCLE13 were ectopically expressed in transgenic roots, nodulation was abolished at the level of the nodulation factor signal transduction, and this inhibition involved long-distance signaling. In addition, composite plants with roots ectopically expressing MtCLE12 or MtCLE13 had elongated petioles. This systemic effect was not observed in transgenic roots ectopically expressing MtCLE12 and MtCLE13 in a sunn-1 mutant background, although nodulation was still strongly reduced. These results suggest multiple roles for CLE signaling in nodulation.

  17. Geometrical characteristics and damage morphology of nodules grown from artificial seeds in multilayer coating.

    Science.gov (United States)

    Shan, Yongguang; He, Hongbo; Wei, Chaoyang; Li, Shuhong; Zhou, Ming; Li, Dawei; Zhao, Yuan'an

    2010-08-01

    Nodules have been planted in an HfO(2)/SiO(2) multilayer system with absorptive gold nanoparticle seeds located on the surface of a substrate. The topography of nodules was scanned by an atomic force microscope and imaged by a scanning electron microscope. The underlying characteristics of nodules were revealed by a focused ion beam. The cross-sectional profiles reveal that nodules grown from small seeds have a continuous boundary and better mechanical stability. A laser-induced damage test shows that nodules decrease the laser-induced damage threshold by up to 3 times. The damage pits are exclusively caused by nodular ejection and triggered by the absorptive seeds. The distribution of electric field and average temperature rise in the nodules were analyzed. Theoretical results met experimental results very well. The strong absorptive seed and microlens effect of the nodule play important roles in laser-induced damage of a planted nodule.

  18. 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.

  19. Crowdsourcing the nodulation gene network discovery environment.

    Science.gov (United States)

    Li, Yupeng; Jackson, Scott A

    2016-05-26

    The Legumes (Fabaceae) are an economically and ecologically important group of plant species with the conspicuous capacity for symbiotic nitrogen fixation in root nodules, specialized plant organs containing symbiotic microbes. With the aim of understanding the underlying molecular mechanisms leading to nodulation, many efforts are underway to identify nodulation-related genes and determine how these genes interact with each other. In order to accurately and efficiently reconstruct nodulation gene network, a crowdsourcing platform, CrowdNodNet, was created. The platform implements the jQuery and vis.js JavaScript libraries, so that users are able to interactively visualize and edit the gene network, and easily access the information about the network, e.g. gene lists, gene interactions and gene functional annotations. In addition, all the gene information is written on MediaWiki pages, enabling users to edit and contribute to the network curation. Utilizing the continuously updated, collaboratively written, and community-reviewed Wikipedia model, the platform could, in a short time, become a comprehensive knowledge base of nodulation-related pathways. The platform could also be used for other biological processes, and thus has great potential for integrating and advancing our understanding of the functional genomics and systems biology of any process for any species. The platform is available at http://crowd.bioops.info/ , and the source code can be openly accessed at https://github.com/bioops/crowdnodnet under MIT License.

  20. Host range, symbiotic effectiveness and nodulation competitiveness ...

    African Journals Online (AJOL)

    SERVER

    2008-04-17

    Apr 17, 2008 ... were surface sterilized according to the method described by. Somasegaran ... ton and Dilworth nitrogen-free nutrient solution (Broughton and. Dilworth ..... the plants and for the use of the experimental facility. ... characteristics, uses and nodulation. ... USG Publishers and distribution, Ludhiana. pp 120-125.

  1. A histochemical study of root nodule development.

    NARCIS (Netherlands)

    Wiel, van de C.C.M.

    1991-01-01

    In cooperation with soil bacteria of the genera Rhizobium , Bradyrhizobium or Azorhizobium , many members of the legume family are able to form specialized organs on their roots, called root nodules. The bacteria, wrapped up inside a plant membrane, are accomodated in large parenchymatic cells locat

  2. The antenna-specific odorant-binding protein AlinOBP13 of the alfalfa plant bug Adelphocoris lineolatus is expressed specifically in basiconic sensilla and has high binding affinity to terpenoids.

    Science.gov (United States)

    Sun, L; Xiao, H-J; Gu, S-H; Zhou, J-J; Guo, Y-Y; Liu, Z-W; Zhang, Y-J

    2014-08-01

    Odorant-binding proteins (OBPs) are crucial in the olfactory pathway of insects. In the present study, the antenna-enriched OBP AlinOBP13 was investigated because of its potential contribution to the peripheral olfactory perception in the alfalfa plant bug Adelphocoris lineolatus. The results of quantitative reverse transcriptase-PCR showed that the transcript level of AlinOBP13 was higher in the adult stage than in the nymph stages. The transcript levels of AlinOBP13 in the male and female antennae significantly increased after 4 and 8 h of starvation, respectively. Fine ultrastructures of different types of chemosensilla in both female and male antennae were investigated using transmission electron microscopy and immunocytochemical labelling. The results revealed that the anti-AlinOBP13 antiserum strongly and specifically labelled short basiconic sensilla; this antiserum was restricted to the inner lumen and the cavities below the sensillum base of the sensilla. By contrast, multiporous sensilla trichodea, medium long sensilla basiconica, and aporous sensilla chaetica were not labelled. The present study is the first to report an OBP showing specific expression in the short basiconic sensilla of a member of the Hemipteran species. The results of a fluorescence displacement binding assay indicated that recombinant AlinOBP13 showed a more specific binding preference to terpenoids than to sex pheromones and other classes of chemicals. This binding ability was dramatically affected by pH; higher binding affinities were displayed at pH 10.0 than at pH 7.4 and 5.0. In addition, the results of dose-dependent electroantennogram recordings from the antennae showed that both female and male adult bugs responded to the terpenoids tested, suggesting an apparent physiological relevance of AlinOBP13 in A. lineolatus chemoreception. The results of this study suggest that AlinOBP13 functions as a specific carrier of terpenoids and provide insights into the mechanism of A

  3. 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.

  4. Effects of Water Stress on The Rooting, Nodulation Potentials

    African Journals Online (AJOL)

    Administrator

    number of nodules/plant, root length/plant (cm) measured at 2 and 4 wks after planting .... height (117cm) was observed from the cowpea plants that received. 500ml of water .... improving gram yield on the semi arid region of Brazil. Biology of.

  5. THE EFFECT OF PRIMING ON GERMINATION AND SEEDLING GROWTH OF ALFALFA (MEDICAGO SATIVA L.) UNDER SALINITY STRESS

    OpenAIRE

    Jorjandi M.; SHARIFI SIRCHI G.R.

    2012-01-01

    Salinity stress is an abiotic stress which has harmful effects on germination, growth and yield of alfalfa (Medicago sativa L.) in many parts of the world. Seed priming is a way of increasing salt tolerance of plants. In this study, the effect of seed priming on germination and growth of alfalfa seedlings, Bami cultivar, under salinity condition was investigated. According to germination percentage of seeds under In vitro conditions, salicylic acid, Vitamin B12 and distilled water were select...

  6. The chemical properties of soil for alfalfa production after biofertiliser application

    OpenAIRE

    Rodrigo Luis Lemes; Cecílio Viega Soares Filho; Manoel Garcia Neto; Reges Heinrichs

    2013-01-01

    The objective of this study was to evaluate the use of biofertilisers for the production of alfalfa shoot, root and nodule dry matter, and also, to evaluate the chemical properties of the soil. This study was conducted in the greenhouse of the Support Department, Animal Production and Health, Faculty of Veterinary Medicine/UNESP, Araçatuba – SP, from May to October 2010. The experimental design was completely randomised with six biofertiliser doses (0, 25, 50, 100, 200 and 400 m3 ha-1) and fi...

  7. GmEXPB2, a Cell Wall β-Expansin, Affects Soybean Nodulation through Modifying Root Architecture and Promoting Nodule Formation and Development1[OPEN

    Science.gov (United States)

    Li, Xinxin; Zhao, Jing; Tan, Zhiyuan; Liao, Hong

    2015-01-01

    Nodulation is an essential process for biological nitrogen (N2) fixation in legumes, but its regulation remains poorly understood. Here, a β-expansin gene, GmEXPB2, was found to be critical for soybean (Glycine max) nodulation. GmEXPB2 was preferentially expressed at the early stage of nodule development. β-Glucuronidase staining further showed that GmEXPB2 was mainly localized to the nodule vascular trace and nodule vascular bundles, as well as nodule cortical and parenchyma cells, suggesting that GmEXPB2 might be involved in cell wall modification and extension during nodule formation and development. Overexpression of GmEXPB2 dramatically modified soybean root architecture, increasing the size and number of cortical cells in the root meristematic and elongation zones and expanding root hair density and size of the root hair zone. Confocal microscopy with green fluorescent protein-labeled rhizobium USDA110 cells showed that the infection events were significantly enhanced in the GmEXPB2-overexpressing lines. Moreover, nodule primordium development was earlier in overexpressing lines compared with wild-type plants. Thereby, overexpression of GmEXPB2 in either transgenic soybean hairy roots or whole plants resulted in increased nodule number, nodule mass, and nitrogenase activity and thus elevated plant N and phosphorus content as well as biomass. In contrast, suppression of GmEXPB2 in soybean transgenic composite plants led to smaller infected cells and thus reduced number of big nodules, nodule mass, and nitrogenase activity, thereby inhibiting soybean growth. Taken together, we conclude that GmEXPB2 critically affects soybean nodulation through modifying root architecture and promoting nodule formation and development and subsequently impacts biological N2 fixation and growth of soybean. PMID:26432877

  8. GmEXPB2, a Cell Wall β-Expansin, Affects Soybean Nodulation through Modifying Root Architecture and Promoting Nodule Formation and Development.

    Science.gov (United States)

    Li, Xinxin; Zhao, Jing; Tan, Zhiyuan; Zeng, Rensen; Liao, Hong

    2015-12-01

    Nodulation is an essential process for biological nitrogen (N2) fixation in legumes, but its regulation remains poorly understood. Here, a β-expansin gene, GmEXPB2, was found to be critical for soybean (Glycine max) nodulation. GmEXPB2 was preferentially expressed at the early stage of nodule development. β-Glucuronidase staining further showed that GmEXPB2 was mainly localized to the nodule vascular trace and nodule vascular bundles, as well as nodule cortical and parenchyma cells, suggesting that GmEXPB2 might be involved in cell wall modification and extension during nodule formation and development. Overexpression of GmEXPB2 dramatically modified soybean root architecture, increasing the size and number of cortical cells in the root meristematic and elongation zones and expanding root hair density and size of the root hair zone. Confocal microscopy with green fluorescent protein-labeled rhizobium USDA110 cells showed that the infection events were significantly enhanced in the GmEXPB2-overexpressing lines. Moreover, nodule primordium development was earlier in overexpressing lines compared with wild-type plants. Thereby, overexpression of GmEXPB2 in either transgenic soybean hairy roots or whole plants resulted in increased nodule number, nodule mass, and nitrogenase activity and thus elevated plant N and phosphorus content as well as biomass. In contrast, suppression of GmEXPB2 in soybean transgenic composite plants led to smaller infected cells and thus reduced number of big nodules, nodule mass, and nitrogenase activity, thereby inhibiting soybean growth. Taken together, we conclude that GmEXPB2 critically affects soybean nodulation through modifying root architecture and promoting nodule formation and development and subsequently impacts biological N2 fixation and growth of soybean.

  9. 超量表达苹果酸脱氢酶基因提高苜蓿对铝毒的耐受性%Transgenic Alfalfa Plants Overexpressing Nodule-enhanced Malate Dehydrogenase Enhances Tolerance to Aluminum Toxicity

    Institute of Scientific and Technical Information of China (English)

    罗小英; 崔衍波; 邓伟; 李德谋; 裴炎

    2004-01-01

    在酸性土壤中,铝毒是许多作物正常生产的主要限制因子.虽可通过施用生石灰来改良土壤酸碱度,但只能改善土壤表层,在实际应用中受到很大限制.紫花苜蓿作为一种非常重要的牧草、饲料,在酸性土壤条件下苜蓿生长缓慢甚至死亡.我国南方水热资源丰富,但土壤多偏酸性,限制了苜蓿南移.有机酸能螯合铝离子,减轻铝对植物根系的危害.苹果酸是游离铝离子的有效螯合剂,而苹果酸脱氢酶(Malate Dehydro-genase,MDH)催化草酰乙酸形成苹果酸.本研究在苜蓿中超量表达苹果酸脱氢酶基因,通过抗生素筛选、组织化学染色和PCR扩增等技术鉴定出转化植株,并对转化株系进行了耐铝胁迫试验,比较分析了转基因株系与非转基因株系的的相对伸长量,转基因株系根相对伸长量比对照高出3.6%~22.5%.说明超量表达neMDH基因可提高了转基因苜蓿对铝毒的耐受性.

  10. Functional implication of β-carotene hydroxylases in soybean nodulation.

    Science.gov (United States)

    Kim, Yun-Kyoung; Kim, Sunghan; Um, Ji-Hyun; Kim, Kyunga; Choi, Sun-Kang; Um, Byung-Hun; Kang, Suk-Woo; Kim, Jee-Woong; Takaichi, Shinichi; Song, Seok-Bo; Lee, Choon-Hwan; Kim, Ho-Seung; Kim, Ki Woo; Nam, Kyoung Hee; Lee, Suk-Ha; Kim, Yul-Ho; Park, Hyang-Mi; Ha, Sun-Hwa; Verma, Desh Pal S; Cheon, Choong-Ill

    2013-07-01

    Legume-Rhizobium spp. symbiosis requires signaling between the symbiotic partners and differential expression of plant genes during nodule development. Previously, we cloned a gene encoding a putative β-carotene hydroxylase (GmBCH1) from soybean (Glycine max) whose expression increased during nodulation with Bradyrhizobium japonicum. In this work, we extended our study to three GmBCHs to examine their possible role(s) in nodule development, as they were additionally identified as nodule specific, along with the completion of the soybean genome. In situ hybridization revealed the expression of three GmBCHs (GmBCH1, GmBCH2, and GmBCH3) in the infected cells of root nodules, and their enzymatic activities were confirmed by functional assays in Escherichia coli. Localization of GmBCHs by transfecting Arabidopsis (Arabidopsis thaliana) protoplasts with green fluorescent protein fusions and by electron microscopic immunogold detection in soybean nodules indicated that GmBCH2 and GmBCH3 were present in plastids, while GmBCH1 appeared to be cytosolic. RNA interference of the GmBCHs severely impaired nitrogen fixation as well as nodule development. Surprisingly, we failed to detect zeaxanthin, a product of GmBCH, or any other carotenoids in nodules. Therefore, we examined the possibility that most of the carotenoids in nodules are converted or cleaved to other compounds. We detected the expression of some carotenoid cleavage dioxygenases (GmCCDs) in wild-type nodules and also a reduced amount of zeaxanthin in GmCCD8-expressing E. coli, suggesting cleavage of the carotenoid. In view of these findings, we propose that carotenoids such as zeaxanthin synthesized in root nodules are cleaved by GmCCDs, and we discuss the possible roles of the carotenoid cleavage products in nodulation.

  11. Visualization of symbiotic tissue in intact root nodules of Vicia tetrasperma using GFP-marked Rhizobium leguminosarum bv. viciae

    National Research Council Canada - National Science Library

    Chovanec, P; Hovorka, O; Novák, K

    2008-01-01

    In rhizobial symbiosis with legume plant hosts, the symbiotic tissue in the root nodules of indeterminate type is localized to the basal part of the nodule where the symbiotic zones contain infected cells (IC...

  12. 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.

  13. VsENOD5, VsENOD12 and VsENOD40 expression during Rhizobium-induced nodule formation on Vicia sativa roots

    DEFF Research Database (Denmark)

    Vijn, I; Yang, W C; Pallisgård, N

    1995-01-01

    We isolated ENOD5, ENOD12 and ENOD40 homologues from Vicia sativa and studied their expression pattern during Rhizobium-induced nodule formation. Comparison of the VsENOD40 nucleotide sequence with the pea, soybean and alfalfa ENOD40 sequences showed that the sequences contain two conserved regions...

  14. VsENOD5, VsENOD12 and VsENOD40 expression during Rhizobium-induced nodule formation on Vicia sativa roots

    DEFF Research Database (Denmark)

    Vijn, I; Yang, W C; Pallisgård, N

    1995-01-01

    We isolated ENOD5, ENOD12 and ENOD40 homologues from Vicia sativa and studied their expression pattern during Rhizobium-induced nodule formation. Comparison of the VsENOD40 nucleotide sequence with the pea, soybean and alfalfa ENOD40 sequences showed that the sequences contain two conserved regio...

  15. Prevalence of single nucleotide polymorphism among 27 diverse alfalfa genotypes as assessed by transcriptome sequencing

    Directory of Open Access Journals (Sweden)

    Li Xuehui

    2012-10-01

    Full Text Available Abstract Background Alfalfa, a perennial, outcrossing species, is a widely planted forage legume producing highly nutritious biomass. Currently, improvement of cultivated alfalfa mainly relies on recurrent phenotypic selection. Marker assisted breeding strategies can enhance alfalfa improvement efforts, particularly if many genome-wide markers are available. Transcriptome sequencing enables efficient high-throughput discovery of single nucleotide polymorphism (SNP markers for a complex polyploid species. Result The transcriptomes of 27 alfalfa genotypes, including elite breeding genotypes, parents of mapping populations, and unimproved wild genotypes, were sequenced using an Illumina Genome Analyzer IIx. De novo assembly of quality-filtered 72-bp reads generated 25,183 contigs with a total length of 26.8 Mbp and an average length of 1,065 bp, with an average read depth of 55.9-fold for each genotype. Overall, 21,954 (87.2% of the 25,183 contigs represented 14,878 unique protein accessions. Gene ontology (GO analysis suggested that a broad diversity of genes was represented in the resulting sequences. The realignment of individual reads to the contigs enabled the detection of 872,384 SNPs and 31,760 InDels. High resolution melting (HRM analysis was used to validate 91% of 192 putative SNPs identified by sequencing. Both allelic variants at about 95% of SNP sites identified among five wild, unimproved genotypes are still present in cultivated alfalfa, and all four US breeding programs also contain a high proportion of these SNPs. Thus, little evidence exists among this dataset for loss of significant DNA sequence diversity from either domestication or breeding of alfalfa. Structure analysis indicated that individuals from the subspecies falcata, the diploid subspecies caerulea, and the tetraploid subspecies sativa (cultivated tetraploid alfalfa were clearly separated. Conclusion We used transcriptome sequencing to discover large numbers of SNPs

  16. Effect of the fluorescence marked rhizobias on alfalfa seedling growth%接种荧光标记根瘤菌对苜蓿幼苗生长的影响

    Institute of Scientific and Technical Information of China (English)

    陈力玉; 张淑卿; 李剑峰; 师尚礼; 霍平慧; 苗阳阳

    2013-01-01

    Fluorescence marked rhizobias (S.LZgn5-cfp6,S.LZgn5-cfp1 6,S.LZgn5-cfp28,S.12531-cfp2,S. 12531-cfp13,S.12531-cfp26,S.LH3436-cfp1,S.LH3436-cfp2,S.LH3436-cfp6)that contained the plasmid with cfp (cyan fluorescent protein)gene were inoculated to Gannong No.5 alfalfa.Biomass,nodule number,nitroge-nase activity and so on were measured.The result showed that the fluorescence marked rhizobias could promote plant growth and biomass accumulation of inoculated alfalfa.Compared with their corresponding original rhizo-bias,the fluorescence marked rhizobias did no harm to alfalfa seedling,and could express stably in alfalfa plant, and had the similar growth promotion ability with original rhizobias.%将含有 cfp(cyan fluorescent protein )基因质粒的荧光标记根瘤菌 S.LZgn5-cfp6、S.LZgn5-cfp16、S.LZgn5-cfp28,S.12531-cfp2、S.12531-cfp13、S.12531-cfp26、S.LH3436-cfp1、S.LH3436-cfp2、S.LH3436-cfp6接种于甘农5号紫花苜蓿,测定苜蓿幼苗生物量、单株结瘤数、固氮酶活性等,结果表明:荧光标记根瘤菌可有效促进植株生长和生物量的积累,有明显促生效果;与出发菌相比,荧光标记根瘤菌未对苜蓿幼苗产生显著的不良影响,能在植物体内稳定表达,具有与出发菌株相近的促生能力。

  17. Effect of Arbuscular Mycorrhizal Fungi Glomus spp. Inoculation on Alfalfa Growth in Soils with Copper Efecto de la Inoculación con Hongos Micorrízicos Arbusculares Glomus spp. sobre el Crecimiento de Alfalfa en Suelos con Cobre

    Directory of Open Access Journals (Sweden)

    Daniela Novoa M

    2010-06-01

    Full Text Available Soils near mining centers usually have high heavy metal (HM levels. It has been found that some plants associated with arbuscular mycorrhizal fungi (AMF improve growth and tolerance to HM in soils. This symbiosis is a biological resource for degraded soil recovery. The objective of this study was to determine the effect of inoculating AMF (Glomus spp. on alfalfa (Medicago sativa L. growth in agricultural soils with different copper (Cu levels for degraded soil recovery. To this effect, alfalfa seeds were grown in soils from the Catemu and Casablanca valleys and inoculated with AMF. Plant height, stem diameter, and number of leaves were measured weekly. Dry matter, mycorrhizal colonization, and Cu concentration in alfalfa plant tissues were measured after 81 days. Inoculation increased plant height by 24%, stem diameter by 11%, and number of leaves by 34%. Inoculation had a significant effect (p ≤ 0.05 on alfalfa plants that were grown in soil with the highest Cu concentration, but had no effect on Cu accumulation in alfalfa plant tissues. A direct relationship was observed between Cu accumulation in alfalfa and Cu concentration in soils. It was concluded that alfalfa inoculated with Glomus spp. is applicable to the soil recovery process whenever soil properties can ensure inoculum effectiveness on alfalfa growth, and avoid toxicity by excessive Cu in alfalfa plant tissues.Los suelos cercanos a centros de actividad minera suelen presentar altos niveles de metales pesados (HM. Se ha encontrado que algunas plantas asociadas a hongos micorrízicos arbusculares (AMF mejoran su crecimiento y tolerancia a los HM presentes en los suelos. Esta simbiosis constituye un recurso biológico para la recuperación de suelos degradados. El objetivo de este estudio fue determinar el efecto de la inoculación con AMF (Glomus spp. sobre el crecimiento de alfalfa (Medicago sativa L. en suelos agrícolas con distintos niveles de cobre (Cu para la recuperación de

  18. Regeneration of foreign genes co-transformed plants of Medicago sativa L by Agrobacterium rhizogenes

    Institute of Scientific and Technical Information of China (English)

    吕德扬; 曹学远; 唐顺学; 田霞

    2000-01-01

    Gene encoding sulphur amino acid-rich protein (HNP) and rol genes were transferred into Medicago sativa L (alfalfa) mediated by Agrobacterium tumafeciens. Regeneration of trans-genie plants was induced successfully from hairy root tissue of cotyledon in alfalfa. Cotyledon tissues were an ideally transformed recipient. There was a negative correlation between age of hairy roots and embryogenesis frequency in alfalfa. Production of co-transformed plants with greater yield and super quality was important for development of new alfalfa varieties.

