WorldWideScience

Sample records for embryogenesis abundant lea

  1. Study of model systems to test the potential function of Artemia group 1 late embryogenesis abundant (LEA) proteins.

    Science.gov (United States)

    Warner, Alden H; Guo, Zhi-hao; Moshi, Sandra; Hudson, John W; Kozarova, Anna

    2016-01-01

    Embryos of the brine shrimp, Artemia franciscana, are genetically programmed to develop either ovoviparously or oviparously depending on environmental conditions. Shortly upon their release from the female, oviparous embryos enter diapause during which time they undergo major metabolic rate depression while simultaneously synthesize proteins that permit them to tolerate a wide range of stressful environmental events including prolonged periods of desiccation, freezing, and anoxia. Among the known stress-related proteins that accumulate in embryos entering diapause are the late embryogenesis abundant (LEA) proteins. This large group of intrinsically disordered proteins has been proposed to act as molecular shields or chaperones of macromolecules which are otherwise intolerant to harsh conditions associated with diapause. In this research, we used two model systems to study the potential function of the group 1 LEA proteins from Artemia. Expression of the Artemia group 1 gene (AfrLEA-1) in Escherichia coli inhibited growth in proportion to the number of 20-mer amino acid motifs expressed. As well, clones of E. coli, transformed with the AfrLEA-1 gene, expressed multiple bands of LEA proteins, either intrinsically or upon induction with isopropyl-β-thiogalactoside (IPTG), in a vector-specific manner. Expression of AfrLEA-1 in E. coli did not overcome the inhibitory effects of high concentrations of NaCl and KCl but modulated growth inhibition resulting from high concentrations of sorbitol in the growth medium. In contrast, expression of the AfrLEA-1 gene in Saccharomyces cerevisiae did not alter the growth kinetics or permit yeast to tolerate high concentrations of NaCl, KCl, or sorbitol. However, expression of AfrLEA-1 in yeast improved its tolerance to drying (desiccation) and freezing. Under our experimental conditions, both E. coli and S. cerevisiae appear to be potentially suitable hosts to study the function of Artemia group 1 LEA proteins under environmentally

  2. CpLEA5, the Late Embryogenesis Abundant Protein Gene from Chimonanthus praecox, Possesses Low Temperature and Osmotic Resistances in Prokaryote and Eukaryotes

    Directory of Open Access Journals (Sweden)

    Yiling Liu

    2015-11-01

    Full Text Available Plants synthesize and accumulate a series of stress-resistance proteins to protect normal physiological activities under adverse conditions. Chimonanthus praecox which blooms in freezing weather accumulates late embryogenesis abundant proteins (LEAs in flowers, but C. praecox LEAs are little reported. Here, we report a group of five LEA genes of C. praecox (CpLEA5, KT727031. Prokaryotic-expressed CpLEA5 was employed in Escherichia coli to investigate bioactivities and membrane permeability at low-temperature. In comparison with the vacant strains, CpLEA5-containing strains survived in a 20% higher rate; and the degree of cell membrane damage in CpLEA5-containing strains was 55% of that of the vacant strains according to a conductivity test, revealing the low-temperature resistance of CpLEA5 in bacteria. CpLEA5 was also expressed in Pichia pastoris. Interestingly, besides low-temperature resistance, CpLEA5 conferred high resistance to salt and alkali in CpLEA5 overexpressing yeast. The CpLEA5 gene was transferred into Arabidopsis thaliana to also demonstrate CpLEA5 actions in plants. As expected, the transgenic lines were more resistant against low-temperature and drought while compared with the wild type. Taken together, CpLEA5-conferred resistances to several conditions in prokaryote and eukaryotes could have great value as a genetic technology to enhance osmotic stress and low-temperature tolerance.

  3. Late Embryogenesis Abundant (LEA) Constitutes a Large and Diverse Family of Proteins Involved in Development and Abiotic Stress Responses in Sweet Orange (Citrus sinensis L. Osb.)

    Science.gov (United States)

    Pedrosa, Andresa Muniz; Martins, Cristina de Paula Santos; Gonçalves, Luana Pereira; Costa, Marcio Gilberto Cardoso

    2015-01-01

    Late Embryogenesis Abundant (LEA) proteins are an ubiquitous group of polypeptides that were first described to accumulate during plant seed dehydration, at the later stages of embryogenesis. Since then they have also been recorded in vegetative plant tissues experiencing water limitation and in anhydrobiotic bacteria and invertebrates and, thereby, correlated with the acquisition of desiccation tolerance. This study provides the first comprehensive study about the LEA gene family in sweet orange (Citrus sinensis L. Osb.), the most important and widely grown fruit crop around the world. A surprisingly high number (72) of genes encoding C. sinensis LEAs (CsLEAs) were identified and classified into seven groups (LEA_1, LEA_2, LEA_3 and LEA_4, LEA_5, DEHYDRIN and SMP) based on their predicted amino acid sequences and also on their phylogenetic relationships with the complete set of Arabidopsis thaliana LEA proteins (AtLEAs). Approximately 60% of the CsLEAs identified in this study belongs to the unusual LEA_2 group of more hydrophobic LEA proteins, while the other LEA groups contained a relatively small number of members typically hydrophilic. A correlation between gene structure and motif composition was observed within each LEA group. Investigation of their chromosomal localizations revealed that the CsLEAs were non-randomly distributed across all nine chromosomes and that 33% of all CsLEAs are segmentally or tandemly duplicated genes. Analysis of the upstream sequences required for transcription revealed the presence of various stress-responsive cis-acting regulatory elements in the promoter regions of CsLEAs, including ABRE, DRE/CRT, MYBS and LTRE. Expression analysis using both RNA-seq data and quantitative real-time RT-PCR (qPCR) revealed that the CsLEA genes are widely expressed in various tissues, and that many genes containing the ABRE promoter sequence are induced by drought, salt and PEG. These results provide a useful reference for further exploration of

  4. Molecular cloning and characterization of the late embryogenesis abundant group 4 (EgLEA4 gene from oil palm (Elaeis guineensis Jacq

    Directory of Open Access Journals (Sweden)

    Watcharasuda Hualkasin

    2013-06-01

    Full Text Available The Late-Embryogenesis Abundant group 4 (LEA4 genes is a group of genes that have been reported to be involved in stress and hormone responses. The completed LEA4 cDNA sequence was first obtained from a set of EST sequences of oil palm (Elaeis guineensis Jacq, named as EgLEA4. The open reading frame is 486 bp in length, encoding a deduced amino acid sequence of 161 residues with a molecular weight of 16.5 kDa and a pI value of about 8.0. Five amino acid motif patterns were found in the EgLEA4 (II, I, III, IV and V and each had a close identity to similar LEA4 patterns of soybean (64%.Comparison of the nucleotide sequences of the cDNA and the genomic DNA demonstrated that the EgLEA4 gene is composed of 2 exons and 1 intron. The 52 untranslated region shows a putative promoter sequence involved in the transcription process, drought stress and hormone responsive elements. RT-PCR analysis showed that the EgLEA4 gene was only expressed in mesocarp, during the late stages of fruit development. It also had a higher expression in induced drought conditions indicating that the EgLEA4 protein may be involved in plant adaptation and stress (drought responsive pathway.

  5. Overexpression Analysis of emv2 gene coding for Late Embryogenesis Abundant Protein from Vigna radiata (Wilczek

    Directory of Open Access Journals (Sweden)

    Rajesh S.

    2008-10-01

    Full Text Available Late embryogenesis abundant (LEA proteins are speculated to protect against water stress deficit in plants. An over expression system for mungbean late embryogenesis abundant protein, emv2 was constructed in a pET29a vector, designated pET-emv2 which is responsible for higher expression under the transcriptional/translational control of T7/lac promoter incorporated in the Escherichia coli BL21 (DE3.Induction protocol was optimized for pET recombinants harboring the target gene. Overexpressed EMV2 protein was purified to homogeneity and the protein profile monitored by SDS-PAGE.

  6. LEAping to conclusions: A computational reanalysis of late embryogenesis abundant proteins and their possible roles

    Directory of Open Access Journals (Sweden)

    Wise Michael J

    2003-10-01

    Full Text Available Abstract Background The late embryogenesis abundant (LEA proteins cover a number of loosely related groups of proteins, originally found in plants but now being found in non-plant species. Their precise function is unknown, though considerable evidence suggests that LEA proteins are involved in desiccation resistance. Using a number of statistically-based bioinformatics tools the classification of a large set of LEA proteins, covering all Groups, is reexamined together with some previous findings. Searches based on peptide composition return proteins with similar composition to different LEA Groups; keyword clustering is then applied to reveal keywords and phrases suggestive of the Groups' properties. Results Previous research has suggested that glycine is characteristic of LEA proteins, but it is only highly over-represented in Groups 1 and 2, while alanine, thought characteristic of Group 2, is over-represented in Group 3, 4 and 6 but under-represented in Groups 1 and 2. However, for LEA Groups 1 2 and 3 it is shown that glutamine is very significantly over-represented, while cysteine, phenylalanine, isoleucine, leucine and tryptophan are significantly under-represented. There is also evidence that the Group 4 LEA proteins are more appropriately redistributed to Group 2 and Group 3. Similarly, Group 5 is better found among the Group 3 LEA proteins. Conclusions There is evidence that Group 2 and Group 3 LEA proteins, though distinct, might be related. This relationship is also evident in the overlapping sets of keywords for the two Groups, emphasising alpha-helical structure and, at a larger scale, filaments, all of which fits well with experimental evidence that proteins from both Groups are natively unstructured, but become structured under stress conditions. The keywords support localisation of LEA proteins both in the nucleus and associated with the cytoskeleton, and a mode of action similar to chaperones, perhaps the cold shock chaperones

  7. Expression Profiles of 12 Late Embryogenesis Abundant Protein Genes from Tamarix hispida in Response to Abiotic Stress

    Directory of Open Access Journals (Sweden)

    Caiqiu Gao

    2014-01-01

    Full Text Available Twelve embryogenesis abundant protein (LEA genes (named ThLEA-1 to -12 were cloned from Tamarix hispida. The expression profiles of these genes in response to NaCl, PEG, and abscisic acid (ABA in roots, stems, and leaves of T. hispida were assessed using real-time reverse transcriptase-polymerase chain reaction (RT-PCR. These ThLEAs all showed tissue-specific expression patterns in roots, stems, and leaves under normal growth conditions. However, they shared a high similar expression patterns in the roots, stems, and leaves when exposed to NaCl and PEG stress. Furthermore, ThLEA-1, -2, -3, -4, and -11 were induced by NaCl and PEG, but ThLEA-5, -6, -8, -10, and -12 were downregulated by salt and drought stresses. Under ABA treatment, some ThLEA genes, such as ThLEA-1, -2, and -3, were only slightly differentially expressed in roots, stems, and leaves, indicating that they may be involved in the ABA-independent signaling pathway. These findings provide a basis for the elucidation of the function of LEA genes in future work.

  8. Functional studies of soybean (Glycine max L.) seed LEA proteins GmPM6, GmPM11, and GmPM30 by CD and FTIR spectroscopy

    NARCIS (Netherlands)

    Hoekstra, F.A.

    2012-01-01

    The protein and mRNA levels of late embryogenesis abundant (LEA) genes may be linked to osmotic stresses. Here, we characterized three soybean hydrophilic LEA proteins – GmPM11 (LEA I), GmPM6 (LEA II), and GmPM30 (LEA III) – by circular dichroism and Fourier transform infrared spectroscopy.

  9. Computational Analysis Of Group 2 Late Embryogenesis Protein (Lea In Different Cultivar Of Bread Wheat (Triticum Aestivum

    Directory of Open Access Journals (Sweden)

    Rasouli Hassan

    2014-12-01

    Full Text Available Late Embryogenesis abundant protein has a crucial role as the cold-acclimation process in the wheat. These proteins encoded by TaWdhn13 gene. This gene is transcriptionally activated and produces the accumulated proteins and metabolites and protection cell structure from freezing damage. The objectives of this study were to isolate the genomic DNA (g-DNA sequence of TaWdhn13, to analyze structure, conserved domains of the gene, and to found a basis for association analysis of the functional sites associated with computational analysis. We here report on the functional assignment to TaWdhn13 gene by computational analysis. The Three-Dimensional (3D model of LEA protein drawing by using the phyre 2 server. For identify the conserved domain and motif of these gene sequence we used the Conserved Domain Database and DNA Motif Searching Database, however, the conserved domains and motif has been recognized. The results showed TaWdhn13 conserved domain incudes: Dehydrin superfamily. Also, motifs structure for this gene includes: 2FE2S_FER_1 Motif, INTEGRIN_BETA Motif, VWFC_1 Motif, EGF_1 Motif and DEFENSIN Motif. Our results reveal that group 2 LEA proteins are most likely to function within the cell nucleus. The analysis of protein property showed that the protein had no trans-membrane domains. The isoelectric point of the protein was 3.41, which was charged with 5.34 negative electrons when pH value of the buffer was 7.0.

  10. Molecular cloning and characterization of a group 3 LEA gene from ...

    African Journals Online (AJOL)

    Late embryogenesis-abundant (LEA) protein is one of the components involved in desiccation tolerance (DT) by maintaining cellular structures in the dry state. In this study, a member of the group 3 LEA, MwLEA1, was cloned from Mongolian wheatgrass (Agropyron mongolium Keng) based on a homologous sequence from ...

  11. The Ubiquitous Distribution of Late Embryogenesis Abundant Proteins across Cell Compartments in Arabidopsis Offers Tailored Protection against Abiotic Stress[C][W][OPEN

    Science.gov (United States)

    Candat, Adrien; Paszkiewicz, Gaël; Neveu, Martine; Gautier, Romain; Logan, David C.; Avelange-Macherel, Marie-Hélène; Macherel, David

    2014-01-01

    Late embryogenesis abundant (LEA) proteins are hydrophilic, mostly intrinsically disordered proteins, which play major roles in desiccation tolerance. In Arabidopsis thaliana, 51 genes encoding LEA proteins clustered into nine families have been inventoried. To increase our understanding of the yet enigmatic functions of these gene families, we report the subcellular location of each protein. Experimental data highlight the limits of in silico predictions for analysis of subcellular localization. Thirty-six LEA proteins localized to the cytosol, with most being able to diffuse into the nucleus. Three proteins were exclusively localized in plastids or mitochondria, while two others were found dually targeted to these organelles. Targeting cleavage sites could be determined for five of these proteins. Three proteins were found to be endoplasmic reticulum (ER) residents, two were vacuolar, and two were secreted. A single protein was identified in pexophagosomes. While most LEA protein families have a unique subcellular localization, members of the LEA_4 family are widely distributed (cytosol, mitochondria, plastid, ER, and pexophagosome) but share the presence of the class A α-helix motif. They are thus expected to establish interactions with various cellular membranes under stress conditions. The broad subcellular distribution of LEA proteins highlights the requirement for each cellular compartment to be provided with protective mechanisms to cope with desiccation or cold stress. PMID:25005920

  12. Abiotic stress in plants: Late Embryogenesis Abundant proteins

    OpenAIRE

    Amara, Imen

    2012-01-01

    [spa] Las proteínas LEA, originalmente fueron descritas en las semillas de algodón; se acumulan en grandes cantidades en estructuras tolerantes a la desecación (semillas, polen) y en tejidos vegetativos sometidos a estrés abiótico, sequía, salinidad y frío. También se hallan en organismos anidrobióticos, en plantas de resurrección, algunos invertebrados y microorganismos. La presencia de proteínas LEA se correlaciona con la adquisición de tolerancia a la desecación. Desde un principio se les ...

  13. The Potential Roles of the G1LEA and G3LEA Proteins in Early Embryo Development and in Response to Low Temperature and High Salinity in Artemia sinica.

    Directory of Open Access Journals (Sweden)

    Wei Zhao

    Full Text Available Late embryogenesis abundant proteins (LEA are stress resistance-related proteins that play crucial roles in protecting against desiccation, cold and high salinity in a variety of animals and plants. However, the expression pattern, distribution and functions of LEA proteins in the post-diapause period of Artemia sinica, and under high salinity and low temperature stresses, remain unknown. In this study, the complete cDNA sequences of the group 1 LEA (As-g1lea and group 3 LEA (As-g3lea genes from A. sinica were cloned. The expression patterns and location of As-G1LEA and As-G1LEA were investigated. The protein abundances of As-G1LEA, As-G3LEA and Trehalase were analyzed during different developmental stages of the embryo and under low temperature and high salinity stresses in A. sinica. The full-length cDNA of As-g1lea was 960 bp, encoding a 182 amino acid protein, and As-g3lea was 2089 bp, encoding a 364 amino acid protein. As-g1lea and As-g3lea showed their highest expressions at 0 h of embryonic development and both showed higher relative expression in embryonic, rather than adult, development stages. The abundances of As-G1LEA, As-G3LEA and trehalose were upregulated under low temperature and downregulated under high salinity stress. These two genes did not show any tissue or organ specific expression. Our results suggested that these LEA proteins might play a pivotal role in stress tolerance in A. sinica.

  14. Enhanced drought tolerance in transgenic Leymus chinensis plants with constitutively expressed wheat TaLEA3.

    Science.gov (United States)

    Wang, Lijuan; Li, Xiaofeng; Chen, Shuangyan; Liu, Gongshe

    2009-02-01

    Leymus chinensis is an important grassland perennial grass. However, its drought tolerance requires to be improved. LEA (late embryogenesis abundant) genes are believed to confer resistance to drought and water deficiency. Using Agrobacterium-mediated transformation, a wheat LEA gene, TaLEA(3), was integrated into L. chinensis. The transgenic lines showed enhanced growth ability under drought stress during which transgenic lines had increased the relative water content, leaf water potential, relative average growth rate, but decreased the malondialdehyde content compared with the non-transgenic plant. Thus, transgenic breeding is an efficient approach to enhance drought tolerance in L. chinensis.

  15. OsLEA1a, a new Em-like protein of cereal plants.

    Science.gov (United States)

    Shih, Ming-Der; Huang, Lin-Tzu; Wei, Fu-Jin; Wu, Ming-Tsung; Hoekstra, Folkert A; Hsing, Yue-Ie C

    2010-12-01

    Proteins abundant in seeds during the late stages of development, late embryogenesis abundant (LEA) proteins, are associated with desiccation tolerance. More than 100 of the group I LEA genes, also termed Em genes, have been identified from plants, bacteria and animals. The wide distribution indicates the functional importance of these genes. In the present study, we characterized a novel Em-like gene, OsLEA1a of rice (Oryza sativa). The encoded OsLEA1a protein has an N-terminal sequence similar to that of other plant Em proteins but lacks a 20-mer motif that is the most significant feature of typical Em proteins. The location of the sole intron indicates that the second exon of OsLEA1a is the mutated product of a typical Em gene. Transcriptome analysis revealed OsLEA1a mainly expressed in embryos, with no or only a few transcripts in osmotic stress-treated vegetative tissues. Structural analysis revealed that the OsLEA1a protein adopts high amounts of disordered conformations in solution and undergoes desiccation-induced conformational changes. Macromolecular interaction studies revealed that OsLEA1a protein interacts with non-reducing sugars and phospholipids but not poly-l-lysine. Thus, although the OsLEA1a protein lost its 20-mer motif, it is still involved in the formation of bioglasses with non-reducing sugars or plasma membrane. However, the protein does not function as a chaperone as do other groups of hydrophilic LEA proteins. The orthologs of the OsLEA1a gene had been identified from various grasses but not in dicot plants. Genetic analysis indicated that rice OsLEA1a locates at a 193 kb segment in chromosome 1 and is conserved in several published cereal genomes. Thus, the ancestor of Em-like genes might have evolved after the divergence of monocot plants.

  16. Complexity of the heat-soluble LEA proteome in Artemia species.

    Science.gov (United States)

    Warner, Alden H; Chakrabortee, Sohini; Tunnacliffe, Alan; Clegg, James S

    2012-09-01

    The brine shrimp Artemia is a well known stress tolerant invertebrate found on most continents. Under certain conditions females produce cysts (encysted gastrulae) that enter diapause, a state of obligate dormancy. During developmental formation of diapause embryos several different types of stress proteins accumulate in large amounts, including the late embryogenesis abundant (LEA) proteins. In this study we used a combination of heterologous group 3 LEA antibodies to demonstrate that the heat-soluble proteome of the cysts contains up to 12 distinct putative group 3 LEA proteins that complement the group 1 LEA proteins found previously. Most antibody-positive, heat-soluble proteins were larger than 50 kDa although antibody positive proteins of 20-38 kDa were also detected. Both nuclei and mitochondria had distinct complements of the putative group 3 LEA proteins. A few small group 3 LEA proteins were induced by cycles of hydration-dehydration along with one protein of about 62 kDa. The expression of group 3 LEA proteins, unlike members of group 1, was not restricted to encysted diapause embryos. Three to five putative group 3 LEA proteins were expressed in gravid females and in larvae. Cysts of different species from various geographic locations had distinct complements of group 3 LEA proteins suggesting rapid evolution of the LEA proteins or differences in the type of group 3 Lea genes expressed. Our results demonstrate the potential importance of group 3 LEA proteins in embryos and other life cycle stages of this animal extremophile. Copyright © 2012 Elsevier Inc. All rights reserved.

  17. ZmLEA3, a multifunctional group 3 LEA protein from maize (Zea mays L.), is involved in biotic and abiotic stresses.

    Science.gov (United States)

    Liu, Yang; Wang, Li; Xing, Xin; Sun, Liping; Pan, Jiaowen; Kong, Xiangpei; Zhang, Maoying; Li, Dequan

    2013-06-01

    Late embryogenesis abundant (LEA) proteins accumulate to high levels during the late stage of seed maturation and in response to water deficit, and are involved in protecting higher plants from damage caused by environmental stresses, especially drought. In the present study, a novel maize (Zea mays L.) group 3 LEA gene, ZmLEA3, was identified and later characterized using transgenic tobacco plants to investigate its functions in abiotic and biotic stresses. Transcript accumulation demonstrated that ZmLEA3 was induced in leaves by high salinity, low temperature, osmotic and oxidative stress as well as by signaling molecules such as ABA, salicylic acid (SA) and methyl jasmonate (MeJA). The transcript of ZmLEA3 could also be induced by pathogens [Pseudomonas syringae pv. tomato DC3000 (pst dc3000)]. ZmLEA3 is located in the cytosol and the nucles. Further study indicated that the ZmLEA3 protein could bind Mn(2+), Fe(3+), Cu(2+) and Zn(2+). Overexpression of ZmLEA3 in transgenic tobacco (Nicotiana tabacum) and yeast (GS115) conferred tolerance to osmotic and oxidative stresses. Interestingly, we also found that overexpression of ZmLEA3 in transgenic tobacco increased the hypersensitive cell death triggered by pst dc3000 and enhanced the expression of PR1a, PR2 and PR4 when compared with the wild type. Thus, we proposed that the ZmLEA3 protein plays a role in protecting plants from damage by protecting protein structure and binding metals under osmotic and oxidative stresses. In addition, ZmLEA3 may also enhance transgenic plant tolerance to biotic stress.

  18. Both plant and animal LEA proteins act as kinetic stabilisers of polyglutamine-dependent protein aggregation.

    Science.gov (United States)

    Liu, Yun; Chakrabortee, Sohini; Li, Ranhui; Zheng, Yizhi; Tunnacliffe, Alan

    2011-02-18

    LEA (late embryogenesis abundant) proteins are intrinsically disordered proteins that contribute to stress tolerance in plants and invertebrates. Here we show that, when both plant and animal LEA proteins are co-expressed in mammalian cells with self-aggregating polyglutamine (polyQ) proteins, they reduce aggregation in a time-dependent fashion, showing more protection at early time points. A similar effect was also observed in vitro, where recombinant LEA proteins were able to slow the rate of polyQ aggregation, but not abolish it altogether. Thus, LEA proteins act as kinetic stabilisers of aggregating proteins, a novel function in protein homeostasis consistent with a proposed role as molecular shields. Copyright © 2011 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  19. Liposomes with diverse compositions are protected during desiccation by LEA proteins from Artemia franciscana and trehalose.