  19. Regeneration of foreign genes co-transformed plants of Medicago sativa L by Agrobacterium rhizogenes

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Gene encoding sulphur amino acid-rich protein (HNP) and rol genes were transferred into Medicago sativa L (alfalfa) mediated by Agrobacterium tumafeciens. Regeneration of transgenic plants was induced successfully from hairy root tissue of cotyledon in alfalfa. Cotyledon tissues were an ideally transformed recipient. There was a negative correlation between age of hairy roots and embryogenesis frequency in alfalfa. Production of co-transformed plants with greater yield and super quality was important for development of new alfalfa varieties.

  20. Transcriptome analysis of resistant and susceptible alfalfa cultivars infected with root-knot nematode Meloidogyne incognita.

    Science.gov (United States)

    Postnikova, Olga A; Hult, Maria; Shao, Jonathan; Skantar, Andrea; Nemchinov, Lev G

    2015-01-01

    Nematodes are one of the major limiting factors in alfalfa production. Root-knot nematodes (RKN, Meloidogyne spp.) are widely distributed and economically important sedentary endoparasites of agricultural crops and they may inflict significant damage to alfalfa fields. As of today, no studies have been published on global gene expression profiling in alfalfa infected with RKN or any other plant parasitic nematode. Very little information is available about molecular mechanisms that contribute to pathogenesis and defense responses in alfalfa against these pests and specifically against RKN. In this work, we performed root transcriptome analysis of resistant (cv. Moapa 69) and susceptible (cv. Lahontan) alfalfa cultivars infected with RKN Meloidogyne incognita, widespread root-knot nematode species and a major pest worldwide. A total of 1,701,622,580 pair-end reads were generated on an Illumina Hi-Seq 2000 platform from the roots of both cultivars and assembled into 45,595 and 47,590 transcripts in cvs Moapa 69 and Lahontan, respectively. Bioinformatic analysis revealed a number of common and unique genes that were differentially expressed in susceptible and resistant lines as a result of nematode infection. Although the susceptible cultivar showed a more pronounced defense response to the infection, feeding sites were successfully established in its roots. Characteristically, basal gene expression levels under normal conditions differed between the two cultivars as well, which may confer advantage to one of the genotypes toward resistance to nematodes. Differentially expressed genes were subsequently assigned to known Gene Ontology categories to predict their functional roles and associated biological processes. Real-time PCR validated expression changes in genes arbitrarily selected for experimental confirmation. Candidate genes that contribute to protection against M. incognita in alfalfa were proposed and alfalfa-nematode interactions with respect to resistance

  1. Transcriptome analysis of resistant and susceptible alfalfa cultivars infected with root-knot nematode Meloidogyne incognita.

    Directory of Open Access Journals (Sweden)

    Olga A Postnikova

    Full Text Available Nematodes are one of the major limiting factors in alfalfa production. Root-knot nematodes (RKN, Meloidogyne spp. are widely distributed and economically important sedentary endoparasites of agricultural crops and they may inflict significant damage to alfalfa fields. As of today, no studies have been published on global gene expression profiling in alfalfa infected with RKN or any other plant parasitic nematode. Very little information is available about molecular mechanisms that contribute to pathogenesis and defense responses in alfalfa against these pests and specifically against RKN. In this work, we performed root transcriptome analysis of resistant (cv. Moapa 69 and susceptible (cv. Lahontan alfalfa cultivars infected with RKN Meloidogyne incognita, widespread root-knot nematode species and a major pest worldwide. A total of 1,701,622,580 pair-end reads were generated on an Illumina Hi-Seq 2000 platform from the roots of both cultivars and assembled into 45,595 and 47,590 transcripts in cvs Moapa 69 and Lahontan, respectively. Bioinformatic analysis revealed a number of common and unique genes that were differentially expressed in susceptible and resistant lines as a result of nematode infection. Although the susceptible cultivar showed a more pronounced defense response to the infection, feeding sites were successfully established in its roots. Characteristically, basal gene expression levels under normal conditions differed between the two cultivars as well, which may confer advantage to one of the genotypes toward resistance to nematodes. Differentially expressed genes were subsequently assigned to known Gene Ontology categories to predict their functional roles and associated biological processes. Real-time PCR validated expression changes in genes arbitrarily selected for experimental confirmation. Candidate genes that contribute to protection against M. incognita in alfalfa were proposed and alfalfa-nematode interactions with

  2. Stimulation of nodulation in field peas (Pisum sativum) by low concentrations of ammonium in hydroponic culture

    Science.gov (United States)

    Waterer, J. G.; Vessey, J. K.; Raper, C. D. Jr; Raper CD, J. r. (Principal Investigator)

    1992-01-01

    Although the inhibitory effects of high concentrations of mineral N (> 1.0 mM) on nodule development and function have often been studied, the effects of low, static concentrations of NH4+ (nodulation are unknown. In the present experiments we examine the effects of static concentrations of NH4+ at 0, 0.1 and 0.5 mM in flowing, hydroponic culture on nodule establishment and nitrogenase activity in field peas [Pisum sativum L. cv. Express (Svalof AB)] for the initial 28 days after planting (DAP). Peas grown in the presence of low concentrations of NH4+ had significantly greater nodule numbers (up to 4-fold) than plants grown without NH4+. Nodule dry weight per plant was significantly higher at 14, 21 and 28 DAP in plants grown in the presence of NH4+, but individual nodule mass was lower than in plants grown without NH4+. The nodulation pattern of the plants supplied with NH4+ was similar to that often reported for supernodulating mutants, however the plants did not express other growth habits associated with supernodulation. Estimates of N2 fixation indicate that the plus-NH4+ peas fixed as much or more N2 than the plants supplied with minus-NH4+ nutrient solution. There were no significant differences in nodule numbers, nodule mass or NH4+ uptake between the plants grown at the two concentrations of NH4+. Nodulation appeared to autoregulate by 14 DAP in the minus-NH4+ treatment. Plant growth and N accumulation in the minus-NH4+ plants lagged behind those of the plus-NH4+ treatments prior to N2 fixation becoming well established in the final week of the experiment. The plus-NH4+ treatments appeared not to elicit autoregulation and plants continued to initiate nodules throughout the experiment.

  3. 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 Improvement1[W][OA

    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

    2011-01-01

    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. PMID:21957014

  4. Enteric methane emission, diet digestibility, and nitrogen excretion from beef heifers fed sainfoin or alfalfa.

    Science.gov (United States)

    Chung, Y-H; Mc Geough, E J; Acharya, S; McAllister, T A; McGinn, S M; Harstad, O M; Beauchemin, K A

    2013-10-01

    Effects of plant-bound condensed tannin (CT)-containing sainfoin vs. CT-free alfalfa (or low-CT alfalfa-sainfoin mixture), plant stage of maturity, and their interaction on enteric methane (CH4) emissions, diet digestibility, and N excretion were studied, using 8 ruminally cannulated beef heifers in 2 sequential short-term experiments (Exp. 1 and 2). In Exp. 1, first growth legumes were harvested daily and offered fresh to heifers. Heifers were assigned to 100% sainfoin or 80% alfalfa:20% sainfoin (as-fed basis). Responses were measured at early (late vegetative to early bud; stage 2 to 3) and late (early flower; stage 5) stage of maturity. In Exp. 2, the same legumes were harvested from second growth (late bud; stage 4) and offered to heifers as hay; 100% sainfoin or 100% alfalfa. In both experiments, heifers were fed once daily at 1× maintenance. When fed as fresh forage (Exp. 1), sainfoin, compared with the alfalfa-sainfoin blend, had greater digestibility of OM (74.7 vs. 70.9%; P = 0.02), yet tended to have lower CP digestibility (73.2 vs. 77.1%; P = 0.059). There was no difference between fresh legumes for CH4 emissions [25.9 g/kg DMI ± 4.02 SE; 8.5% of gross energy intake (GEI) ± 1.26 SE; or 36.8 g/kg digested OM ± 1.75 SE]. The fresh legumes were more digestible at early, rather than at late, maturity and, consequently, enteric CH4 (27.4 vs. 24.4 g/kg DMI; P emissions (22.4 g/kg DMI ± 1.29 SD and 7.1% GEI ± 0.40 SD). However, on the basis of OM digested, CH4 emissions were lower for sainfoin than alfalfa hay (44.3 vs. 59.0 g/kg; P = 0.008). Percentage of total N excretion in urine was less for sainfoin compared with alfalfa, both for fresh legumes in Exp. 1 (74 vs. 78%; P = 0.017) or hay in Exp. 2 (64 vs. 72%; P emissions from beef cattle fed at maintenance as compared with feeding either 80% alfalfa:20% sainfoin (fresh forages) or 100% alfalfa (hay). Feeding fresh legumes harvested between the late vegetative to early bud stage, compared with

  5. Role of lupeol synthase in Lotus japonicus nodule formation.

    Science.gov (United States)

    Delis, Costas; Krokida, Afrodite; Georgiou, Sofia; Peña-Rodríguez, Luis M; Kavroulakis, Nektarios; Ioannou, Efstathia; Roussis, Vassilios; Osbourn, Anne E; Papadopoulou, Kalliope K

    2011-01-01

    • Triterpenes are plant secondary metabolites, derived from the cyclization of 2,3-oxidosqualene by oxidosqualene cyclases (OSCs). Here, we investigated the role of lupeol synthase, encoded by OSC3, and its product, lupeol, in developing roots and nodules of the model legume Lotus japonicus. • The expression patterns of OSC3 in different developmental stages of uninfected roots and in roots infected with Mesorhizobium loti were determined. The tissue specificity of OSC3 expression was analysed by in situ hybridization. Functional analysis, in which transgenic L. japonicus roots silenced for OSC3 were generated, was performed. The absence of lupeol in the silenced plant lines was determined by GC-MS. • The expression of ENOD40, a marker gene for nodule primordia initiation, was increased significantly in the OSC3-silenced plant lines, suggesting that lupeol influences nodule formation. Silenced plants also showed a more rapid nodulation phenotype, consistent with this. Exogenous application of lupeol to M. loti-infected wild-type plants provided further evidence for a negative regulatory effect of lupeol on the expression of ENOD40. • The synthesis of lupeol in L. japonicus roots and nodules can be solely attributed to OSC3. Taken together, our data suggest a role for lupeol biosynthesis in nodule formation through the regulation of ENOD40 gene expression.

  6. Milker′s nodule

    Directory of Open Access Journals (Sweden)

    Kaviarasan P

    2009-01-01

    Full Text Available Milker′s nodule virus is a parapox virus that is endemic in cattle. The virus can be grown in tissue culture of both bovine and human cell lines. We report a 53-year-old male, milker who presented with multiple painful nodules on both hands for one month. The nodules were tender and showed central umblication and surrounding rim of erythema. Both epitrochlear and axillary lymph nodes were enlarged on both sides and were non-tender. The skin biopsy revealed marked edema in the epidermis with monouclear inflammatory cell infilitration in the upper dermis. The diagnosis was made on clinical basis. As there was no specific treatment the patient was managed only with antibiotics for secondary infection. The lesions partially resolved after two weeks. This case in reported for its occupational importance.

  7. Alfalfa contains substantial 9-hydroperoxide lyase activity and a 3Z:2E-enal isomerase

    NARCIS (Netherlands)

    Vliegenthart, J.F.G.; Noordermeer, M.A.; Veldink, G.A.

    1999-01-01

    Fatty acid hydroperoxides formed by lipoxygenase can be cleaved by hydroperoxide lyase resulting in the formation of short-chain aldehydes and omega-oxo acids. Plant hydroperoxide lyases use 13- or 9-hydroperoxy linoleic and linolenic acid as substrates. Alfalfa (Medicago sativa L.) has been reporte

  8. First report of 16SrII subgroup D phytoplasma in alfalfa in Sudan

    Science.gov (United States)

    In 2016, alfalfa (Medicago sativa) plants from four commercial fields in Sudan, each about 60 ha, were observed with leaf yellowing symptoms and stunted growth. Total nucleic acid was extracted from leaves of 14 symptomatic samples by a CTAB protocol and used as the template in PCR assays with the u...

  9. Alfalfa interseeded into silage corn can enhance productivity and soil and water conservation

    Science.gov (United States)

    Alfalfa and corn silage are widely planted for dairy forage production systems throughout the northern regions of the USA, accounting for about 0.8 and 1.9 million hectares per year, respectively. Much of this area could benefit from strategies to reduce soil erosion and nutrient losses. Because the...

  10. Alfalfa contains substantial 9-hydroperoxide lyase activity and a 3Z:2E-enal isomerase

    NARCIS (Netherlands)

    Vliegenthart, J.F.G.; Noordermeer, M.A.; Veldink, G.A.

    1999-01-01

    Fatty acid hydroperoxides formed by lipoxygenase can be cleaved by hydroperoxide lyase resulting in the formation of short-chain aldehydes and omega-oxo acids. Plant hydroperoxide lyases use 13- or 9-hydroperoxy linoleic and linolenic acid as substrates. Alfalfa (Medicago sativa L.) has been

  11. Variable salinity responses of 12 alfalfa genotypes and comparative expression analyses of salt-response genes

    Science.gov (United States)

    Twelve alfalfa genotypes that were selected for biomass under salinity, differences in Na and Cl concentrations in shoots and K/Na ratio were evaluated in this long-term salinity experiment. The selected plants were cloned to reduce genetic variability within each genotype. Salt tolerance (ST) index...

  12. Enhancing forage yields and soil conservation by interseeding alfalfa into silage corn

    Science.gov (United States)

    Recent field studies have identified prohexadione-calcium (PHD) as an effective plant growth regulator for enhancing the establishment of alfalfa interseeded into corn as a dual-purpose cover and forage crop. Foliar applications of PHD on seedlings doubled or tripled stand survival of interseeded al...

  13. Pythium and Fusarium species causing seed rot and damping-off of alfalfa

    Science.gov (United States)

    Seed rot and damping-off is an important disease of alfalfa, severely affecting stand establishment when conditions favor the disease. This disease may have been overlooked as a cause of poor stand establishment and reduced vigor of adult plants. Globally, 15 Pythium species have been found to cause...

  14. Alfalfa contains substantial 9-hydroperoxide lyase activity and a 3Z:2E-enal isomerase

    NARCIS (Netherlands)

    Vliegenthart, J.F.G.; Noordermeer, M.A.; Veldink, G.A.

    1999-01-01

    Fatty acid hydroperoxides formed by lipoxygenase can be cleaved by hydroperoxide lyase resulting in the formation of short-chain aldehydes and omega-oxo acids. Plant hydroperoxide lyases use 13- or 9-hydroperoxy linoleic and linolenic acid as substrates. Alfalfa (Medicago sativa L.) has been reporte

  15. Nodules size: An important factor in nodule mining?

    Digital Repository Service at National Institute of Oceanography (India)

    Valsangkar, A.B.

    A study of about 850 different sized nodules from 234 sites in the Central Indian Basin (CIB) showed a clear inverse relationship between size and grade of nodules. Among the different sized nodules, only the small (less than 2 cm) and medium (2...

  16. Evolutionary relationship of alfalfa mosaic virus with cucumber mosaic virus and brome mosaic virus

    OpenAIRE

    Savithri, HS; Murthy, MRN

    1983-01-01

    The amino acid sequences of the non-structural protein (molecular weight 35,000; 3a protein) from three plant viruses - cucumber mosaic, brome mosaic and alfalfa mosaic have been systematically compared using the partial genomic sequences for these three viruses already available. The 3a protein of cucumber mosaic virus has an amino acid sequence homology of 33.7% with the corresponding protein of brome mosaic virus. A similar protein from alfalfa mosaic virus has a homology of 18.2% and 14.2...

  17. Antioxidant effects of alfalfa can improve iron oxide nanoparticle damage: Invivo and invitro studies.

    Science.gov (United States)

    Sadeghi, Leila; Tanwir, Farzeen; Yousefi Babadi, Vahid

    2016-11-01

    Medicago sativa Linn. or alfalfa (Leguminosae) has been used traditionally as an effective cure for CNS, heart and metabolic disorders and digestive aids. Alfalfa is a resistant plant against stress due to small antioxidant molecules and enzymes. Our previous work demonstrated that 100 μg/ml of 50 nm size Fe2O3-NP causes harsh oxidative stress in HepG2 cells and 100 mg/kg of same nanoparticle causes extreme damage in rat's liver. Therefore it can be used as a useful model for invivo and invitro studies of oxidative stress. This study assessed the effects of two concentration of alfalfa on the mentioned invivo and invitro damage. Our results showed alfalfa reduced reactive oxygen species (ROS) production and enhanced reduced glutathione (GSH) that cause reduction of DNA fragmentation and prevent apoptosis pathway so improve viability of the cells. Results also showed alfalfa decreased hepatic enzymes penetrating and lipid peroxidation in rat's liver. Note that Fe2O3-NP potentially has widespread biological application but its usage is limited due to bio incompatibility. A suitable antioxidant compound that reduce nanoparticle side effects can be used as an effective adjuvant with iron oxide nanoparticle (and may be other nanomaterials) in biological applications. Copyright © 2016 Elsevier Inc. All rights reserved.

  18. Proteome analysis of alfalfa roots in response to water deifcit stress

    Institute of Scientific and Technical Information of China (English)

    Rahman Md Atikur; Kim Yong-Goo; AlamIftekhar; LIU Gong-she; Lee Hyoshin; Lee Jeung Joo; Lee Byung-Hyun

    2016-01-01

    To evaluate the response of alfalfa to water deifcit (WD) stress, WD-induced candidates were investigated through a proteomic approach. Alfalfa seedlings were exposed to WD stress for 12 and 15 days respectively, folowed by 3 days re-watering. Water deifcit increased H2O2content, lipid peroxidation, DPPH (1,1-diphenyl-2-picrylhydrazyl)-radical scavenging activity, and the free proline level in alfalfa roots. Root proteins were extracted and separated by two-dimentional polyacrylamide gel electrophoresis (2-DE). A total of 49 WD-responsive proteins were identiifed in alfalfa roots; 25 proteins were reproducibly found to be up-regulated and 24 were down-regulated. Two proteins, namely cytosolic ascorbate peroxidase (APx2) and putative F-box protein were newly detected on 2-DE maps of WD-treated plants. We identiifed several proteins including agamous-like 65, albumin b-32, inward rectifying potassium channel, and auxin-independent growth promoter. The identiifed proteins are involved in a variety of celular functions including calcium signaling, abacisic acid (ABA) biosynthesis, reactive oxygen species (ROS) regulation, transcription/ translation, antioxidant/detoxiifcation/stress defense, energy metabolism, signal transduction, and storage. These results indicate the potential candidates were responsible for adaptive response in alfalfa roots.

  19. The Role of Proanthocyanidins Complex in Structure and Nutrition Interaction in Alfalfa Forage

    Directory of Open Access Journals (Sweden)

    Arjan Jonker

    2016-05-01

    Full Text Available Alfalfa (Medicago sativa L. is one of the main forages grown in the world. Alfalfa is a winter hardy, drought tolerant, N-fixing legume with a good longevity, high yield, high nutrient levels, high digestibility, unique structural to non-structural components ratio, high dry matter intake, and high animal productivity per hectare. However, its main limitation is its excessively rapid initial rate of protein degradation in the rumen, which results in pasture bloat and inefficient use of protein with consequent excessive excretions of nitrogen into the environment. Proanthocyanidins are secondary plant metabolites that can bind with protein and thereby reduce the rate and extent of ruminal protein degradation. However, these secondary metabolites do not accumulate in alfalfa. This review aims to firstly describe the events involved in the rapid release of protein from alfalfa and its effect on ruminant nutrition, environmental pollution, and pasture bloat; secondly, to describe occurrence, structure, functions and benefits of moderate amounts of proanthocyanidin; and finally, to describe the development of alfalfa which accumulates moderate amounts of proanthocyanidins. The emphasis of this review focuses on the role of proanthocyanidins compounds in structure and nutrition interaction in ruminant livestock systems.

  20. The Role of Proanthocyanidins Complex in Structure and Nutrition Interaction in Alfalfa Forage.

    Science.gov (United States)

    Jonker, Arjan; Yu, Peiqiang

    2016-05-23

    Alfalfa (Medicago sativa L.) is one of the main forages grown in the world. Alfalfa is a winter hardy, drought tolerant, N-fixing legume with a good longevity, high yield, high nutrient levels, high digestibility, unique structural to non-structural components ratio, high dry matter intake, and high animal productivity per hectare. However, its main limitation is its excessively rapid initial rate of protein degradation in the rumen, which results in pasture bloat and inefficient use of protein with consequent excessive excretions of nitrogen into the environment. Proanthocyanidins are secondary plant metabolites that can bind with protein and thereby reduce the rate and extent of ruminal protein degradation. However, these secondary metabolites do not accumulate in alfalfa. This review aims to firstly describe the events involved in the rapid release of protein from alfalfa and its effect on ruminant nutrition, environmental pollution, and pasture bloat; secondly, to describe occurrence, structure, functions and benefits of moderate amounts of proanthocyanidin; and finally, to describe the development of alfalfa which accumulates moderate amounts of proanthocyanidins. The emphasis of this review focuses on the role of proanthocyanidins compounds in structure and nutrition interaction in ruminant livestock systems.

  1. Nodulin gene expression in the developing pea root nodule.

    NARCIS (Netherlands)

    Govers, F.

    1987-01-01

    Infection of leguminous plants with bacteria of the genus Rhizobium results in a symbiotic interaction which brings about the development of an entirely new organ on the plant, the root nodule. Within this organ about half of the plant cells are inhabited by bacteroids, the endosymblotic form

  2. 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.

  3. Strategy for polymetallic nodule mining

    Digital Repository Service at National Institute of Oceanography (India)

    Sharma, R.

    , geological, technical and economic conditions, a singe polymetallic nodule mining operation can be carried out for a period of lime 4 • There are several factors for selection of a mine sites sucha~: Nodule distribution, grade and abundance Distribution... (Ol As = (C) (E) (M) Where As= annual nodule recovery rate (dmllycar) D = d'urdtion of mining operation (years) . C =average nodule abundance in mineable area (dmt/sq.km) E =-overall mining efficiency (perccnt) M =proportion of mineable area...

  4. Alfalfa production using saline drainage water

    Science.gov (United States)

    A three year study investigated the use of saline (alfalfa in the presence of shallow saline groundwater. The irrigation treatments included; irrigating twice between cuttings with non-saline water, 2) irrigating with moderately saline water...

  5. 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.

  6. A functional-structural modelling approach to autoregulation of nodulation.

    Science.gov (United States)

    Han, Liqi; Gresshoff, Peter M; Hanan, Jim

    2011-04-01

    Autoregulation of nodulation is a long-distance shoot-root signalling regulatory system that regulates nodule meristem proliferation in legume plants. However, due to the intricacy and subtleness of the signalling nature in plants, molecular and biochemical details underlying mechanisms of autoregulation of nodulation remain largely unknown. The purpose of this study is to use functional-structural plant modelling to investigate the complexity of this signalling system. There are two major challenges to be met: modelling the 3D architecture of legume roots with nodulation and co-ordinating signalling-developmental processes with various rates. Soybean (Glycine max) was chosen as the target legume. Its root system was observed to capture lateral root branching and nodule distribution patterns. L-studio, a software tool supporting context-sensitive L-system modelling, was used for the construction of the architectural model and integration with the internal signalling. A branching pattern with regular radial angles was found between soybean lateral roots, from which a root mapping method was developed to characterize the laterals. Nodules were mapped based on 'nodulation section' to reveal nodule distribution. A root elongation algorithm was then developed for simulation of root development. Based on the use of standard sub-modules, a synchronization algorithm was developed to co-ordinate multi-rate signalling and developmental processes. The modelling methods developed here not only allow recreation of legume root architecture with lateral branching and nodulation details, but also enable parameterization of internal signalling to produce different regulation results. This provides the basis for using virtual experiments to help in investigating the signalling mechanisms at work.

  7. Effect of replacing alfalfa with panicled-tick clover or sericea lespedeza in corn-alfalfa-based substrates on in vitro ruminal methane production.

    Science.gov (United States)

    Naumann, H D; Lambert, B D; Armstrong, S A; Fonseca, M A; Tedeschi, L O; Muir, J P; Ellersieck, M R

    2015-06-01

    Methane emissions from ruminant livestock contribute to total anthropogenic greenhouse gas emissions and reduce metabolizable energy intake by the animal. Condensed tannins (CT) are polyphenolic plant secondary compounds commonly produced by some perennial forage legumes that characteristically bind to protein, carbohydrates, and minerals. The degree to which CT may affect ruminant nutrition depends upon the concentration, structural composition, and biological activity of the CT. The objective of our experiment was to determine the effect of replacing alfalfa in a corn-alfalfa-based substrate with a legume containing CT on in vitro CH4 production and the dynamics of fermentation using an in vitro gas production technique. All fermented substrates contained 50% ground corn as the energy concentrate portion, whereas the forage portion (50%) of each diet was comprised of alfalfa (control) or some combination of alfalfa and sericea lespedeza (SL) or panicled-tick clover (PTC). Our treatments consisted of PTC or SL 15, 30, and 45, which corresponded with 15, 30, or 45% replacement of the diet (alfalfa component) with either PTC or SL. Substrates containing 45% PTC or SL reduced in vitro CH4 production. Treatments did not affect total gas production as compared with that of the control. Replacement of alfalfa with SL or PTC increased fermentable organic matter (FOM). The PTC treatment increased FOM by as much as 1.8% at the 45% replacement level, whereas FOM of SL 45 was increased by less than 1%. The replacement of alfalfa with PTC increased substrate nutritive value greater than replacement with SL. There were no correlations between any physicochemical constituent of the substrates and CH4 production. A combination of factors associated with the inclusion of PTC and SL contributed to the in vitro CH4 production, and CT in these forages was likely a major contributing factor. Further confirmation of these results on in situ or in vivo animal systems is required. If

  8. Identification of genes induced during Medicago sativa nodule development by using the cDNA-AFLP technique

    Institute of Scientific and Technical Information of China (English)

    XIE Rong; HE Zhishui; LIU Fanghua; ZOU Huasong; ZHU Jiabi; YU Guanqiao

    2006-01-01

    A variety of genes involved in nodule development and function were induced during symbiotic root nodule development in alfalfa. We identified those genes by comparing the cDNA-amplified fragment length polymorphism (cDNA-AFLP) patterns of infected nodules and uninfected roots. Seventy-one transcript-derived fragments (TDFs) were isolated from about 3000 TDFs, which are enhanced or specifically expressed in nodule. The differential expression patterns of 61 genes were confirmed by reverse Northern hybridization or Northern hybridization analyses. Among these, 31 exhibited significant similarities to characterized database entries. These results suggest that the genes corresponding to the 31 TDFs may play important roles in signal transduction, gene expression and regulation, substance transportation or auto-regulation of nodulation during nodule development. The Northern hybridization analysis also indicates that the gene corresponding to TDF RX89 was not expressed in the nodule elicited by bacA mutant, presumably it is specially induced at the stage of nodule development when bacteroids begin to be differentiated.

  9. IDENTIFICATION OF SOIL FUNGI ISOLATED FROM ALFALFA (Medicago sativa L TO FIND SPECIFIC FUNGI WHICH IMPROVED THE GROWTH OF ALFALFA

    Directory of Open Access Journals (Sweden)

    T. Yudiarti

    2014-10-01

    Full Text Available Objective of the study was to identify all kinds of fungi which can life in the alfalfa plantation inBaturaden Purwokerto-Central Java. Fungi used in this study was 38 isolates. All fungi have been takenfrom the isolation of soil and root of diseased plant. Macroscopic and microscopic methods were usedfor identification. Potato Dextrose Agar (PDA medium was used to grow the fungi. All fungi wereidentified using book identification of fungi. The results showed that from 38 isolates, six species wasdetermined and one was unidentifed. Those species identified were Cuninghammela sp, Trichoderma sp,Vertilicium sp, Eupenicillium sp, Pythium sp, Aspergillus sp.

  10. Solitary pulmonary nodule

    Science.gov (United States)

    ... Chest x-ray Granulomatosis with polyangiitis Histoplasmosis Lung cancer - small cell Pulmonary tuberculosis Skin nodules Valley fever Review Date 8/1/2015 Updated by: Yi-Bin Chen, MD, Leukemia/Bone Marrow Transplant Program, ... Cancer Lung Diseases Browse the Encyclopedia A.D.A. ...

  11. NODUL TIROID SOLITER

    Directory of Open Access Journals (Sweden)

    I Gusti Ayu Prema Yani Sidemen

    2014-10-01

    Full Text Available Latar Belakang: Nodul tiroid adalah pembengkakan atau massa pada kelenjar tiroid.Pembedahan merupakan pilihan terapi utama, namun pembedahan yang tidak dilakukandengan baik berisiko tinggi mencederai 2 struktur penting, yakni kelenjar paratiroid dannervus rekuren laringeal. Risiko nodul tiroid akan meningkat seiring pertambahan usiadan kurangnya asupan sodium.Insiden terjadinya nodul tiroid lebih sering ditemukanpada laki-laki dan usia di atas 50 tahun, namun pada kasus ini nodul tiroid soliterditemukan pada seorang perempuan dewasa muda sehingga melatar belakangi penulisuntuk mendiskusikan kasus ini dalam laporan kasus.Kasus: Seorang pasien perempuan 32 tahun dengan keluhan tumbuh benjolan padaleher kiri sejak ± 1 tahun yang lalu. Benjolan tersebut dirasakan semakin membesar.Pasien tidak mengeluhkan rasa nyeri di daerah benjolan. Gangguan pada saat makanataupun minum, riwayat sesak, dada berdebar-debar, sering berkeringat, ataupunpenurunan berat badan disangkal. Pada pemeriksaan fisik didapatkan keadaan umumdalam batas normal.  Pada pemeriksaan status lokalis pada region colli anterior lateralsinistra ditemukan massa lunak yang mobile dengan batas tegas dan ukuran ± 3,2 cm x3,4 cm.Hasil: Hasil pemeriksaan patologi anatomi menunjukkan terdapat degenerasi kistikpada massa tiroid tersebut. Pasien kemudian menjalani tindakan pembedahan, yaknitiroidektomi dengan isthmolobectomy. Tiroidektomi dengan teknik diseksi kapsularpada pasien ini berhasil dibuktikan dengan tidak terdapatnya kerusakan pita suara ataukejang pasca operasi.Kesimpulan: Kondisi pasca tiroidektomi dengan teknik diseksi kapsular pada pasien inibaik, tidak terdapat komplikasi kerusakan pita suara dan nervus rekuren laringeal,pasien hidup, dan luka operasi terawat baik.

  12. 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.

  13. Gene Expression Analysis of Alfalfa Seedlings Response to Acid-Aluminum

    Directory of Open Access Journals (Sweden)

    Peng Zhou

    2016-01-01

    Full Text Available Acid-Aluminum (Al is toxic to plants and greatly affects crop production worldwide. To understand the responses of plants to acid soils and Aluminum toxicity, we examined global gene expression using microarray data in alfalfa seedlings with the treatment of acid-Aluminum. 3,926 genes that were identified significantly up- or downregulated in response to Al3+ ions with pH 4.5 treatment, 66.33% of which were found in roots. Their functional categories were mainly involved with phytohormone regulation, reactive oxygen species, and transporters. Both gene ontology (GO enrichment and KEGG analysis indicated that phenylpropanoid biosynthesis, phenylalanine metabolism, and flavonoid biosynthesis played a critical role on defense to Aluminum stress in alfalfa. In addition, we found that transcription factors such as the MYB and WRKY family proteins may be also involved in the regulation of reactive oxygen species reactions and flavonoid biosynthesis. Thus, the finding of global gene expression profile provided insights into the mechanisms of plant defense to acid-Al stress in alfalfa. Understanding the key regulatory genes and pathways would be advantageous for improving crop production not only in alfalfa but also in other crops under acid-Aluminum stress.

  14. Study of phenanthrene utilizing bacterial consortia associated with cowpea (Vigna unguiculata) root nodules.

    Science.gov (United States)

    Sun, Ran; Crowley, David E; Wei, Gehong

    2015-02-01

    Many legumes have been selected as model plants to degrade organic contaminants with their special associated rhizosphere microbes in soil. However, the function of root nodules during microbe-assisted phytoremediation is not clear. A pot study was conducted to examine phenanthrene (PHE) utilizing bacteria associated with root nodules and the effects of cowpea root nodules on phytoremediation in two different types of soils (freshly contaminated soil and aged contaminated soil). Cowpea nodules in freshly-contaminated soil showed less damage in comparison to the aged-contaminated soil, both morphologically and ultra-structurally by scanning electron microscopy. The study of polycyclic aromatic hydrocarbon (PAH) attenuation conducted by high performance liquid chromatography revealed that more PAH was eliminated from liquid culture around nodulated roots than nodule-free roots. PAH sublimation and denaturation gradient gel electrophoresis were applied to analyze the capability and diversity of PAH degrading bacteria from the following four parts of rhizo-microzone: bulk soil, root surface, nodule surface and nodule inside. The results indicated that the surface and inside of cowpea root nodules were colonized with bacterial consortia that utilized PHE. Our results demonstrated that root nodules not only fixed nitrogen, but also enriched PAH-utilizing microorganisms both inside and outside of the nodules. Legume nodules may have biotechnological values for PAH degradation.

  15. Stimulation of nodulation in field peas (Pisum sativum) by low concentrations of ammonium in hydroponic culture

    Science.gov (United States)

    Waterer, J. G.; Vessey, J. K.; Raper, C. D. Jr; Raper CD, J. r. (Principal Investigator)

    1992-01-01

    Although the inhibitory effects of high concentrations of mineral N (> 1.0 mM) on nodule development and function have often been studied, the effects of low, static concentrations of NH4+ (hydroponic culture on nodule establishment and nitrogenase activity in field peas [Pisum sativum L. cv. Express (Svalof AB)] for the initial 28 days after planting (DAP). Peas grown in the presence of low concentrations of NH4+ had significantly greater nodule numbers (up to 4-fold) than plants grown without NH4+. Nodule dry weight per plant was significantly higher at 14, 21 and 28 DAP in plants grown in the presence of NH4+, but individual nodule mass was lower than in plants grown without NH4+. The nodulation pattern of the plants supplied with NH4+ was similar to that often reported for supernodulating mutants, however the plants did not express other growth habits associated with supernodulation. Estimates of N2 fixation indicate that the plus-NH4+ peas fixed as much or more N2 than the plants supplied with minus-NH4+ nutrient solution. There were no significant differences in nodule numbers, nodule mass or NH4+ uptake between the plants grown at the two concentrations of NH4+. Nodulation appeared to autoregulate by 14 DAP in the minus-NH4+ treatment. Plant growth and N accumulation in the minus-NH4+ plants lagged behind those of the plus-NH4+ treatments prior to N2 fixation becoming well established in the final week of the experiment. The plus-NH4+ treatments appeared not to elicit autoregulation and plants continued to initiate nodules throughout the experiment.

  16. RNA-seq transcriptome profiling reveals that Medicago truncatula nodules acclimate N2 fixation before emerging P deficiency reaches the nodules

    Science.gov (United States)

    Cabeza, Ricardo A.; Liese, Rebecca; Lingner, Annika; von Stieglitz, Ilsabe; Neumann, Janice; Salinas-Riester, Gabriela; Pommerenke, Claudia; Dittert, Klaus; Schulze, Joachim

    2014-01-01

    Legume nodules are plant tissues with an exceptionally high concentration of phosphorus (P), which, when there is scarcity of P, is preferentially maintained there rather than being allocated to other plant organs. The hypothesis of this study was that nodules are affected before the P concentration in the organ declines during whole-plant P depletion. Nitrogen (N2) fixation and P concentration in various organs were monitored during a whole-plant P-depletion process in Medicago truncatula. Nodule gene expression was profiled through RNA-seq at day 5 of P depletion. Until that point in time P concentration in leaves reached a lower threshold but was maintained in nodules. N2-fixation activity per plant diverged from that of fully nourished plants beginning at day 5 of the P-depletion process, primarily because fewer nodules were being formed, while the activity of the existing nodules was maintained for as long as two weeks into P depletion. RNA-seq revealed nodule acclimation on a molecular level with a total of 1140 differentially expressed genes. Numerous genes for P remobilization from organic structures were increasingly expressed. Various genes involved in nodule malate formation were upregulated, while genes involved in fermentation were downregulated. The fact that nodule formation was strongly repressed with the onset of P deficiency is reflected in the differential expression of various genes involved in nodulation. It is concluded that plants follow a strategy to maintain N2 fixation and viable leaf tissue as long as possible during whole-plant P depletion to maintain their ability to react to emerging new P sources (e.g. through active P acquisition by roots). PMID:25151618

  17. Overexpressing Arabidopsis ABF3 increases tolerance to multiple abiotic stresses and reduces leaf size in alfalfa.

    Science.gov (United States)

    Wang, Zhi; Su, Guoxia; Li, Min; Ke, Qingbo; Kim, Soo Young; Li, Hongbing; Huang, Jin; Xu, Bingcheng; Deng, Xi-Ping; Kwak, Sang-Soo

    2016-12-01

    Arabidopsis ABSCISIC ACID-RESPONSIVE ELEMENT-BINDING FACTOR 3 (ABF3), a bZIP transcription factor, plays an important role in regulating multiple stress responses in plants. Overexpressing AtABF3 increases tolerance to various stresses in several plant species. Alfalfa (Medicago sativa L.), one of the most important perennial forage crops worldwide, has high yields, high nutritional value, and good palatability and is widely distributed in irrigated and semi-arid regions throughout the world. However, drought and salt stress pose major constraints to alfalfa production. In this study, we developed transgenic alfalfa plants (cv. Xinjiang Daye) expressing AtABF3 under the control of the sweetpotato oxidative stress-inducible SWPA2 promoter (referred to as SAF plants) via Agrobacterium tumefaciens-mediated transformation. After drought stress treatment, we selected two transgenic lines with high expression of AtABF3, SAF5 and SAF6, for further characterization. Under normal conditions, SAF plants showed smaller leaf size compared to non-transgenic (NT) plants, while no other morphological changes were observed. Moreover, SAF plants exhibited enhanced drought stress tolerance and better growth under drought stress treatment, which was accompanied by a reduced transpiration rate and lower reactive oxygen species contents. In addition, SAF plants showed an increased tolerance to salt and oxidative stress. Therefore, these transgenic AtABF3 alfalfa plants might be useful for breeding forage crops with enhanced tolerance to environmental stress for use in sustainable agriculture on marginal lands. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  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 Carbon and Nitrogen Sequestration Patterns and Effects of Temperature and Precipitation in Three Agro-Pastoral Ecotones of Northern China

    Science.gov (United States)

    Wang, Xiaoya; Zhang, Yingjun; Xie, Yue

    2012-01-01

    Alfalfa (Medicago sativa L.) is a primary and widely cultivated forage crop in China. As a perennial leguminous grass, continuous planted alfalfa may influence carbon and nitrogen sequestration in soils. We evaluated the effect of alfalfa, planted for different lengths of time, and temperature and precipitation on soil organic carbon (SOC) and total nitrogen (TN) contents, and estimated soil SOC and TN inventories from 0–60 cm in three agro-pastoral ecotones of northern China. Alfalfa SOC and TN storage patterns were significantly different with increasing soil depths between the three regions of northern China. Continuous alfalfa grassland planted had a positive effect on accumulation of both SOC and TN in the Northwest region, whereas SOC storage peaked 6–7 years after planting in the Northeast and North region. Moreover, relatively higher TN storage appeared 7 years after planting in the Northeast and North regions. This study controlled as many factors as possible, but we caution that such temporal inferences could be artifacts of site selection. The regression analysis indicated that SOC and TN accumulation was mainly dependent on temperature (≥10°C of effective total accumulated temperature) in the North region. Precipitation in the growing season was the main limiting factor for SOC storage in the Northwest region and TN accumulation in the North regions. Therefore, the different climate factors affecting SOC and TN sequestration in alfalfa occurred at a regional scale. PMID:23209775

  20. Alfalfa carbon and nitrogen sequestration patterns and effects of temperature and precipitation in three agro-pastoral ecotones of northern China.