    Science.gov (United States)

    Moore, Daniel S; Hansen, Richard; Hand, Steven C

    2016-01-01

    Intracellular accumulation of Late Embryogenesis Abundant (LEA) proteins and the disaccharide trehalose is associated with cellular desiccation tolerance in a number of animal species. Two LEA proteins from anhydrobiotic embryos of the brine shrimp Artemia franciscana were tested for the ability to protect liposomes of various compositions against desiccation-induced damage in the presence and absence of trehalose. Damage was assessed by carboxyfluorescein leakage after drying and rehydration. Further, using a cytoplasmic-localized (AfrLEA2) and a mitochondrial-targeted (AfrLEA3m) LEA protein allowed us to evaluate whether each may preferentially stabilize membranes of a particular lipid composition based on the protein's subcellular location. Both LEA proteins were able to offset damage during drying of liposomes that mimicked the lipid compositions of the inner mitochondrial membrane (with cardiolipin), outer mitochondrial membrane, and the inner leaflet of the plasma membrane. Thus liposome stabilization by AfrLEA3m or AfrLEA2 was not dependent on lipid composition, provided physiological amounts of bilayer and non-bilayer-forming lipids were present (liposomes with a non-biological composition of 100% phosphatidylcholine were not protected by either protein). Additive protection by LEA proteins plus trehalose was dependent on the lipid composition of the target membrane. Minimal additional damage occurred to liposomes stored at room temperature in the dried state for one week compared to liposomes rehydrated after 24h. Consistent with the ability to stabilize lipid bilayers, molecular modeling of the secondary structures for AfrLEA2 and AfrLEA3m revealed bands of charged amino acids similar to other amphipathic proteins that interact directly with membranes. Copyright © 2015. Published by Elsevier B.V.

  20. Isolation and functional characterization of a Medicago sativa L. gene, MsLEA3-1.

    Science.gov (United States)

    Bai, Yongqin; Yang, Qingchuan; Kang, Junmei; Sun, Yan; Gruber, Margaret; Chao, Yuehui

    2012-03-01

    A full-length cDNA of 1,728 nt, called MsLEA3-1, was cloned from alfalfa by rapid amplification of cDNA ends from an expressed sequence tag homologous to soybean pGmPM10 (accession No. AAA91965.1). MsLEA3-1, encodes a deduced protein of 436 amino acids, a calculated molecular weight of 47.0 kDa, a theoretical isoelectric point of 5.18, and closest homology with late embryogenesis abundant proteins in soybean. Sequence homology suggested a signal peptide in the N terminus, and subcellular localization with GFP revealed that MsLEA3-1 was localized preferentially to the nucleolus. The transcript titre of MsLEA3-1 was strongly enriched in leaves compared with roots and stems of mature alfalfa plants. Gene expression of MsLEA3-1 was strongly induced when seedlings were treated with NaCl and ABA. Expression of the MsLEA3-1 transgenic was detected in transgenic tobacco. Malondialdehyde content and, electrical conductivity content were reduced and electrical conductivity and proline content were increased in transgenic tobacco compared with non-transgenic tobacco under salt stress. The results showed that accumulation of the MsLEA3-1 protein in the vegetative tissues of transgenic plants enhanced their tolerance to salt stress. These results demonstrate a role for the MsLEA3-1 protein in stress protection and suggest the potential of the MsLEA3-1 gene for genetic engineering of salt tolerance.

  1. Ectopic Expression of an Atypical Hydrophobic Group 5 LEA Protein from Wild Peanut, Arachis diogoi Confers Abiotic Stress Tolerance in Tobacco.

    Directory of Open Access Journals (Sweden)

    Akanksha Sharma

    Full Text Available Late embryogenesis abundant (LEA proteins are a group of hydrophilic proteins, which accumulate in plants under varied stress conditions like drought, salinity, extreme temperatures and oxidative stress suggesting their role in the protection of plants against these stresses. A transcript derived fragment (TDF corresponding to LEA gene, which got differentially expressed in wild peanut, Arachis diogoi against the late leaf spot pathogen, Phaeoisariopsis personata was used in this study. We have cloned its full length cDNA by RACE-PCR, which was designated as AdLEA. AdLEA belongs to the atypical Group 5C of LEA protein family as confirmed by sequence analysis. Group 5C LEA protein subfamily contains Pfam LEA_2 domain and is highly hydrophobic. In native conditions, expression of AdLEA was upregulated considerably upon hormonal and abiotic stress treatments emphasizing its role in abiotic stress tolerance. Subcellular localization studies showed that AdLEA protein is distributed in both nucleus and cytosol. Ectopic expression of AdLEA in tobacco resulted in enhanced tolerance of plants to dehydration, salinity and oxidative stress with the transgenic plants showing higher chlorophyll content and reduced lipid peroxidation as compared to wild type plants. Overexpressed AdLEA tobacco plants maintained better photosynthetic efficiency under drought conditions as demonstrated by chlorophyll fluorescence measurements. These plants showed enhanced transcript accumulation of some stress-responsive genes. Our study also elucidates that ROS levels were significantly reduced in leaves and stomatal guard cells of transgenic plants upon stress treatments. These results suggest that AdLEA confers multiple stress tolerance to plants, which make it a potential gene for genetic modification in plants.

  2. OsLEA3-2, an abiotic stress induced gene of rice plays a key role in salt and drought tolerance.

    Directory of Open Access Journals (Sweden)

    Jianli Duan

    Full Text Available Late embryogenesis abundant (LEA proteins are involved in tolerance to drought, cold and high salinity in many different organisms. In this report, a LEA protein producing full-length gene OsLEA3-2 was identified in rice (Oryza sativa using the Rapid Amplification of cDNA Ends (RACE method. OsLEA3-2 was found to be only expressed in the embryo and can be induced by abiotic stresses. The coding protein localizes to the nucleus and overexpression of OsLEA3-2 in yeast improved growth performance compared with control under salt- and osmotic-stress conditions. OsLEA3-2 was also inserted into pHB vector and overexpressed in Arabidopsis and rice. The transgenic Arabidopsis seedlings showed better growth on MS media supplemented with 150 mM mannitol or 100 mM NaCl as compared with wild type plants. The transgenic rice also showed significantly stronger growth performance than control under salinity or osmotic stress conditions and were able to recover after 20 days of drought stress. In vitro analysis showed that OsLEA3-2 was able to protect LDH from aggregation on freezing and inactivation on desiccation. These results indicated that OsLEA3-2 plays an important role in tolerance to abiotic stresses.

  3. Group 1 LEA proteins contribute to the desiccation and freeze tolerance of Artemia franciscana embryos during diapause.

    Science.gov (United States)

    Toxopeus, Jantina; Warner, Alden H; MacRae, Thomas H

    2014-11-01

    Water loss either by desiccation or freezing causes multiple forms of cellular damage. The encysted embryos (cysts) of the crustacean Artemia franciscana have several molecular mechanisms to enable anhydrobiosis-life without water-during diapause. To better understand how cysts survive reduced hydration, group 1 late embryogenesis abundant (LEA) proteins, hydrophilic unstructured proteins that accumulate in the stress-tolerant cysts of A. franciscana, were knocked down using RNA interference (RNAi). Embryos lacking group 1 LEA proteins showed significantly lower survival than control embryos after desiccation and freezing, or freezing alone, demonstrating a role for group 1 LEA proteins in A. franciscana tolerance of low water conditions. In contrast, regardless of group 1 LEA protein presence, cysts responded similarly to hydrogen peroxide (H2O2) exposure, indicating little to no function for these proteins in diapause termination. This is the first in vivo study of group 1 LEA proteins in an animal and it contributes to the fundamental understanding of these proteins. Knowing how LEA proteins protect A. franciscana cysts from desiccation and freezing may have applied significance in aquaculture, where Artemia is an important feed source, and in the cryopreservation of cells for therapeutic applications.

  4. Late Embryogenesis Abundant Proteins

    NARCIS (Netherlands)

    Shih, M.D.; Hoekstra, F.A.; Hsing, Y.I.C.

    2008-01-01

    During the late maturation stage of seed development, water content decreases greatly. One of the most striking characteristics of mature orthodox seeds is their ability to withstand severe desiccation. Mechanisms of plant drought/desiccation tolerance have been studied by numerous groups, and a

  5. Overexpression of TaLEA gene from Tamarix androssowii improves salt and drought tolerance in transgenic poplar (Populus simonii × P. nigra.

    Directory of Open Access Journals (Sweden)

    Weidong Gao

    Full Text Available Late embryogenesis abundant (LEA genes were confirmed to confer resistance to drought and water deficiency. An LEA gene from Tamarixandrossowii (named TaLEA was transformed into Xiaohei poplar (Populussimonii × P. nigra via Agrobacterium. Twenty-five independent transgenic lines were obtained that were resistant to kanamycin, and 11 transgenic lines were randomly selected for further analysis. The polymerase chain reaction (PCR and ribonucleic acid (RNA gel blot indicated that the TaLEA gene had been integrated into the poplar genome. The height growth rate, malondialdehyde (MDA content, relative electrolyte leakage and damages due to salt or drought to transgenic and non-transgenic plants were compared under salt and drought stress conditions. The results showed that the constitutive expression of the TaLEA gene in transgenic poplars could induce an increase in height growth rate and a decrease in number and severity of wilted leaves under the salt and drought stresses. The MDA content and relative electrolyte leakage in transgenic lines under salt and drought stresses were significantly lower compared to those in non-transgenic plants, indicating that the TaLEA gene may enhance salt and drought tolerance by protecting cell membranes from damage. Moreover, amongst the lines analyzed for stress tolerance, the transgenic line 11 (T11 showed the highest tolerance levels under both salinity and drought stress conditions. These results indicated that the TaLEA gene could be a salt and drought tolerance candidate gene and could confer a broad spectrum of tolerance under abiotic stresses in poplars.

  6. Computational and statistical analyses of amino acid usage and physico-chemical properties of the twelve late embryogenesis abundant protein classes.

    Directory of Open Access Journals (Sweden)

    Emmanuel Jaspard

    Full Text Available Late Embryogenesis Abundant Proteins (LEAPs are ubiquitous proteins expected to play major roles in desiccation tolerance. Little is known about their structure - function relationships because of the scarcity of 3-D structures for LEAPs. The previous building of LEAPdb, a database dedicated to LEAPs from plants and other organisms, led to the classification of 710 LEAPs into 12 non-overlapping classes with distinct properties. Using this resource, numerous physico-chemical properties of LEAPs and amino acid usage by LEAPs have been computed and statistically analyzed, revealing distinctive features for each class. This unprecedented analysis allowed a rigorous characterization of the 12 LEAP classes, which differed also in multiple structural and physico-chemical features. Although most LEAPs can be predicted as intrinsically disordered proteins, the analysis indicates that LEAP class 7 (PF03168 and probably LEAP class 11 (PF04927 are natively folded proteins. This study thus provides a detailed description of the structural properties of this protein family opening the path toward further LEAP structure - function analysis. Finally, since each LEAP class can be clearly characterized by a unique set of physico-chemical properties, this will allow development of software to predict proteins as LEAPs.

  7. DrwH, a novel WHy domain-containing hydrophobic LEA5C protein from Deinococcus radiodurans, protects enzymatic activity under oxidative stress.

    Science.gov (United States)

    Jiang, Shijie; Wang, Jin; Liu, Xiaoli; Liu, Yingying; Guo, Cui; Zhang, Liwen; Han, Jiahui; Wu, Xiaoli; Xue, Dong; Gomaa, Ahmed E; Feng, Shuai; Zhang, Heng; Chen, Yun; Ping, Shuzhen; Chen, Ming; Zhang, Wei; Li, Liang; Zhou, Zhengfu; Zuo, Kaijing; Li, Xufeng; Yang, Yi; Lin, Min

    2017-08-24

    Water stress and hypersensitive response (WHy) domain is typically found as a component of atypical late embryogenesis abundant (LEA) proteins closely associated with resistance to multiple stresses in numerous organisms. Several putative LEA proteins have been identified in Deinococcus bacteria; however their precise function remains unclear. This work reports the characterization of a Deinococcus-specific gene encoding a novel WHy domain-containing hydrophobic LEA5C protein (named DrwH) in D. radiodurans R1. The expression of the drwH gene was induced by oxidative and salinity stresses. Inactivation of this gene resulted in increased sensitivity to oxidative and salinity stresses as well as reduced activities of antioxidant enzymes. The WHy domain of the DrwH protein differs structurally from that of a previously studied bacterial LEA5C protein, dWHy1, identified as a gene product from an Antarctic desert soil metagenome library. Further analysis indicated that in E. coli, the function of DrwH is related to oxidative stress tolerance, whereas dWHy1 is associated with freezing-thawing stress tolerance. Under oxidative stress induced by H2O2, DrwH protected the enzymatic activities of malate dehydrogenase (MDH) and lactate dehydrogenase (LDH). These findings provide new insight into the evolutionary and survival strategies of Deinococcus bacteria under extreme environmental conditions.

  8. Lea protein expression during cold-induced dehydration in the Arctic collembola Megaphorura arctica

    Directory of Open Access Journals (Sweden)

    Popović Ž.D.

    2011-01-01

    Full Text Available The Arctic springtail Megaphorura arctica (Tullberg, 1876 employs a strategy known as cryoprotective dehydration to survive winter temperatures as low as -25°C. During cryoprotective dehydration, water is lost from the animal to ice in its surroundings as a result of the difference in vapour pressure between the animal’s supercooled body fluids and ice (Worland et al., 1998; Holmstrup and Somme, 1998. This mechanism ensures that as the habitat temperature falls, the concentration of solutes remains high enough to prevent freezing (Holmstrup et al., 2002. In M. arctica, accumulation of trehalose, a cryo/anhydro protectant, occurs in parallel with dehydration. Recent studies have identified a number of genes and cellular processes involved in cryoprotective dehydration in M. arctica (Clark et al., 2007; Clark et al., 2009; Purać et al., 2011. One of them includes late embryogenesis abundant (LEA proteins. This study, together with that of Bahrndorff et al. (2008, suggests that LEA proteins may be involved in protective dehydration in this species.

  9. Two novel heat-soluble protein families abundantly expressed in an anhydrobiotic tardigrade.

    Directory of Open Access Journals (Sweden)

    Ayami Yamaguchi

    Full Text Available Tardigrades are able to tolerate almost complete dehydration by reversibly switching to an ametabolic state. This ability is called anhydrobiosis. In the anhydrobiotic state, tardigrades can withstand various extreme environments including space, but their molecular basis remains largely unknown. Late embryogenesis abundant (LEA proteins are heat-soluble proteins and can prevent protein-aggregation in dehydrated conditions in other anhydrobiotic organisms, but their relevance to tardigrade anhydrobiosis is not clarified. In this study, we focused on the heat-soluble property characteristic of LEA proteins and conducted heat-soluble proteomics using an anhydrobiotic tardigrade. Our heat-soluble proteomics identified five abundant heat-soluble proteins. All of them showed no sequence similarity with LEA proteins and formed two novel protein families with distinct subcellular localizations. We named them Cytoplasmic Abundant Heat Soluble (CAHS and Secretory Abundant Heat Soluble (SAHS protein families, according to their localization. Both protein families were conserved among tardigrades, but not found in other phyla. Although CAHS protein was intrinsically unstructured and SAHS protein was rich in β-structure in the hydrated condition, proteins in both families changed their conformation to an α-helical structure in water-deficient conditions as LEA proteins do. Two conserved repeats of 19-mer motifs in CAHS proteins were capable to form amphiphilic stripes in α-helices, suggesting their roles as molecular shield in water-deficient condition, though charge distribution pattern in α-helices were different between CAHS and LEA proteins. Tardigrades might have evolved novel protein families with a heat-soluble property and this study revealed a novel repertoire of major heat-soluble proteins in these anhydrobiotic animals.

  10. Tekstiilikonverents Hispaanias / Lea Pruuli

    Index Scriptorium Estoniae

    Pruuli, Lea

    1998-01-01

    8. - 16. märtsini Barcelonas ja Madridis European Textile Networki konverentsist. Eesti tekstiilikunstnike delegatsiooni kuulusid Katrin Pere, Signe Kivi, Lylian Meister, Merike Männi, Aune Taamal, Aet Ollisaar, Anne Tootma ja Lea Pruuli. Eesti tekstiilikunstist kõnelesid Signe Kivi, Katrin Pere. K. Pere valiti ETN juhatuse vaatlejaliikmeks.

  11. Molecular dynamics simulations and CD spectroscopy reveal hydration-induced unfolding of the intrinsically disordered LEA proteins COR15A and COR15B from Arabidopsis thaliana.

    Science.gov (United States)

    Navarro-Retamal, Carlos; Bremer, Anne; Alzate-Morales, Jans; Caballero, Julio; Hincha, Dirk K; González, Wendy; Thalhammer, Anja

    2016-10-07

    The LEA (late embryogenesis abundant) proteins COR15A and COR15B from Arabidopsis thaliana are intrinsically disordered under fully hydrated conditions, but obtain α-helical structure during dehydration, which is reversible upon rehydration. To understand this unusual structural transition, both proteins were investigated by circular dichroism (CD) and molecular dynamics (MD) approaches. MD simulations showed unfolding of the proteins in water, in agreement with CD data obtained with both HIS-tagged and untagged recombinant proteins. Mainly intramolecular hydrogen bonds (H-bonds) formed by the protein backbone were replaced by H-bonds with water molecules. As COR15 proteins function in vivo as protectants in leaves partially dehydrated by freezing, unfolding was further assessed under crowded conditions. Glycerol reduced (40%) or prevented (100%) unfolding during MD simulations, in agreement with CD spectroscopy results. H-bonding analysis indicated that preferential exclusion of glycerol from the protein backbone increased stability of the folded state.

  12. Retinoid signalling during embryogenesis

    NARCIS (Netherlands)

    Pijnappel, W.W.M.; Hendriks, H.F.J.; Durston, A.J.

    1996-01-01

    Conclusion: Retinoids are suspected to have multiple functions during embryogenesis, which are carried out via various different signal transduction pathways involving active retinoids and nuclear retinoid receptors. Research focuses on the identification of the retinoid signal transduction

  13. Meditsiinidoktor Lea Pehme / Alan Altraja

    Index Scriptorium Estoniae

    Altraja, Alan, 1965-

    2008-01-01

    7. sept. 2007 kaitses Tartu Ülikooli arstiteaduskonna nõukogu ees doktoriväitekirja "Tuberkuloosi epidemioloogiline olukord Eestis 1991-2003 rõhuasetusega kopsuvälisele tuberkuloosile ja kopsutuberkuloosi diagnoosimise viivitust mõjutavatele teguritele" Lea Pehme

  14. 34 CFR 200.71 - LEA eligibility.

    Science.gov (United States)

    2010-07-01

    ... least five percent of the LEA's total population ages 5 to 17 years, inclusive. (d) Education finance... 34 Education 1 2010-07-01 2010-07-01 false LEA eligibility. 200.71 Section 200.71 Education Regulations of the Offices of the Department of Education OFFICE OF ELEMENTARY AND SECONDARY EDUCATION...

  15. Lea blood group antigen on human platelets

    Energy Technology Data Exchange (ETDEWEB)

    Dunstan, R.A.; Simpson, M.B.; Rosse, W.F.

    1985-01-01

    One- and two-stage radioligand assays were used to determine if human platelets possess the Lea antigen. Goat IgG anti-Lea antibody was purified by multiple adsorptions with Le(a-b-) human red blood cells, followed by affinity chromatography with synthetic Lea substance and labeling with /sup 125/I. Human IgG anti-Lea antibody was used either in a two stage radioassay with /sup 125/I-labeled mouse monoclonal IgG anti-human IgG as the second antibody or, alternatively, purified by Staph protein A chromatography, labeled with /sup 125/I, and used in a one-stage radioassay. Platelets from donors of appropriate red blood cell phenotypes were incubated with the antisera, centrifuged through phthalate esters, and assayed in a gamma scintillation counter. Dose response and saturation curve analysis demonstrate the presence of Lewis a antigen on platelets from Lea+ donors. Furthermore, platelets from an Le(a-b-) donor incubated in Le (a+b-) plasma adsorb Lea antigen in a similar manner to red blood cells. The clinical significance of these antigens in platelet transfusion remains undefined.

  16. Isolation and expression analysis of LEA genes in peanut (Arachis ...

    Indian Academy of Sciences (India)

    Cloning and classification of peanut LEA protein genes. Three AhLEA1 genes with full-length ORFs were identified. These genes encoded three different proteins, namely, AhLEA1-1 (96 aa, 10.50 kDa, pI 5.53), AhLEA1-2 (94 aa, 10.14 kDa, pI 6.03) and AhLEA1-3 (98 aa, 10.70 kDa, pI 6.04). The alignment of the identified ...

  17. Application of Somatic Embryogenesis in Woody Plants.

    Science.gov (United States)

    Guan, Yuan; Li, Shui-Gen; Fan, Xiao-Fen; Su, Zhen-Hong

    2016-01-01

    Somatic embryogenesis is a developmental process where a plant somatic cell can dedifferentiate to a totipotent embryonic stem cell that has the ability to give rise to an embryo under appropriate conditions. This new embryo can further develop into a whole plant. In woody plants, somatic embryogenesis plays a critical role in clonal propagation and is a powerful tool for synthetic seed production, germplasm conservation, and cryopreservation. A key step in somatic embryogenesis is the transition of cell fate from a somatic cell to embryo cell. Although somatic embryogenesis has already been widely used in a number of woody species, propagating adult woody plants remains difficult. In this review, we focus on molecular mechanisms of somatic embryogenesis and its practical applications in economic woody plants. Furthermore, we propose a strategy to improve the process of somatic embryogenesis using molecular means.

  18. Somatic Embryogenesis: Identified Factors that Lead to Embryogenic Repression. A Case of Species of the Same Genus

    Science.gov (United States)

    Nic-Can, Geovanny I.; Galaz-Ávalos, Rosa M.; De-la-Peña, Clelia; Alcazar-Magaña, Armando; Wrobel, Kazimierz; Loyola-Vargas, Víctor M.

    2015-01-01

    Somatic embryogenesis is a powerful biotechnological tool for the mass production of economically important cultivars. Due to the cellular totipotency of plants, somatic cells under appropriate conditions are able to develop a complete functional embryo. During the induction of somatic embryogenesis, there are different factors involved in the success or failure of the somatic embryogenesis response. Among these factors, the origin of the explant, the culture medium and the in vitro environmental conditions have been the most studied. However, the secretion of molecules into the media has not been fully addressed. We found that the somatic embryogenesis of Coffea canephora, a highly direct embryogenic species, is disrupted by the metabolites secreted from C. arabica, a poorly direct embryogenic species. These metabolites also affect DNA methylation. Our results show that the abundance of two major phenolic compounds, caffeine and chlorogenic acid, are responsible for inhibiting somatic embryogenesis in C. canephora. PMID:26038822

  19. Optimization of somatic embryogenesis procedure for commercial ...

    African Journals Online (AJOL)

    Tuoyo Aghomotsegin

    2016-09-07

    Sep 7, 2016 ... from the Mars Center for Cocoa Science (MCCS) germplasm collection. Evaluation of a pilot scale production of plantlets from nine commercial cacao clones by somatic embryogenesis. Primary somatic embryogenesis procedures. Immature cacao flowers (approximately 900 in total), 6-8 mm in length, from ...

  20. Somatic Embryogenesis in Juniperus Procera using Juniperus ...

    African Journals Online (AJOL)

    The study of somatic embryogenesis in Juniperus communis has been conducted as a preliminary study for the further development of somatic embryogenesis, micropropagation and long-term conservation/cryopreservation in Juniperus procera, which is economically and ecologically important and endangered forest ...

  1. Somatic Embryogenesis in Juniperus Procera using Juniperus ...

    African Journals Online (AJOL)

    ABSTRACT. The study of somatic embryogenesis in Juniperus communis has been conducted as a preliminary study for the further development of somatic embryogenesis, micropropagation and long-term conservation/cryopreservation in Juniperus procera, which is economically and ecologically important and ...

  2. Sulphur depletion altered somatic embryogenesis in Theobroma ...

    African Journals Online (AJOL)

    Sulphur depletion altered somatic embryogenesis in Theobroma cacao L. Biochemical difference related to sulphur metabolism between embryogenic and non embryogenic calli. Minyaka Emile, Niemenak Nicolas, Issali Emmanuel Auguste, Sangare Abdourahamane, Denis Ndoumou Omokolo ...

  3. Zygotic embryogenesis in Anthurium (Araceae).

    Science.gov (United States)

    Matsumoto, T K; Kuehnle, A R; Webb, D T

    1998-11-01

    Morphological, anatomical, and histochemical aspects of zygotic embryogenesis by Anthurium andraeanum Lind. were investigated from 4 to 24 wk postpollination. Anatomical features were correlated with morphology of the spadix and capacity of embryos to germinate in vitro. Development from a single-cell zygote to fully mature seed takes 24 wk. The suspensor was two ranked and obvious during the early stages of embryogeny. It was apparent by week 8, substantial until week 14, and diminished rapidly until its absence by week 22. Differentiation of the shoot apex, cotyledon, and protoderm occurs at 14 wk. The embryo starts to derive nutrition from the endosperm at this time, and germination of cultured ovules reached 56%. By 20 wk the shoot apex had visible leaf primordia and the root apex was clearly defined. The cotyledon was well developed and surrounded the shoot tip. The storage of protein and starch was at its greatest in the endosperm and embryo. Furthermore, 100% germination of cultured ovules and embryos occurred at 20 wk and thereafter. Fully mature embryos at 24 wk are green and contain protoxylem elements.

  4. New theory of uterovaginal embryogenesis.

    Science.gov (United States)

    Makiyan, Zograb

    2016-01-02

    The explanation of uterine and vaginal embryogenesis in humans still poses many controversies, because it is difficult to assess early stages of an embryo. The literature review revealed many disagreements in Mullerian theory, inciting some authors to propose new embryological hypotheses. In the original Mullerian theory: the paramesonephral ducts form the Fallopian tubes, uterus and vagina; the mesonephral ducts regress in female embryos. The aim of this article is to investigate the development of Mullerian ducts in humans, using comparative analysis of fundamental embryological theory and various utero-vaginal anomalies. Between 1998 and 2015, 434 patients with various uterovaginal malformations had been operated on at the Scientific Centre of Obstetrics Gynaecology and Perynatology in Moscow. The anatomies of the uterovaginal malformations in these patients were diagnosed with ultrasound and MRI and then verified during surgical correction by laparoscopy. A systematic comparison of uterovaginal malformations to those in the literature has allowed us to formulate a new theory of embryonic morphogenesis. The new theory is significantly different: ovary, ovarian ligamentum proprium, and ligamentum teres uteri derive from gonadal ridges; Fallopian tubes and vagina completely develop from mesonephral ducts. The uterus develops in the area of intersection between the mesonephral ducts with gonadal ridges by the fusion of the two. The new theory may to induce future embryological studies. The hypothetic possibility that the ovary and endometrium derive from the gonadal ridges could be the key to understanding the enigmatic aetiologies of extragenital and ovarian endometriosis.

  5. Missugune tuleb X Academica? / Lea Kivi, Lauri Randveer

    Index Scriptorium Estoniae

    Kivi, Lea

    2006-01-01

    Vastavad Lea Kivi ja Lauri Randveer välissuhete osakonnast 23. - 27. septembrini toimuvast Saksa-Eesti akadeemilisest nädalast teemal "Viimane aastakümme Eestis ja Saksamaal: kuhu me oleme jõudnud?"

  6. Some ethylene biosynthesis and AP2/ERF genes reveal a specific pattern of expression during somatic embryogenesis in Hevea brasiliensis.

    Science.gov (United States)

    Piyatrakul, Piyanuch; Putranto, Riza-Arief; Martin, Florence; Rio, Maryannick; Dessailly, Florence; Leclercq, Julie; Dufayard, Jean-François; Lardet, Ludovic; Montoro, Pascal

    2012-12-26

    Ethylene production and signalling play an important role in somatic embryogenesis, especially for species that are recalcitrant in in vitro culture. The AP2/ERF superfamily has been identified and classified in Hevea brasiliensis. This superfamily includes the ERFs involved in response to ethylene. The relative transcript abundance of ethylene biosynthesis genes and of AP2/ERF genes was analysed during somatic embryogenesis for callus lines with different regeneration potential, in order to identify genes regulated during that process. The analysis of relative transcript abundance was carried out by real-time RT-PCR for 142 genes. The transcripts of ERFs from group I, VII and VIII were abundant at all stages of the somatic embryogenesis process. Forty genetic expression markers for callus regeneration capacity were identified. Fourteen markers were found for proliferating calli and 35 markers for calli at the end of the embryogenesis induction phase. Sixteen markers discriminated between normal and abnormal embryos and, lastly, there were 36 markers of conversion into plantlets. A phylogenetic analysis comparing the sequences of the AP2 domains of Hevea and Arabidopsis genes enabled us to predict the function of 13 expression marker genes. This first characterization of the AP2/ERF superfamily in Hevea revealed dramatic regulation of the expression of AP2/ERF genes during the somatic embryogenesis process. The gene expression markers of proliferating callus capacity to regenerate plants by somatic embryogenesis should make it possible to predict callus lines suitable to be used for multiplication. Further functional characterization of these markers opens up prospects for discovering specific AP2/ERF functions in the Hevea species for which somatic embryogenesis is difficult.

  7. Embryogenesis in Oak species. A review

    Directory of Open Access Journals (Sweden)

    Aranzazu Gomez-Garay

    2014-08-01

    Full Text Available Aim of study: A review on the propagation methods of four Quercus species, namely Q. suber, Q. robur, Q. ilex and Q. canariensis, through somatic embryogenesis and anther embryogenesis are presented.Area of study: The study comprises both Mediterranean and Atlantic oak species located in Spain.Material and Methods: Somatic embryogenesis was induced on immature zygotic embryos of diverse oak species, permitting the multiplication of half-sib families. Induction of haploid embryos and doubled haploids was assayed in both Q. suber and Q. ilex by temperature stress treatments of anthers containing late vacuolated microspores. The haploid origin of the anther embryos has been evaluated by quantitative nuclear DNA analysis through flow cytometry and by DNA microsatellite markers. Genetic transformation of cork oak has also been performed by means of Agrobacterium tumefaciens vectors. Proteomic analysis has been conducted to screen the diverse protein profiles followed by in vitro derived embryos during their development.Research highlights: Successful plant regeneration from both somatic and haploid embryos has been achieved. In the particular case of cork oak, doubled-haploid plants were obtained. Plantlets regenerated from selected parent trees through somatic embryogenesis were acclimated in the greenhouse and in the nursery, and were planted in an experimental plot in the field. Preliminary evaluation of the cork quality of the plants showed a good heritability correlation with the parent trees. This article revises the work of and is dedicated to Dr. M.A. Bueno, who devoted much of her professional life to the research on Biotechnology and Genetics of forest species, leading the Laboratory of Forest Biotechnology at the Spanish Institute of Agronomic Research (INIA.Key words: anther embryogenesis; microspore; pollen; Quercus canariensis; Quercus ilex; Quercus robur; Quercus suber; somatic embryogenesis

  8. Temperature manipulation during layer chick embryogenesis

    NARCIS (Netherlands)

    Walstra, I.; Napel, ten J.; Kemp, B.; Brand, van den H.

    2010-01-01

    The current study investigated the effects of temperature manipulation (TM) during late embryogenesis on temperature preference, response to high environmental temperature, behavior, and performance in young layer chicks. Control (CC) embryos (n = 96) were incubated at 37.8°C eggshell temperature

  9. Somatic embryogenesis and plant regeneration of recalcitrant ...

    African Journals Online (AJOL)

    Many restrictive factors still remain in cotton tissue culture such as long duration, unpredictability and a high degree of genotype dependence. The main objective of this study was to develop a protocol allowing consistent somatic embryogenesis and plant regeneration from five recalcitrant cotton cultivars. Our results ...

  10. Optimization of somatic embryogenesis procedure for commercial ...

    African Journals Online (AJOL)

    Optimization of somatic embryogenesis procedure for commercial clones of Theobroma cacao L. ... Overall, flower petals performed better than staminodes, and our best performing genotype yielded an average of 7-10 embryos produced in brown callus explants with embryogenic response during primary somatic ...

  11. Chitinases and arabinogalactan proteins in somatic embryogenesis

    NARCIS (Netherlands)

    Hengel, van A.J.

    1998-01-01

    In vitro cultured carrot suspension cells can function as starting material for the generation of somatic embryos. Compounds secreted by suspension cells can influence the process of somatic embryogenesis. One class of such compounds, the secreted EP3 endochitinases,

  12. Somatic Embryogenesis and Plant Regeneration in Eggplant ...

    African Journals Online (AJOL)

    DR SIDHU

    2013-02-20

    Feb 20, 2013 ... Two as well as three way interactions of three eggplant genotypes, media compositions and explants. (hypocotyl, cotyledon and ... hypocotyl of BR-16 was not able to induce somatic embryogenesis on MS media fortified with 1.5 mgl-1 IBA + 1.0 mgl-1 BAP. ..... Manipulation of Plants. Oxford University Press ...

  13. Comparative proteomic analysis of early somatic and zygotic embryogenesis in Theobroma cacao L.

    Science.gov (United States)

    Noah, Alexandre Mboene; Niemenak, Nicolas; Sunderhaus, Stephanie; Haase, Christin; Omokolo, Denis Ndoumou; Winkelmann, Traud; Braun, Hans-Peter

    2013-01-14

    Somatic embryogenesis can efficiently foster the propagation of Theobroma cacao, but the poor quality of resulted plantlet hinders the use of this technique in the commercial scale. The current study has been initiated to systematically compare the physiological mechanisms underlying somatic and zygotic embryogenesis in T. cacao on the proteome level. About 1000 protein spots per fraction could be separated by two-dimensional isoelectric focusing/SDS PAGE. More than 50 of the protein spots clearly differed in abundance between zygotic and somatic embryos: 33 proteins spots were at least 3-fold higher in abundance in zygotic embryos and 20 in somatic embryos. Analyses of these protein spots differing in volume by mass spectrometry resulted in the identification of 68 distinct proteins. Many of the identified proteins are involved in genetic information processing (21 proteins), carbohydrate metabolism (11 proteins) and stress response (7 proteins). Somatic embryos especially displayed many stress related proteins, few enzymes involved in storage compound synthesis and an exceptional high abundance of endopeptidase inhibitors. Phosphoenolpyruvate carboxylase, which was accumulated more than 3-fold higher in zygotic embryos, represents a prominent enzyme in the storage compound metabolism in cacao seeds. Implications on the improvement of somatic embryogenesis in cacao are discussed. Copyright © 2012 Elsevier B.V. All rights reserved.

  14. Localization and identification of phenolic compounds in Theobroma cacao L. somatic embryogenesis.

    Science.gov (United States)

    Alemanno, L; Ramos, T; Gargadenec, A; Andary, C; Ferriere, N

    2003-10-01

    Cocoa breeders and growers continue to face the problem of high heterogeneity between individuals derived from one progeny. Vegetative propagation by somatic embryogenesis could be a way to increase genetic gains in the field. Somatic embryogenesis in cocoa is difficult and this species is considered as recalcitrant. This study was conducted to investigate the phenolic composition of cocoa flowers (the explants used to achieve somatic embryogenesis) and how it changes during the process, by means of histochemistry and conventional chemical techniques. In flowers, all parts contained polyphenolics but their locations were specific to the organ considered. After placing floral explants in vitro, the polyphenolic content was qualitatively modified and maintained in the calli throughout the culture process. Among the new polyphenolics, the three most abundant were isolated and characterized by 1H- and 13C-NMR. They were hydroxycinnamic acid amides: N-trans-caffeoyl-l-DOPA or clovamide, N-trans-p-coumaroyl-l-tyrosine or deoxiclovamide, and N-trans-caffeoyl-l-tyrosine. The same compounds were found also in fresh, unfermented cocoa beans. The synthesis kinetics for these compounds in calli, under different somatic embryogenesis conditions, revealed a higher concentration under non-embryogenic conditions. Given the antioxidant nature of these compounds, they could reflect the stress status of the tissues.

  15. 34 CFR 300.815 - Subgrants to LEAs.

    Science.gov (United States)

    2010-07-01

    ... 34 Education 2 2010-07-01 2010-07-01 false Subgrants to LEAs. 300.815 Section 300.815 Education Regulations of the Offices of the Department of Education (Continued) OFFICE OF SPECIAL EDUCATION AND REHABILITATIVE SERVICES, DEPARTMENT OF EDUCATION ASSISTANCE TO STATES FOR THE EDUCATION OF CHILDREN WITH...

  16. 34 CFR 300.817 - Reallocation of LEA funds.

    Science.gov (United States)

    2010-07-01

    ... 34 Education 2 2010-07-01 2010-07-01 false Reallocation of LEA funds. 300.817 Section 300.817 Education Regulations of the Offices of the Department of Education (Continued) OFFICE OF SPECIAL EDUCATION AND REHABILITATIVE SERVICES, DEPARTMENT OF EDUCATION ASSISTANCE TO STATES FOR THE EDUCATION OF...

  17. 34 CFR 300.816 - Allocations to LEAs.

    Science.gov (United States)

    2010-07-01

    ... 34 Education 2 2010-07-01 2010-07-01 false Allocations to LEAs. 300.816 Section 300.816 Education Regulations of the Offices of the Department of Education (Continued) OFFICE OF SPECIAL EDUCATION AND REHABILITATIVE SERVICES, DEPARTMENT OF EDUCATION ASSISTANCE TO STATES FOR THE EDUCATION OF CHILDREN WITH...

  18. Isolation and expression analysis of LEA genes in peanut

    Indian Academy of Sciences (India)

    In order to isolate peanut genes, an expressed sequence tag (EST) sequencing project was carried out using a peanut seed cDNA library. From 6258 ESTs, 19 LEA-encoding genes were identified and could be classified into eight distinct groups. Expression of these genes in seeds at different developmental stages and in ...

  19. Efficient Hardware Implementation of the Lightweight Block Encryption Algorithm LEA

    Directory of Open Access Journals (Sweden)

    Donggeon Lee

    2014-01-01

    Full Text Available Recently, due to the advent of resource-constrained trends, such as smartphones and smart devices, the computing environment is changing. Because our daily life is deeply intertwined with ubiquitous networks, the importance of security is growing. A lightweight encryption algorithm is essential for secure communication between these kinds of resource-constrained devices, and many researchers have been investigating this field. Recently, a lightweight block cipher called LEA was proposed. LEA was originally targeted for efficient implementation on microprocessors, as it is fast when implemented in software and furthermore, it has a small memory footprint. To reflect on recent technology, all required calculations utilize 32-bit wide operations. In addition, the algorithm is comprised of not complex S-Box-like structures but simple Addition, Rotation, and XOR operations. To the best of our knowledge, this paper is the first report on a comprehensive hardware implementation of LEA. We present various hardware structures and their implementation results according to key sizes. Even though LEA was originally targeted at software efficiency, it also shows high efficiency when implemented as hardware.

  20. Direct somatic embryogenesis in Coffea canephora.

    Science.gov (United States)

    Quiroz-Figueroa, Francisco; Monforte-González, Miriam; Galaz-Avalos, Rosa M; Loyola-Vargas, Victor M

    2006-01-01

    Somatic embryogenesis (SE) provides a useful model to study embryo development in plants. In contrast to zygotic embryogenesis, SE can easily be observed, the culture conditions can be controlled, and large quantities of embryos can be easily obtained. In Coffea spp several model systems have been reported for in vitro SE induction. SE for coffee was first reported in Coffea canephora. Several systems have been developed since then, including SE from callus cultures derived from leaf explants; a two-phase experimental protocol for SE from leaves of Coffea arabica; and from leaf explants of Arabusta or C. arabica using a medium with cytokinins. Here we report a protocol using young leaves from in vitro seedling pre-conditioned with growth regulators. This is a simplified method to obtain a faster and more efficient protocol to produce direct somatic embryos in C. canephora.

  1. Optimization of somatic embryogenesis procedure for commercial ...

    African Journals Online (AJOL)

    Tuoyo Aghomotsegin

    2016-09-07

    Sep 7, 2016 ... de mejoramientogénetico del CATIE,” M.S. thesis, Escuela de. Posgrado, Programa de educación para el desarrollo y la conservación del Centro Agronómico Tropical de Investigación y. Enseñanza, Turrialba, ON Costa Rica. pp. 1-92. Sondahl M R, Liu S, Bellato C (1993). Cacao somatic embryogenesis.

  2. Transcriptome analysis of zebrafish embryogenesis using microarrays.

    Directory of Open Access Journals (Sweden)

    Sinnakaruppan Mathavan

    2005-08-01

    Full Text Available Zebrafish (Danio rerio is a well-recognized model for the study of vertebrate developmental genetics, yet at the same time little is known about the transcriptional events that underlie zebrafish embryogenesis. Here we have employed microarray analysis to study the temporal activity of developmentally regulated genes during zebrafish embryogenesis. Transcriptome analysis at 12 different embryonic time points covering five different developmental stages (maternal, blastula, gastrula, segmentation, and pharyngula revealed a highly dynamic transcriptional profile. Hierarchical clustering, stage-specific clustering, and algorithms to detect onset and peak of gene expression revealed clearly demarcated transcript clusters with maximum gene activity at distinct developmental stages as well as co-regulated expression of gene groups involved in dedicated functions such as organogenesis. Our study also revealed a previously unidentified cohort of genes that are transcribed prior to the mid-blastula transition, a time point earlier than when the zygotic genome was traditionally thought to become active. Here we provide, for the first time to our knowledge, a comprehensive list of developmentally regulated zebrafish genes and their expression profiles during embryogenesis, including novel information on the temporal expression of several thousand previously uncharacterized genes. The expression data generated from this study are accessible to all interested scientists from our institute resource database (http://giscompute.gis.a-star.edu.sg/~govind/zebrafish/data_download.html.

  3. Spaceflight reduces somatic embryogenesis in orchardgrass (Poaceae)

    Science.gov (United States)

    Conger, B. V.; Tomaszewski, Z. Jr; McDaniel, J. K.; Vasilenko, A.

    1998-01-01

    Somatic embryos initiate and develop from single mesophyll cells in in vitro cultured leaf segments of orchard-grass (Dactylis glomerata L.). Segments were plated at time periods ranging from 21 to 0.9 d (21 h) prior to launch on an 11 d spaceflight (STS-64). Using a paired t-test, there was no significant difference in embryogenesis from preplating periods of 14 d and 21 d. However, embryogenesis was reduced by 70% in segments plated 21 h before launch and this treatment was significant at P=0.0001. The initial cell divisions leading to embryo formation would be taking place during flight in this treatment. A higher ratio of anticlinal:periclinal first cell divisions observed in the flight compared to the control tissue suggests that microgravity affects axis determination and embryo polarity at a very early stage. A similar reduction in zygotic embryogenesis would reduce seed formation and have important implications for long-term space flight or colonization where seeds would be needed either for direct consumption or to grow another generation of plants.

  4. How microspores transform into haploid embryos: changes associated with embryogenesis induction and microspore-derived embryogenesis.

    Science.gov (United States)

    Seguí-Simarro, José M; Nuez, Fernando

    2008-09-01

    Microspore embryogenesis is the most powerful androgenic pathway to produce haploid and doubled haploid plants. To deviate a microspore toward embryogenesis, a number of factors, different for each species, must concur at the same time and place. Once induced, the microspore undergoes numerous changes at different levels, from overall morphology to gene expression. Induction of microspore embryogenesis not only implies the expression of an embryogenic program, but also a stress-related cellular response and a repression of the gametophytic program to revert the microspore to a totipotent status. In this review, we compile the most recent advances in the understanding of the changes undergone by the induced microspore to readapt to the new developmental scenario. We devote special attention to the efforts made to uncover changes in the transcriptome of the induced microspore and microspore-derived embryo (MDE). Finally, we discuss the influence that an in vitro environment exerts over the MDE, as compared with its zygotic counterpart.

  5. Lea Endrikson - läheksin Venemaale tagasi, kui tööle kutsuks Gazprom või Lukoil / Lea Endrikson ; interv. Aive Antsov

    Index Scriptorium Estoniae

    Endrikson, Lea, 1964-

    2007-01-01

    Ilukaupluste keti TKM Beauty juht ning endine Venemaa Stockmanni kaubamajade turundusjuht Lea Endrikson vastab juhitööd, oma karjääri ning tööpakkumisi puudutavatele küsimustele. Lisa: TKM Beauty; Lea Endrikson

  6. Embryogenesis of brassica rapa l. under clinorotation

    Science.gov (United States)

    Popova, A.; Ivanenko, G.

    Investigation of reproductive development of higher plants in spaceflight represents scientific interest first of all with the necessity to work out the plant space technologies for creation of controlled life-support systems. In such systems mainly the higher plants are considered to be an important component that makes it necessary to obtain the several generations of higher plants with their full ontogenesis. As a rule, seeds obtained in three species of the higher plants in a series of experiments differ from the control by some parameters (Merkis, Laurinavichius, 1983; Musgrave et al., 1998; 2000; Levinskikh et all. 1999; Stankovich et al., 2002). It was shown, that immature embryos generated in microgravity were at a range of developmental stage, while the ground control embryos had all reached the premature stage of development (Kuang et al., 2003). Besides, the distinctions in a degree of nutrient substances accumulation in them were revealed (Kuang et al., 2000). Therefore, the elucidation of the possible reasons for distortion of plant reproduction in microgravity demands the further research. In this study we examined embryogenesis of higher plant Brassica rapa L. with an application of slow horizontal clinostats, that allows to deprive the plants the opportunity to perceive the gravitational stimulus. Some plants were clinorotated from the moment sowing of seeds; in other series the experiment plants were placed on clinostats after formation of flower buds. Temporal fixation of the material was used in these experiments, which allow to obtain material for studying of consecutive stages of embryogenesis. The development of 2-21 day-old embryos was studied. Comparative embryological analysis has shown a similarity in the main of process of embryo differentiation produced under clinorotation and in the stationary control. At the early stages of embryogenesis, the distortion in suspensor formation was observed more frequently. Embryos generated in

  7. Storia di Lea Garofalo e di sua figlia Denise

    Directory of Open Access Journals (Sweden)

    Anna Lisa Tota

    2017-11-01

    Full Text Available Questo articolo analizza il caso di Lea Garofalo e le modalità, attraverso cui la sua memoria è stata iscritta nel discorso pubblico italiano. In particolare, focalizza l’attenzione sul film “Lea” di Marco Tullio Giordana, dedicato appunto alla storia del femminicidio di Lea Garofalo e alla storia di sua figlia Denise Cosco. Questo caso è emblematico, in quanto mette a tema il destino e le difficoltà che incontrano le donne, quando decidono di opporsi e resistere alla cultura mafiosa e criminale delle famiglie, di cui fanno parte. Inoltre, esso sfida quegli stereotipi, ancora largamente diffusi nel senso comune, secondo i quali questi fenomeni di criminalità organizzata sarebbero diffusi prevalentemente nel sud del nostro paese. Di fatto questo femminicidio dell’n’drangheta avviene a Milano e riguarda attività criminali organizzate nel capoluogo lombardo. Parole chiave: Memoria pubblica, femminicidio, lotta alla criminalità organizzata, testimoni di giustizia   This article deals with Lea Garofalo’s case and the forms that have shaped its cultural memory in the Italian public discourse. Its main focus is related to “Lea”, the movie by the Italian filmmaker Marco Tullio Giordana on the story of this femicide and the relation between Lea Garofalo and her daughter named Denise Cosco. This case is exemplary, because it concerns the destine and the difficulties encountered by women, when they decide to contrast and resist against the mafiosi culture of the families, to which they belong. Moreover, it challenges those stereotypes, still very common, according to which mafia’s and camorra’s criminal activities are overspread mainly in South Italy. This femicide has been committed by n’drangheta in the centre of Milan and it is caused through criminal activities that  took place in the North part of the country. Keywords: Public memory, femicide, fight against organized crime, witnesses

  8. Comparison of callus induction and somatic embryogenesis of some ...

    African Journals Online (AJOL)

    The Kerman genotype did not show embryogenesis. In the histological studies, the different development stages of the embryos (globular, heart, torpedo and cotyledonary) together with callus cells were showed. Key words: Hypocotyl explants, somatic embryo, in vitro regeneration, germination, somatic embryogenesis ...