    Directory of Open Access Journals (Sweden)

    Shujuan Chang

    Full Text Available Alfalfa (Medicago sativa L. is a primary and widely cultivated forage crop in China. As a perennial leguminous grass, continuous planted alfalfa may influence carbon and nitrogen sequestration in soils. We evaluated the effect of alfalfa, planted for different lengths of time, and temperature and precipitation on soil organic carbon (SOC and total nitrogen (TN contents, and estimated soil SOC and TN inventories from 0-60 cm in three agro-pastoral ecotones of northern China. Alfalfa SOC and TN storage patterns were significantly different with increasing soil depths between the three regions of northern China. Continuous alfalfa grassland planted had a positive effect on accumulation of both SOC and TN in the Northwest region, whereas SOC storage peaked 6-7 years after planting in the Northeast and North region. Moreover, relatively higher TN storage appeared 7 years after planting in the Northeast and North regions. This study controlled as many factors as possible, but we caution that such temporal inferences could be artifacts of site selection. The regression analysis indicated that SOC and TN accumulation was mainly dependent on temperature (≥10°C of effective total accumulated temperature in the North region. Precipitation in the growing season was the main limiting factor for SOC storage in the Northwest region and TN accumulation in the North regions. Therefore, the different climate factors affecting SOC and TN sequestration in alfalfa occurred at a regional scale.

  1. [Efficiency of alfalfa and reed in the phytoremediation of hydrocarbon polluted soil].

    Science.gov (United States)

    Muratova, A Iu; Turkovskaia, O V; Hubner, T; Kuschk, P

    2003-01-01

    The efficacy of plants as means of decontaminating hydrocarbon-polluted soil has been studied. Ditch reed (Phragmites australis) and alfalfa (Medicago sativa) markedly intensified processes of pollutant destruction, the effect being particularly pronounced in the case of polycyclic aromatic hydrocarbons (PAHs). Comparative analysis of microflora in soils (including those devoid of plants and rhizosphere) demonstrated that, in addition to preventing the pollutant-induced decrease in the amount of heterotrophic microorganisms, the plants stimulated their development, significantly increasing the population of destructors. Effects of plants on major physiological groups of soil microorganisms under conditions of pollution were ambiguous. The rhizosphere consortium of alfalfa was less susceptible to effects of pollutants than that of reed.

  2. Transcription reprogramming during root nodule development in Medicago truncatula.

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    Sandra Moreau

    Full Text Available Many genes which are associated with root nodule development and activity in the model legume Medicago truncatula have been described. However information on precise stages of activation of these genes and their corresponding transcriptional regulators is often lacking. Whether these regulators are shared with other plant developmental programs also remains an open question. Here detailed microarray analyses have been used to study the transcriptome of root nodules induced by either wild type or mutant strains of Sinorhizobium meliloti. In this way we have defined eight major activation patterns in nodules and identified associated potential regulatory genes. We have shown that transcription reprogramming during consecutive stages of nodule differentiation occurs in four major phases, respectively associated with (i early signalling events and/or bacterial infection; plant cell differentiation that is either (ii independent or (iii dependent on bacteroid differentiation; (iv nitrogen fixation. Differential expression of several genes involved in cytokinin biosynthesis was observed in early symbiotic nodule zones, suggesting that cytokinin levels are actively controlled in this region. Taking advantage of databases recently developed for M. truncatula, we identified a small subset of gene expression regulators that were exclusively or predominantly expressed in nodules, whereas most other regulators were also activated under other conditions, and notably in response to abiotic or biotic stresses. We found evidence suggesting the activation of the jasmonate pathway in both wild type and mutant nodules, thus raising questions about the role of jasmonate during nodule development. Finally, quantitative RT-PCR was used to analyse the expression of a series of nodule regulator and marker genes at early symbiotic stages in roots and allowed us to distinguish several early stages of gene expression activation or repression.

  3. 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 (13)Clabeling procedure was used to label C photoassimilates of pods and leaves in alfalfa, and the Δ(13)C 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 (13)C 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.

  4. 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

    2017-01-01

    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. PMID:28169330

  5. Transcriptome responses in alfalfa associated with tolerance to intensive animal grazing

    Science.gov (United States)

    Wang, Junjie; Zhao, Yan; Ray, Ian; Song, Mingzhou

    2016-01-01

    Tolerance of alfalfa (Medicago sativa L.) to animal grazing varies widely within the species. However, the molecular mechanisms influencing the grazing tolerant phenotype remain uncharacterized. The objective of this study was to identify genes and pathways that control grazing response in alfalfa. We analyzed whole-plant de novo transcriptomes from grazing tolerant and intolerant populations of M. sativa ssp. falcata subjected to grazing by sheep. Among the Gene Ontology terms which were identified as grazing responsive in the tolerant plants and differentially enriched between the tolerant and intolerant populations (both grazed), most were associated with the ribosome and translation-related activities, cell wall processes, and response to oxygen levels. Twenty-one grazing responsive pathways were identified that also exhibited differential expression between the tolerant and intolerant populations. These pathways were associated with secondary metabolite production, primary carbohydrate metabolic pathways, shikimate derivative dependent pathways, ribosomal subunit composition, hormone signaling, wound response, cell wall formation, and anti-oxidant defense. Sequence polymorphisms were detected among several differentially expressed homologous transcripts between the tolerant and intolerant populations. These differentially responsive genes and pathways constitute potential response mechanisms for grazing tolerance in alfalfa. They also provide potential targets for molecular breeding efforts to develop grazing-tolerant cultivars of alfalfa. PMID:26763747

  6. 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.

  7. Nodule Initiation Involves the Creation of a New Symplasmic Field in Specific Root Cells of Medicago Species

    Science.gov (United States)

    Complainville, Arnaud; Brocard, Lysiane; Roberts, Ian; Dax, Edna; Sever, Noa; Sauer, Norbert; Kondorosi, Adam; Wolf, Shmuel; Oparka, Karl; Crespi, M.

    2003-01-01

    The organogenesis of nitrogen-fixing nodules in legume plants is initiated in specific root cortical cells and regulated by long-distance signaling and carbon allocation. Here, we explore cell-to-cell communication processes that occur during nodule initiation in Medicago species and their functional relevance using a combination of fluorescent tracers, electron microscopy, and transgenic plants. Nodule initiation induced symplasmic continuity between the phloem and nodule initials. Macromolecules such as green fluorescent protein could traffic across short or long distances from the phloem into these primordial cells. The created symplasmic field was regulated throughout nodule development. Furthermore, Medicago truncatula transgenic plants expressing a viral movement protein showed increased nodulation. Hence, the establishment of this symplasmic field may be a critical element for the control of nodule organogenesis. PMID:14615602

  8. Alfalfa -- a sustainable crop for biomass energy production

    Science.gov (United States)

    Alfalfa (Medicago sativa) has the potential to be a significant contributor to America's renewable energy future. In an alfalfa biomass energy production system, alfalfa forage would be separated into stem and leave fractions. The stems would be processed to produce energy, and the leaves would be s...

  9. Comparative drought response in eleven diverse alfalfa accessions

    Science.gov (United States)

    Alfalfa (Medicago sativa L.) production is often negatively affected by drought stress. This is particularly true for alfalfa that is cultivated on rangeland. Thus, the development of drought-tolerant alfalfa cultivars is of great significance. A greenhouse study was conducted to evaluate 11 alfa...

  10. Cash in on N credits when corn follows alfalfa

    Science.gov (United States)

    When alfalfa is killed, some of the accumulated N in the soil and in alfalfa leaves, stems, and roots becomes available to subsequent crops. This increased N supply is known as the “alfalfa N credit,” which is the amount of fertilizer or available manure N a grower can save, resulting in higher net ...

  11. The Undergraduate ALFALFA Team: Collaborative Research Projects

    Science.gov (United States)

    Cannon, John M.; Koopmann, Rebecca A.; Haynes, Martha P.; Undergraduate ALFALFA Team, ALFALFA Team

    2016-01-01

    The NSF-sponsored Undergraduate ALFALFA (Arecibo Legacy Fast ALFA) Team (UAT) has allowed faculty and students from a wide range of public and private colleges and especially those with small astronomy programs to learn how science is accomplished in a large collaboration while contributing to the scientific goals of a legacy radio astronomy survey. The UAT has achieved this through close collaboration with ALFALFA PIs to identify research areas accessible to undergraduates. In this talk we will summarize the main research efforts of the UAT, including multiwavelength followup observations of ALFALFA sources, the UAT Collaborative Groups Project, the Survey of HI in Extremely Low-mass Dwarfs (SHIELD), and the Arecibo Pisces-Perseus Supercluster Survey. This work has been supported by NSF grants AST-0724918/0902211, AST-075267/0903394, AST-0725380, and AST-1211005.

  12. Ethylene, a Hormone at the Center-Stage of Nodulation.

    Science.gov (United States)

    Guinel, Frédérique C

    2015-01-01

    Nodulation is the result of a beneficial interaction between legumes and rhizobia. It is a sophisticated process leading to nutrient exchange between the two types of symbionts. In this association, within a nodule, the rhizobia, using energy provided as photosynthates, fix atmospheric nitrogen and convert it to ammonium which is available to the plant. Nodulation is recognized as an essential process in nitrogen cycling and legume crops are known to enrich agricultural soils in nitrogenous compounds. Furthermore, as they are rich in nitrogen, legumes are considered important as staple foods for humans and fodder for animals. To tightly control this association and keep it mutualistic, the plant uses several means, including hormones. The hormone ethylene has been known as a negative regulator of nodulation for almost four decades. Since then, much progress has been made in the understanding of both the ethylene signaling pathway and the nodulation process. Here I have taken a large view, using recently obtained knowledge, to describe in some detail the major stages of the process. I have not only reviewed the steps most commonly covered (the common signaling transduction pathway, and the epidermal and cortical programs), but I have also looked into steps less understood (the pre-infection step with the plant defense response, the bacterial release and the formation of the symbiosome, and nodule functioning and senescence). After a succinct review of the ethylene signaling pathway, I have used the knowledge obtained from nodulation- and ethylene-related mutants to paint a more complete picture of the role played by the hormone in nodule organogenesis, functioning, and senescence. It transpires that ethylene is at the center of this effective symbiosis. It has not only been involved in most of the steps leading to a mature nodule, but it has also been implicated in host immunity and nodule senescence. It is likely responsible for the activation of other hormonal

  13. 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.

  14. Salt stress alters DNA methylation levels in alfalfa (Medicago spp).

    Science.gov (United States)

    Al-Lawati, A; Al-Bahry, S; Victor, R; Al-Lawati, A H; Yaish, M W

    2016-02-26

    Modification of DNA methylation status is one of the mechanisms used by plants to adjust gene expression at both the transcriptional and posttranscriptional levels when plants are exposed to suboptimal conditions. Under abiotic stress, different cultivars often show heritable phenotypic variation accompanied by epigenetic polymorphisms at the DNA methylation level. This variation may provide the raw materials for plant breeding programs that aim to enhance abiotic stress tolerance, including salt tolerance. In this study, methylation-sensitive amplified polymorphism (MSAP) analysis was used to assess cytosine methylation levels in alfalfa (Medicago spp) roots exposed to increasing NaCl concentrations (0.0, 8.0, 12.0, and 20.0 dS/m). Eleven indigenous landraces were analyzed, in addition to a salt-tolerant cultivar that was used as a control. There was a slight increase in DNA methylation upon exposure to high levels of soil salinity. Phylogenetic analysis using MSAP showed epigenetic variation within and between the alfalfa landraces when exposed to saline conditions. Based on MSAP and enzyme-linked immunosorbent assay results, we found that salinity increased global DNA methylation status, particularly in plants exposed to the highest level of salinity (20 dS/m). Quantitative reverse transcription-polymerase chain reaction indicated that this might be mediated by the overexpression of methyltransferase homolog genes after exposure to saline conditions. DNA demethylation using 5-azacytidine reduced seedling lengths and dry and fresh weights, indicating a possible decrease in salinity tolerance. These results suggest that salinity affects DNA methylation flexibility.

  15. Nodule carbohydrate catabolism is enhanced in the Medicago truncatula A17-Sinorhizobium medicae WSM419 symbiosis

    Directory of Open Access Journals (Sweden)

    Estibaliz eLarrainzar

    2014-08-01

    Full Text Available The symbiotic association between Medicago truncatula and Sinorhizobium meliloti is a well-established model system in the legume-Rhizobium community. Despite its wide use, the symbiotic efficiency of this model has been recently questioned and an alternative microsymbiont, S. medicae, has been proposed. However, little is known about the physiological mechanisms behind the higher symbiotic efficiency of S. medicae WSM419. In the present study, we inoculated M. truncatula Jemalong A17 with either S. medicae WSM419 or S. meliloti 2011 and compared plant growth, photosynthesis, N2-fixation rates, and plant nodule carbon and nitrogen metabolic activities in the two systems. M. truncatula plants in symbiosis with S. medicae showed increased biomass and photosynthesis rates per plant. Plants grown in symbiosis with S. medicae WSM419 also showed higher N2-fixation rates, which were correlated with a larger nodule biomass, while nodule number was similar in both systems. In terms of plant nodule metabolism, M. truncatula-S. medicae WSM419 nodules showed increased sucrose-catabolic activity, mostly associated with sucrose synthase, accompanied by a reduced starch content, whereas nitrogen-assimilation activities were comparable to those measured in nodules infected with S. meliloti 2011. Taken together, these results suggest that S. medicae WSM419 is able to enhance plant carbon catabolism in M. truncatula nodules, which allows for the maintaining of high symbiotic N2-fixation rates, better growth and improved general plant performance.

  16. 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.

  17. Short communication. Nitrogen content of residual alfalfa taproots under irrigation

    Directory of Open Access Journals (Sweden)

    S. Cela

    2013-04-01

    Full Text Available The decomposition of alfalfa (Medicago sativa L. residues can provide significant amounts of N to subsequent crops, but most of the data on this subject has been obtained from 1-2 year old alfalfa stands. The objective of this study was to determine the biomass of alfalfa taproots and their N content in irrigated alfalfa stands that are more than 2 years old. Twenty-two commercial irrigated alfalfa fields were evaluated in the Ebro Valley (Northeast Spain from 2006 to 2010. The taproot biomass in the arable layer (0 to 30 cm depth ranged from 1.8 to 10.1 Mg ha-1 and averaged 4.8 Mg ha-1. In contrast, the N concentration in alfalfa taproots was constant among fields and averaged 24.6 g N kg-1. The total amount of N contained in alfalfa taproots (0-30 cm depth ranged from 47 to 96 kg N ha-1 in 55% of the fields, ranged from 97 to 200 kg N ha-1 in 22% of the fields, and exceeded 200 kg N ha-1 in 23% of the fields. The N content of the irrigated alfalfa taproots studied here is in the upper range previously reported in other areas, mainly with younger alfalfa stands. Based on the current finding, a classification of the quality of irrigated alfalfa stands is proposed to improve the estimates of the residual-N effects of alfalfa on subsequent crops.

  18. Expression of anthocyanins and proanthocyanidins after transformation of alfalfa with maize Lc.

    Science.gov (United States)

    Ray, Heather; Yu, Min; Auser, Patricia; Blahut-Beatty, Laureen; McKersie, Brian; Bowley, Steve; Westcott, Neil; Coulman, Bruce; Lloyd, Alan; Gruber, Margaret Y

    2003-07-01

    Three anthocyanin regulatory genes of maize (Zea mays; Lc, B-Peru, and C1) were introduced into alfalfa (Medicago sativa) in a strategy designed to stimulate the flavonoid pathway and alter the composition of flavonoids produced in forage. Lc constructs included a full-length gene and a gene with a shortened 5'-untranslated region. Lc RNA was strongly expressed in Lc transgenic alfalfa foliage, but accumulation of red-purple anthocyanin was observed only under conditions of high light intensity or low temperature. These stress conditions induced chalcone synthase and flavanone 3-hydroxylase expression in Lc transgenic alfalfa foliage compared with non-transformed plants. Genotypes containing the Lc transgene construct with a full-length 5'-untranslated region responded more quickly to stress conditions and with a more extreme phenotype. High-performance liquid chromatography analysis of field-grown tissue indicated that flavone content was reduced in forage of the Lc transgenic plants. Leucocyanidin reductase, the enzyme that controls entry of metabolites into the proanthocyanidin pathway, was activated both in foliage and in developing seeds of the Lc transgenic alfalfa genotypes. Proanthocyanidin polymer was accumulated in the forage, but (+)-catechin monomers were not detected. B-Peru transgenic and C1 transgenic populations displayed no visible phenotypic changes, although these transgenes were expressed at detectable levels. These results support the emerging picture of Lc transgene-specific patterns of expression in different recipient species. These results demonstrate that proanthocyanidin biosynthesis can be stimulated in alfalfa forage using an myc-like transgene, and they pave the way for the development of high quality, bloat-safe cultivars with ruminal protein bypass.

  19. Flue gas desulfurization products as sulfur sources for alfalfa and soybean

    Energy Technology Data Exchange (ETDEWEB)

    Chen, L.M.; Dick, W.A.; Nelson, S. [Ohio State University, Wooster, OH (US). School of Natural Resources

    2005-02-01

    Sulfur deficiencies in soil are expected to increase due to growth of high-yielding crop varieties, use of S-free fertilizers, and removal of S from industrial emissions. Flue gas desulfurization (FGD) products, created when coal is burned and SO{sub 2} is removed from the flue gases, may serve as efficient S sources. However, there are few reports on their use for the enhancement of crop growth. Agricultural gypsum and two types of FGD products, that contain either vermiculite or perlite, were applied at 0, 16, and 67 kg S ha{sup -1} to an agricultural soil (Wooster silt loam, Typic Fragiudalf). Dry weight of a new planting of alfalfa (Medicago sativa L.) was increased up to 40% by the treatments of FGD products or gypsum compared with the untreated control. Gypsum and FGD products were also applied at 0, 8,16, and 24 kg S ha{sup -1} to five established alfalfa stands in different Ohio regions. Mean alfalfa yield was significantly (P {le} 0.05) increased by approximately 5.0% in 2001 and 6.0% in 2002 with the S treatments of FGD products or gypsum compared with the untreated control. Alfalfa yields for FGD products and gypsum treatments were similar. A slight positive yield response was observed for soybean (Glycine max L.) when soils were treated with S-containing materials. Soil and plant analyses were made to assess potential adverse environmental impacts and none were observed. Thus, these FGD products can be safely applied to agricultural soils as S sources and can improve alfalfa yields in S-deficient soils.

  20. Multiple pulmonary rheumatoid nodules.

    Science.gov (United States)

    Sargin, Gokhan; Senturk, Taskin

    2015-01-01

    We present a case of 45-year-old female patient with the diagnosis of seropositive rheumatoid arthritis, who was admitted to our rheumatology department with exacerbation of the disease. The patient's disease activity score (DAS 28) was 6.9. Physical examination revealed changes in the lung auscultation as a rough breathing sound at the middle and lower lobe of the right lung. Chest X-ray revealed multiple nodular densities in both lungs. Lung biopsy was performed for the diagnosis and revealed necrotizing granulomas with central fibrinoid necrosis surrounded by epithelioid cells. Such a histopathological picture is typical for rheumatoid nodules. Finally the patient was treated with rituximab, with significant improvement.

  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. Allelopathic effects of redroot pigweed (Amaranthus retroflexus L. on germination & growth of cucumber, alfalfa, common bean and bread wheat

    Directory of Open Access Journals (Sweden)

    Hamideh BAKHSHAYESHAN-AGDAM

    2015-11-01

    Full Text Available Allelopathy is one of the important interactions among plants. Weeds can reduce crops productions in farms by their allelopathic effects. Redroot pigweed (Amaranthus retroflexus L. is the most common weed in Iran with well-known allelopathic potential. In the presented experiment, the allelopathic effects of redroot pigweed on germination and growth of four important crop species including cucumber (Cucumis sativus L., alfalfa (Medicago sativa L., common bean (Phaseulus vulgaris L. and bread wheat (Triticum aestivum L. was studied. The effect of different concentrations of redroot pigweed leachate on seed germination and seedlings growth parameters of tested plants was significant, but not same in all studied species. Bread wheat and cucumber were more resistance in seed germination stage in comparison to common bean and alfalfa. Except alfalfa, all plant species showed certain rate of resistance in the most measured parameters. According to the obtained results, bread wheat and common bean were the most resistant species, cucumber was resistant at low concentration but sensitive at high concentration, and alfalfa was the most sensitive species to the redroot pigweed leachate treatments. Therefore, the cultivation of resistant plant species (such as bread wheat and common bean plants in the regions with redroot pigweed’s invasion is appropriate way in management of the farms.

  3. Identification of Molecular Marker Linked to Salt Tolerance Gene in Alfalfa

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    The study has established the F2 offspring obtained by crossing salt-tolerant with salt-sensitive alfalfa, and appraised the salt-tolerant F2 offspring seedling was evaluated in pot culture. With the F2 segregated population, the research has obtained a molecular marker linked with salt-tolerant genes of alfalfa using the improved BSA combined with RAPD. The RAPD PCR products were excised from the agarose gel and purified using a kit, then were mixed with pMD-18T vector and sequenced. Sequencing result indicated the RAPD marker was 1 438 bp in length. Similarity researches using blast in Genbank indicated that the nucleotide sequence of the RAPD marker showed 93% and 91% similarity with mth2-6el8 gene fragment (347 bp) and mth2-33122 gene fragment (334 bp) of Medicago truncatula respectively. Medicago truncatula is a close relative of alfalfa and Mth2-6e18 is a molecular marker of the gene coding for a cysteine protease which was salt inducible in some plants. These results indicated the RAPD marker was possibly related to cysteine protease genes in alfalfa.

  4. 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.

  5. 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.

  6. Uptake and translocation of sulfamethazine by alfalfa grown under hydroponic conditions.

    Science.gov (United States)

    Kurwadkar, Sudarshan; Struckhoff, Garrett; Pugh, Kameron; Singh, Om

    2017-03-01

    Antibiotics are routinely used in intensive animal agriculture operations collectively known as Concentrated Animal Feed Operations (CAFO) which include dairy, poultry and swine farms. Wastewater generated by CAFOs often contains low levels of antibiotics and is typically managed in an anaerobic lagoon. The objective of this research is to investigate the uptake and fate of aqueous sulfamethazine (SMN) antibiotic by alfalfa (Medicago sativa) grass grown under hydroponic conditions. Uptake studies were conducted using hydroponically grown alfalfa in a commercially available nutrient solution supplemented with 10mg/L of SMN antibiotic. Analysis of alfalfa sap, root zone, middle one-third, and top portion of the foliage showed varying uptake rate and translocation of SMN. The highest average amount of SMN (8.58μg/kg) was detected in the root zone, followed by the top portion (1.89μg/kg), middle one-third (1.30μg/kg), and sap (0.38μg/kg) samples, indicating a clear distribution of SMN within the sampled regions. The ultraviolet (UV) spectra of parent SMN and translocated SMN identified in different parts of the plant present the possibility of metabolization during the uptake process. Uptake of SMN using alfalfa grown under hydroponic conditions has potential as a promising remediation technology for removal of similar antibiotics from wastewater lagoons. Copyright © 2016. Published by Elsevier B.V.

  7. Grafting analysis indicates that malfunction of TRICOT in the root causes a nodulation-deficient phenotype in Lotus japonicus.

    Science.gov (United States)

    Suzaki, Takuya; Kawaguchi, Masayoshi

    2013-03-01

    Leguminous plants develop root nodules in symbiosis with soil rhizobia. Nodule formation occurs following rhizobial infection of the host root that induces dedifferentiation of some cortical cells and the initiation of a new developmental program to form nodule primordia. In a recent study, we identified a novel gene, TRICOT (TCO), that acts as a positive regulator of nodulation in Lotus japonicus. In addition to its role in nodulation, tco mutant plants display pleiotropic defects including abnormal shoot apical meristem formation. Here, we investigated the effect of the tco mutation on nodulation using a grafting approach. The results strongly indicate that the nodulation-deficient phenotype of the mutant results from malfunction of the TCO gene in the root.

  8. Partitioning and mobilization of photoassimilate in alfalfa subjected to water deficits. [Medicago sativa L

    Energy Technology Data Exchange (ETDEWEB)

    Hall, M.H. (Univ. of Idaho, Moscow (USA)); Sheaffer, C.C.; Heichel, G.H. (Univ. of Minnesota, St. Paul (USA))

    Faster regrowth of a stressed alfalfa (Medicago sativa L.) crop compared to an unstressed crop after rewatering has been reported. The bases of this compensatory response are unknown, but they may be important to understanding adaptation to water stress and to developing crop water management strategies. The authors objectives was to determine the effect of stress induced by water deficit on photoassimilate partitioning and the utilization of stored assimilates during regrowth of alfalfa. Field and greenhouse experiments were conducted using cultivars differing in winterhardiness. Plants were subjected to water stress, pulse-labeled with {sup 14}CO{sub 2}, and sampled following 0, 1, 14, 21, and 28-d translocation periods. Following the 14-d sampling, herbage was harvested and water stress was removed. Cultivars contrasting in winterhardiness responded similarly to water stress. Stressed plant roots contained 73 and 114% more total plant radioactivity (TPR) than the control at the 1 and 14-d translocation periods, respectively. Water stress significantly increased root starch and TPR percentage in the starch fraction, but had much smaller effects on root soluble-sugar concentration and TPR percentage of the root sugar fraction. Herbage regrowth mass following harvest and rewatering of the water-stressed plants was similar to that of the control. Compared to the control, water-stressed alfalfa has greater total nonstructural carbohydrates in the roots, apparently due to increased photoassimilate partitioning to the roots. However, the greater root carbohydrate concentrations did not result in compensatory herbage regrowth following rewatering.

  9. Association of AFLP and SCAR markers with common leafspot resistance in autotetraploid alfalfa (Medicago sativa).

    Science.gov (United States)

    Wang, Y; Bi, B; Yuan, Q H; Li, X L; Gao, J M

    2012-03-14

    To identify amplified fragment length polymorphism (AFLP) markers associated with resistance or susceptibility of alfalfa to common leafspot (CLS) caused by the fungus Pseudopeziza medicaginis (Dermateaceae), bulked segregant analysis was conducted based on an F(1(M × M)) population of 93 plants and a BC(1)S population of 91 plants. Three AFLP markers, ACTCAA(R206), TAGCAC(R185), and GGACTA(S264), were found to be associated with CLS resistance or susceptibility. All three markers were found at significantly different frequencies (71.9, 80.3 and 91.8%) compared to resistant or susceptible plants in the original population. Subsequently, these three AFLP markers were converted into three SCAR markers, ACTCAA(R136), TAGCAC(R128) and GGACTA(S254), which are easier to employ in breeding programs. The three SCAR markers were used in a randomly selected population with 50% resistance; the probability of finding one resistant plant was increased to 67.3, 66.7 and 90.0% with markers ACTCAA(R136), TAGCAC(R128) and GGACTA(S254), independently. If two of the SCAR markers were used simultaneously, the probability would be higher than 89%. The three SCAR markers identified in this study would be applicable for selection for CLS resistance in alfalfa breeding programs. Moreover, the genetic analysis indicated that CLS resistance in alfalfa is conferred by a single dominant gene.

  10. ECONOMIC ANALYSIS OF ALFALFA INTEGRATED MANAGEMENT PRACTICES

    OpenAIRE

    Ward, Clement E.; Dowdy, Alan K.; Berberet, Richard C.; Stritzke, Jimmie F.

    1990-01-01

    Integrated pest management (IMP) initially focused on insect pest control. More recently, IPM encompasses a broader concept of management, one which crosses several disciplinary boundaries. This article reports results of research dealing with four integrated management decisions for alfalfa (cultivar selection, inset control, weed control, and end-of-season harvest options.

  11. Overexpression of GsZFP1 enhances salt and drought tolerance in transgenic alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Tang, Lili; Cai, Hua; Ji, Wei; Luo, Xiao; Wang, Zhenyu; Wu, Jing; Wang, Xuedong; Cui, Lin; Wang, Yang; Zhu, Yanming; Bai, Xi

    2013-10-01

    GsZFP1 encodes a Cys2/His2-type zinc-finger protein. In our previous study, when GsZFP1 was heterologously expressed in Arabidopsis, the transgenic Arabidopsis plants exhibited enhanced drought and cold tolerance. However, it is still unknown whether GsZFP1 is also involved in salt stress. GsZFP1 is from the wild legume Glycine soja. Therefore, the aims of this study were to further elucidate the functions of the GsZFP1 gene under salt and drought stress in the forage legume alfalfa and to investigate its biochemical and physiological functions under these stress conditions. Our data showed that overexpression of GsZFP1 in alfalfa resulted in enhanced salt tolerance. Under high salinity stress, greater relative membrane permeability and malondialdehyde (MDA) content were observed and more free proline and soluble sugars accumulated in transgenic alfalfa than in the wild-type (WT) plants; in addition, the transgenic lines accumulated less Na(+) and more K(+) in both the shoots and roots. Overexpression of GsZFP1 also enhanced the drought tolerance of alfalfa. The fold-inductions of stress-responsive marker gene expression, including MtCOR47, MtRAB18, MtP5CS, and MtRD2, were greater in transgenic alfalfa than those of WT under drought stress conditions. In conclusion, the transgenic alfalfa plants generated in this study could be used for farming in salt-affected as well as arid and semi-arid areas. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  12. Morphogenesis of root nodules in white clover. II. The effect of mutation in genes nod IJ of the microsymbiont upon the nodule structure

    Directory of Open Access Journals (Sweden)

    Barbara Łotocka

    2014-01-01

    Full Text Available Morphogenesis of ineffective root nodules initiated on the roots of white clover 'Astra' by the Rhizobium leguminosarum biovar. trifolii strains ANU261 (Tn5 insertion in nod 1 gene and ANU262 (Tn5 insertion in nod J gene was investigated. Following changes were observed, as compared to the wild-type nodulation: the exaggerated, not delayed reaction of root hairs; the delay in nodulation with the number of nodules the same as in plants inoculated with a wild strain; the formation and organization of the nodule primordium not changed in comparison with the wild-type nodules; infection threads abnormally branched and diffusing with bacteria deprived of light zone and enriched with storage material; infected cells of bacteroidal tissue abnormally strongly osmiophilic and only slightly vacuolated; symbiosomes with very narrowed peribacteroidal space, subject to premature degradation; abnormal accumulation of starch in the nodule tissues; nodule development blocked at the stage of laterally situated meristem and single nodule bundle; inhibition of divisions in the meristem and vacuolation of its cells; the appearance of single cells with colonies of saprophytic rhizobia embedded in the fibrillar matrix in the old, degraded regions of the bacteroidal tissue.

  13. Water-tolerant legume nodulation.

    Science.gov (United States)

    Capoen, Ward; Goormachtig, Sofie; Holsters, Marcelle

    2010-03-01

    Water-tolerant nodulation is an adaptation of legumes that grow in wet or temporarily flooded habitats. This nodulation mode takes place at lateral root bases via intercellular bacterial invasion in cortical infection pockets. The tropical legume Sesbania rostrata has become a model for the study of the molecular basis of crack entry nodulation compared with root hair curl nodulation. For intercellular invasion, Nodulation Factor (NF) signalling recruits an ethylene-dependent, common Sym gene-independent pathway, leading to local cell death. The NF structure requirements are less stringent than for intracellular invasion in root hairs, which is correlated with a very specific NF-induced calcium spiking signature, presumably necessary for correct gene expression to assemble a functional entry complex in the epidermis.

  14. Genetic constraints that determine rhizobium-root nodule formation in Parasponia andersonii

    NARCIS (Netherlands)

    Seifi Kalhor, M.

    2016-01-01

    Bacteria of the genus Rhizobium play a very important role in agriculture by inducing nitrogen-fixing nodules on the roots of legumes. Root nodule symbiosis enables nitrogen‐fixing bacteria (Rhizobium) to convert atmospheric nitrogen into a form that is directly available for plant growth.

  15. Genetic constraints that determine rhizobium-root nodule formation in Parasponia andersonii

    NARCIS (Netherlands)

    Seifi Kalhor, M.

    2016-01-01

    Bacteria of the genus Rhizobium play a very important role in agriculture by inducing nitrogen-fixing nodules on the roots of legumes. Root nodule symbiosis enables nitrogen‐fixing bacteria (Rhizobium) to convert atmospheric nitrogen into a form that is directly available for plant growth.

  16. Medicago LYK3, an entry receptor in rhizobial nodulation factor signaling

    NARCIS (Netherlands)

    Smit, P.; Limpens, E.H.M.; Geurts, R.; Fedorova, E.; Dolgikh, E.; Gough, C.; Bisseling, T.

    2007-01-01

    Rhizobia secrete nodulation (Nod) factors, which set in motion the formation of nitrogen-fixing root nodules on legume host plants. Nod factors induce several cellular responses in root hair cells within minutes, but also are essential for the formation of infection threads by which rhizobia enter

  17. Epigenetic control of root and nodule development : the role of plant-specific histone deacetylases and LHP1 in root cell reprogramming

    NARCIS (Netherlands)

    Schilderink, S.

    2012-01-01

    In plants, unlike in animals, most organs develop post embryonically. These organs originate from clusters of undifferentiated dividing cells that form so-called meristems. Differentiated cells can be re-activated to enter the cell cycle and to ultimately give rise to new meristems. These differenti

  18. Epigenetic control of root and nodule development : the role of plant-specific histone deacetylases and LHP1 in root cell reprogramming

    NARCIS (Netherlands)

    Schilderink, S.

    2012-01-01

    In plants, unlike in animals, most organs develop post embryonically. These organs originate from clusters of undifferentiated dividing cells that form so-called meristems. Differentiated cells can be re-activated to enter the cell cycle and to ultimately give rise to new meristems. These

  19. Soil decontamination of 2,4,6- trinitrotoluene by alfalfa (Medicago sativa).

    Science.gov (United States)

    Fallahi, S; Habibi-Rezaei, M; Khayami, M; Heydari, R

    2007-12-15

    Present study investigate the toxicity effect of 2,4,6-trinitrotoluene (TNT) on a terrestrial plant, alfalfa (Medicago sativa) in artificial soils. In this study, TNT toxicity assessment was performed on spiked silica with this nitroaromatic compound by determination of the percent of emergence and shoots and roots biomasses at the concentration range of 3.2-10000 mg kg(-1) Dry Weight (DW). The emergence was reduced by 22-32% after 5 days of exposure at TNT concentrations up to 100 mg kg(-1) DW; shoot and root biomasses were reduced by 48-50 and 63-74%, respectively after 30 days exposure at TNT concentrations alfalfa cultivation for 30 days, TNT was partially transformed at the extent of 15-27%. This transformation decreased at higher TNT soil concentrations. TNT is taken up and metabolized by plants to its downstream derivatives.

  20. Nodulation of Mimosa spp. by the beta-proteobacterium Ralstonia taiwanensis.

    Science.gov (United States)

    Chen, Wen-Ming; James, Euan K; Prescott, Alan R; Kierans, Martin; Sprent, Janet I

    2003-12-01

    Several beta-proteobacteria have been isolated from legume root nodules and some of these are thought to be capable of nodulating and fixing N2. However, in no case has there been detailed studies confirming that they are the active symbionts. Here, Ralstonia taiwanensis LMG19424, which was originally isolated from Mimosa pudica nodules, was transformed to carry the green fluorescent protein (gfp) reporter gene before being used to inoculate axenically-grown seedlings of M. pudica and M. diplotricha. Plants were harvested at various intervals for 56 days after inoculation, then examined for evidence of infection and nodule formation. Nodulation of both Mimosa spp. was abundant, and acetylene reduction assays confirmed that nodules had nitrogenase activity. Confocal laser scanning microscopy (CLSM) showed that fresh M. pudica nodules with nitrogenase activity had infected cells containing bacteroids expressing gfp. In parallel, fixed and embedded nodules from both Mimosa spp. were sectioned for light and electron microscopy, followed by immunogold labeling with antibodies raised against gfp and nitrogenase Fe (nifH) protein. Significant immunolabeling with these antibodies confirmed that R. taiwanensis LMG19424 is an effective N2-fixing symbiont of Mimosa spp. Both species were infected via root hairs and, in all respects, the nodule ontogeny and development was similar to that described for other mimosoid legumes. The nodules were indeterminate with a persistent meristem, an invasion zone containing host cells being invaded via prominent infection threads, and an N2-fixing zone with infected cells containing membrane-bound symbiosomes.

  1. 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%.

  2. Nodulation and mycorrhization of transgenic soybean after glyphosate application

    Directory of Open Access Journals (Sweden)

    Aloisio Freitas Chagas Junior

    2013-12-01

    Full Text Available The sensitivity of nitrogen-fixing bacteria like Bradyrhizobium spp with in relation to glyphosate can be influenced by the concentration of the herbicide. This study aims to evaluate the effects of two formulations of glyphosate, in two application periods, on nodulation and mycorrhization in a soybean of variety RR P98Y11, using greenhouse, in pots with capacity of 4 L with savanna soils. Six treatments were utilized using Roundup Ultra and Roundup Original with one and two applications, witness without herbicide and witness only with mineral nitrogen, in a completely randomized design. Four evaluations were done, every 15 days, to determine the plant height, shoot and root dry weight, number of nodules, nodule dry weigh and mycorrhizae. It was observed an increase on the number of nodules, with the use of Roundup Ultra until 15 days after the first application. After this period, the witness only with inoculants showed a larger number of nodules. Roundup Ultra had positive influences on the dry weight of nodules, regardless the number of applications. There was no influence of glyphosate formulation considering the mycorrhizal colonization.

  3. ECOLOGICAL RISK ASSESSMENT OF ALFALFA (MEDICAGO VARIA L.) GENETICLALY ENGINEERED TO EXPRESS A HUMAN METALLOTHIONEIN (HMT) GENE

    Science.gov (United States)

    The objectives of these studies were two-fold: (1) to determine efficacy of low and high expression hMT gene constructs by assessing accumulation of Cu in shoots of parental and transgenic plants of alfalfa (Medicago varia L.) exposed to different concentrations of CuSO4 by addit...

  4. ECOLOGICAL RISK ASSESSMENT OF ALFALFA (MEDICAGO VARIA L.) GENETICLALY ENGINEERED TO EXPRESS A HUMAN METALLOTHIONEIN (HMT) GENE

    Science.gov (United States)

    The objectives of these studies were two-fold: (1) to determine efficacy of low and high expression hMT gene constructs by assessing accumulation of Cu in shoots of parental and transgenic plants of alfalfa (Medicago varia L.) exposed to different concentrations of CuSO4 by addit...

  5. 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.

  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. Final Report: Regulation and Function of Two Cell Wall protein Genes in Me Dicago Roots and Root Nodules, August 1, 1995 - January 31, 1999

    Energy Technology Data Exchange (ETDEWEB)

    Cooper, James B.

    2000-05-08

    During the period of DOE funding we synthesized several PRP peptides, generated rabbit antisera against two PRP repeats found in early nodulin PRPs, and developed confocal microscopy methods for root immunohistochemistry. Using the antibodies, we completed extensive descriptive studies of PRP deposition in medic and alfalfa roots showing that PRPs deposition is developmentally regulated in roots and spatially restricted within the walls of specific root tissues. Domain-specific antibodies were isolated from polyclonal sera using peptide affinity chromatography and were then used to demonstrate that nodule-specific epitopes are shared by several nodule-specific proteins. The following provides a more detailed summary of this work.

  8. 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.