  9. Boron-Mediated Plant Somatic Embryogenesis: A Provocative Model

    Directory of Open Access Journals (Sweden)

    Dhananjay K. Pandey

    2012-01-01

    Full Text Available A central question in plant regeneration biology concerns the primary driving forces invoking the acquisition of somatic embryogenesis. Recently, the role of micronutrient boron (B in the initiation and perpetuation of embryogenesis has drawn considerable attention within the scientific community. This interest may be due in part to the bewildering observation that the system-wide induction of embryogenic potential significantly varied in response to a minimal to optimal supply of B (minimal ≤ 0.1 mM, optimal = 0.1 mM. At the cellular level, certain channel proteins and cell wall-related proteins important for the induction of embryogenesis have been shown to be transcriptionally upregulated in response to minimal B supply suggesting the vital role of B in the induction of embryogenesis. At the molecular level, minimal to no B supply increased the endogenous level of auxin, which subsequently influenced the auxin-inducible somatic embryogenesis receptor kinases, suggesting the role of B in the induction of embryogenesis. Also, minimal B concentration may “turn on” other genetic and/or cellular transfactors reported earlier to be essential for cell-restructuring and induction of embryogenesis. In this paper, both the direct and indirect roles of B in the induction of somatic embryogenesis are highlighted and suggested for future validation.

  10. The use of somatic embryogenesis for plant propagation in cassava

    NARCIS (Netherlands)

    Raemakers, K.; Jacobsen, E.; Visser, R.

    2000-01-01

    In cassava, somatic embryogenesis starts with the culture of leaf explants on solid Murashige and Skoog-based medium supplemented with auxins. Mature somatic embryos are formed within 6 wk. The cotyledons of the primary somatic embryos are used as explants for a new cycle of somatic embryogenesis.

  11. Microspore embryogenesis: reprogramming cell fate from pollen to embryo development

    NARCIS (Netherlands)

    Hui Li,

    2014-01-01

    Microspore embryogenesis is an expression of plant cell totipotency that leads to the production of haploid embryos. Besides being a widely exploited plant breeding tool, microspore embryogenesis is also a fascinating system that can be used to obtain a deeper mechanistic understanding of plant

  12. Direct somatic embryogenesis in Swietenia macrophylla King

    Directory of Open Access Journals (Sweden)

    Raúl Collado

    2006-04-01

    Full Text Available Swietenia macrophylla King is difficult to be propagated by tissue culture and there is not an efficient system via organogenesis, due to problems of microbial contamination, phenolic oxidation and death of tissue in the phase of in vitro establishment of explants. In order to establish a protocol for obtaining somatic embryos, zygotic embryos were used as initial plant material. Three combinations of 2,4-D with kinetin were studied, to obtain the formation of somatic embryos. After six weeks of culture, the number of explants with high and low somatic embryogenesis frequency were determined. So that the somatic embryos in globular stage reach the final stages of torpedo and cotyledonal, these were placed in three treatments with 6-BAP (0.2, 0.4 y 0.6 mg.l-1. The number of somatic embryos that reached the torpedo and cotyledonal stages were evaluated after 30 days of culture. Results demonstrated that direct somatic embryogenesis from immature zygotic embryos is obtained in the culture medium composed by MS salts with 4.0 mg.l-1 of 2,4-D and 1.0 mg.l-1 of kinetin. Higher percentage of somatic embryos in cotiledonal stage (91.7 %, was obtained with 0.4 mg.l-1 of 6-BAP. Key word: forestry, growth regulator, mahogany, somatic embryo, tissue culture

  13. Studies for Somatic Embryogenesis in Sweet Potato

    Science.gov (United States)

    Bennett, J. Rasheed; Prakash, C. S.

    1997-01-01

    The purpose of this study was to improve the somatic embryo (SE) system for plant production of sweet potato (Ipomoea batatas L(Lam)). Explants isolated from SE-derived sweet potato plants were compared with control (non SE-derived) plants for their competency for SE production. Leaf explants were cultured on Murashige-Skoog (MS) medium with 2,4-dichlorophenoxy acetic acid (0.2 mg/L) and 6-benzylaminopurine (2.5 mg/L) for 2 weeks in darkness and transferred to MS medium with abscisic acid (2.5 mg/L). Explants isolated from those plants developed through somatic embryogenesis produced new somatic embryos rapidly and in higher frequency than those isolated from control plants They also appeared to grow faster in tissue culture than the control plants. Current studies in the laboratory are examining whether plants derived from a cyclical embryogenesis system (five cycles) would have any further positive impact on the rapidity and frequency of somatic embryo development. More detailed studies using electron microscopy are expected to show the point of origin of the embryos and to allow determination of their quality throughout the cyclical process. This study may facilitate improved plant micropropagation, gene transfer and germplasm conservation in sweet potato.

  14. Rape embryogenesis. V. Accumulation of lipid bodies

    Directory of Open Access Journals (Sweden)

    Teresa Tykarska

    2014-01-01

    Full Text Available In embryo of winter rape var. Górczański lipid bodies have been observed in the light microscope starting from two-celled proembryo. Their number and size increase in the course of embryogenesis, especially since greening of endosperm and embryo. Lipid bodies, either single or in clusters, are present in all embryo cells, the clusters occur in various parts of the cytoplasm. During embryo maturation lipid bodies move and locate in a number of layers by the cell wall. At the same time their appearance change. Typical storage lipids originate. Lipid bodies are unevenly distributed within the embryo: their number and size decrease along the embryo axis from its part below cotyledons towards root apex. Moreover, they are histologically diversified: the biggest are located in epidermis and cortex, whereas the smallest -in central cylinder. In columella there are fewest lipid bodies.

  15. Rape embryogenesis. III. Embryo development in time

    Directory of Open Access Journals (Sweden)

    Teresa Tykarska

    2014-01-01

    Full Text Available It was found that the growth curve of the rape embryo axis is of triple sigmoid type. Embryo growth occurs in 3 phases corresponding to 3 different periods of development. Phase I includes growth of the apical cell up to it's division into two layers of octants. Phase II comprises the increase of the spherical proembryo to the change of its symmetry from radial to bilateral. Phase III includes, growth of the embryo from the heart stage up to the end of embryogenesis. In each phase the relative growth rate increases drastically and then diminishes. The differences in growth intensity during the same phase are several-fold. The growth intensity maximum of the embryo axis occurs in phase II. The phasic growth intensity maxima occur: in phase I during apical cell elongation, :before its division, and in phases II and III in the periods of cell division ;growth in globular and torpedo-shaped -shaped embryos.

  16. Large impact of the apoplast on somatic embryogenesis in Cyclamen persicum offers possibilities for improved developmental control in vitro

    Directory of Open Access Journals (Sweden)

    Zimmer Andreas D

    2010-04-01

    Full Text Available Abstract Background Clonal propagation is highly desired especially for valuable horticultural crops. The method with the potentially highest multiplication rate is regeneration via somatic embryogenesis. However, this mode of propagation is often hampered by the occurrence of developmental aberrations and non-embryogenic callus. Therefore, the developmental process of somatic embryogenesis was analysed in the ornamental crop Cyclamen persicum by expression profiling, comparing different developmental stages of embryogenic cell cultures, zygotic vs. somatic embryos and embryogenic vs. non-embryogenic cell cultures. Results The analysis was based on a cDNA microarray representing 1,216 transcripts and was exemplarily validated by realtime PCR. For this purpose relative transcript abundances of homologues of a putative receptor kinase, two different glutathione S-transferases (GST, a xyloglucan endotransglycosylase (XET and a peroxidase (POX were quantitatively measured by realtime PCR for three different comparisons. In total, 417 genes were found to be differentially expressed. Gene Ontology annotation revealed that transcripts coding for enzymes that are active in the extracellular compartment (apoplast were significantly overrepresented in several comparisons. The expression profiling results are underpinned by thorough histological analyses of somatic and zygotic embryos. Conclusions The putative underlying physiological processes are discussed and hypotheses on improvement of the protocol for in vitro somatic embryogenesis in Cyclamen persicum are deduced. A set of physiological markers is proposed for efficient molecular control of the process of somatic embryogenesis in C. persicum. The general suitability of expression profiling for the development and improvement of micropropagation methods is discussed.

  17. The LEA's Role in a Decentralized School System: The School Principals' View

    Science.gov (United States)

    Addi-Raccah, Audrey; Gavish, Yakov

    2010-01-01

    Since the wave of school reform decentralization, schools now maintain a more dynamic and diverse relationship with their environment than they did in the past. School principals' relationships with the local educational authority (LEA) are a prominent example of this change in Israel. LEAs try to gain more pedagogic influence over schools while…

  18. Developmental localization and methylesterification of pectin epitopes during somatic embryogenesis of banana (Musa spp. AAA).

    Science.gov (United States)

    Xu, Chunxiang; Zhao, Lu; Pan, Xiao; Samaj, Jozef

    2011-01-01

    The plant cell walls play an important role in somatic embryogenesis and plant development. Pectins are major chemical components of primary cell walls while homogalacturonan (HG) is the most abundant pectin polysaccharide. Developmental regulation of HG methyl-esterification degree is important for cell adhesion, division and expansion, and in general for proper organ and plant development. Developmental localization of pectic homogalacturonan (HG) epitopes and the (1→4)-β-D-galactan epitope of rhamnogalacturonan I (RG-I) and degree of pectin methyl-esterification (DM) were studied during somatic embryogenesis of banana (Musa spp. AAA). Histological analysis documented all major developmental stages including embryogenic cells (ECs), pre-globular, globular, pear-shaped and cotyledonary somatic embryos. Histochemical staining of extracellularly secreted pectins with ruthenium red showed the most intense staining at the surface of pre-globular, globular and pear-shaped somatic embryos. Biochemical analysis revealed developmental regulation of galacturonic acid content and DM in diverse embryogenic stages. Immunodots and immunolabeling on tissue sections revealed developmental regulation of highly methyl-esterified HG epitopes recognized by JIM7 and LM20 antibodies during somatic embryogenesis. Cell walls of pre-globular/globular and late-stage embryos contained both low methyl-esterified HG epitopes as well as partially and highly methyl-esterified ones. Extracellular matrix which covered surface of early developing embryos contained pectin epitopes recognized by 2F4, LM18, JIM5, JIM7 and LM5 antibodies. De-esterification of cell wall pectins by NaOH caused a decrease or an elimination of immunolabeling in the case of highly methyl-esterified HG epitopes. However, immunolabeling of some low methyl-esterified epitopes appeared stronger after this base treatment. These data suggest that both low- and highly-methyl-esterified HG epitopes are developmentally

  19. Developmental localization and methylesterification of pectin epitopes during somatic embryogenesis of banana (Musa spp. AAA.

    Directory of Open Access Journals (Sweden)

    Chunxiang Xu

    Full Text Available The plant cell walls play an important role in somatic embryogenesis and plant development. Pectins are major chemical components of primary cell walls while homogalacturonan (HG is the most abundant pectin polysaccharide. Developmental regulation of HG methyl-esterification degree is important for cell adhesion, division and expansion, and in general for proper organ and plant development.Developmental localization of pectic homogalacturonan (HG epitopes and the (1→4-β-D-galactan epitope of rhamnogalacturonan I (RG-I and degree of pectin methyl-esterification (DM were studied during somatic embryogenesis of banana (Musa spp. AAA. Histological analysis documented all major developmental stages including embryogenic cells (ECs, pre-globular, globular, pear-shaped and cotyledonary somatic embryos. Histochemical staining of extracellularly secreted pectins with ruthenium red showed the most intense staining at the surface of pre-globular, globular and pear-shaped somatic embryos. Biochemical analysis revealed developmental regulation of galacturonic acid content and DM in diverse embryogenic stages. Immunodots and immunolabeling on tissue sections revealed developmental regulation of highly methyl-esterified HG epitopes recognized by JIM7 and LM20 antibodies during somatic embryogenesis. Cell walls of pre-globular/globular and late-stage embryos contained both low methyl-esterified HG epitopes as well as partially and highly methyl-esterified ones. Extracellular matrix which covered surface of early developing embryos contained pectin epitopes recognized by 2F4, LM18, JIM5, JIM7 and LM5 antibodies. De-esterification of cell wall pectins by NaOH caused a decrease or an elimination of immunolabeling in the case of highly methyl-esterified HG epitopes. However, immunolabeling of some low methyl-esterified epitopes appeared stronger after this base treatment.These data suggest that both low- and highly-methyl-esterified HG epitopes are

  20. Indirect somatic embryogenesis in cassava for genetic modification purposes.

    Science.gov (United States)

    Raemakers, Krit; Pereira, Isolde; van Putten, Herma Koehorst; Visser, Richard

    2006-01-01

    In cassava both direct and indirect somatic embryogenesis is described. Direct somatic embryogenesis starts with the culture of leaf explants on Murashige and Skoog (MS) medium supplemented with auxins. Somatic embryos undergo secondary somatic embryogenesis when cultured on the same medium. Indirect somatic embryogenesis is initiated by subculture of directly induced embryogenic tissue on auxin-supplemented medium with Gresshoff and Doy salts and vitamins. A very fine friable embryogenic callus (FEC) is formed after a few rounds of subculture and stringent selection. This FEC is maintained by subculture on auxin supplemented medium. Lowering of the auxin concentration allows the FEC to form mature somatic embryos that develop into plants when transferred to a cytokinin-supplemented medium.

  1. REGENERATION OF Pimpinella pruatjan THROUGH SOMATIC EMBRYOGENESIS

    Directory of Open Access Journals (Sweden)

    I. Roostika

    2016-10-01

    Full Text Available Pruatjan (Pimpinella pruatjan Molk. is an Indonesian endangered plant which has various medicinal properties such as aphrodisiac, diuretic, and tonic. The plant is commonly harvested from its natural habitat, therefore it becomes endangered. Regeneration of pruatjan through organogenesis has been studied, but its shoot multiplication was very low (5 shoots per explant. The study aimed to investigate the best regeneration technique of pruatjan through somatic embryogenesis. This research was conducted at the tissue culture laboratory, Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development in 2004-2005. Callus formation of pruatjan was induced from the petioles and leaves in Driver and Kuniyaki’s (DKW based medium containing 2,4-D combined with picloram at the level of 0.5, 1.0, 1.5, and 1.5 ppm. Embryogenic calli were then transferred into embryo development medium in two ways. First, they were directly transferred into media containing IBA/NAA at the level of 0.5, 1.0, and 1.5 ppm. Second, they were indirectly transferred into media containing 2.0 ppm 2,4-D and 0.3% casein hydrolysate prior to the IBA/ NAA media. Parameters evaluated were fresh weight, dry weight, time initiation of embryogenic callus formation, and total number of embryos. The result showed that calli of pruatjan were successfully induced from the petioles and leaves. The best calli were induced from the leaves in the DKW medium containing 2.0 ppm 2,4-D and 0.5 ppm picloram. Embryo development of the calli was best if they were first grown in the media containing 2.0 ppm 2,4-D and 0.3% casein hydrolysate then transferred to the IBA/NAA media. The total number of somatic embryos was counted up to 103 on the medium containing 1.5 ppm IBA. This study indicated that pruatjan somatic embryogenesis regeneration required three different media, i.e. for callus induction, development and maturation, and for

  2. Somatic Embryogenesis of Abies cephalonica Loud.

    Science.gov (United States)

    Krajňáková, Jana; Häggman, Hely

    2016-01-01

    Greek fir (Abies cephalonica Loudon) belongs to the Mediterranean fir species and is widely distributed in the mountains of Central and Southern Greece. Considering a climatic scenario, infestation by pathogens or insects and fire episodes, it has been proposed that Mediterranean firs could be in danger in some parts of their present range but, on the other hand, could also replace other species in more northern zones with temperate humid climates (e.g., silver fir, Abies alba Mill.). As fir species are generally highly productive and therefore important for commercial forestry, they have traditionally been involved in conventional tree improvement programs. A lot of effort has been put into the development of vegetative propagation methods for firs, in order to rapidly gain the benefits of traditional breeding to be utilized in reforestation. The present paper provides up to date information on protocols for somatic embryogenesis (i.e., the most promising in vitro method for vegetative propagation) of Greek fir. Moreover, the protocols for cryopreservation and long-term storage of embryogenic material are described as well.

  3. Environmental magnetic fields: Influences on early embryogenesis

    Energy Technology Data Exchange (ETDEWEB)

    Cameron, I.L.; Hardman, W.E.; Winters, W.D.; Zimmerman, S.; Zimmerman, A.M. (Univ. of Texas Health Science Center, San Antonio (United States))

    1993-04-01

    A 10-mG, 50 to 60-Hz magnetic field is in the intensity and frequency range that people worldwide are often exposed to in homes and in the workplace. Studies about the effects of 50- to 100-Hz electromagnetic fields on various species of animal embryos (fish, chick, fly, sea urchin, rat, and mouse) indicate that early stages of embryonic development are responsive to fluctuating magnetic fields. Chick, sea urchin, and mouse embryos are responsive to magnetic field intensities of 10-100 mG. Results from studies on sea urchin embryos indicate that exposure to conditions of rotating 60-Hz magnetic fields, e.g., similar to those in our environment, interferes with cell proliferation at the morula stage in a manner dependent on field intensity. The cleavage stages, prior to the 64-cell stage, were not delayed by this rotating 60-Hz magnetic field suggesting that the ionic surges, DNA replication, and translational events essential for early cleavage stages were not significantly altered. Studies of histone synthesis in early sea urchin embryos indicated that the rotating 60-Hz magnetic field decreased zygotic expression of early histone genes at the morula stage and suggests that this decrease in early histone production was limiting to cell proliferation. Whether these comparative observations from animal development studies will be paralleled by results from studies of human embryogenesis, as suggested by some epidemiology studies, has yet to be established. 38 refs.

  4. Trahv? Herned klassinurgas? Või hoopis ihunuhtlus? / Pilme, Lea; Kivistik, Ly; Sirgmets, Raili; Part, Aivar

    Index Scriptorium Estoniae

    2008-01-01

    Suhtumisest koolijuhtidele trahvimisõiguse andmise ideesse vestlevad Rakvere Põhikooli direktor Lea Pilme, Uhtna Põhikooli direktor Ly Kivistik, Väike-Maarja Gümnaasiumi direktor Raili Sirgmets ja Rakvere Gümnaasiumi direktor Aivar Part

  5. DNA methylation during sexual embryogenesis and implications on the induction of somatic embryogenesis in Castanea sativa Miller.

    Science.gov (United States)

    Viejo, M; Rodríguez, R; Valledor, L; Pérez, M; Cañal, M J; Hasbún, R

    2010-12-01

    From anthesis to mature seed formation, burrs from cross-pollinated adult Castanea sativa Miller trees were characterized and seven developmental stages defined based on macro and micromorphological traits. In order to get an insight into the involvement of epigenetic mechanisms in sexual embryogenesis and to define somatic embryogenesis induction capability, global DNA methylation and the somatic embryogenic competence were quantified. On cross-pollinated trees once fertilization takes place, at least one ovule per ovary becomes dominant, and transient DNA demethylation occurs coinciding with the start of the sexual embryogenic programme. Unfertilized ovules from the same cluster, which maintain their prior size, increase their methylation level and undergo degeneration. These results were validated using non-cross-pollinated trees and the asynchrony of flower receptivity. When testing in vitro somatic embryogenesis response of isolated dominant ovules and axes from zygotic embryos under cross-pollinated conditions, the highest competence was found for reaching seed maturity. Thus, a "developmental window" of somatic embryogenesis in chestnut has been characterized. It includes from fertilization to embryo maturity, and a transient decrease in methylation is necessary after fertilization for the development of the somatic embryogenesis response.

  6. Systematic identification of long noncoding RNAs expressed during zebrafish embryogenesis

    DEFF Research Database (Denmark)

    Pauli, Andrea; Valen, Eivind; Lin, Michael F.

    2012-01-01

    vertebrate embryogenesis has been elusive. To identify lncRNAs with potential functions in vertebrate embryogenesis, we performed a time series of RNA-Seq experiments at eight stages during early zebrafish development. We reconstructed 56,535 high-confidence transcripts in 28,912 loci, recovering the vast...... overlapping lncRNAs, and precursors for small RNAs (sRNAs). Zebrafish lncRNAs share many of the characteristics of their mammalian counterparts: relatively short length, low exon number, low expression, and conservation levels comparable to introns. Subsets of lncRNAs carry chromatin signatures characteristic...

  7. Embryogenesis and plant regeneration from unpollinated ovaries of ...

    African Journals Online (AJOL)

    The system of somatic embryogenesis of Amorphophallus konjac had been built through unpollinated ovaries. The embryogenic calli were induced on Murashige and Skoog (MS) basal medium supplemented with 9.0 μM 6- benzylaminopurine (BA), 0.4 μM 2,4dichlorophenoxyacetic acid (D), 1.0 μM naphthaleneacetic acid ...

  8. Embryogenesis and plant regeneration from unpollinated ovaries of ...

    African Journals Online (AJOL)

    Yomi

    glucomannan crop, which has been widely used in food, medicine, chemistry and agriculture industries (Cescutti et al., 2002). Konjac mainly reproduces asexually by .... trehalose of Oncidium 'Gower Ramsey' (Fang et al.,. 2006). The system of the konjac somatic embryogenesis can be useful not only for germchits or seed ...

  9. Optimization of somatic embryogenesis induction in Iranian melon ...

    African Journals Online (AJOL)

    In this study, regeneration of Cucumis melo cv. Khatooni: one specific Iranian melon- via somatic embryogenesis was investigated. Three different types of explant (cotyledon, hypocotyl and cotyledonary petiole) from six days old seedlings were used. Growth regulator treatments were two levels of 6-benzyl aminopurine (0 ...

  10. Effect of explant age, hormones on somatic embryogenesis and ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-02-18

    Feb 18, 2009 ... from Solanum melongena L. (eggplant) cotyledon explants is associated with hormone-modulated enhancement of polyamine biosynthesis and conjugation. Protoplasma. 211:51-63. Sharma P, Rajam MV (1995). GenoType, explant and position effects on organogenesis and somatic embryogenesis in ...

  11. Optimization of somatic embryogenesis induction in Iranian melon ...

    African Journals Online (AJOL)

    Jane

    2011-07-11

    Jul 11, 2011 ... flexuosus. Afr. J. Biotechnol., 8(22): 6228–6232. Deakin JR, Bohn GW, Whitaker TW (1971). Interspecific hybridization in. Cucumis. Econ. Bot., 25: 195-211. Debeaujon I, Branchard M (1992). Induction of somatic embryogenesis and caulogenesis from cotyledon and leaf protoplast derived colonies of melon ...