  9. Prognosis of Thyroid Nodules in Individuals Living in the Zhitomir Region of Ukraine

    Science.gov (United States)

    Hayashida, Naomi; Sekitani, Yui; Takahashi, Jumpei; Kozlovsky, Alexander A.; Gutevych, Oleksandr K.; Saiko, Aleksey S.; Nirova, Nina V.; Petrova, Anjela A.; Rafalskiy, Ruslan M.; Chorny, Sergey A.; Daniliuk, Valery V.; Anami, Masanobu; Yamashita, Shunichi; Takamura, Noboru

    2012-01-01

    Objective After the accident at the Chernobyl Nuclear Power Plant (CNPP), the incidence of thyroid cancer increased among children. Recently, a strong relationship between solid thyroid nodules and the incidence of thyroid cancer was shown in atomic bomb survivors. To assess the prognosis of benign thyroid nodules in individuals living in the Zhitomir region of Ukraine, around the CNPP, we conducted a follow-up investigation of screening data from 1991 to 2000 in the Ukraine. Patients and Methods Participants of this study were 160 inhabitants with thyroid nodules (nodule group) and 160 inhabitants without thyroid nodules (normal control group) intially identified by ultrasonography from 1991 to 2000. All participants were aged 0 to 10 years old and lived in the same area at the time of the accident. We performed follow-up screening of participants and assessed thyroid nodules by fine needle aspiration biopsy. Results Among the nodule group participants, the number and size of nodules were significantly increased at the follow-up screening compared with the initial screening. No thyroid nodules were observed among the normal control group participants. The prevalence of thyroid abnormality, especially nodules that could be cancerous (malignant or suspicious by fine needle aspiration biopsy), was 7.5% in the nodule group and 0% in the normal control group (P<0.001). Conclusions Our study indicated that a thyroid nodule in childhood is a prognostic factor associated with an increase in the number and size of nodules in individuals living in the Zhitomir region of Ukraine. PMID:23209797

  10. Prognosis of thyroid nodules in individuals living in the Zhitomir region of Ukraine.

    Directory of Open Access Journals (Sweden)

    Naomi Hayashida

    Full Text Available OBJECTIVE: After the accident at the Chernobyl Nuclear Power Plant (CNPP, the incidence of thyroid cancer increased among children. Recently, a strong relationship between solid thyroid nodules and the incidence of thyroid cancer was shown in atomic bomb survivors. To assess the prognosis of benign thyroid nodules in individuals living in the Zhitomir region of Ukraine, around the CNPP, we conducted a follow-up investigation of screening data from 1991 to 2000 in the Ukraine. PATIENTS AND METHODS: Participants of this study were 160 inhabitants with thyroid nodules (nodule group and 160 inhabitants without thyroid nodules (normal control group intially identified by ultrasonography from 1991 to 2000. All participants were aged 0 to 10 years old and lived in the same area at the time of the accident. We performed follow-up screening of participants and assessed thyroid nodules by fine needle aspiration biopsy. RESULTS: Among the nodule group participants, the number and size of nodules were significantly increased at the follow-up screening compared with the initial screening. No thyroid nodules were observed among the normal control group participants. The prevalence of thyroid abnormality, especially nodules that could be cancerous (malignant or suspicious by fine needle aspiration biopsy, was 7.5% in the nodule group and 0% in the normal control group (P<0.001. CONCLUSIONS: Our study indicated that a thyroid nodule in childhood is a prognostic factor associated with an increase in the number and size of nodules in individuals living in the Zhitomir region of Ukraine.

  11. CLE Peptides Control Medicago truncatula Nodulation Locally and Systemically1[C][W][OA

    Science.gov (United States)

    Mortier, Virginie; Den Herder, Griet; Whitford, Ryan; Van de Velde, Willem; Rombauts, Stephane; D'haeseleer, Katrien; Holsters, Marcelle; Goormachtig, Sofie

    2010-01-01

    The CLAVATA3/embryo-surrounding region (CLE) peptides control the fine balance between proliferation and differentiation in plant development. We studied the role of CLE peptides during indeterminate nodule development and identified 25 MtCLE peptide genes in the Medicago truncatula genome, of which two genes, MtCLE12 and MtCLE13, had nodulation-related expression patterns that were linked to proliferation and differentiation. MtCLE13 expression was up-regulated early in nodule development. A high-to-low expression gradient radiated from the inner toward the outer cortical cell layers in a region defining the incipient nodule. At later stages, MtCLE12 and MtCLE13 were expressed in differentiating nodules and in the apical part of mature, elongated nodules. Functional analysis revealed a putative role for MtCLE12 and MtCLE13 in autoregulation of nodulation, a mechanism that controls the number of nodules and involves systemic signals mediated by a leucine-rich repeat receptor-like kinase, SUNN, which is active in the shoot. When MtCLE12 and MtCLE13 were ectopically expressed in transgenic roots, nodulation was abolished at the level of the nodulation factor signal transduction, and this inhibition involved long-distance signaling. In addition, composite plants with roots ectopically expressing MtCLE12 or MtCLE13 had elongated petioles. This systemic effect was not observed in transgenic roots ectopically expressing MtCLE12 and MtCLE13 in a sunn-1 mutant background, although nodulation was still strongly reduced. These results suggest multiple roles for CLE signaling in nodulation. PMID:20348212

  12. microRNA160 dictates stage-specific auxin and cytokinin sensitivities and directs soybean nodule development.

    Science.gov (United States)

    Nizampatnam, Narasimha Rao; Schreier, Spencer John; Damodaran, Suresh; Adhikari, Sajag; Subramanian, Senthil

    2015-10-01

    Legume nodules result from coordinated interactions between the plant and nitrogen-fixing rhizobia. The phytohormone cytokinin promotes nodule formation, and recent findings suggest that the phytohormone auxin inhibits nodule formation. Here we show that microRNA160 (miR160) is a key signaling element that determines the auxin/cytokinin balance during nodule development in soybean (Glycine max). miR160 appears to promote auxin activity by suppressing the levels of the ARF10/16/17 family of repressor ARF transcription factors. Using quantitative PCR assays and a fluorescence miRNA sensor, we show that miR160 levels are relatively low early during nodule formation and high in mature nodules. We had previously shown that ectopic expression of miR160 in soybean roots led to a severe reduction in nodule formation, coupled with enhanced sensitivity to auxin and reduced sensitivity to cytokinin. Here we show that exogenous cytokinin restores nodule formation in miR160 over-expressing roots. Therefore, low miR160 levels early during nodule development favor cytokinin activity required for nodule formation. Suppression of miR160 levels using a short tandem target mimic (STTM160) resulted in reduced sensitivity to auxin and enhanced sensitivity to cytokinin. In contrast to miR160 over-expressing roots, STTM160 roots had increased nodule formation, but nodule maturation was significantly delayed. Exogenous auxin partially restored proper nodule formation and maturation in STTM160 roots, suggesting that high miR160 activity later during nodule development favors auxin activity and promotes nodule maturation. Therefore, miR160 dictates developmental stage-specific sensitivities to auxin and cytokinin to direct proper nodule formation and maturation in soybean.

  13. Simplification of a haemolytic micromethod for toxic saponin quantification in alfalfa

    Directory of Open Access Journals (Sweden)

    Piotr M. Górski

    2013-12-01

    Full Text Available A simplification of a haemolytic micromethod is presented. In the orginal method, alfalfa (Medicago media Pers. leaf sap is spotted on the plate covered with a blood-gelatine suspension. In the presented method, the mashed alfalfa pulp is used instead of sap. Due to saponin diffusion and the reaction with erythrocytes, a haemolytic ring appears, which has a width proportional to the concentration of toxic saponins. It is shown that the width of haemolytic ring does not depend on the sample weight ranging from 20 to 100 mg. This allows for the omission of laborious sap pressing and sample quantification. Individual alfalfa plants with different saponin contents were tested using leaf sap and leaf pulp for analyses. Good agreement was found with sap and leaf pulp methods. The correlation obtained by both methods was high, r = 0.87. The modified method requires only a small amount of plant material and makes the analyses of large numbers of individual plants per day possible. The method is especially recommended for breeding purposes.

  14. Soybean nodule-enhanced CLE peptides in roots act as signals in GmNARK-mediated nodulation suppression.

    Science.gov (United States)

    Lim, Chae Woo; Lee, Young Woo; Hwang, Cheol Ho

    2011-09-01

    The number of nodules formed in the roots of leguminous plants is systemically controlled by autoregulation of nodulation (AON). This study characterized two of the CLAVATA3/endosperm-surrounding region (CLE) genes involved in AON signal transduction. The GmRIC1 and GmRIC2 genes initiated expression solely in the roots at approximately 3 days after inoculation (DAI) with Nod factor-producing rhizobia, corresponding to the time point of AON, and the expression was up-regulated by cytokinins. Levels of GmRIC1 and GmRIC2 gene expression were much higher in the supernodulation mutant, SS2-2, than in wild-type (WT) soybeans during nodule development, even after initiation of nitrogen fixation. At 3 DAI, GmRIC2 was induced in the cells of the pericycle and the outer cortex, which undergo cell division to form nodule primordia and spreads from the central region to the whole nodule as it develops. Overexpression of GmRIC1 and GmRIC2 strongly suppressed the nodulation of WT roots as well as transgenic hairy roots in a GmNARK-dependent manner. This systemic suppression of nodulation was caused by the secretion of two CLE proteins into the extracellular space. Double grafting between WT and SS2-2 soybeans showed that signal Q is larger in SS2-2 than in WT roots during nodulation. The results of this study suggest that GmRIC1 and GmRIC2 are good candidates for root-derived signal Q in AON signal transduction.

  15. Rhizobium nod factors reactivate the cell cycle during infection and nodule primordium formation, but the cycle is only completed in primordium formation.

    NARCIS (Netherlands)

    Yang, W.C.; Blank, de C.; Meskiene, I.; Hirt, H.; Bakker, J.; Kammen, van A.; Franssen, H.; Bisseling, T.

    1994-01-01

    Rhizobia induce the formation of root nodules on the roots of leguminous plants. In temperate legumes, nodule organogenesis starts with the induction of cell divisions in regions of the root inner cortex opposite protoxylem poles, resulting in the formation of nodule primordia. It has been postulate

  16. Distribution and utilization of nitrogenated compounds explanted by the soybean nodules by plants during seeds developing; Distribuicao e utilizacao dos compostos nitrogenados exportados pelos nodulos de soja em plantas em fase de enchimento de graos

    Energy Technology Data Exchange (ETDEWEB)

    Alencar, Severino Matias de [Universidade Federal Rural do Rio de Janeiro, RJ (Brazil). Dept. de Solos; Neves, Maria Cristina Prata; Rumjaneck, Norma Gouvea [Empresa Brasileira de Pesquisa Agropecuaria, Seropedica, RJ (Brazil). E-mail: agrob@cnps.embrapa.br

    1997-07-01

    An experiment was carried out, using radioisotopes, for evaluation of the leaf, schuck and seeds areas and, examination of the pattern which is used by the nitrogenated compounds explanted by the soybean nodules.

  17. A legume genetic framework controls infection of nodules by symbiotic and endophytic bacteria.

    Science.gov (United States)

    Zgadzaj, Rafal; James, Euan K; Kelly, Simon; Kawaharada, Yasuyuki; de Jonge, Nadieh; Jensen, Dorthe B; Madsen, Lene H; Radutoiu, Simona

    2015-06-01

    Legumes have an intrinsic capacity to accommodate both symbiotic and endophytic bacteria within root nodules. For the symbionts, a complex genetic mechanism that allows mutual recognition and plant infection has emerged from genetic studies under axenic conditions. In contrast, little is known about the mechanisms controlling the endophytic infection. Here we investigate the contribution of both the host and the symbiotic microbe to endophyte infection and development of mixed colonised nodules in Lotus japonicus. We found that infection threads initiated by Mesorhizobium loti, the natural symbiont of Lotus, can selectively guide endophytic bacteria towards nodule primordia, where competent strains multiply and colonise the nodule together with the nitrogen-fixing symbiotic partner. Further co-inoculation studies with the competent coloniser, Rhizobium mesosinicum strain KAW12, show that endophytic nodule infection depends on functional and efficient M. loti-driven Nod factor signalling. KAW12 exopolysaccharide (EPS) enabled endophyte nodule infection whilst compatible M. loti EPS restricted it. Analysis of plant mutants that control different stages of the symbiotic infection showed that both symbiont and endophyte accommodation within nodules is under host genetic control. This demonstrates that when legume plants are exposed to complex communities they selectively regulate access and accommodation of bacteria occupying this specialized environmental niche, the root nodule.

  18. Abscisic acid coordinates nod factor and cytokinin signaling during the regulation of nodulation in Medicago truncatula.

    Science.gov (United States)

    Ding, Yiliang; Kalo, Peter; Yendrek, Craig; Sun, Jongho; Liang, Yan; Marsh, John F; Harris, Jeanne M; Oldroyd, Giles E D

    2008-10-01

    Nodulation is tightly regulated in legumes to ensure appropriate levels of nitrogen fixation without excessive depletion of carbon reserves. This balance is maintained by intimately linking nodulation and its regulation with plant hormones. It has previously been shown that ethylene and jasmonic acid (JA) are able to regulate nodulation and Nod factor signal transduction. Here, we characterize the nature of abscisic acid (ABA) regulation of nodulation. We show that application of ABA inhibits nodulation, bacterial infection, and nodulin gene expression in Medicago truncatula. ABA acts in a similar manner as JA and ethylene, regulating Nod factor signaling and affecting the nature of Nod factor-induced calcium spiking. However, this action is independent of the ethylene signal transduction pathway. We show that genetic inhibition of ABA signaling through the use of a dominant-negative allele of ABSCISIC ACID INSENSITIVE1 leads to a hypernodulation phenotype. In addition, we characterize a novel locus of M. truncatula, SENSITIVITY TO ABA, that dictates the sensitivity of the plant to ABA and, as such, impacts the regulation of nodulation. We show that ABA can suppress Nod factor signal transduction in the epidermis and can regulate cytokinin induction of the nodule primordium in the root cortex. Therefore, ABA is capable of coordinately regulating the diverse developmental pathways associated with nodule formation and can intimately dictate the nature of the plants' response to the symbiotic bacteria.

  19. Manganese nanoparticles: impact on non-nodulated plant as a potent enhancer in nitrogen metabolism and toxicity study both in vivo and in vitro.

    Science.gov (United States)

    Pradhan, Saheli; Patra, Prasun; Mitra, Shouvik; Dey, Kushal Kumar; Jain, Sneha; Sarkar, Samapd; Roy, Shuvrodeb; Palit, Pratip; Goswami, Arunava

    2014-09-03

    Mung bean plants were grown under controlled conditions and supplemented with macro- and micronutrients. The objective of this study was to determine the response of manganese nanoparticles (MnNP) in nitrate uptake, assimilation, and metabolism compared with the commercially used manganese salt, manganese sulfate (MS). MnNP was modulated to affect the assimilatory process by enhancing the net flux of nitrogen assimilation through NR-NiR and GS-GOGAT pathways. This study was associated with toxicological investigation on in vitro and in vivo systems to promote MnNP as nanofertilizer and can be used as an alternative to MS. MnNP did not impart any toxicity to the mice brain mitochondria except in the partial inhibition of complex II-III activity in ETC. Therefore, mitochondrial dysfunction and neurotoxicity, which were noted by excess usage of elemental manganese, were prevented. This is the first attempt to highlight the nitrogen uptake, assimilation, and metabolism in a plant system using a nanoparticle to promote a biosafe nanomicronutrient-based crop management.

  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. Determining the Site of Action of Strigolactones during Nodulation.

    Science.gov (United States)

    McAdam, Erin L; Hugill, Cassandra; Fort, Sebastien; Samain, Eric; Cottaz, Sylvain; Davies, Noel W; Reid, James B; Foo, Eloise

    2017-09-01

    Strigolactones (SLs) influence the ability of legumes to associate with nitrogen-fixing bacteria. In this study, we determine the precise stage at which SLs influence nodulation. We show that SLs promote infection thread formation, as a null SL-deficient pea (Pisum sativum) mutant forms significantly fewer infection threads than wild-type plants, and this reduction can be overcome by the application of the synthetic SL GR24. We found no evidence that SLs influence physical events in the plant before or after infection thread formation, since SL-deficient plants displayed a similar ability to induce root hair curling in response to rhizobia or Nod lipochitooligosaccharides (LCOs) and SL-deficient nodules appear to fix nitrogen at a similar rate to those of wild-type plants. In contrast, an SL receptor mutant displayed no decrease in infection thread formation or nodule number, suggesting that SL deficiency may influence the bacterial partner. We found that this influence of SL deficiency was not due to altered flavonoid exudation or the ability of root exudates to stimulate bacterial growth. The influence of SL deficiency on infection thread formation was accompanied by reduced expression of some early nodulation genes. Importantly, SL synthesis is down-regulated by mutations in genes of the Nod LCO signaling pathway, and this requires the downstream transcription factor NSP2 but not NIN This, together with the fact that the expression of certain SL biosynthesis genes can be elevated in response to rhizobia/Nod LCOs, suggests that Nod LCOs may induce SL biosynthesis. SLs appear to influence nodulation independently of ethylene action, as SL-deficient and ethylene-insensitive double mutant plants display essentially additive phenotypes, and we found no evidence that SLs influence ethylene synthesis or vice versa. © 2017 American Society of Plant Biologists. All Rights Reserved.

  2. Nodulation gene regulation and quorum sensing control density-dependent suppression and restriction of nodulation in the Bradyrhizobium japonicum-soybean symbiosis.

    Science.gov (United States)

    Jitacksorn, Siriluck; Sadowsky, Michael J

    2008-06-01

    The nodulation of Glycine max cv. Lambert and the nodulation-restricting plant introduction (PI) genotype PI 417566 by wild-type Bradyrhizobium japonicum USDA110 is regulated in a population-density-dependent manner. Nodulation on both plant genotypes was suppressed (inhibited) when plants received a high-density inoculum (10(9) cells/ml) of strain USDA110 grown in complex medium, and more nodules were produced on plants receiving a low-cell-density inoculum (10(5) cells/ml). Since cell-free supernatants from strain USDA110 grown to high cell density in complex medium decreased the expression of an nodY-lacZ fusion, this phenomenon was attributed to bradyoxetin-induced repression of nod gene expression. Inoculation of either the permissive soybean genotype (cv. Lambert) or PI 417566 with 10(9) cells/ml of the nodD2, nolA, nodW, and nwsB mutants of USDA110 enhanced nodulation (up to 24%) relative to that seen with inoculations done with 10(5) cells/ml of the mutants or the wild-type strain, indicating that these genes are involved in population-density-dependent nodulation of soybeans. In contrast, the number of nodules produced by an nodD1 mutant on either soybean genotype was less than those seen with the wild-type strain inoculated at a low inoculum density. The nodD2 mutant outcompeted B. japonicum strain USDA123 for nodulation of G. max cv. Lambert at a high or low inoculum density, and the results of root-tip-marking and time-to-nodulate studies indicated that the nolA and nodD2 mutants nodulated this soybean genotype faster than wild-type USDA110. Taken together, the results from these studies indicate that the nodD2 mutant of B. japonicum may be useful to enhance soybean nodulation at high inoculum densities and that NodD2 is a key repressor influencing host-controlled restriction of nodulation, density-dependent suppression of nodulation, perception of bradyoxetin, and competitiveness in the soybean-B. japonicum symbiosis.

  3. Truncated hemoglobins in actinorhizal nodules of Datisca glomerata

    NARCIS (Netherlands)

    Pawlowski, K.; Jacobsen, K.R.; Alloisio, N.; Denison, R.F.; Klein, M.; Tjepkema, J.D.; Winzer, T.; Sirrenberg, A.; Guan, C.; Berry, A.M.

    2007-01-01

    Three types of hemoglobins exist in higher plants, symbiotic, non-symbiotic, and truncated hemoglobins. Symbiotic (class II) hemoglobins play a role in oxygen supply to intracellular nitrogen-fixing symbionts in legume root nodules, and in one case ( Parasponia Sp.), a non-symbiotic (class I) hemogl

  4. Nodulation genes and type III secretion systems in rhizobia

    Science.gov (United States)

    For establishment of symbiosis, rhizobia and legumes have to communicate. Specific signaling starts with the release of flavonoids by the plant. All rhizobia encode at least one NodD protein, which responds to the presence of specific flavonoids by activation of nodulation genes. In Bradyrhizobium j...

  5. Truncated hemoglobins in actinorhizal nodules of Datisca glomerata

    NARCIS (Netherlands)

    Pawlowski, K.; Jacobsen, K.R.; Alloisio, N.; Denison, R.F.; Klein, M.; Tjepkema, J.D.; Winzer, T.; Sirrenberg, A.; Guan, C.; Berry, A.M.