  12. Plant regeneration via somatic embryogenesis from root explants of ...

    African Journals Online (AJOL)

    A system for induction of callus and plant regeneration via somatic embryogenesis from root explants of Hevea brasiliensis Muell. Arg. clone Reyan 87-6-62 was evaluated. The influence of plant growth regulators (PGRs) including 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (6-BA) and kinetin (KT) on ...

  13. Improvements of cyclic somatic embryogenesis of cassava (Manihot esculenta Crantz).

    NARCIS (Netherlands)

    Raemakers, C.J.J.M.; Schavemaker, C.M.; Jacobsen, E.; Visser, R.G.F.

    1993-01-01

    In cassava a cyclic system of somatic embryogenesis was developed. Primary (torpedo shaped or germinated) embryos, originating from leaf lobes, could only be obtained after culture on solid medium. Cyclic embryos, originating from embryos, could be obtained in both liquid and on solid medium. The

  14. Influence of plant growth regulators on somatic embryogenesis ...

    African Journals Online (AJOL)

    Generating somatic embryos from the inner teguments of hevea seeds is difficult. Like other ligneous plants, the rubber-tree is generally considered to be recalcitrant with regard to somatic embryogenesis. In this study, the ability of callus from inner integument explants to develop embryogenic callus lines was highlighted.

  15. Somatic embryogenesis from leaf explants of hermaphrodite Carica ...

    African Journals Online (AJOL)

    In culture medium supplemented with 2,4-dichlorophenoxyacetic, FEC overgrew into a yellowish friable mass that fully covered the leaf explants. The somatic embryogenesis process occurred asynchronously, with new globular embryos continuously forming from the FEC. Torpedo and early cotyledonary somatic embryos ...

  16. Somatic embryogenesis from immature male inflorescences of East ...

    African Journals Online (AJOL)

    With regard to the pest and disease problems of the East African highland banana, these findings underscore the fact that somatic embryogenesis is essential in the development of an In vitro regeneration system and is a critical step for development of resistant varieties through genetic transformation. Key Words: Cell ...

  17. Optimization of embryogenic-callus induction and embryogenesis of ...

    African Journals Online (AJOL)

    ONOS

    2010-09-06

    Sep 6, 2010 ... Chunhua Fu1#, Cheng Lei1,2#, Lu Gan1, Maoteng Li1,YingYang1 and Longjiang Yu1*. Institute of Resource Biology and Biotechnology, College of Life Science and ..... Varisai MS, Wang CS, Thiruvengadam M, Jayabalan N (2004). In vitro plant regeneration via somatic embryogenesis through cell ...

  18. Somatic embryogenesis and plant regeneration from leaf explants of ...

    African Journals Online (AJOL)

    An attempt was made to study the somatic embryogenesis and plant regeneration from the in vitro leaf explants of Rumex vesicarius L. a renowned medicinal plant, which belongs to polygonaceae family. Effective in vitro regeneration of R. vesicarius was achieved via young leaf derived somatic embryo cultures.

  19. Somatic embryogenesis and plant regeneration of recalcitrant cottons

    African Journals Online (AJOL)

    STORAGESEVER

    2009-02-04

    Feb 4, 2009 ... regeneration through somatic embryogenesis for cotton cultivars with Fusarium and/or Verticillium wilts resistance. MATERIALS AND METHODS. Plant materials. Five upland cotton (G. hirsutum) cultivars (Shann724, Zhong6331,. CCRI18, Liaomian12 and Jinmian14), provided by Cotton Genetics.

  20. Effect of age on somatic embryogenesis from immature zygotic ...

    African Journals Online (AJOL)

    Triticale is an important cereal crop grown throughout the world. The study reports somatic embryogenesis from immature zygotic embryos of 5 Turkish triticale genotypes. The explants were initially cultured on MS medium supplemented with 2 mg dm-3 2,4-D, 500 mg dm-3 glutamine, 100 mg dm-3 casein hydrolysate, 2% ...

  1. Evaluation of somatic embryogenesis and plant regeneration in ...

    African Journals Online (AJOL)

    Optimization of tissue culture conditions for Sorghum bicolor L. through somatic embryogenesis from immature embryos is important for the genetic manipulation and improvement of this agronomically valuable crop. In an attempt to develop a successfully reproducible in vitro regeneration protocol for a group of diverse ...

  2. Somatic embryogenesis from zygotic embryos and thin cell layers ...

    African Journals Online (AJOL)

    Oil palm hybrid BRS Manicoré is important for plantations in the north of Brazil, as it is resistant to fatal yellowing and is compact. Seed germination is slow and reduced, so somatic embryogenesis is a promising alternative for its propagation. Two kinds of starting explants were used: Zygotic embryos (ZE) and thin cell layers ...

  3. Induction of plant somatic embryogenesis in liquid medium

    NARCIS (Netherlands)

    Kreuger, M.

    1996-01-01


    The large scale propagation of plants via somatic embryogenesis, has so far been difficult to achieve. In this thesis research is described leading to embryogenic cell lines that can be maintained for a long period, without loss of genetic stability. It is also described how embryogenic

  4. Somatic embryogenesis for efficient micropropagation of guava (Psidium guajava L.).

    Science.gov (United States)

    Akhtar, Nasim

    2013-01-01

    Guava (Psidium guajava L.) is well known for edible fruit, environment friendly pharmaceutical and commercial products for both national and international market. The conventional propagation and in vitro organogenesis do not meet the demand for the good quality planting materials. Somatic embryogenesis for efficient micropropagation of guava (P. guajava L.) has been developed to fill up the gap. Somatic embryogenesis and plantlets regeneration are achieved from 10-week post-anthesis zygotic embryo explants by 8-day inductive treatment with different concentrations of 2,4-dichlorophenoxy acetic acid (2,4-D) on MS agar medium containing 5% sucrose. Subsequent development and maturation of somatic embryos occur after 8 days on MS basal medium supplemented with 5% sucrose without plant growth regulator. The process of somatic embryogenesis shows the highest relative efficiency in 8-day treatment of zygotic embryo explants with 1.0 mg L(-1) 2,4-D. High efficiency germination of somatic embryos and plantlet regeneration takes place on half strength semisolid MS medium amended with 3% sucrose within 2 weeks of subculture. Somatic plantlets are grown for additional 2 weeks by subculturing in MS liquid growth medium containing 3% sucrose. Well-grown plantlets from liquid medium have survived very well following 2-4 week hardening process. The protocol of somatic embryogenesis is optimized for high efficiency micropropagation of guava species.

  5. Induction of somatic embryogenesis in saffron using thidiazuron (TDZ).

    Science.gov (United States)

    Sheibani, M; Azghandi, A V; Nemati, S H

    2007-10-15

    In vitro propagation of saffron either through somatic embryogenesis or cormogenesis is considered to be an efficient alternative method for large-scale propagation of pathogen-free corms. In order to develop an efficient protocol for in vitro propagation of saffron, a factorial experiment was carried out based on completely randomized design to investigate the effects of various concentrations of TDZ (0, 0.1, 0.25 and 0.5 mg L(-1)) on somatic embryogenesis induction from 5 different types of corm explants (terminal or axillary buds, upper or lower parts of the corm tissue and terminal buds from pre-treated corms at 4 degrees C for 2 weeks). The results revealed that TDZ concentrations affected the induction of somatic embryogenesis significantly while different types of corm explants showed no significant effect on this process. Among TDZ concentrations used, 0.5 mg L(-1) was the most effective treatment for embryogenesis induction. Embryogenic calli (globular stage) proliferated well when subcultured into MS medium supplemented with 0.25 mg L(-1) TDZ before transferring to hormone-free MS medium containing 6% sucrose for maturation (scutellar or horn-shape stage). Matured embryos were transferred to half strength MS medium without growth regulators for further development, from which microcorms were produced at the basal part after 3 months.

  6. An efficient somatic embryogenesis based plant regeneration from ...

    African Journals Online (AJOL)

    An efficient and rapid somatic embryogenesis based plant regeneration from hypocotyls of three Catharanthus roseus cultivars, Pacifica cherry red (PCR), Heatwave mix color (HWMC), and Mediterranean Rose Red (MRR) was standardized. Hypocotyl-derived primary calluses (HPC) were formed on callus induction ...

  7. LEA Detection and Tracking Method for Color-Independent Visual-MIMO.

    Science.gov (United States)

    Kim, Jai-Eun; Kim, Ji-Won; Kim, Ki-Doo

    2016-07-02

    Communication performance in the color-independent visual-multiple input multiple output (visual-MIMO) technique is deteriorated by light emitting array (LEA) detection and tracking errors in the received image because the image sensor included in the camera must be used as the receiver in the visual-MIMO system. In this paper, in order to improve detection reliability, we first set up the color-space-based region of interest (ROI) in which an LEA is likely to be placed, and then use the Harris corner detection method. Next, we use Kalman filtering for robust tracking by predicting the most probable location of the LEA when the relative position between the camera and the LEA varies. In the last step of our proposed method, the perspective projection is used to correct the distorted image, which can improve the symbol decision accuracy. Finally, through numerical simulation, we show the possibility of robust detection and tracking of the LEA, which results in a symbol error rate (SER) performance improvement.

  8. Ubiquitous LEA29Y Expression Blocks T Cell Co-Stimulation but Permits Sexual Reproduction in Genetically Modified Pigs.

    Science.gov (United States)

    Bähr, Andrea; Käser, Tobias; Kemter, Elisabeth; Gerner, Wilhelm; Kurome, Mayuko; Baars, Wiebke; Herbach, Nadja; Witter, Kirsti; Wünsch, Annegret; Talker, Stephanie C; Kessler, Barbara; Nagashima, Hiroshi; Saalmüller, Armin; Schwinzer, Reinhard; Wolf, Eckhard; Klymiuk, Nikolai

    2016-01-01

    We have successfully established and characterized a genetically modified pig line with ubiquitous expression of LEA29Y, a human CTLA4-Ig derivate. LEA29Y binds human B7.1/CD80 and B7.2/CD86 with high affinity and is thus a potent inhibitor of T cell co-stimulation via this pathway. We have characterized the expression pattern and the biological function of the transgene as well as its impact on the porcine immune system and have evaluated the potential of these transgenic pigs to propagate via assisted breeding methods. The analysis of LEA29Y expression in serum and multiple organs of CAG-LEA transgenic pigs revealed that these animals produce a biologically active transgenic product at a considerable level. They present with an immune system affected by transgene expression, but can be maintained until sexual maturity and propagated by assisted reproduction techniques. Based on previous experience with pancreatic islets expressing LEA29Y, tissues from CAG-LEA29Y transgenic pigs should be protected against rejection by human T cells. Furthermore, their immune-compromised phenotype makes CAG-LEA29Y transgenic pigs an interesting large animal model for testing human cell therapies and will provide an important tool for further clarifying the LEA29Y mode of action.

  9. 34 CFR 76.792 - How does an SEA allocate funds to eligible charter school LEAs under a covered program in which...

    Science.gov (United States)

    2010-07-01

    ... charter school LEA receives at least a pro rata portion of the proportionate amount of funds for which the charter school LEA is eligible under each covered program. The pro rata amount must be based on the number... SEA may implement procedures to provide the charter school LEA with a pro rata portion of the...

  10. Somatic Embryogenesis in Two Orchid Genera (Cymbidium, Dendrobium).

    Science.gov (United States)

    da Silva, Jaime A Teixeira; Winarto, Budi

    2016-01-01

    The protocorm-like body (PLB) is the de facto somatic embryo in orchids. Here we describe detailed protocols for two orchid genera (hybrid Cymbidium Twilight Moon 'Day Light' and Dendrobium 'Jayakarta', D. 'Gradita 31', and D. 'Zahra FR 62') for generating PLBs. These protocols will most likely have to be tweaked for different cultivars as the response of orchids in vitro tends to be dependent on genotype. In addition to primary somatic embryogenesis, secondary (or repetitive) somatic embryogenesis is also described for both genera. The use of thin cell layers as a sensitive tissue assay is outlined for hybrid Cymbidium while the protocol outlined is suitable for bioreactor culture of D. 'Zahra FR 62'.

  11. Cracking the egg: virtual embryogenesis of real robots.

    Science.gov (United States)

    Cussat-Blanc, Sylvain; Pollack, Jordan

    2014-01-01

    All multicellular living beings are created from a single cell. A developmental process, called embryogenesis, takes this first fertilized cell down a complex path of reproduction, migration, and specialization into a complex organism adapted to its environment. In most cases, the first steps of the embryogenesis take place in a protected environment such as in an egg or in utero. Starting from this observation, we propose a new approach to the generation of real robots, strongly inspired by living systems. Our robots are composed of tens of specialized cells, grown from a single cell using a bio-inspired virtual developmental process. Virtual cells, controlled by gene regulatory networks, divide, migrate, and specialize to produce the robot's body plan (morphology), and then the robot is manually built from this plan. Because the robot is as easy to assemble as Lego, the building process could be easily automated.

  12. La eminencia intercondílea normal y patológica

    OpenAIRE

    Quiles Casas, Carlos

    2017-01-01

    INTRODUCCIÓN.- La eminencia intercondílea normal suele describirse en los libros de texto de forma sucinta. OBJETIVOS.- Describir en detalle la anatomía normal del área intercondílea, comparándola con los cambios producidos por la artrosis avanzada, incluyendo estudios de imagen e histológicos. MATERIAL Y MÉTODOS.- Se utilizaron catorce platillos tibiales de donantes y cien superficies tibiales de enfermos con artrosis de rodilla. En las piezas anatómicas se usó reconstrucción d...

  13. Histology of somatic embryogenesis in rice (Oryza sativa cv. 5272)

    OpenAIRE

    Vega, Rafael; Vásquez, Nelly; Ana M Espinoza; Andrés M Gatica; Valdez-Melara, Marta

    2015-01-01

    Rice (Oryza sativa cv. 5272) embryogenic calli were obtained from mature zygotic embryos culture on Murashige & Skoog (1962) medium supplemented with 2.5 mg/l 2,4- dichlorophenoxyacetic acid. Histological analysis of somatic embryogenesis revealed that after two weeks of culture of explants on the callus induction medium, somatic embryo development began with a cluster of proembryogenic cells in the peripheral region of the calli. The outer cell layer of embryogenic calli consisted of small a...

  14. POLYPHENOLS DISTRIBUTION AND RESERVE SUBSTANCES ANALYSIS IN CACAO SOMATIC EMBRYOGENESIS

    OpenAIRE

    Adriana María Gallego Rúa; Ana María Henao Ramírez; Aura Inés Urrea Trujillo; Lucía Atehortúa Garcés

    2016-01-01

    ABSTRACTIn order to understand the causes of lack of regeneration in cacao somatic embryos, two cacao varieties with different responses to regeneration potential were described based on their capacity to store different compounds. It is well known that seed reserves play a central role in the regenerative capability of somatic embryos; thus, we followed histochemical changes and reserve fluctuations of proteins, polysaccharides and polyphenols during somatic embryogenesis (SE) in the two cac...

  15. Whole transcriptome profiling of maize during early somatic embryogenesis reveals altered expression of stress factors and embryogenesis-related genes.

    Directory of Open Access Journals (Sweden)

    Stella A G D Salvo

    Full Text Available Embryogenic tissue culture systems are utilized in propagation and genetic engineering of crop plants, but applications are limited by genotype-dependent culture response. To date, few genes necessary for embryogenic callus formation have been identified or characterized. The goal of this research was to enhance our understanding of gene expression during maize embryogenic tissue culture initiation. In this study, we highlight the expression of candidate genes that have been previously regarded in the literature as having important roles in somatic embryogenesis. We utilized RNA based sequencing (RNA-seq to characterize the transcriptome of immature embryo explants of the highly embryogenic and regenerable maize genotype A188 at 0, 24, 36, 48, and 72 hours after placement of explants on tissue culture initiation medium. Genes annotated as functioning in stress response, such as glutathione-S-transferases and germin-like proteins, and genes involved with hormone transport, such as PINFORMED, increased in expression over 8-fold in the study. Maize genes with high sequence similarity to genes previously described in the initiation of embryogenic cultures, such as transcription factors BABY BOOM, LEAFY COTYLEDON, and AGAMOUS, and important receptor-like kinases such as SOMATIC EMBRYOGENESIS RECEPTOR LIKE KINASES and CLAVATA, were also expressed in this time course study. By combining results from whole genome transcriptome analysis with an in depth review of key genes that play a role in the onset of embryogenesis, we propose a model of coordinated expression of somatic embryogenesis-related genes, providing an improved understanding of genomic factors involved in the early steps of embryogenic culture initiation in maize and other plant species.

  16. Packaging of Post Acclimatized Somatic Embryogenesis Cocoa Plantlet (Theobroma cacao L.)

    OpenAIRE

    Soedarsianto Soedarsianto; Teguh Iman Santoso

    2009-01-01

    Clonal plants that produced by somatic embryogenesis technique is one of the best choice to produce supperior clonal cacao (Theobroma cacao L.) planting materials. The somatic embryogenesis technique is a possible way for massive propagation, the outcome is true to type plants, the architecture similarity that the seedlings but there is not segregation like seedlings plants. At present mass production started of plantlets production until post-acclimatized plantlets of somatic embryogenesis c...

  17. Localization and Identification of Phenolic Compounds in Theobroma cacao L. Somatic Embryogenesis

    OpenAIRE

    ALEMANNO, L.; Ramos, T.; GARGADENEC, A.; Andary, C.; FERRIERE, N.

    2003-01-01

    Cocoa breeders and growers continue to face the problem of high heterogeneity between individuals derived from one progeny. Vegetative propagation by somatic embryogenesis could be a way to increase genetic gains in the field. Somatic embryogenesis in cocoa is difficult and this species is considered as recalcitrant. This study was conducted to investigate the phenolic composition of cocoa flowers (the explants used to achieve somatic embryogenesis) and how it changes during the process, by m...

  18. 45 CFR 618.420 - Access to schools operated by LEAs.

    Science.gov (United States)

    2010-10-01

    ... 45 Public Welfare 3 2010-10-01 2010-10-01 false Access to schools operated by LEAs. 618.420 Section 618.420 Public Welfare Regulations Relating to Public Welfare (Continued) NATIONAL SCIENCE... such recipient otherwise makes available to such person, pursuant to the same policies and criteria of...

  19. Phenotypic Characterization of LEA Rat: A New Rat Model of Nonobese Type 2 Diabetes

    Directory of Open Access Journals (Sweden)

    Tadashi Okamura

    2013-01-01

    Full Text Available Animal models have provided important information for the genetics and pathophysiology of diabetes. Here we have established a novel, nonobese rat strain with spontaneous diabetes, Long-Evans Agouti (LEA rat derived from Long-Evans (LE strain. The incidence of diabetes in the males was 10% at 6 months of age and 86% at 14 months, while none of the females developed diabetes. The blood glucose level in LEA male rats was between 200 and 300 mg/dl at 120 min according to OGTT. The glucose intolerance in correspondence with the impairment of insulin secretion was observed in male rats, which was the main cause of diabetes in LEA rats. Histological examination revealed that the reduction of β-cell mass was caused by progressive fibrosis in pancreatic islets in age-dependent manner. The intracytoplasmic hyaline droplet accumulation and the disappearance of tubular epithelial cell layer associated with thickening of basement membrane were evident in renal proximal tubules. The body mass index and glycaemic response to exogenous insulin were comparable to those of control rats. The unique characteristics of LEA rat are a great advantage not only to analyze the progression of diabetes, but also to disclose the genes involved in type 2 diabetes mellitus.

  20. Phenotypic Characterization of LEA Rat: A New Rat Model of Nonobese Type 2 Diabetes

    Science.gov (United States)

    Okamura, Tadashi; Pei, Xiang Yuan; Miyoshi, Ichiro; Shimizu, Yukiko; Takanashi-Yanobu, Rieko; Mototani, Yasumasa; Kanai, Takao; Satoh, Jo; Kimura, Noriko; Kasai, Noriyuki

    2013-01-01

    Animal models have provided important information for the genetics and pathophysiology of diabetes. Here we have established a novel, nonobese rat strain with spontaneous diabetes, Long-Evans Agouti (LEA) rat derived from Long-Evans (LE) strain. The incidence of diabetes in the males was 10% at 6 months of age and 86% at 14 months, while none of the females developed diabetes. The blood glucose level in LEA male rats was between 200 and 300 mg/dl at 120 min according to OGTT. The glucose intolerance in correspondence with the impairment of insulin secretion was observed in male rats, which was the main cause of diabetes in LEA rats. Histological examination revealed that the reduction of β-cell mass was caused by progressive fibrosis in pancreatic islets in age-dependent manner. The intracytoplasmic hyaline droplet accumulation and the disappearance of tubular epithelial cell layer associated with thickening of basement membrane were evident in renal proximal tubules. The body mass index and glycaemic response to exogenous insulin were comparable to those of control rats. The unique characteristics of LEA rat are a great advantage not only to analyze the progression of diabetes, but also to disclose the genes involved in type 2 diabetes mellitus. PMID:23691528

  1. 24 CFR 3.420 - Access to schools operated by LEAs.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 1 2010-04-01 2010-04-01 false Access to schools operated by LEAs. 3.420 Section 3.420 Housing and Urban Development Office of the Secretary, Department of Housing and Urban Development NONDISCRIMINATION ON THE BASIS OF SEX IN EDUCATION PROGRAMS OR ACTIVITIES RECEIVING...

  2. [Lea Kõiv. Paul Johansen und das Stadtarchiv Reval/Tallinn] / Paul Kaegbein

    Index Scriptorium Estoniae

    Kaegbein, Paul

    2008-01-01

    Arvustus: Lea Kõiv. Paul Johansen und das Stadtarchiv Reval/Tallinn. - Nordosteuropa als Geschichtsregion. Helsinki : Aue-Stiftung; Lübeck : Schmict-Röhmild, 2006. Lk. 45-59. Artiklis on kasutatud seni avalikkusele vähetuntud materjale ja osaliselt käsitletud avalikke vaidlusi, kuhu olid segatud nii Paul Johansen kui järgmine linnaarhivaar Rudolf Kenkmaa

  3. 13 CFR 113.420 - Access to schools operated by LEAs.

    Science.gov (United States)

    2010-01-01

    ... 13 Business Credit and Assistance 1 2010-01-01 2010-01-01 false Access to schools operated by LEAs. 113.420 Section 113.420 Business Credit and Assistance SMALL BUSINESS ADMINISTRATION NONDISCRIMINATION... operated by such recipient; or (b) Any other school or educational unit operated by such recipient, unless...

  4. Epithelial self-organization in fruit fly embryogenesis

    Science.gov (United States)

    Hutson, M. Shane

    2010-03-01

    During fruit fly embryogenesis, there are several morphogenetic events in which sheets of epithelial cells expand, contract and bend due to coordinated intra- and intercellular forces. This tissue-level reshaping is accompanied by changes in the shape and arrangement of individual cells -- changes that can be measured quantitatively and dynamically using modern live-cell imaging techniques. Such data sets represent rich targets for computational modeling of self-organization; however, reproducing the observed cell- and tissue-level reshaping is not enough. The inverse problem of using cell shape changes to determine cell-level forces is ill-posed -- yielding non-unique solutions that cannot discriminate between active changes in cell shape and passive deformation. These non-unique solutions can be tested experimentally using in vivo laser-microsurgery -- i.e., cutting a targeted region of an epithelium and carefully tracking the temporal and spatial dependence of the subsequent strain relaxation. This technique uses a variety of incisions (hole, line or closed curve) to probe different aspects of epithelial mechanics: the local mesoscopic strain; the distribution of intracellular forces; changes in the cell-level power-law rheology; and the question of active versus passive deformation. I will discuss my group's work using laser-microsurgery to investigate two morphogenetic events in fruit fly embryogenesis: germband retraction and dorsal closure. In both cases, we find a substantial active mechanical role for the amnioserosa -- an epithelium that undergoes apoptosis near the end of embryogenesis and makes no part of the fly larva -- in reshaping an adjacent epithelium that becomes the larval epidermis. In these examples, self-organization of the fly embryo relies not only on self-organization of individual tissues, but also on the mechanical interactions between tissues.