    2007-01-01

    Three types of hemoglobins exist in higher plants, symbiotic, non-symbiotic, and truncated hemoglobins. Symbiotic (class II) hemoglobins play a role in oxygen supply to intracellular nitrogen-fixing symbionts in legume root nodules, and in one case ( Parasponia Sp.), a non-symbiotic (class I)

  6. Legume nodule senescence: roles for redox and hormone signalling in the orchestration of the natural aging process.

    Science.gov (United States)

    Puppo, Alain; Groten, Karin; Bastian, Fabiola; Carzaniga, Raffaella; Soussi, Mariam; Lucas, M Mercedes; de Felipe, Maria Rosario; Harrison, Judith; Vanacker, Hélène; Foyer, Christine H

    2005-03-01

    Research on legume nodule development has contributed greatly to our current understanding of plant-microbe interactions. However, the factors that orchestrate root nodule senescence have received relatively little attention. Accumulating evidence suggests that redox signals contribute to the establishment of symbiosis and senescence. Although degenerative in nature, nodule senescence is an active process programmed in development in which reactive oxygen species (ROS), antioxidants, hormones and proteinases have key roles. Nodules have high levels of the redox buffers, ascorbate and glutathione, which are important in the nodulation process and in senescence. These metabolites decline with N-fixation as the nodule ages but the resultant decrease in redox buffering capacity does not necessarily lead to enhanced ROS or oxidative stress. We propose models by which ROS and antioxidants interact with hormones such as abscisic acid in the orchestration of nodule senescence.

  7. The Class II trehalose 6-phosphate synthase gene PvTPS9 modulates trehalose metabolism in Phaseolus vulgaris nodules.

    Directory of Open Access Journals (Sweden)

    Aarón Barraza

    2016-11-01

    Full Text Available Legumes form symbioses with rhizobia, producing nitrogen-fixing nodules on the roots of the plant host. The network of plant signaling pathways affecting carbon metabolism may determine the final number of nodules. The trehalose biosynthetic pathway regulates carbon metabolism and plays a fundamental role in plant growth and development, as well as in plant-microbe interactions. The expression of genes for trehalose synthesis during nodule development suggests that this metabolite may play a role in legume-rhizobia symbiosis. In this work, PvTPS9, which encodes a Class II trehalose-6-phosphate synthase (TPS of common bean (Phaseolus vulgaris, was silenced by RNA interference in transgenic nodules. The silencing of PvTPS9 in root nodules resulted in a reduction of 85% (± 1% of its transcript, which correlated with a 30% decrease in trehalose contents of transgenic nodules and in untransformed leaves. Composite transgenic plants with PvTPS9 silenced in the roots showed no changes in nodule number and nitrogen fixation, but a severe reduction in plant biomass and altered transcript profiles of all Class II TPS genes. Our data suggest that PvTPS9 plays a key role in modulating trehalose metabolism in the symbiotic nodule and, therefore, in the whole plant.

  8. Urgency of further developing alfalfa industry in China%我国苜蓿产业亟待振兴

    Institute of Scientific and Technical Information of China (English)

    孙启忠; 玉柱; 徐春城

    2012-01-01

    resources, which would benefit the establishment of alfalfa industry belt and increased the comprehensively productive ability and supply ability by optimizing regional distribution, planting scale, exercising mechanization and rationally managing with standard.

  9. Genetic characterization of fast-growing rhizobia able to nodulate Prosopis alba in North Spain.

    Science.gov (United States)

    Iglesias, Olga; Rivas, Raúl; García-Fraile, Paula; Abril, Adriana; Mateos, Pedro F; Martinez-Molina, Eustoquio; Velázquez, Encarna

    2007-12-01

    Prosopis is a Mimosaceae legume tree indigenous to South America and not naturalized in Europe. In this work 18 rhizobial strains nodulating Prosopis alba roots were isolated from a soil in North Spain that belong to eight different randomly amplified polymorphic DNA groups phylogenetically related to Sinorhizobium medicae, Sinorhizobium meliloti and Rhizobium giardinii according to their intergenic spacer and 16S rRNA gene sequences. The nodC genes of isolates close to S. medicae and S. meliloti were identical to those of S. medicae USDA 1,037(T) and S. meliloti LMG 6,133(T) and accordingly all these strains were able to nodulate both alfalfa and Prosopis. These nodC genes were phylogenetically divergent from those of the isolates close to R. giardinii that were identical to that of R. giardinii H152(T) and therefore all these strains formed nodules in common beans and Prosopis. The nodC genes of the strains isolated in Spain were phylogenetically divergent from that carried by Mesorhizobium chacoense Pr-5(T) and Sinorhizobium arboris LMG 1,4919(T) nodulating Prosopis in America and Africa, respectively. Therefore, Prosopis is a promiscuous host which can establish symbiosis with strains carrying very divergent nodC genes and this promiscuity may be an important advantage for this legume tree to be used in reforestation.

  10. PENINGKATAN PRODUKSI DAN KECERNAAN BAHAN KERING ALFALFA DENGAN PEMUPUKAN FOSFAT

    Directory of Open Access Journals (Sweden)

    - Sunarni

    2014-06-01

    Full Text Available Alfalfa (Medicago    sativa    L. as one of legume forage crops for ruminant livestock is very sensitive to the phosphorus deficiency. Phosphorus fertilizers have been used widely to overcome those problem. However, the high cost of superphosphate (SP is now focusing attention on cheaper rock phosphate (RP fertilizer. A field experiment was conducted on a latosolic soil (low pH, and low available Bray II extractable P. The objective of the research is to evaluate the effect of phosphorus fertilizer from difference sources on dry matter production, and in    vitro dry matter digestibility of alfalfa. Completely randomized block design with 7 treatments in 3 replicates was used in this field experiment. The treatments were T0 (control, T1 (SP, 100 kg P2O5/ha, T2 (SP, 200 kg P2O5/ha, T3 (SP, 300 kg P2O5/ha, T4 (RP, 100 kg P2O5/ha, T5 (RP, 200 kg P2O5/ha, T6 (RP, 300 kg P2O5/ha. Size of each plot was 2 m x 2 m, and fertilized with P fertilizer (SP, RP according to the assigned treatment. All plot received basal fertilizer of urea (50 kg N/ha, KCl (100 kg K2O/ha and poultry manure (2 ton/ha. Medicago    sativa was defoliated on 12 weeks after planting and analyzed for dry matter (DM production and in    vitro DM digestibility. Result showed that DM production significantly influenced by the treatments. Superphosphate fertilization resulted DM production significantly higher compared to control and RP, except on RP 300 kg P2O5/ha was non-significant difference. In    vitro dry matter digestibility (IVDMD of alfalfa was not affected by the treatment. There was non-significant difference in IVDMD between the treatments. Therefore, RP (300 kg P2O5/ha could replace SP to increased DM production and not affected for IVDMD of Medicago    sativa in acid latosolic soil.

  11. Novel paralogous gene families with potential function in legume nodules and seeds.

    Science.gov (United States)

    Silverstein, Kevin A T; Graham, Michelle A; VandenBosch, Kathryn A

    2006-04-01

    Within the plant kingdom, legumes are unusual in their ability to form nitrogen-fixing nodules in symbiosis with certain bacteria in the family Rhizobiaceae (rhizhobia). Genes that are required for signaling between plant and symbiont, and for the development and maintenance of the nodule, were either created de novo or adopted from other plant pathways. Only in recent years have genome-scale sequence data from legumes made it possible to identify large, novel families of genes probably evolved to function in nodulation. Members of these novel families are expressed in seeds or nodules, and are homologous to defense-related proteins. Perhaps the most striking example is a large family (of more than 340 members) of cysteine cluster proteins that have homology to plant defensins.

  12. Study of root para-nodules formation in wheat (Triticum durum ...

    African Journals Online (AJOL)

    djemel

    2013-08-28

    Aug 28, 2013 ... Frankia strains can induce N2-fixing root nodules on certain non-leguminous plants. ... glauca and Alnus glutinosa, and the soil bacteria Frankia that lead to the .... morphology of nitrogen fixing paranodules on wheat roots.

  13. The integral membrane protein SEN1 is required for symbiotic nitrogen fixation in Lotus japonicus nodules.

    Science.gov (United States)

    Hakoyama, Tsuneo; Niimi, Kaori; Yamamoto, Takeshi; Isobe, Sawa; Sato, Shusei; Nakamura, Yasukazu; Tabata, Satoshi; Kumagai, Hirotaka; Umehara, Yosuke; Brossuleit, Katja; Petersen, Thomas R; Sandal, Niels; Stougaard, Jens; Udvardi, Michael K; Tamaoki, Masanori; Kawaguchi, Masayoshi; Kouchi, Hiroshi; Suganuma, Norio

    2012-01-01

    Legume plants establish a symbiotic association with bacteria called rhizobia, resulting in the formation of nitrogen-fixing root nodules. A Lotus japonicus symbiotic mutant, sen1, forms nodules that are infected by rhizobia but that do not fix nitrogen. Here, we report molecular identification of the causal gene, SEN1, by map-based cloning. The SEN1 gene encodes an integral membrane protein homologous to Glycine max nodulin-21, and also to CCC1, a vacuolar iron/manganese transporter of Saccharomyces cerevisiae, and VIT1, a vacuolar iron transporter of Arabidopsis thaliana. Expression of the SEN1 gene was detected exclusively in nodule-infected cells and increased during nodule development. Nif gene expression as well as the presence of nitrogenase proteins was detected in rhizobia from sen1 nodules, although the levels of expression were low compared with those from wild-type nodules. Microscopic observations revealed that symbiosome and/or bacteroid differentiation are impaired in the sen1 nodules even at a very early stage of nodule development. Phylogenetic analysis indicated that SEN1 belongs to a protein clade specific to legumes. These results indicate that SEN1 is essential for nitrogen fixation activity and symbiosome/bacteroid differentiation in legume nodules.

  14. The MicroRNA390/TAS3 Pathway Mediates Symbiotic Nodulation and Lateral Root Growth.

    Science.gov (United States)

    Hobecker, Karen Vanesa; Reynoso, Mauricio Alberto; Bustos-Sanmamed, Pilar; Wen, Jiangqi; Mysore, Kirankumar S; Crespi, Martín; Blanco, Flavio Antonio; Zanetti, María Eugenia

    2017-08-01

    Legume roots form two types of postembryonic organs, lateral roots and symbiotic nodules. Nodule formation is the result of the interaction of legumes with rhizobia and requires the mitotic activation and differentiation of root cells as well as an independent, but coordinated, program that allows infection by rhizobia. MicroRNA390 (miR390) is an evolutionarily conserved microRNA that targets the Trans-Acting Short Interference RNA3 (TAS3) transcript. Cleavage of TAS3 by ARGONAUTE7 results in the production of trans-acting small interference RNAs, which target mRNAs encoding AUXIN RESPONSE FACTOR2 (ARF2), ARF3, and ARF4. Here, we show that activation of the miR390/TAS3 regulatory module by overexpression of miR390 in Medicago truncatula promotes lateral root growth but prevents nodule organogenesis, rhizobial infection, and the induction of two key nodulation genes, Nodulation Signaling Pathway1 (NSP1) and NSP2 Accordingly, inactivation of the miR390/TAS3 module, either by expression of a miR390 target mimicry construct or mutations in ARGONAUTE7, enhances nodulation and rhizobial infection, alters the spatial distribution of the nodules, and increases the percentage of nodules with multiple meristems. Our results revealed a key role of the miR390/TAS3 pathway in legumes as a modulator of lateral root organs, playing opposite roles in lateral root and nodule development. © 2017 American Society of Plant Biologists. All Rights Reserved.

  15. ALFALFA: HI Cosmology in the Local Universe

    CERN Document Server

    Giovanelli, Riccardo

    2007-01-01

    For the last 25 years, the 21 cm line has been used productively to investigate the large-scale structure of the Universe, its peculiar velocity field and the measurement of cosmic parameters. In February 2005 a blind HI survey that will cover 7074 square degrees of the high latitude sky was started at Arecibo, using the 7-beam feed L-band feed array (ALFA). Known as the Arecibo Legacy Fast ALFA (ALFALFA) Survey, the program is producing a census of HI-bearing objects over a cosmologically significant volume of the local Universe. With respect to previous blind HI surveys, ALFALFA offers an improvement of about one order of magnitude in sensitivity, 4 times the angular resolution, 3 times the spectral resolution, and 1.6 times the total bandwidth of HIPASS. ALFALFA can detect 7 X 10**4 D**2 solar masses of HI, where D is the source distance in Mpc. As of mid 2007, 44% of the survey observations and 15% of the source extraction are completed. We discuss the status of the survey and present a few preliminary re...

  16. Characterization of Feeding Injuries Caused by Ceresa nigripectus Remes Lenicov (Hemiptera: Membracidae) on Alfalfa Stems.

    Science.gov (United States)

    Grosso, T P; Mercado, M I; Ponessa, G I; Conci, L R; Virla, E G

    2016-04-01

    Piercing-sucking insects cause mechanical and physiological injury to plants. Ceresa nigripectus Remes Lenicov is a pest of alfalfa in subtropical regions of South America and a carrier of the ArAWB phytoplasma. The aim of this study was to determine the feeding habits of this treehopper and to describe the effects of the feeding injuries on stem vascular tissues in alfalfa. Adults and nymphs of C. nigripectus inserted their stylets repeatedly girdling the stem. One week after feeding, alfalfa stems exhibited numerous feeding canals with salivary deposits, most of which reached the phloem. Two weeks after feeding, cortex and phloem cells next to the salivary sheath collapsed, mature tracheal elements became sparse and appeared with an increased cross-section area, and phenolic compounds increased in cells and cell walls compared to undamaged plants. Three weeks after feeding, an annular callus, formed by abnormal cell division and hypertrophy of preexisting cortex and vascular cambium cells, appeared immediately above the stem girdle. Parenchyma cells from the outer layers of the callus differentiated to form secondary anomalous amphicribal bundles in the wound. The aerial parts above the stem girdle eventually withered and died.

  17. 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.

  18. Expression of a complete soybean leghemoglobin gene in root nodules of transgenic Lotus corniculatus.

    Science.gov (United States)

    Stougaard, J; Petersen, T E; Marcker, K A

    1987-08-01

    The complete soybean leghemoglobin lbc(3) gene was transferred into the legume Lotus corniculatus using an Agrobacterium rhizogenes vector system. Organ-specific expression of the soybean gene was observed in root nodules formed on regenerated transgenic plants after infection with Rhizobium loti. The primary transcript was processed in the same way as in soybean nodules and the resulting mRNA was translated into Lbc(3) protein. Quantitative determination of the Lbc(3) protein in nodules of transgenic plants indicated that the steady-state level of the soybean protein is comparable to that of endogenous Lotus leghemoglobin.

  19. Nitric oxide (NO): a key player in the senescence of Medicago truncatula root nodules.

    Science.gov (United States)

    Cam, Yvan; Pierre, Olivier; Boncompagni, Eric; Hérouart, Didier; Meilhoc, Eliane; Bruand, Claude

    2012-10-01

    Nitric oxide (NO) is a signalling and defence molecule involved in diverse plant developmental processes, as well as in the plant response to pathogens. NO has also been detected at different steps of the symbiosis between legumes and rhizobia. NO is required for an optimal establishment of the Medicago truncatula-Sinorhizobium meliloti symbiotic interaction, but little is known about the role of NO in mature nodules. Here, we investigate the role of NO in the late steps of symbiosis. Genetic and pharmacological approaches were conducted to modulate the NO level inside root nodules, and their effects on nitrogen fixation and root nodule senescence were monitored. An increase in endogenous NO levels led to a decrease in nitrogen fixation and early nodule senescence, characterized by cytological modifications of the nodule structure and the early expression of a specific senescence marker. By contrast, a decrease in NO levels led to a delay in nodule senescence. Together, our results strongly suggest that NO is a signal in developmental as well as stress-induced nodule senescence. In addition, this work demonstrates the pivotal role of the bacterial NO detoxification response in the prevention of early nodule senescence, and hence the maintenance of efficient symbiosis.

  20. Genome Wide Identification and Expression Profiling of Ethylene Receptor Genes during Soybean Nodulation.

    Science.gov (United States)

    Wang, Youning; Yuan, Jinhong; Yang, Wei; Zhu, Lin; Su, Chao; Wang, Xiaodi; Wu, Haiyan; Sun, Zhengxi; Li, Xia

    2017-01-01

    It has long been known that the gaseous plant hormone ethylene plays a key role in nodulation in legumes. The perception of ethylene by a family of five membrane-localized receptors is necessary to trigger the ethylene signaling pathway, which regulates various biological responses in Arabidopsis. However, a systematic analysis of the ethylene receptors in leguminous plants and their roles in nodule development is lacking. In this study, we performed a characterization of ethylene receptor genes based on the latest Glycine max genome sequence and a public microarray database. Eleven ethylene receptor family genes were identified in soybean through homology searches, and they were divided into two subgroups. Exon-intron analysis showed that the gene structures are highly conserved within each group. Further analysis of their expression patterns showed that these ethylene receptor genes are differentially expressed in various soybean tissues and organs, including functional nodules. Notably, the ethylene receptor genes showed different responses to rhizobial infection and Nod factors, suggesting a possible role for ethylene receptors and ethylene signaling in rhizobia-host cell interactions and nodulation in soybean. Together, these data indicate the functional divergence of ethylene receptor genes in soybean, and that some of these receptors mediate nodulation, including rhizobial infection, nodule development, and nodule functionality. These findings provide a foundation for further elucidation of the molecular mechanism by which the ethylene signaling pathway regulates nodulation in soybean, as well as other legumes.

  1. Expression of a fungal ferulic acid esterase in alfalfa modifies cell wall digestibility

    Science.gov (United States)

    2014-01-01

    Background Alfalfa (Medicago sativa) is an important forage crop in North America owing to its high biomass production, perennial nature and ability to fix nitrogen. Feruloyl esterase (EC 3.1.1.73) hydrolyzes ester linkages in plant cell walls and has the potential to further improve alfalfa as biomass for biofuel production. Results In this study, faeB [GenBank:AJ309807] was synthesized at GenScript and sub-cloned into a novel pEACH vector containing different signaling peptides to target type B ferulic acid esterase (FAEB) proteins to the apoplast, chloroplast, endoplasmic reticulum and vacuole. Four constructs harboring faeB were transiently expressed in Nicotiana leaves, with FAEB accumulating at high levels in all target sites, except chloroplast. Stable transformed lines of alfalfa were subsequently obtained using Agrobacterium tumefaciens (LBA4404). Out of 136 transgenic plants regenerated, 18 independent lines exhibited FAEB activity. Subsequent in vitro digestibility and Fourier transformed infrared spectroscopy (FTIR) analysis of FAEB-expressing lines showed that they possessed modified cell wall morphology and composition with a reduction in ester linkages and elevated lignin content. Consequently, they were more recalcitrant to digestion by mixed ruminal microorganisms. Interestingly, delignification by alkaline peroxide treatment followed by exposure to a commercial cellulase mixture resulted in higher glucose release from transgenic lines as compared to the control line. Conclusion Modifying cell wall crosslinking has the potential to lower recalcitrance of holocellulose, but also exhibited unintended consequences on alfalfa cell wall digestibility due to elevated lignin content. The combination of efficient delignification treatment (alkaline peroxide) and transgenic esterase activity complement each other towards efficient and effective digestion of transgenic lines. PMID:24650274

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

    Directory of Open Access Journals (Sweden)

    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.

  3. [Root Nodule Bacteria Sinorhizobium meliloti: Tolerance to Salinity and Bacterial Genetic Determinants].

    Science.gov (United States)

    Roumiantseva, M L; Muntyan, V S

    2015-01-01

    The theoretical and experimental data on salt tolerance of root nodule bacteria Sinorhizobium meliloti (Ensifer meliloti), an alfalfa symbiont, and on genetic determination of this feature are reviewed. Extensive data on the genes affecting adaptation of proteobacteria are provided, as well as on the groups of genes with activity depending on the osmolarity of the medium. Structural and functional polymorphism of the bet genes involved in betaine synthesis and transport in S. meliloti is discussed. The phenotypic and. genotypic polymorphism in 282 environmental rhizobial strains isolated from the centers of alfalfa diversity affected by aridity and salinity is discussed. The isolates from the Aral Sea area and northern Caucasus were shown to possess the betC gene represented by two types of alleles: the dominant A-type allele found in Rm 1021 and the less common divergent E-type allele, which was revealed in regions at the frequencies at the frequencies of 0.35 and 0.48, respectively. In the isolates with the salt-tolerant phenotype, which were isolated from root nodules and subsequently formed less effective symbioses with alfalfa, the frequency of E-type alleles was 2.5 times higher. Analysis of the nucleotide and amino acid sequences of the E-type allele of the betC gene revealed that establishment of this allele in the population was a result of positive selection. It is concluded that diversification of the functionally diverse bet genes occurring in S. meliloti affects the salt tolerance and symbiotic effectivity of rhizobia.