  5. SOMATIC EMBRYOGENESIS AND MORPHOANATOMY OF Ocotea porosa SOMATIC EMBRYOS

    Directory of Open Access Journals (Sweden)

    Luciana Luiza Pelegrini

    2013-12-01

    Full Text Available http://dx.doi.org/10.5902/1980509812343Ocotea porosa seeds have strong tegument dormancy, recalcitrant behavior, low and irregular germinationand that makes its natural propagation difficult. The aim of this study was to establish a protocol ofregeneration of Ocotea porosa from somatic embryogenesis. Immature embryonic axes were inoculatedon WPM culture medium supplemented with 2.4-D (200 μM combined or not with hydrolyzed casein orglutamine (0.5 or 1 g l-1, during 90 days. The repetitive embryogenesis was induced on medium with 2.4-D(22.62 μM combined with 2-iP (2.46 μM followed by transfer to culture medium with hydrolyzed caseinor glutamine (1 g l-1 during 90 days. The maturation of somatic embryos was tested in culture mediumcontaining NAA (0.5 μM and 2-iP (5; 10 and 20 μM. The highest percentage of somatic embryos induction(8.3% was observed in WPM culture medium containing 200 μM 2.4-D and 1 g L-1 hydrolyzed casein andthe development of somatic embryos occurred indirectly. Repetitive somatic embryogenesis was promotedin WPM medium containing hydrolyzed casein or glutamine. However, the culture medium containinghydrolyzed casein promoted the maintenance of embryogenic capacity for more than two years. Duringthe maturity phase, there was a low progression of globular embryos to cordiform and torpedo stages.The different ontogenetic stages of somatic embryos of Ocotea porosa were characterized by histologicalstudies.

  6. SOMATIC EMBRYOGENESIS AND MORPHOANATOMY OF Ocotea porosa SOMATIC EMBRYOS

    Directory of Open Access Journals (Sweden)

    Luciana Luiza Pelegrini

    2013-01-01

    Full Text Available Ocotea porosa seeds have strong tegument dormancy, recalcitrant behavior, low and irregular germination and that makes its natural propagation difficult. The aim of this study was to establish a protocol of regeneration of Ocotea porosa from somatic embryogenesis. Immature embryonic axes were inoculated on WPM culture medium supplemented with 2.4-D (200 μM combined or not with hydrolyzed casein or glutamine (0.5 or 1 g l-1, during 90 days. The repetitive embryogenesis was induced on medium with 2.4-D (22.62 μM combined with 2-iP (2.46 μM followed by transfer to culture medium with hydrolyzed casein or glutamine (1 g l-1 during 90 days. The maturation of somatic embryos was tested in culture medium containing NAA (0.5 μM and 2-iP (5; 10 and 20 μM. The highest percentage of somatic embryos induction (8.3% was observed in WPM culture medium containing 200 μM 2.4-D and 1 g L-1 hydrolyzed casein and the development of somatic embryos occurred indirectly. Repetitive somatic embryogenesis was promoted in WPM medium containing hydrolyzed casein or glutamine. However, the culture medium containing hydrolyzed casein promoted the maintenance of embryogenic capacity for more than two years. During the maturity phase, there was a low progression of globular embryos to cordiform and torpedo stages. The different ontogenetic stages of somatic embryos of Ocotea porosa were characterized by histological studies.

  7. Somatic embryogenesis from zygotic embryos of Euterpe oleracea Mart.

    OpenAIRE

    Ledo Ana da Silva; Lameira Osmar Alves; Benbadis Abdellatif Kemaleddine; Menezes Ilmarina Campos de; Oliveira Maria do Socorro Padilha de; Medeiros Filho Sebastião

    2002-01-01

    The aim of this work was to study the morphogenetic responses of zygotic embryos of açai palm (Euterpe oleracea Mart.) submitted to several conditions of in vitro culture. Several research experiments were conducted, in laboratory, using vegetable material collected from açai palm plants at Embrapa Amazon Oriental, Belém-PA, Brazil. It was possible to verify the expression of a direct, repetitive and no-synchronized model of somatic embryogenesis in mature zygotic embryos cultivated in primar...

  8. Lipid Rafts: Keys to Sperm Maturation, Fertilization, and Early Embryogenesis

    Directory of Open Access Journals (Sweden)

    Natsuko Kawano

    2011-01-01

    Full Text Available Cell membranes are composed of many different lipids and protein receptors, which are important for regulating intracellular functions and cell signaling. To orchestrate these activities, the cell membrane is compartmentalized into microdomains that are stably or transiently formed. These compartments are called “lipid rafts”. In gamete cells that lack gene transcription, distribution of lipids and proteins on these lipid rafts is focused during changes in their structure and functions such as starting flagella movement and membrane fusion. In this paper, we describe the role of lipid rafts in gamete maturation, fertilization, and early embryogenesis.

  9. Effect of Salicylic Acid on Somatic Embryogenesis and Plant Regeneration in Hedychium bousigonianum

    Science.gov (United States)

    The objective of this study was to induce somatic embryogenesis in Hedychium bousigonianum Pierre ex Gagnepain and assess the influence of salicylic acid (S) on somatic embryogenesis. Somatic embryos and subsequently regenerated plants were successfully obtained 30 days after transfer of embryogenic...

  10. Subcellular localization and oligomerization of the Arabidopsis thaliana somatic embryogenesis receptor kinase 1 protein

    NARCIS (Netherlands)

    Shah, K.; Gadella, T.W.J.; Erp, van H.; Hecht, V.; Vries, de S.C.

    2001-01-01

    The Arabidopsis thaliana somatic embryogenesis receptor kinase 1 (AtSERK1) gene is expressed in developing ovules and early embryos. AtSERK1 is also transiently expressed during somatic embryogenesis. The predicted AtSERK1 protein contains an extracellular domain with a leucine zipper motif followed

  11. Subcellular localization and oligomerization of the Aradopsis thaliana somatic embryogenesis receptor kinase 1 protein

    NARCIS (Netherlands)

    Shah, K.; Gadella, Th.W.J.; van Erp, H.; Hecht, V.; de Vries, S.C.

    2001-01-01

    The Arabidopsis thaliana somatic embryogenesis receptor kinase 1 (AtSERK1) gene is expressed in developing ovules and early embryos. AtSERK1 is also transiently expressed during somatic embryogenesis. The predicted AtSERK1 protein contains an extracellular domain with a leucine zipper motif followed

  12. Somatic embryogenesis and plant regeneration in Anthurium andraeanum hybrids.

    Science.gov (United States)

    Kuehnle, A R; Chen, F C; Sugii, N

    1992-08-01

    A method for the production of somatic embryos and subsequent plant regeneration for Anthurium andraeanum Linden ex André (Monocotyledonae) hybrids is described. Whole leaf blade explants, derived from plantlets grown in vitro, formed translucent embryogénic calli at their basal ends within one month of culture in the dark. Secondary somatic embryos formed frequently and without an intervening callus on surfaces of primary embryos. Embryogenesis was induced with three genotypes using a modified half-strength Murashige and Skoog (MS) medium supplemented with 1.0 to 4.0 mg l(-1) 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.33 to 1.0 mg l(-1) kinetin. A combination of 2% sucrose with 1% glucose in the medium favored embryogenesis over 3% sucrose alone. Whole leaf blades on medium solidified with 0.18% Gelrite produced more somatic embryos than leaves on medium with 0.7% Bacto-agar. Within two to three months after culture initiation, embryos were transferred to modified MS medium containing 0.2 mg l(-1) 6-benzyladenine (BA) and 2% sucrose and placed in the light for conversion into plantlets. Rooted plantlets were recovered and transferred into pots with tree fern fiber medium and grown in the greenhouse.

  13. The role of chromatin modifications in somatic embryogenesis in plants

    Directory of Open Access Journals (Sweden)

    Clelia eDe-la-Peña

    2015-08-01

    Full Text Available Somatic embryogenesis (SE is a powerful tool for plant genetic improvement, when used in combination with agricultural traditional techniques, and it is being used to understand the different processes that occur during the development of plant embryogenesis. SE onset depends on a complex network of interactions among plant growth regulators, mainly auxins and cytokinins, during the proembryogenic early stages, and ethylene, gibberellic and abscisic acids later in the development of the somatic embryos. These growth regulators control spatial and temporal regulation of multiple genes in order to initiate the change in the genetic program of the somatic cells, as well as the transition among embryo developmental stages. In recent years, epigenetic mechanisms have emerged as critical factors during SE. Some early reports indicate that auxins modify the levels of DNA methylation in embryogenic cells. The changes in DNA methylation patterns are associated with the regulation of several genes involved in SE, such as WUS, BBM1, LEC, and several others. In this review, we highlight the more recent discoveries in the role of epigenetic regulation of SE. In addition, we include a survey of novel approaches to the study of SE, and new opportunities to focus SE studies.

  14. Lethality in PARP-1/Ku80 double mutant mice reveals physiologicalsynergy during early embryogenesis

    Energy Technology Data Exchange (ETDEWEB)

    Henrie, Melinda S.; Kurimasa, Akihiro; Burma, Sandeep; Menissier-de Murcia, Josiane; de Murcia, Gilbert; Li, Gloria C.; Chen,David J.

    2002-09-24

    Ku is an abundant heterodimeric nuclear protein, consisting of 70-kDa and 86-kDa tightly associated subunits that comprise the DNA binding component of DNA-dependent protein kinase. Poly(ADP)ribose polymerase-1 (PARP-1) is a 113-kDa protein that catalyzes the synthesis of poly(ADP-ribose) on target proteins. Both Ku and PARP-1 recognize and bind to DNA ends. Ku functions in the non-homologous end joining (NHEJ) repair pathway whereas PARP-1 functions in the single strand break repair and base excision repair (BER) pathways. Recent studies have revealed that PARP-1 and Ku80 interact in vitro. To determine whether the association of PARP-1 and Ku80 has any physiological significance or synergistic function in vivo, mice lacking both PARP-1 and Ku80 were generated. The resulting offspring died during embryonic development displaying abnormalities around the gastrulation stage. In addition, PARP-1-/-Ku80-/- cultured blastocysts had an increased level of apoptosis. These data suggest that the functions of both Ku80 and PARP-1 are essential for normal embryogenesis and that a loss of genomic integrity leading to cell death through apoptosis is likely the cause of the embryonic lethality observed in these mice.

  15. P comp Phytoc osition hemic ns of fo al, pro our lea Easte oximate ...

    African Journals Online (AJOL)

    SAM

    y of minera e, 2003). Lea nt were Oc ngronema ng. Tel: +2348 e Creative Com al of Biote utrient ed in S rence I. gu State, Nige uations just diets of South ate and ant e scent leaf ema latifolium minary phyto strongly pres hest tannin c ochemicals gnificant va ibre (33.52% y clearly indi xtensive use iable amount en though the.

  16. Somatic embryogenesis in cassava genotypes from the northeast of Brazil

    Directory of Open Access Journals (Sweden)

    Terezinha Feitosa

    2007-03-01

    Full Text Available A method for the induction of somatic embryogenesis in eight cassava genotypes from northeastern Brazil is described. The explants used were shoot apexes isolated both from in vitro grown plants and from shoots that sprouted from stem cuttings. Somatic embryogenesis was achieved in high frequencies by the addition in the induction medium of the auxin picloram over a wide range of concentrations. Green cotyledons of primary somatic embryos were used as explants to induce somatic (cyclic secondary embryogenesis in an inducing medium supplemented with picloram at 12 mg/L. The method could be used not only for the mass production of plants of the cassava genotypes, but also to generate explants (green cotyledons of somatic embryos as themselves excellent targets for genetic transformation.Um método para a indução de embriogênese somática em oito genótipos de mandioca cultivados no Nordeste brasileiro foi desenvolvido. A indução de embriogênese somática foi feita utilizando como explantes ápices caulinares isolados de plantas cultivadas in vitro e ápices caulinares isolados a partir de brotações induzidas em casa-de-vegetação em manivas de plantas adultas. Em todos os genótipos a auxina picloram, em uma ampla faixa de concentrações, foi capaz de induzir embriogênese somática em altas freqüências e com um grande número de embriões por explante. Foi mostrado também, que é possível induzir embriogênese somática secundária (cíclica a partir de cotilédones verdes de embriões somáticos maduros, utilizando picloram no meio de indução. O método aqui apresentado poderá ser utilizado para a produção em massa de plantas dos genótipos utilizados. A alta freqüência de embriogênese somática secundária obtida quando cotilédones verdes de embriões somáticos são utilizados como explantes, mostra que tais cotilédones podem se constituir em excelentes alvos para a transformação genética e posterior obtenção de

  17. Anatomy of somatic embryogenesis in Carica papaya L.

    Directory of Open Access Journals (Sweden)

    Juliana A. Fernando

    2001-09-01

    Full Text Available Mature zygotic embryos of Carica papaya L. ‘Sunrise Solo’ were used as explants for embryogenesis induction. The explants were inoculated on Murashige and Skoog culture medium supplemented with 2 mg.L-1 2,4-dichlorophenoxyacetic acid and incubated in darkness at 25+2°C. Histological analysis of callogenesis and somatic embryogenesis indicated occurrence of direct and indirect somatic embryogenesis development. Direct somatic embryo formation was observed from hypocotyledonary epidermic cells only from explant 18 days after inoculation. Somatic embryos formed indirectly were originated from embryogenic superficial cells of pre-embryonic complexes located on peripherical and on internal cell layers of callus 49 days after inoculation. Diverse morphological differences including disformed embryos were observed among the somatic embryos.Embriões zigóticos maduros de Carica papaya L. ‘Sunrise Solo’ foram utilizados como explantes para indução da embriogênese. Estes explantes foram inoculados em meio de cultura de Murashige & Skoog suplementado com 2,0 mg.L-1 de 2,4 ácido diclorofenoxiacético (2,4-D e mantidos no escuro em câmara de crescimento à temperatura de 21°C por período de tempo variável. Através da análise histológica foi possível verificar que os primeiros embriões somáticos formaram-se diretamente a partir de células únicas da epiderme hipocotiledonar do explante após o 18º dia de cultura. Porém, os demais embriões somáticos originaram-se indiretamente a partir de células superficiais de complexos pré-embriônicos presentes nas camadas periféricas e internas do calo após o 49º dia de cultura. Foram detectadas algumas diferenças morfológicas entre os embriões somáticos obtidos.

  18. LEA: An Algorithm to Estimate the Level of Location Exposure in Infrastructure-Based Wireless Networks

    Directory of Open Access Journals (Sweden)

    Francisco Garcia

    2017-01-01

    Full Text Available Location privacy in wireless networks is nowadays a major concern. This is due to the fact that the mere fact of transmitting may allow a network to pinpoint a mobile node. We consider that a first step to protect a mobile node in this situation is to provide it with the means to quantify how accurately a network establishes its position. To achieve this end, we introduce the location-exposure algorithm (LEA, which runs on the mobile terminal only and whose operation consists of two steps. In the first step, LEA discovers the positions of nearby network nodes and uses this information to emulate how they estimate the position of the mobile node. In the second step, it quantifies the level of exposure by computing the distance between the position estimated in the first step and its true position. We refer to these steps as a location-exposure problem. We tested our proposal with simulations and testbed experiments. These results show the ability of LEA to reproduce the location of the mobile node, as seen by the network, and to quantify the level of exposure. This knowledge can help the mobile user decide which actions should be performed before transmitting.

  19. Oil palm (Elaeis guineensis Jacq. tissue culture ESTs: Identifying genes associated with callogenesis and embryogenesis

    Directory of Open Access Journals (Sweden)

    Ooi Leslie CL

    2008-05-01

    Full Text Available Abstract Background Oil palm (Elaeis guineensis Jacq. is one of the most important oil bearing crops in the world. However, genetic improvement of oil palm through conventional breeding is extremely slow and costly, as the breeding cycle can take up to 10 years. This has brought about interest in vegetative propagation of oil palm. Since the introduction of oil palm tissue culture in the 1970s, clonal propagation has proven to be useful, not only in producing uniform planting materials, but also in the development of the genetic engineering programme. Despite considerable progress in improving the tissue culture techniques, the callusing and embryogenesis rates from proliferating callus cultures remain very low. Thus, understanding the gene diversity and expression profiles in oil palm tissue culture is critical in increasing the efficiency of these processes. Results A total of 12 standard cDNA libraries, representing three main developmental stages in oil palm tissue culture, were generated in this study. Random sequencing of clones from these cDNA libraries generated 17,599 expressed sequence tags (ESTs. The ESTs were analysed, annotated and assembled to generate 9,584 putative unigenes distributed in 3,268 consensi and 6,316 singletons. These unigenes were assigned putative functions based on similarity and gene ontology annotations. Cluster analysis, which surveyed the relatedness of each library based on the abundance of ESTs in each consensus, revealed that lipid transfer proteins were highly expressed in embryogenic tissues. A glutathione S-transferase was found to be highly expressed in non-embryogenic callus. Further analysis of the unigenes identified 648 non-redundant simple sequence repeats and 211 putative full-length open reading frames. Conclusion This study has provided an overview of genes expressed during oil palm tissue culture. Candidate genes with expression that are modulated during tissue culture were identified. However

  20. Oil palm (Elaeis guineensis Jacq.) tissue culture ESTs: identifying genes associated with callogenesis and embryogenesis.

    Science.gov (United States)

    Low, Eng-Ti L; Alias, Halimah; Boon, Soo-Heong; Shariff, Elyana M; Tan, Chi-Yee A; Ooi, Leslie Cl; Cheah, Suan-Choo; Raha, Abdul-Rahim; Wan, Kiew-Lian; Singh, Rajinder

    2008-05-29

    Oil palm (Elaeis guineensis Jacq.) is one of the most important oil bearing crops in the world. However, genetic improvement of oil palm through conventional breeding is extremely slow and costly, as the breeding cycle can take up to 10 years. This has brought about interest in vegetative propagation of oil palm. Since the introduction of oil palm tissue culture in the 1970s, clonal propagation has proven to be useful, not only in producing uniform planting materials, but also in the development of the genetic engineering programme. Despite considerable progress in improving the tissue culture techniques, the callusing and embryogenesis rates from proliferating callus cultures remain very low. Thus, understanding the gene diversity and expression profiles in oil palm tissue culture is critical in increasing the efficiency of these processes. A total of 12 standard cDNA libraries, representing three main developmental stages in oil palm tissue culture, were generated in this study. Random sequencing of clones from these cDNA libraries generated 17,599 expressed sequence tags (ESTs). The ESTs were analysed, annotated and assembled to generate 9,584 putative unigenes distributed in 3,268 consensi and 6,316 singletons. These unigenes were assigned putative functions based on similarity and gene ontology annotations. Cluster analysis, which surveyed the relatedness of each library based on the abundance of ESTs in each consensus, revealed that lipid transfer proteins were highly expressed in embryogenic tissues. A glutathione S-transferase was found to be highly expressed in non-embryogenic callus. Further analysis of the unigenes identified 648 non-redundant simple sequence repeats and 211 putative full-length open reading frames. This study has provided an overview of genes expressed during oil palm tissue culture. Candidate genes with expression that are modulated during tissue culture were identified. However, in order to confirm whether these genes are suitable as

  1. Improvements of cyclic somatic embryogenesis of cassava (Manihot esculenta Crantz).

    Science.gov (United States)

    Raemakers, C J; Schavemaker, C M; Jacobsen, E; Visser, R G

    1993-02-01

    In cassava a cyclic system of somatic embryogenesis was developed. Primary (torpedo shaped or germinated) embryos, originating from leaf lobes, could only be obtained after culture on solid medium. Cyclic embryos, originating from embryos, could be obtained in both liquid and on solid medium. The production of embryos in liquid medium was distinctly higher, faster and more synchronized than on solid medium. Lower densities and fragmentation of starting embryos improved the production significantly. The highest production found was 32.1 embryos per initial embryo. In all treatments the explants initiated multiple embryos. The production of single embryos was achieved by pressing starting embryos through a fine meshed sieve, indicating that embryos can be produced from a piece of tissue with a restricted number of cells. The shoot conversion rate of embryos from liquid medium was comparable with that of embryos from solid medium.

  2. Somatic embryogenesis and plant regeneration of Capsicum baccatum L.

    Directory of Open Access Journals (Sweden)

    Peddaboina Venkataiah

    2016-06-01

    Full Text Available A plant regeneration protocol via somatic embryogenesis was achieved in cotyledon and leaf explants of Capsicum baccatum, when cultured on MS medium supplemented with various concentrations of 2,4-dichlorophenoxy acetic acid (2,4-D, 0.5–5.0 mg l−1 in combination with Kinetin (Kn, 0.5 mg l−1 and 3% sucrose. Various stages were observed during the development of somatic embryos, including globular, heart, and torpedo-stages. Torpedo stage embryos were separated from the explants and subcultured on medium supplemented with various concentrations of different plant growth regulators for maturation. Maximum percentage (55% of somatic embryo germination and plantlet formation was found at 1.0 mg l−1 BA. Finally, about 68% of plantlets were successfully established under field conditions. The regenerated plants were morphologically normal, fertile and able to set viable seeds.

  3. Current insights into hormonal regulation of microspore embryogenesis

    Directory of Open Access Journals (Sweden)

    Iwona eŻur

    2015-06-01

    Full Text Available Plant growth regulator (PGR crosstalk and interaction with the plant’s genotype and environmental factors play a crucial role in microspore embryogenesis (ME, controlling microspore-derived embryo differentiation and development as well as haploid/doubled haploid plant regeneration. The complexity of the PGR network which could exist at the level of biosynthesis, distribution, gene expression or signaling pathways, renders the creation of an integrated model of ME-control crosstalk impossible at present. However, the analysis of the published data together with the results received recently with the use of modern analytical techniques brings new insights into hormonal regulation of this process. This review presents a short historical overview of the most important milestones in the recognition of hormonal requirements for effective ME in the most important crop plant species and complements it with new concepts that evolved over the last decade of ME studies.

  4. Current insights into hormonal regulation of microspore embryogenesis

    Science.gov (United States)

    Żur, Iwona; Dubas, Ewa; Krzewska, Monika; Janowiak, Franciszek

    2015-01-01

    Plant growth regulator (PGR) crosstalk and interaction with the plant’s genotype and environmental factors play a crucial role in microspore embryogenesis (ME), controlling microspore-derived embryo differentiation and development as well as haploid/doubled haploid plant regeneration. The complexity of the PGR network which could exist at the level of biosynthesis, distribution, gene expression or signaling pathways, renders the creation of an integrated model of ME-control crosstalk impossible at present. However, the analysis of the published data together with the results received recently with the use of modern analytical techniques brings new insights into hormonal regulation of this process. This review presents a short historical overview of the most important milestones in the recognition of hormonal requirements for effective ME in the most important crop plant species and complements it with new concepts that evolved over the last decade of ME studies. PMID:26113852

  5. [Direct and indirect somatic embryogenesis in Freesia refracta].

    Science.gov (United States)

    Wang, L; Duan, X G; Hao, S

    1999-06-01

    Somatic embryogenesis can be induced in tissue cultures of Freesia refracta either directly from the epidermal cells of explant, or indirectly via intervening callus. In direct pathway, somatic embryos were in contact with maternal tissue in a suspensor-like structure. In indirect pathway, the explants first proliferacted to give rise to calluses before embryoids were induced. The two sorts of calluses were defined to embryogenic callus and non-embryogenic callus according to producing of somatic embryos. An indirect somatic embryo is developed from a pre-embryogenically determined cell. This kind of somatic embryo has no suspensor structure instead of a complex with maternal tissue. Somatic embryos have their own vascular tissues, and can develop new plantlets independently.

  6. Transcriptional identification and characterization of differentially expressed genes associated with embryogenesis in radish (Raphanus sativus L.).

    Science.gov (United States)

    Zhai, Lulu; Xu, Liang; Wang, Yan; Zhu, Xianwen; Feng, Haiyang; Li, Chao; Luo, Xiaobo; Everlyne, Muleke M; Liu, Liwang

    2016-02-23

    Embryogenesis is an important component in the life cycle of most plant species. Due to the difficulty in embryo isolation, the global gene expression involved in plant embryogenesis, especially the early events following fertilization are largely unknown in radish. In this study, three cDNA libraries from ovules of radish before and after fertilization were sequenced using the Digital Gene Expression (DGE) tag profiling strategy. A total of 5,777 differentially expressed transcripts were detected based on pairwise comparison in the three libraries (0_DAP, 7_DAP and 15_DAP). Results from Gene Ontology (GO) and pathway enrichment analysis revealed that these differentially expressed genes (DEGs) were implicated in numerous life processes including embryo development and phytohormones biosynthesis. Notably, some genes encoding auxin response factor (ARF ), Leafy cotyledon1 (LEC1) and somatic embryogenesis receptor-like kinase (SERK ) known to be involved in radish embryogenesis were differentially expressed. The expression patterns of 30 genes including LEC1-2, AGL9, LRR, PKL and ARF8-1 were validated by qRT-PCR. Furthermore, the cooperation between miRNA and mRNA may play a pivotal role in the radish embryogenesis process. This is the first report on identification of DEGs profiles related to radish embryogenesis and seed development. These results could facilitate further dissection of the molecular mechanisms underlying embryogenesis and seed development in radish.

  7. Somatic embryogenesis from bud and leaf explants of date palm (Phoenix dactylifera L.) cv. Najda.

    Science.gov (United States)

    Mazri, Mouaad Amine; Belkoura, Ilham; Meziani, Reda; Mokhless, Boutaïna; Nour, Souad

    2017-05-01

    An efficient regeneration system through somatic embryogenesis was developed for date palm cv. Najda. Adventitious bud and proximal leaf segments cultured on Murashige and Skoog (MS) medium supplemented with various combinations of auxins and cytokinins induced embryogenesis after at least 6 months of culture. Somatic embryogenesis induction seemed correlated with the type of the explant, the induction period and the auxin used. The highest rate of somatic embryogenesis (86.0%) was obtained on bud explants cultured on MS medium supplemented with 45.0 µM 2,4-dichlorophenoxyacetic acid (2,4-D), and 4.5 µM kinetin or 4.5 µM 6-(dimethylallylamino) purine (2iP). Whereas, low levels of embryogenesis were obtained on media supplemented with 1-naphthalene acetic acid (NAA) or 2-naphthoxyacetic acid (NOA). Proximal leaf segments showed somatic embryogenesis only when cultured on media supplemented with 2,4-D or picloram. Statistical analysis revealed significant effects of explant type and plant growth regulators (PGRs) combination on somatic embryogenesis. Somatic embryos were germinated successfully on PGR-free MS medium with or without activated charcoal (50.0-60.0 and 26.6-36.6%, respectively), and 80.0% of plantlets survived after transferring to a glasshouse for 6 months. Our results will be useful for large-scale propagation of date palm cv. Najda, characterized by high fruit quality and bayoud disease resistance.

  8. Dynamic nucleosome organization at hox promoters during zebrafish embryogenesis.

    Directory of Open Access Journals (Sweden)

    Steven E Weicksel

    Full Text Available Nucleosome organization at promoter regions plays an important role in regulating gene activity. Genome-wide studies in yeast, flies, worms, mammalian embryonic stem cells and transformed cell lines have found well-positioned nucleosomes flanking a nucleosome depleted region (NDR at transcription start sites. This nucleosome arrangement depends on DNA sequence (cis-elements as well as DNA binding factors and ATP-dependent chromatin modifiers (trans-factors. However, little is understood about how the nascent embryonic genome positions nucleosomes during development. This is particularly intriguing since the embryonic genome must undergo a broad reprogramming event upon fusion of sperm and oocyte. Using four stages of early embryonic zebrafish development, we map nucleosome positions at the promoter region of 37 zebrafish hox genes. We find that nucleosome arrangement at the hox promoters is a progressive process that takes place over several stages. At stages immediately after fertilization, nucleosomes appear to be largely disordered at hox promoter regions. At stages after activation of the embryonic genome, nucleosomes are detectable at hox promoters, with positions becoming more uniform and more highly occupied. Since the genomic sequence is invariant during embryogenesis, this progressive change in nucleosome arrangement suggests that trans-factors play an important role in organizing nucleosomes during embryogenesis. Separating hox genes into expressed and non-expressed groups shows that expressed promoters have better positioned and occupied nucleosomes, as well as distinct NDRs, than non-expressed promoters. Finally, by blocking the retinoic acid-signaling pathway, we disrupt early hox gene transcription, but observe no effect on nucleosome positions, suggesting that active hox transcription is not a driving force behind the arrangement of nucleosomes at the promoters of hox genes during early development.

  9. Reproduction of the Medicinal Plant Pelargonium sidoides via Somatic Embryogenesis.

    Science.gov (United States)

    Duchow, Stefanie; Blaschek, Wolfgang; Classen, Birgit

    2015-08-01

    The medicinal plant Pelargonium sidoides DC. (Geraniaceae) was traditionally used for the treatment of the common cold and cough in South Africa. Today an aequous-ethanolic root extract from this plant is approved for the treatment of acute bronchitis and is globally marketed also as an immunostimulant. The increasing demand of the plant material for the industrial production indicates the need of new effective methods for the propagation of P. sidoides. Here we report somatic embryogenesis and in vitro plantlet regeneration from somatic cells of inflorescence shoots and petioles of P. sidoides. A one-week cultivation of explants in media containing different concentrations of thidiazuron (1, 2.2, 3, and 4 mg/L) followed by a cultivation period without phytohormones resulted in the induction of somatic embryos within 2-4 weeks. After 2-4 months, the embryos generated roots and could be transferred into a greenhouse, where flower formation took place and the development of seeds occurred with high germination rates. The root umckalin concentration, determined by high-performance thin-layer chromatography, was comparable to that of seed-cultivated plants (100 ± 6 vs. 113 ± 10 µg umckalin/g dried roots). For the first time, direct somatic embryogenesis has been established as an appropriate cultivation method for P. sidoides plants used as raw material in the pharmaceutical industry. Moreover, genetically identical plants (chemical races) can be easily generated by this procedure. Georg Thieme Verlag KG Stuttgart · New York.

  10. Ontogenic appearance of MHC class I (B-F) antigens during chicken embryogenesis

    DEFF Research Database (Denmark)

    Dunon, D; Salomonsen, J; Skjødt, K

    1990-01-01

    .5 of embryogenesis. B-F cell-surface expression first becomes detectable in hemopoietic organs by day 10-12 of embryogenesis and somewhat later in nonhemopoietic organs. Flow cytometry analysis of hemopoietic cells throughout embryogenesis revealed B-Fhi and B-Flo cell populations. The percentage of B-F+ cells......Expression of chicken MHC class I (B-F) antigens during ontogeny was determined by binding of anticlass I antibody and appearance of B-F transcripts by Northern blotting in chicken organs during embryogenesis until 2 weeks after hatching. MHC class I transcripts first become detectable in day 6...... in spleen and bone marrow decreased around hatching, which could reflect either cell flows in these organs during this period or the sensitivity of hemopoietic cells to hatching stress. Udgivelsesdato: 1990-null...

  11. Possible role of light and polyamines in the onset of somatic embryogenesis of Coffea canephora.

    Science.gov (United States)

    De-la-Peña, Clelia; Galaz-Avalos, Rosa M; Loyola-Vargas, Víctor M

    2008-07-01

    The concentration of free and bound polyamines was studied during the somatic embryogenesis induction process in Coffea canephora explants. In the present study we show that when the induction of somatic embryogenesis in C. canephora is carried out under light conditions and in the presence of the plant growth regulator, benzylaminopurine, a cytokinin, a faster response to induction is obtained. In the darkness, the response is delayed for more than 20 days, and the number of embryos is smaller. In the absence of benzylaminopurine no embryogenic response was observed. The pronounced changes in the levels of putrescine, spermidine, and spermine, both free and bound, found in C. canephora suggest that a close correlation exists between polyamine biosynthesis and somatic embryogenesis in C. canephora during a period of cellular differentiation associated with the induction of somatic embryogenesis.

  12. The periodization of wheat embryogenesis on the base of anatomical, morphological and time criterions

    Directory of Open Access Journals (Sweden)

    Natalia N. Kruglova

    2013-04-01

    Full Text Available The periodization of wheat embryogenesis on the base of anatomy-morphological and temporal criterions has been proposed. The stages of non-differentiated embryo, embryo differentiation and differentiated embryo were described.

  13. Transcriptional identification and characterization of differentially expressed genes associated with embryogenesis in radish (Raphanus sativus L.)

    OpenAIRE

    Zhai, Lulu; Xu, Liang; Wang, Yan; Zhu, Xianwen; Feng, Haiyang; Li, Chao; Luo, Xiaobo; Everlyne, Muleke M.; Liu, Liwang

    2016-01-01

    Embryogenesis is an important component in the life cycle of most plant species. Due to the difficulty in embryo isolation, the global gene expression involved in plant embryogenesis, especially the early events following fertilization are largely unknown in radish. In this study, three cDNA libraries from ovules of radish before and after fertilization were sequenced using the Digital Gene Expression (DGE) tag profiling strategy. A total of 5,777 differentially expressed transcripts were det...

  14. Cell dedifferentiation, callus induction and somatic embryogenesis in Crataegus spp.

    Science.gov (United States)

    Taimori, N; Kahrizi, D; Abdossi, V; Papzan, A H

    2016-09-30

    The present study describes the effects of light conditions, different kinds and concentrations of auxins [Naphthylacetic acid (NAA) and dichlorophenoxyacetic acid (2,4-D)] with cytokinin (Kin) in MS medium on callus induction and embryogenesis in Crataegus pseudoheterophylla, C. aronia and C.meyeri. At first leave explants sections were cultured on different combinations of plant growth regulators in dark and light for callus initiation and light conditions to evaluation the percentage and duration of survival, callus diameter, callus fresh weight and dry. Results of effects of plant growth regulators and light conditions on callus initiation revealed that highest percentage of callus initiation leaves in treatment (0.5 mg/l 2.4-D+0.5 mg/l KIN) for species C.pseudoheterophylla in dark conditions (100%). Dark conditions (100%) were more effective on callogenesis than light conditions (Photoperiodicity of 16-h and at light intensity of 40 µmol m-2 s-1). The callus induction of in vitro (64-100%) leaves was better than the ex vitro ones (0-100%). The combination of 2,4-D and Kin of in vitro leaves callogenesis has been indicated faster (one weeks) than the other combinations. The results also showed that the highest percentage (100%) and survival duration (6 months) was found in species C. pseudoheterophylla and C. meyeri in 0.1 mg/l 2,4.D + 0.5 mg/l KIN and 0.5 mg/l 2,4.D + 0.5 mg/l Kin. The minimum survival (0%) was absorbed in species C. aronia in 1 mg/l NAA. Maximum callus (10.63 and 10.00 mm respectively) was shown in 0.1 mg/l 2,4.D + 0.5 mg/l Kin and 0.5 mg/l 2,4.D + 0.5 mg/l Kin and was not significant differences after five week among species. The results showed that the highest fresh (1081.49 mg) and dry weight (506.88 and 506.98 mg respectively) was absorbed in species C. pseudoheterophylla in 0.1 mg/l 2,4.D + 0.5 mg/l Kin and 0.5 mg/l 2,4.D + 0.5 mg/l Kin. The embryogenesis was not occurred in any plant growth regulator combinations and species. The

  15. DIRECT AND INDIRECT SOMATIC EMBRYOGENESIS ON ARABICA COFFEE (Coffea arabica

    Directory of Open Access Journals (Sweden)

    Meynarti Sari Dewi Ibrahim

    2013-10-01

    Full Text Available Propagation of Coffea arabica L. through direct and indirect somatic embryogenesis technique is promising for producing large number of coffee seedlings. The objectives of the research were to evaluate methods for direct and indirect somatic embryo-genesis induction of C. arabica var. Kartika. The explants were the youngest fully expanded leaves of arabica coffee. The evalu-ated medium was modified Murashige and Skoog (MS medium supplemented with a combination of 2.26 µM 2,4-D + 4.54 or 9.08 µM thidiazuron; 4.52 µM 2,4-D + 4.54 or 9.08 µM thidiazuron; or 9.04 µM 2,4-D + 9.08 µM thidiazuron. Both calli (100 mg and pre-embryos developed from the edge of leaf explants were subcultured into regeneration medium (half strength MS with modified vitamin, supplemented with kinetine 9.30 µM and adenine sulfate 40 mg L-1. The results showed coffee leaf explant cultured on medium containing 2.26 µM 2,4-D + 4.54 or 9.08 µM thidiazuron to induce direct somatic embriogenesis from explant, while that of 4.52 or 9.04 µM 2,4-D + 9.08 µM thidiazuron to induced indirect somatic embrio-genesis. The medium for calli induction from coffee by explants was medium supplemented with 4.52 or 9.04 µM 2,4-D in combination with 9.08 µM thidiazuron. On the other hand, the best medium for activation of induction of somatic embryos was MS medium supplemented with 9.04 µM 2,4-D + 9.08 µM thidiazuron. Based on this results, the first step for developing micropropagation for coffee has been resolved. The subsequent studies will be directed to evaluate agronomic performance of the derived planting materials.

  16. DOUGLAS LEA MEMORIAL LECTURE: From targets to genes: a brief history of radiosensitivity

    Science.gov (United States)

    Steel, G. Gordon

    1996-02-01

    The biological work of Douglas Lea spanned the period from 1934 to his early death in 1947, and during this short period he made important contributions to the theory of radiation action. He interpreted experimental data relating to the effects of radiation on viruses, bacteria, bean roots, etc in terms of the inactivation of discrete targets, which he identified with cellular genes. He thus laid the foundation of much subsequent research. It is now well recognized that mammalian cells differ substantially in radiosensitivity, especially in the low-dose region of the survival curve. The dependence of radiosensitivity on dose rate has been widely studied; this has practical significance for clinical radiotherapy as well as mechanistic implications. Since Lea's time there have been a number of efforts to describe models that can relate cell killing to radiation dose, dose rate, and track structure. So far these have not led to a comprehensive and widely accepted picture. Microdosimetric considerations lead to the concept of differing severity of lesions induced in DNA. Much is known about the sequence of processes that subsequently lead to cell inactivation: this can be divided into phases of induction, processing, and manifestation. Chromosomal events are currently attracting much attention, as they did in Lea's time. Considerable progress has also been made in identifying genes that control the repair of radiation damage. It has been found that mutation is frequently associated with the loss of a large segment of the genome around the damage site and this will have important implications for interactive processes between particle tracks.

  17. Petrophysical and geotechnical properties of Lea Park Formation shale from Weyburn, Saskatchewan

    Energy Technology Data Exchange (ETDEWEB)

    Chan, T.K. [Saskatchewan Univ., Saskatoon, SK (Canada). Environmental Engineering; Hawkes, C.D. [Saskatchewan Univ., Saskatoon, SK (Canada). Dept. of Civil and Geological Engineering

    2005-07-01

    The petrophysical and geotechnical properties of the shale rock found within the Leak Park Formation were characterized by testing preserved core samples from a well in a mature oil field in Weyburn Saskatchewan. EnCana Corporation has been using carbon dioxide (CO{sub 2}) injection at the well site since 2000 for enhanced oil recovery. The site is also the focus of a major international research project on carbon storage and monitoring. The objective of the testing program was to determine the potential for this formation to act as a barrier to carbon dioxide (CO{sub 2}) leakage, in the event that CO{sub 2} injected into the Midale unit was to leak through the primary caprock that overlies the injection zone. The core samples had water contents of approximately 17 per cent by weight and a porosity of 22 per cent. Plastic and liquid limits were measured to be 22.5 and 94.7 per cent respectively. The shale consists mainly of quartz, mixed-layer illite-montmorillonite, and illite. Results indicate that the Lea Park Formation is a relatively uniform, ultra low-permeability aquitard that should be effective for mitigating upwards migration of CO{sub 2}. The results of this testing program will also be useful in hydrogeological investigations of field-scale flow patterns in the Weyburn region, the design of drilling programs and drilling fluids for wells that penetrate the Lea Park Formation, and for geotechnical engineering projects that might use the Lea Park Formation as a construction material. 8 refs., 4 tabs., 7 figs.

  18. Raccoon abundance inventory report

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — This report summarizes the results of a raccoon abundance inventory on Clarence Cannon National Wildlife Refuge in 2012. Determining raccoon abundance allows for...

  19. Genetic variants regulating expression levels and isoform diversity during embryogenesis.

    Science.gov (United States)

    Cannavò, Enrico; Koelling, Nils; Harnett, Dermot; Garfield, David; Casale, Francesco P; Ciglar, Lucia; Gustafson, Hilary E; Viales, Rebecca R; Marco-Ferreres, Raquel; Degner, Jacob F; Zhao, Bingqing; Stegle, Oliver; Birney, Ewan; Furlong, Eileen E M

    2017-01-19

    Embryonic development is driven by tightly regulated patterns of gene expression, despite extensive genetic variation among individuals. Studies of expression quantitative trait loci (eQTL) indicate that genetic variation frequently alters gene expression in cell-culture models and differentiated tissues. However, the extent and types of genetic variation impacting embryonic gene expression, and their interactions with developmental programs, remain largely unknown. Here we assessed the effect of genetic variation on transcriptional (expression levels) and post-transcriptional (3' RNA processing) regulation across multiple stages of metazoan development, using 80 inbred Drosophila wild isolates, identifying thousands of developmental-stage-specific and shared QTL. Given the small blocks of linkage disequilibrium in Drosophila, we obtain near base-pair resolution, resolving causal mutations in developmental enhancers, validated transcription-factor-binding sites and RNA motifs. This fine-grain mapping uncovered extensive allelic interactions within enhancers that have opposite effects, thereby buffering their impact on enhancer activity. QTL affecting 3' RNA processing identify new functional motifs leading to transcript isoform diversity and changes in the lengths of 3' untranslated regions. These results highlight how developmental stage influences the effects of genetic variation and uncover multiple mechanisms that regulate and buffer expression variation during embryogenesis.

  20. Embryogenesis, hatching and larval development of Artemia during orbital spaceflight

    Science.gov (United States)

    Spooner, B. S.; Debell, L.; Armbrust, L.; Guikema, J. A.; Metcalf, J.; Paulsen, A.

    1994-08-01

    Developmental biology studies, using gastrula-arrested cysts of the brine shrimp Artemia franciscana, were conducted during two flights of the space shuttle Atlantis (missions STS-37 and STS-43) in 1991. Dehydrated cysts were activated, on orbit, by addition of salt water to the cysts, and then development was terminated by the addition of fixative. Development took place in 5 ml syringes, connected by tubing to activation syringes, containing salt water, and termination syringes, containing fixative. Comparison of space results with simultaneous ground control experiments showed that equivalent percentages of naupliar larvae hatched in the syringes (40%). Thus, reactivation of development, completion of embryogenesis, emergence and hatching took place, during spaceflight, without recognizable alteration in numbers of larvae produced. Post-hatching larval development was studied in experiments where development was terminated, by intrduction of fixative, 2 days, 4 days, and 8 days after reinitiation of development. During spaceflight, successive larval instars or stages, interrupted by molts, occurred, generating brine shrimp at appropriate larval instars. Naupliar larvae possessed the single naupliar eye, and development of the lateral pair of adult eyes also took place in space. Transmission electron microscopy revealed extensive differentiation, including skeletal muscle and gut endoderm, as well as the eye tissues. These studies demonstrate the potential value of Artemia for developmental biology studies during spaceflight, and show that extensive degress of development can take place in this microgravity environment.

  1. Four queries concerning the metaphysics of early human embryogenesis.

    Science.gov (United States)

    Howsepian, A A

    2008-04-01

    In this essay, I attempt to provide answers to the following four queries concerning the metaphysics of early human embryogenesis. (1) Following its first cellular fission, is it coherent to claim that one and only one of two "blastomeric" twins of a human zygote is identical with that zygote? (2) Following the fusion of two human pre-embryos, is it coherent to claim that one and only one pre-fusion pre-embryo is identical with that postfusion pre-embryo? (3) Does a live human being come into existence only when its brain comes into existence? (4) At implantation, does a pre-embryo become a mere part of its mother? I argue that either if things have quidditative properties or if criterialism is false, then queries (1) and (2) can be answered in the affirmative; that in light of recent developments in theories of human death and in light of a more "functional" theory of brains, query (3) can be answered in the negative; and that plausible mereological principles require a negative answer to query (4).

  2. The effects of microgravity on gametogenesis, fertilization, and early embryogenesis

    Science.gov (United States)

    Tan, X.

    Gametogenesis fertilization and early embryogenesis are crucial periods for normal development afterwards In past three decades many experiments have been conducted in space and in simulated weightlessness induced by clinostats to elucidate the issue Different animal species including Drosophila wasp shrimp fish amphibian mouse rats etc have been used for the study Oogenesis and spermatogenesis are affected by microgravity in different ways Some researches found that microgravity condition perturbed the process of oogenesis in many species A significant increased frequency of chromosomal non-disjunction was found in Drosophila females resulting the loss of chromosomes during meiosis and inhibition of cell division Studies on wasp showed a decreased hatchability and accumulation of unhatched eggs when the insects were exposed to spaceflight at different stages of oogenesis For experiments conducted on vertebrate animal models the results are somehow different however Microgravity has no significant effect for fish Medaka etc amphibian South African clawed toad Xenopus laevis or mammals mouse Spermatogenesis on the other hand is more significantly affected by microgravity condition Some researches indicated sperm are sensitive to changes in gravitational force and this sensitivity affects the ability of sperm to fertilize eggs Sperm swim with higher velocity in microgravity which is coupled with altered protein phosphorylation level in sperm under microgravity condition Microgravity also induced activation of the

  3. Excess caffeine exposure impairs eye development during chick embryogenesis

    Science.gov (United States)

    Ma, Zheng-lai; Wang, Guang; Cheng, Xin; Chuai, Manli; Kurihara, Hiroshi; Lee, Kenneth Ka Ho; Yang, Xuesong

    2014-01-01

    Caffeine has been an integral component of our diet and medicines for centuries. It is now known that over consumption of caffeine has detrimental effects on our health, and also disrupts normal foetal development in pregnant mothers. In this study, we investigated the potential teratogenic effect of caffeine over-exposure on eye development in the early chick embryo. Firstly, we demonstrated that caffeine exposure caused chick embryos to develop asymmetrical microphthalmia and induced the orbital bone to develop abnormally. Secondly, caffeine exposure perturbed Pax6 expression in the retina of the developing eye. In addition, it perturbed the migration of HNK-1+ cranial neural crest cells. Pax6 is an important gene that regulates eye development, so altering the expression of this gene might be the cause for the abnormal eye development. Thirdly, we found that reactive oxygen species (ROS) production was significantly increased in eye tissues following caffeine treatment, and that the addition of anti-oxidant vitamin C could rescue the eyes from developing abnormally in the presence of caffeine. This suggests that excess ROS induced by caffeine is one of the mechanisms involved in the teratogenic alterations observed in the eye during embryogenesis. In sum, our experiments in the chick embryo demonstrated that caffeine is a potential teratogen. It causes asymmetrical microphthalmia to develop by increasing ROS production and perturbs Pax6 expression. PMID:24636305

  4. Changes of gentiopicroside synthesis during somatic embryogenesis in Gentiana macrophylla.

    Science.gov (United States)

    Chen, Li-Yu; Chen, Qian-Liang; Xu, Dan; Hao, Jian-Guo; Schläppi, Michael; Xu, Zi-Qin

    2009-12-01

    IN VITRO plant regeneration of Gentiana macrophylla Pall. and determination of gentiopicroside content during somatic embryogenesis are described in the present work. The highest percentage of embryogenic callus formation was observed in Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg/L 6-benzylaminopurine (BA). Calli were subcultured on MS medium containing 1.0 mg/L 2,4-D, 1.0 mg/L BA and 500 mg/L lactalbumin hydrolysate (LH) at intervals of 25 days. A higher frequency of somatic embryo maturation was achieved on MS medium containing B5 vitamins (MB) supplemented with different concentrations of 1-naphthaleneacetic acid (NAA) and BA than with a combination of NAA and kinetin (KT). Addition of AgNO(3) improved maturation of somatic embryos while thidiazuron (TDZ) promoted vitrification. The gentiopicroside contents of embryogenic calli and globular-, heart-, torpedo-, and cotyledon-shaped embryoids were determined by high-performance liquid chromatography (HPLC). Gentiopicroside was not detectable in embryogenic calli, but in all types of somatic embryos. The highest gentiopicroside content was observed in cotyledon-shaped embryoids, reaching more than 12 mg/g dry weight. Georg Thieme Verlag KG Stuttgart. New York.