  4. Transcription of the soybean leghemoglobin genes during nodule development

    DEFF Research Database (Denmark)

    Marcker, Anne; Ø Jensen, Erik; Marcker, Kjeld A

    1984-01-01

    mechanism as is the case for vertebrate globin genes. Concomitantly with the increase in Lb gene transcription some of the other nodule specific plant genes are activated. These specific changes in the activities of the Lb and nodulin genes precede the activation of the bacterial nitrogenase gene. Thus......During the early stages of soybean nodule development the leghemoglobin (Lb) genes are activated sequentially in the opposite order to which they are arranged in the soybean genome. At a specific stage after the initial activation of all the Lb genes, a large increment occurs in the transcription...... of the Lb(c1), Lb(c3) and Lb(a) genes while the transcription of the Lb(c2) gene is not amplified to a similar extent. All the Lb genes retain significant activity for a long period during the lifetime of a nodule. Consequently the soybean Lb genes are not regulated by a developmental gene switching...

  5. The nodule conductance to O₂ diffusion increases with phytase activity in N₂-fixing Phaseolus vulgaris L.

    Science.gov (United States)

    Lazali, Mohamed; Drevon, Jean Jacques

    2014-07-01

    To understand the relationship between phosphorus use efficiency (PUE) and respiration for symbiotic nitrogen fixation (SNF) in legume nodules, six recombinant inbred lines of common bean (RIL Phaseolus vulgaris L.), contrasting in PUE for SNF, were inoculated with Rhizobium tropici CIAT899, and grown under hydroaeroponic culture with sufficient versus deficient P supply (250 versus 75 μmol P plant(-1) week(-1)). At the flowering stage, the biomass of plants and phytase activity in nodules were analyzed after measuring O2 uptake by nodulated roots. Our results show that the P-deficiency significantly increased the phytase activity in nodules of all RILs though with highest extent for RILs 147, 29 and 83 (ca 45%). This increase in phytase activity was associated with an increase in nodule respiration (ca 22%) and in use of the rhizobial symbiosis (ca 21%). A significant correlation was found under P-deficiency between nodule O2 permeability and phytase activity in nodules for RILs 104, 34 and 115. This observation is to our knowledge the first description of a correlation between O2 permeability and phytase activity of a legume nodule. It is concluded that the variation of phytase activity in nodules can increase the internal utilization of P and might be involved in the regulation of nodule permeability for the respiration linked with SNF and the adaptation to P-deficiency. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  6. Molecular cloning and characterization of three isoprenyl diphosphate synthase genes from alfalfa.

    Science.gov (United States)

    Sun, Yan; Long, Ruicai; Kang, Junmei; Zhang, Tiejun; Zhang, Ze; Zhou, He; Yang, Qingchuan

    2013-02-01

    Isoprenoid is the precursor for the biosynthesis of saponins, abscisic acid, gibberellins, chlorophylls and many other products in plants. Saponins are an important group of bioactive plant natural products. The alfalfa (Medicago sativa L.) saponins are glycosides of different triterpene aglycones and possess many biological activities. We isolated three genes (MsFPPS, MsGPPS and MsGGPPS) encoding isoprenyl diphosphate synthases (IDS) from alfalfa via a homology-based PCR approach. The enzyme activity assay of purified recombined MsFPPS and MsGGPPS expressed in Escherichia coli indicated that they all had IDS activity. Expression analysis of the three genes in different alfalfa tissues using real time PCR displayed that they were expressed in all tissues although they had a different expression patterns. MsFPPS and MsGPS displayed a significant increase in transcript level in response to methyl jasmonate, but the transcript level of MsGGPPS decreased obviously. To elucidate the functions of the three IDSs, their overexpression driven by a constitutive cauliflower mosaic virus-35S promoter in tobacco plants was applied and analyzed. The T(0) transgenic plants of MsFPPS showed high levels of squalene content when compared with control. However, no differences were detected in T(0) transgenic plants of MsGPPS and MsGGPPS. In addition, the overexpression of MsFPPS induced senescence response in transgenic plant leaves. This result may indicate that MsFPPS performs a role not only in phytosterol and triterpene biosynthesis, but also in growth regulation.

  7. A translationally controlled tumor protein gene Rpf41 is required for the nodulation of Robinia pseudoacacia.

    Science.gov (United States)

    Chou, Minxia; Xia, Congcong; Feng, Zhao; Sun, Yali; Zhang, Dehui; Zhang, Mingzhe; Wang, Li; Wei, Gehong

    2016-03-01

    Translationally controlled tumor protein (TCTP) is fundamental for the regulation of development and general growth in eukaryotes. Its multiple functions have been deduced from its involvement in several cell pathways, but its potential involvement in symbiotic nodulation of legumes cannot be suggested a priori. In the present work, we identified and characterized from the woody leguminous tree Robinia pseudoacacia a homolog of TCTP, Rpf41, which was up-regulated in the infected roots at 15 days post-inoculation but decreased in the matured nodules. Subcellular location assay showed that Rpf41 protein was located in the plasma membrane, cytoplasm, nucleus, and also maybe in cytoskeleton. Knockdown of Rpf41 via RNA interference (RNAi) resulted in the impaired development of both nodule and root hair. Compared with wild plants, the root and stem length, fresh weight and nodule number per plant was decreased dramatically in Rpf41 RNAi plants. The number of ITs or nodule primordia was also significantly reduced in the Rpf41 RNAi roots. The analyses of nodule ultrastructure showed that the infected cell development in Rpf41 RNAi nodules remained in zone II, which had fewer infected cells. Furthermore, the symbiosomes displayed noticeable shrinkage of bacteroid and peribacteroid space enlargement in the infected cells of Rpf41 RNAi nodules. In the deeper cell layers, a more remarkable aberration of the infected cell ultrastructure was observed, and electron-transparent lesions in the bacteroid cytoplasm were detected. These results identify TCTP as an important regulator of symbiotic nodulation in legume for the first time, and it may be involved in symbiotic cell differentiation and preventing premature aging of the young nodules in R. pseudoacacia.

  8. Endophytic and epiphytic hydrocarbon-utilizing bacteria associated with root nodules of legumes

    Energy Technology Data Exchange (ETDEWEB)

    Dashti, N.; Khanafer, M.; Radwan, S.S. [Kuwait Univ., (Kuwait). Dept. of Biological Sciences, Faculty of Sciences

    2005-07-01

    During their withdrawal from Kuwait in 1991, the Iraqi forces damaged and set fire to approximately 700 oil wells. Oil gushed from the wells for a period of 7 months, resulting in oil lakes which covered about 50 square km of the Kuwaiti desert and posing an environmental problem. Most of the crude oil has been pumped out, leaving the lake bottoms polluted with oil to depths reaching 20 to 25 cm. The oily areas have been mediated through indigenous hydrocarbon-utilizing microorganisms, but recovery is slow. Rhizospheres of crop plants, including legumes, are rich in oil-utilizing bacteria. Cultivation of broad beans in oily desert samples has enhanced oil biodegradation. This paper discussed the evidence that rhizobium strains inside the nodules on roots of broad beans are active in hydrocarbon utilization, and that the nodules are also colonized on their entire surfaces with oil-utilizing bacteria. Nodule-associated hydrocarbon utilizers appear to contribute together with rhizospheric hydrocarbon utilizers to the phytoremediation of oily soil. Broad beans were removed from soil and their root surfaces were sterilized to eliminate rhizospheric microorganisms. Plants with intact nodules were tested for their potential of attenuating to crude oil in water. Plants were divided into 2 groups: control plants in which all nodules were removed; and experimental plants which were used directly without further treatment. To isolate rhizobium from inside the nodules, fresh nodules were washed, sterilized and homogenized in sterile water. Bacterial strains were tested for their hydrocarbon utilization potential by streaking cell suspensions on the surface of sterile inorganic mediums containing 1 per cent of crude oil or of individual pure aliphatic and aromatic test hydrocarbons. All bacterial isolates were tested for growth on a solid Ashbery's nitrogen free medium. Results indicated that hydrocarbons were more efficiently eliminated from water supporting disinfected

  9. Performance of Three Adelphocoris spp. (Hemiptera:Miridae) on Flowering and Non-lfowering Cotton and Alfalfa

    Institute of Scientific and Technical Information of China (English)

    GAO Zhen; PAN Hong-sheng; LIU Bing; LU Yan-hui; LIANG Ge-mei

    2014-01-01

    The genus Adelphocoris (Hemiptera:Miridae) is a group of key insect pests in cotton ifelds in China that includes three dominant species:A. suturalis (Jakovlev), A. lineolatus (Goeze) and A. fasciaticollis (Reuter). Previous ifeld surveys have found that adults of these Adelphocoris species usually move onto speciifc host plants when the plants enter the lfowering stage. To determine the potential trade-offs for this host-plant preference behavior, the performance of these three Adelphocoris spp. on lfowering and non-lfowering cotton and alfalfa were compared in the laboratory. The results showed that Adelphocoris spp. had signiifcantly higher nymphal developmental and survival rates, along with increased adult longevity and fecundity on lfowering cotton and alfalfa than on non-lfowering plants of either species. In addition, compared with cotton plants, alfalfa generally promoted better performance of these three Adelphocoris species, especially for A. lineolatus. Simple correlation analysis indicated that female adult longevity was positively correlated to male adult longevity and female fecundity, and female fecundity was positively correlated to nymphal development and survival rates. This study demonstrated a positive correlation between adult preference and offspring/adult performance for all three Adelphocoris species, with no evidence of any trade-offs for this preference for lfowering host plants. These ifndings support the hypothesis that hemimetabolous insects such as mirid bugs have positive adult preference-adult/nymphal performance relationships, which is likely due to the similar feeding habits and nutritional requirements of adults and nymphs.

  10. Identification and Selection for Salt Tolerance in Alfalfa (Medicago sativa L.) Ecotypes via Physiological Traits

    OpenAIRE

    Monirifar, Hassan; Maryam BARGHI

    2009-01-01

    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 greenhou...

  11. Reductions in potash for last-year alfalfa production

    Science.gov (United States)

    Applying the correct amount of potassium (K) fertilizer is critical for alfalfa yield and stand persistence; however, excess K fertilizer will decrease profits and increase the risk of milk fever when alfalfa is fed to fresh cows. Stand persistence is often not a major concern in the last year of al...

  12. Alfalfa virus S, a new species in the family Alphaflexiviridae

    Science.gov (United States)

    A new species of the family Alphaflexiviridae provisionally named alfalfa virus S (AVS) was discovered 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 high throughput sequenc...

  13. Practices that support coexistence: A survey of Alfalfa growers

    Science.gov (United States)

    The alfalfa industry has worked hard to foster the coexistence of genetically-engineered (GE) and conventional alfalfa production by developing a set of best management practices that aim to limit adventitious-presence (AP) of GE traits in conventional seed. The general goal is to minimize transgene...

  14. Lessons learned in managing alfalfa-grass mixtures

    Science.gov (United States)

    Grass-alfalfa mixtures have a number of benefits that make them attractive to producers. However, they can be problematic to establish and maintain. Research programs have made progress in understanding the benefits and challenges of alfalfa-grass mixtures. Mixtures may have greater winter survival ...

  15. Yield and forage nutritive value of reduced lignin alfalfa

    Science.gov (United States)

    Reduced lignin alfalfa (Medicago sativa L.) cultivars have the potential to increase the feeding value of alfalfa for livestock by improving the forage fiber digestibility and to increase harvest management flexibility. The objectives were to compare the yield and forage nutritive value of reduced ...

  16. Complete genome sequence of the alfalfa latent virus

    Science.gov (United States)

    Alfalfa latent virus (ALV) is a member of the carlavirus group and occurs symptomlessly in alfalfa (Medicago sativa). In the US it is prevalent in Nebraska and Wisconsin. The virus is recognized as a strain of Pea streak virus (PeSV) So far, no complete genomic sequence of PSV or ALV is availab...

  17. Effects of alfalfa germplasm and stage of maturity on digestive process and productive response of dairy cows fed alfalfa hay-based diets

    Directory of Open Access Journals (Sweden)

    Igino Andrighetto

    2010-01-01

    Full Text Available The effects on the digestive process and the productive performances of dairy cows fed diets containing alfalfa hay from  different germplasms and maturity were assessed in the present study. Three different lots of first-cut alfalfa hay were  used in the study: the improved variety “Boreal”, harvested at two consecutive maturity stages (early flowering and full  flowering and the ecotype “Vogherese”, at full flowering. Cutting the plant at an earlier stage of maturity improved hay  quality in comparison with the more mature forages (crude protein: 21.4 vs 16.5% DM; P  DM; P  er lignin content for the latter (8.6 vs 8.2% DM; P  tating cows were formulated using an equal amount of corn silage and the maximum inclusion of one of the tested hays  as forage base. The better quality of the early cut hay made it possible to increase its inclusion in the diet up to 44% of  total dietary DM, while the two more mature hays covered only 36% of total DM of the respective diets. According to a  3 x 3 Latin square design, the diets were fed to 3 Italian Brown cows (initial average days in milk 121 ± 24 and milk  yield 20 kg ± 1.3 fitted with ruminal cannula in 3 consecutive periods of 28 d each. Alfalfa stage of maturity and  germplasm did not affect dietary DM intake (average 16 kg/d. Degradability parameters of dietary DM, calculated by in  situ nylon bags technique, showed similar kinetics of rumen disappearance for all diets. No differences were noticed in  the ruminal rate of passage of the solid phase among diets, while the liquid phase showed a slower rate of passage for  the early flowering hay diet. Consistent with the degradation process, the stage of maturity or the alfalfa germplasm did  not affect the rumen fluid data or the in vivo digestibility coefficients of the diets. Milk yield did not show any change due  to either alfalfa maturity or cultivar, while milk protein was lowered when cows received the early cut hay

  18. Alfalfa variety development. Minnesota Agripower Project, Task II research report

    Energy Technology Data Exchange (ETDEWEB)

    Lamb, J.F.S.; Samac, D.A.; Sheaffer, C.C.

    1997-10-30

    This report briefly summarizes preliminary results from crossbreeding alfalfa to develop desirable characteristics for a dedicated biomass feed stock. The varieties development is part of a larger project which includes preparation and gasification of the alfalfa stems for energy production, and use of the co-product alfalfa leaves in livestock feed. The desired alfalfa traits include winter hardiness, resistance to major pathogens, resistance to foliar disease complexes, many thick, tall, solid, non-lodging stems with high lignin content, delayed flowering, and high quality leaves retained through harvest. Currently no alfalfa varieties meet these criteria. Three crosses were made using old European varieties, with thick stems, and modern resistant varieties. The crossbreeds showed some resistance to diseases, but increased resistance is needed to maximize leaf and steam yield. 1 tab.

  19. A rhizobia strain isolated from root nodule of gymno-sperm Podocarpus macrophyllus

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Symbiotic nitrogen fixation of rhizobia and leguminous plants is considered as the most important biologic nitrogen fixation system on earth. Symbiotic nodulation of gymnosperm Podocarpus macro-phyllus and rhizobia has never been reported. In this study, 11 endophytic bacteria strains were isolated from root nodules of P. macrophyllus and its variation P. macrophyllus var. maki. The plant infection tests on these strains indicated that the isolated strains could be nodulated on P. macrophyllus plants, and weak nitrogenase activity of nodules was found in acetylene reduction method. According to the physiological and biochemical characteristics of the 11 strains, GXLO 02 was selected as the representative strain. 16S rDNA full-length sequence analysis of GXLO 02 confirmed that the representative strain GXLO 02 belongs to Rhizobium sp.

  20. Alteration of enod40 expression modifies medicago truncatula root nodule development induced by sinorhizobium meliloti

    Science.gov (United States)

    Charon, C; Sousa, C; Crespi, M; Kondorosi, A

    1999-01-01

    Molecular mechanisms involved in the control of root nodule organogenesis in the plant host are poorly understood. One of the nodulin genes associated with the earliest phases of this developmental program is enod40. We show here that transgenic Medicago truncatula plants overexpressing enod40 exhibit accelerated nodulation induced by Sinorhizobium meliloti. This resulted from increased initiation of primordia, which was accompanied by a proliferation response of the region close to the root tip and enhanced root length. The root cortex of the enod40-transformed plants showed increased sensitivity to nodulation signals. T(1) and T(2) descendants of two transgenic lines with reduced amounts of enod40 transcripts (probably from cosuppression) formed only a few and modified nodulelike structures. Our results suggest that induction of enod40 is a limiting step in primordium formation, and its function is required for appropriate nodule development. PMID:10521525

  1. Effects of diets containing alfalfa hay or barley flour mixed alfalfa silage on feeding behavior, productivity, rumen fermentation and blood metabolites in lactating cows.

    Science.gov (United States)

    Khadem, Ali-Akbar; Sharifi, Majid; Afzalzadeh, Ahmad; Rezaeian, Mohammad

    2009-08-01

    The effects of barley flour on the fermentation parameters of alfalfa silage and on the productivity of dairy cows were investigated. Alfalfa forage was ensiled either with or without barley flour. Barley flour was soaked in water for 24 h before being mixed with alfalfa (12 kg: 100 kg dry matter bases) at ensiling. Eighteen multi-parous cows were assigned to three equal treatment groups using a completely randomized design. Three isocaloric and isonitrogenous total mixed rations containing alfalfa hay, ordinary alfalfa silage or barley flour mixed alfalfa silage were then prepared. The concentrations of ammonia nitrogen, acetic acid and butyric acid were lower in barley flour mixed alfalfa silage compared to that in ordinary alfalfa silage but the concentration of lactic acid was lower in the ordinary alfalfa silage. Feeding behavior, milk yield and composition, ruminal fermentation and blood metabolites were measured. Although dry matter intake and milk production were not affected, the effect of preparation of alfalfa influenced feeding behavior and rumen fermentation parameters. Cows on alfalfa silage diets spent longer ruminating compared to those fed alfalfa hay. The ruminal ammonia nitrogen and blood urea were affected by ensiling (alfalfa hay versus alfalfa silages) while both parameters were lower in cows fed on barley flour mixed alfalfa silage than those fed on ordinary silage. Although similar blood glucose was recorded for cows fed on alfalfa silages, it was higher in cows fed on alfalfa hay. It is concluded that the addition of barely flour when making alfalfa silage may improve both the fermentation process during ensilage and the ruminal ammonia nitrogen utilization with no significant effects on productivity.

  2. Liming influence on soil chemical properties, nutritional status and yield of alfalfa grown in acid soil Influência da calagem nas propriedades químicas do solo, estado nutricional e produção da alfafa em um solo ácido

    Directory of Open Access Journals (Sweden)

    Adônis Moreira

    2010-08-01

    Full Text Available Alfalfa is an important forage crop with high nutritive value, although highly susceptible to soil acidity. Liming is one of the most efficient and prevailing practices to correct soil acidity and improve alfalfa yield. The objective of this study was to evaluate response to liming of alfalfa grown in a greenhouse on a Typic Quartzipsamment soil. The treatments consisted of four lime rates (0, 3.8, 6.6 and 10.3 Mg ha-1 and two cuts. Alfalfa dry matter increased quadratically with increasing lime rates. In general, dry matter yield was maximized by a lime rate of 8.0 Mg ha-1. Except for the control, the dry matter nutrient contents in the treatments were adequate. The positive linear correlation between root and nodule dry matter with lime rates indicated improvement of these plant traits with decreasing soil acidity. The soil acidity indices pH, base saturation, Ca2+ concentration, Mg2+ concentration, and H + Al were relevant factors in the assessment of alfalfa yield. The magnitude of influence of these soil acidity indices on yield as determined by the coefficient of determination (R² varied and decreased in the order: base saturation, H + Al, pH, Ca and Mg concentrations. Optimum values of selected soil chemical properties were defined for maximum shoot dry matter; these values can serve as a guideline for alfalfa liming to improve the yield of this forage on acid soils.A alfafa é uma importante forrageira com alto valor nutritivo, porém é altamente suscetível à acidez do solo. A calagem é uma das mais eficientes práticas para corrigir esse problema e melhorar a produtividade da alfafa. O objetivo deste trabalho foi avaliar a resposta à calagem da alfafa cultivada em Neossolo Quartzarênico em casa de vegetação. Os tratamentos consistiram de quatro doses de calcário (0; 3,8; 6,6 e 10,3 Mg ha-1 e duas épocas de corte. A produção de matéria seca foi significativamente aumentada com a adição de calcário, com o m

  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. Metal biosorption capability of Cupriavidus taiwanensis and its effects on heavy metal removal by nodulated Mimosa pudica.