  5. Clonal propagation of Cyclamen persicum via somatic embryogenesis.

    Science.gov (United States)

    Winkelmann, Traud

    2010-01-01

    Cyclamen (Cyclamen persicum) is an economically important ornamental pot plant with local use as cut flower as well. Traditionally, it is propagated via seeds, but interest is given in vegetative propagation of parental lines as well as superior single plants. Somatic embryogenesis is an efficient in vitro propagation method for many cyclamen cultivars. Starting from ovules of unpollinated flowers, callus is induced and propagated in a medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-(gamma,gamma-dimethylallylamino)purine (2iP). Transfer to hormone-free medium results in the differentiation of somatic embryos, which afterwards germinate on the same medium. These first culture stages take about 6-7 months and are carried out in complete darkness. Two to four months after the transfer to light, plantlets develop which can be acclimatized in the greenhouse. The regenerated plants are characterized by low percentages of somaclonal variation. This protocol has proven useful not only for clonal propagation, but also for artificial seed preparation, cryopreservation, genetic transformation and protoplast regeneration.

  6. Somatic embryogenesis from zygotic embryos of Euterpe oleracea Mart.

    Directory of Open Access Journals (Sweden)

    Ledo Ana da Silva

    2002-01-01

    Full Text Available The aim of this work was to study the morphogenetic responses of zygotic embryos of açai palm (Euterpe oleracea Mart. submitted to several conditions of in vitro culture. Several research experiments were conducted, in laboratory, using vegetable material collected from açai palm plants at Embrapa Amazon Oriental, Belém-PA, Brazil. It was possible to verify the expression of a direct, repetitive and no-synchronized model of somatic embryogenesis in mature zygotic embryos cultivated in primary MS medium supplemented with 2,4-D (339.36 muM and transferred to a secondary MS medium in the presence of NAA (0.537 muM and 2iP (12.30 muM. The conversion of somatic embryos into seedlings was reached after 210 days with the transfer of the cultures to a third medium with sucrose and mineral salts concentrations reduced to a half, without growth regulators.

  7. Evaluation from an LEA Perspective: Proceedings from an AERA Symposium. Annual Meeting of the American Educational Research Association (66th, New York, New York, March 19-23, 1982).

    Science.gov (United States)

    Thompson, Bruce; And Others

    The views of local education agency (LEA) evaluators tend to be underrepresented in the evaluation literature. This symposium was organized to fill this gap. The non-LEA symposium participants (Carol H. Weiss and Bruce Thompson) were asked to pose questions to the LEA participants (Steven Frankel, Montgomery County (MD) Public Schools; Freda…

  8. Comparison of Lea Symbols chart and Sheridan Gardiner chart in assessing vision screening among pre-school children: a Malaysia perspective.

    Science.gov (United States)

    Omar, Rokiah; Hussin, Duratul Ain; Knight, Victor Feizal

    2012-03-01

    Compare the performance of Lea Symbols and Sheridan Gardiner charts against the standard test chart used to determine reduced VA during vision screening among pre-schoolers. Seven hundred seventy five pre-school participated where 389 subjects were boys and 386 subjects were girls were tested using the Lea Symbols chart and the Sheridan Gardiner chart. The mean age of the pre-school children was 5.3 +/- 0.7 years old. The Lea Symbols chart showed a higher sensitivity (97.5%) compared to the Sheridan Gardiner chart (57.1%). While the Sheridan Gardiner chart showed higher specificity (92.0%) compared to the Lea Symbols chart (45.0%). The Lea Symbols chart offers a better pick-up rate of visual impairment during a vision-screening program especially cases of amblyopia among pre-school children compared to the Sheridan Gardiner chart in Malaysia.

  9. Rheumatic fever in Ireland: the role of Dr Monica Lea Wilson (1889-1971).

    Science.gov (United States)

    Ward, O Conor

    2013-02-01

    In 1869 William Stokes pointed out that the severity of rheumatic fever in Dublin had declined over recent decades. Similar worldwide decline led to the closure of many internationally famous rheumatic fever centres. The discovery by Robert Collis that rheumatic fever was a sequel to haemolytic streptococcal infection and the subsequent discovery of penicillin accelerated the decline. St Gabriel's Hospital in Dublin opened in 1951 under the clinical direction of Dr Monica Lea Wilson. Contrary to contemporary medical opinion a regimen of very prolonged bed rest was enforced. From 1961 the family doctors became concerned at the adverse psychological effects of the unnecessarily prolonged hospital stay. Twenty-seven of the 56 inpatients were re-assessed. None of them showed any evidence of active rheumatic fever and their parents took them home. The hospital closed in 1968. Dr Lea Wilson distanced herself from mainstream medicine and she is best remembered for having presented an unrecognized Caravaggio painting to the Jesuit Order in recognition of their pastoral support at the time of the controversial assassination in 1920 of her husband Percival, an Inspector in the Royal Irish Constabulary.

  10. Anatomical Study of Somatic Embryogenesis in Glycine max (L. Merrill

    Directory of Open Access Journals (Sweden)

    Juliana Aparecida Fernando

    2002-09-01

    Full Text Available A comparative anatomical analysis of somatic embryogenesis in two soybean (Glycine max (L. Merrill genotypes was carried out. The somatic embryos were originated from cotyledonary explants obtained from immature zygotic embryos. The medium used for somatic embryogenesis induction was Murashige and Skoog, 1962, salts and Gamborg et al., 1968, vitamins (MSB supplemented with 0.8 mg.L-1 of 2,4-D for genotype PI 123439 and 40 mg.L-1 of 2,4-D for ‘Williams 82’. Globular structures, constituted by meristematic cells, originated from subepidermal cell divisions of the cotyledonary mesophyll. In PI 123439, the globular structures presented tracheary differentiation among meristematic cells and they could follow distinct morphogenetic process depending on their location along the explant. For ‘Williams 82’ it was observed globular structures along the cotyledonary explant surface. They gave rise to somatic embryos. These embryos showed different morphologies and they were classified based on their shape and number of cotyledons. The ability of these morphological types to convert to plantlets was discussed.Realizou-se uma análise anatômica comparativa da embriogênese somática em dois genótipos de soja (Glycine max (L. Merrill. Os embriões somáticos foram obtidos a partir de explantes cotiledonares excisados de embriões zigóticos imaturos do genótipo PI 123439, adaptado às condições tropicais, e ‘Williams 82’. O meio utilizado para indução da embriogênese somática constituiu-se de sais de Murashige e Skoog,1962, e vitaminas de Gamborg et al., 1968 (MSB suplementado com 0,8 mg.L-1 de 2,4-D (PI 123439 e 40 mg.L-1 (‘Williams 82’. Estruturas globulares originaram-se a partir de divisões celulares nas camadas subepidérmicas do mesofilo cotiledonar e foram constituídas por células meristemáticas. No genótipo PI 123439, as estruturas globulares apresentaram diferenciação traqueal entre as células meristemáticas e

  11. Effects of light quality on somatic embryogenesis in Araujia sericifera.

    Science.gov (United States)

    Torné, Josep M.; Moysset, Luisa; Santos, Mireya; Simón, Esther

    2001-03-01

    The effects of photoperiod, light quality and end-of-day (EOD) phytochrome photoconversion on somatic embryogenesis (SE) of Araujia sericifera petals have been studied. Petals from immature flowers were cultured under 8- and 16-h photoperiods using Gro-lux fluorescent lamps. The photon fluence rate was 90-100 µmol m-2 s-1 and the red (R):far-red (FR) ratio was 98. R, FR, R followed by FR (R-FR) and FR followed by R (FR-R) light treatments were applied for 3 weeks at the end of the photoperiods. In a set of experiments, DL-alpha-difluoromethylarginine (DFMA) or methylglyoxal bis(guanylhydrazone) (MGBG), both inhibitors of polyamine biosynthesis, were added to the culture medium in order to study the involvement of polyamine metabolism. The level of SE was the same in long (LD) and short (SD) days. Thus, the light effect was accomplished after 8 h. All EOD treatments that decreased the Pfr level inhibited SE when applied after SD, but not after LD. The FR-R treatment after LD caused an additional stimulatory effect on SE, even in the presence of polyamine inhibitors. DFMA inhibited SE in both SD and LD, but MGBG did not modify SE in either SD or LD. The R, FR and R-FR treatments did not alter the level of SE when applied after LD in the presence of DFMA or MGBG. However, these treatments decreased SE after SD when the medium contained polyamine inhibitors. Our results suggest that Gro-lux lamps, which produce an extremely high R:FR ratio, promote SE in A. sericifera and a timing response to phytochrome photoconversion during photoperiodic induction. Thus, our data corroborate the involvement of phytochromes and polyamines in SE in A. sericifera, which responded as a light-dominant long-day plant.

  12. Altered A-to-I RNA editing in human embryogenesis.

    Directory of Open Access Journals (Sweden)

    Ronit Shtrichman

    Full Text Available Post-transcriptional events play an important role in human development. The question arises as to whether Adenosine to Inosine RNA editing, catalyzed by the ADAR (Adenosine Deaminase acting on RNA enzymes, differs in human embryogenesis and in adulthood. We tested the editing of various target genes in coding (FLNA, BLCAP, CYFIP2 and non-coding sequences at their Alu elements (BRCA1, CARD11, RBBP9, MDM4, FNACC, as well as the transcriptional levels of the ADAR1 enzymes. This analysis was performed on five fetal and adult human tissues: brain, heart, liver, kidney, and spleen, as well as on human embryonic stem cells (hESCs, which represent the blastocyst stage in early human development. Our results show substantially greater editing activity for most adult tissue samples relative to fetal ones, in six of the eight genes tested. To test the effect of reduced A-to-I RNA editing activity in early human development we used human embryonic stem cells (hESCs as a model and tried to generate hESC clones that overexpress the ADAR1-p110 isoform. We were unable to achieve overexpression of ADAR1-p110 by either transfection or lentiviral infection, though we easily generated hESC clones that expressed the GFP transgene and overexpressed ADAR1-p110 in 293T cells and in primary human foreskin fibroblast (HFF cells. Moreover, in contrast to the expected overexpression of ADAR1-p110 protein following its introduction into hESCs, the expression levels of this protein decreased dramatically 24-48 hr post infection. Similar results were obtained when we tried to overexpress ADAR1-p110 in pluripotent embryonal carcinoma cells. This suggests that ADAR1 protein is substantially regulated in undifferentiated pluripotent hESCs. Overall, our data suggest that A-to-I RNA editing plays a critical role during early human development.

  13. Cloning and expression of embryogenesis-regulating genes in Araucaria angustifolia (Bert. O. Kuntze (Brazilian Pine

    Directory of Open Access Journals (Sweden)

    Paulo Sérgio Schlögl

    2012-01-01

    Full Text Available Angiosperm and gymnosperm plants evolved from a common ancestor about 300 million years ago. Apart from morphological and structural differences in embryogenesis and seed origin, a set of embryogenesis-regulating genes and the molecular mechanisms involved in embryo development seem to have been conserved alike in both taxa. Few studies have covered molecular aspects of embryogenesis in the Brazilian pine, the only economically important native conifer in Brazil. Thus eight embryogenesis-regulating genes, viz.,ARGONAUTE 1, CUP-SHAPED COTYLEDON 1, WUSCHEL-related WOX, S-LOCUS LECTIN PROTEIN KINASE, SCARECROW-like, VICILIN 7S, LEAFY COTYLEDON 1, and REVERSIBLE GLYCOSYLATED POLYPEPTIDE 1, were analyzed through semiquantitative RT-PCR during embryo development and germination. All the eight were found to be differentially expressed in the various developmental stages of zygotic embryos, seeds and seedling tissues. To our knowledge, this is the first report on embryogenesis-regulating gene expression in members of the Araucariaceae family, as well as in plants with recalcitrant seeds.

  14. Fractura supracondílea de codo en extensión en niños

    National Research Council Canada - National Science Library

    Vega Fernández, Enrique; Tórrez Hernández, Manuel E; Martínez Mesa, Javier

    2006-01-01

    La fractura supracondílea de húmero en niños es muy frecuente en la primera década de vida y necesita un tratamiento preciso, en aras de obtener un resultado satisfactorio en un hueso que está en formación...

  15. Feng shui väärtus on vaimujõus / Marika Tomberg ; komment. Lea Eermann, komment. Rein Weber

    Index Scriptorium Estoniae

    Tomberg, Marika

    1999-01-01

    Feng shui järgi korraldatud Lea Eermanni kodust Nõmmel. Feng shui- ja geobioloogia-alase nõustamisega tegelev arhitekt Rein Weber keskkonna mõjust tervisele, magamistoast. Enda ja maja kaitsmisest halbade mõjude eest. 10 illustratsiooni

  16. 34 CFR 611.47 - What are a scholarship recipient's reporting responsibilities upon the close of the LEA's...

    Science.gov (United States)

    2010-07-01

    ... 34 Education 3 2010-07-01 2010-07-01 false What are a scholarship recipient's reporting... EDUCATION TEACHER QUALITY ENHANCEMENT GRANTS PROGRAM Scholarships § 611.47 What are a scholarship recipient...'s academic year, a scholarship recipient whose LEA reports under § 611.46(a) that he or she is...

  17. Inimesed otsivad kirikust lähedust / Lea Altnurme ; küsitlenud Bianca Mikovitš

    Index Scriptorium Estoniae

    Altnurme, Lea

    2006-01-01

    Vestlus Tartu Ülikooli usuteaduskonna teaduri Lea Altnurmega, kes uuris oma doktoritöös, kuidas on muutunud eestlaste usulised suundumused eelmise sajandi teisel poolel ning leidis, et tänapäeva inimesed tahavad oma usus olla sõltumatud ja vabad

  18. Lea's Pies

    Science.gov (United States)

    1996-01-01

    The Technology Transfer Office at Stennis Space Center worked with a pie company owner to develop an inexpensive container that would protect pies and keep them in a near frozen condition for shipping in 48 hours. A NASA engineer made a thermal barrier envelope from a metalized mylar called 'space blanket material,' developed during the Apollo era. The envelope protects the pies from heat transfer. Pictured, a NASA engineer removes the temperature logger from a pecan pie shipped to him in a prototype envelope.

  19. Comparison of visual acuity results in preschool children with lea symbols and Bailey-Lovie E chart.

    Science.gov (United States)

    Sanker, Nijil; Dhirani, Sayyeda; Bhakat, Premjit

    2013-01-01

    To compare visual acuity with two visual acuity charts in preschool children. Visual acuity measurement with Lea symbols and Bailey-Lovie tumbling E chart was performed on children between 3 and 6 years of age. Visual acuity data from the two charts were analyzed with Bland-Altman plot to determine the limits of agreement. The Wilcoxon signed test was performed in children aged 3-4 years and in children aged 5-6 years separately to evaluate the influence of age. The inter-eye difference between the two charts were further analyzed with the paired t-test. A p value > 0.05 was considered statistically significant. A total of 47 children were enrolled for the study. The average logarithm of the Minimum Angle of Resolution (LogMAR) monocular visual acuity with Lea symbols (0.17 ± 0.13) was better than the Bailey-Lovie tumbling E chart (0.22 ± 0.14). The mean difference between Bailey-Lovie tumbling E chart and Lea symbol chart was 0.05 ± 0.12 in logMAR units. A second analysis eliminating outliers showed the same result but lower differences (n = 43, 0.05 ± 0.05 logMAR units). Visual acuity results between the two charts in children aged 3-4 years showed a significant difference (p = 0.000), but not for children aged 5-6 years (p = 0.059). Inter-eye differences between the two charts was not statistically significant (p = 0.77). Bailey-Lovie tumbling E chart is comparable to the Lea symbols chart in pre-school children. But preference should be given to Lea symbols for children aged 3-4 years as the symbols are more familiar than a directional test for this age group.

  20. Somatic Embryogenesis in Broad-Leaf Woody Plants: What We Can Learn from Proteomics.

    Science.gov (United States)

    Correia, Sandra I; Alves, Ana C; Veríssimo, Paula; Canhoto, Jorge M

    2016-01-01

    Proteomic approaches have been used to understand several regulatory aspects of plant development. Somatic embryogenesis is one of those developmental pathways that have beneficiated from the integration of proteomics data to the understanding of the molecular mechanisms that control embryogenic competence acquisition, somatic embryo development and conversion into viable plants. Nevertheless, most of the results obtained are based on the traditional model systems, very often not easily compared with the somatic embryogenesis systems of economical relevant woody species. The aim of this work is to summarize some of the applications of proteomics in the understanding of particular aspects of the somatic embryogenesis process in broad-leaf woody plants (model and non-model systems).

  1. Sex determination by PCR in Carica papaya L. plants obtained via somatic embryogenesis

    Directory of Open Access Journals (Sweden)

    Laisyn Posada Pérez

    2015-02-01

    Full Text Available For the molecular identification of the sex of papaya plants (Carica papaya L. have been used in field crop plants, however it has not been determined in vitro plants. The objective of this research was to identify hermaphrodite papaya plants var. `Maradol Red' obtained via somatic embryogenesis from immature zygotic embryos. For this, a multiplex PCR enables simultaneous amplification of two fragments (1300 and 800 bp for hermaphrodite and one fragment (1300 bp to female plants was used. DNA was extracted from leaves of in vitro plants by alkaline lysis with NaOH. PCR amplification of DNA extracted from leaf samples, produced fragments of the expected size. In 20 papaya plants lines regenerated through somatic embryogenesis it was achieved differentiate hermaphrodite plants and female plants. This is the first report on sex determination in plants obtained by somatic embryogenesis in the variety `Red Maradol'. Key words: hermaphrodite plants, multiple PCR, zygotic embryos

  2. Arabinogalactans and arabinogalactan-proteins induce embryogenesis in wheat (Triticum aestivum L.) microspore culture.

    Science.gov (United States)

    Letarte, Jocelyne; Simion, Ecaterina; Miner, Mai; Kasha, Ken J

    2006-01-01

    The objective of this study was to improve induction of embryogenesis in wheat microspore culture in order to obtain a high number of regenerable embryos. The arabinogalactan (AG) Larcoll and the arabinogalactan-protein (AGP) from gum arabic were tested on two spring genotypes to see if they could increase microspore viability and induce embryogenesis in the microspore culture. Adding Larcoll significantly decreased microspore mortality in both genotypes regardless of the presence or absence of ovaries in the culture. Similarly, gum arabic had a strong effect on the number of embryos produced and regenerated green plants. In fact, by using only gum arabic we were able to obtain green plants from wheat microspore cultures without the presence of ovaries. In addition to preventing a high mortality rate of the cells, our results show that the induction of embryogenesis in wheat microspore cultures is strongly affected by the use of both AG or AGP.

  3. Osteosíntesis y artrolisis en el tratamiento de las fracturas supra e intercondíleas graves del codo en el adulto

    OpenAIRE

    Fernández Sabaté, Alfons; Ferrer Escobar, Humberto; Alvarez Gonzalez, A.; Latorre Marti, J.; Orduña Serra, Modesto

    1982-01-01

    Los autores estudian la evoluciónen el tratamiento de las fracturas graves de la extremidad distal del hiimero hasta llegar a la técnica actual de la osteosíntesis estable. Presentan y analizan una serie de 21 fracturas condíleas, supracondíleas y supraintercondíleas tratadas quirtirgicamente mediante abordaje por vía transolecraniana, osteosíntesis con placas y tornillos y fijación del olécranon con compresión. La movilización precoz postoperatoria favorece la obtención de un buen arco móvil...

  4. POLYPHENOLS DISTRIBUTION AND RESERVE SUBSTANCES ANALYSIS IN CACAO SOMATIC EMBRYOGENESIS

    Directory of Open Access Journals (Sweden)

    Adriana María Gallego Rúa

    2016-04-01

    Full Text Available ABSTRACTIn order to understand the causes of lack of regeneration in cacao somatic embryos, two cacao varieties with different responses to regeneration potential were described based on their capacity to store different compounds. It is well known that seed reserves play a central role in the regenerative capability of somatic embryos; thus, we followed histochemical changes and reserve fluctuations of proteins, polysaccharides and polyphenols during somatic embryogenesis (SE in the two cacao varieties. The study showed that, in somatic embryos of the regenerating variety, polyphenols were localized mainly in the periphery of the embryo (epidermal cells and proteins were the main storage substance in the embryo expression medium, while the non-regenerating variety had a high presence of polysaccharides with random distribution of polyphenols at the end of the embryo induction step.Análisis de distribución de polifenoles y sustancias de reserva en embriogénesis somática de cacaoRESUMENDos variedades de cacao con diferentes respuestas a la regeneración fueron descritas en función de su capacidad para almacenar diferentes compuestos, con el fin de aproximarse al entendimiento de las causas de la falta de regeneración en embriones somáticos de cacao. Es bien sabido que las reservas de semillas desempeñan un papel central en la capacidad de regeneración de embriones somáticos; por tanto, se realizó un seguimiento de cambios histoquímicos y fluctuaciones de reserva de proteínas, polisacáridos y polifenoles durante la embriogénesis somática (SE en dos variedades de cacao. El estudio mostró que, en los embriones somáticos de la variedad regenerante, los polifenoles se localizaron principalmente en la periferia del embrión (células de la epidermis y las proteínas fueron el componente principal de almacenamiento en el medio de expresión de embriones, mientras que la variedad no regenerante tenía una alta presencia de polisac

  5. Zebrafish foxo3b negatively regulates canonical Wnt signaling to affect early embryogenesis.

    Directory of Open Access Journals (Sweden)

    Xun-wei Xie

    Full Text Available FOXO genes are involved in many aspects of development and vascular homeostasis by regulating cell apoptosis, proliferation, and the control of oxidative stress. In addition, FOXO genes have been showed to inhibit Wnt/β-catenin signaling by competing with T cell factor to bind to β-catenin. However, how important of this inhibition in vivo, particularly in embryogenesis is still unknown. To demonstrate the roles of FOXO genes in embryogenesis will help us to further understand their relevant physiological functions. Zebrafish foxo3b gene, an orthologue of mammalian FOXO3, was expressed maternally and distributed ubiquitously during early embryogenesis and later restricted to brain. After morpholino-mediated knockdown of foxo3b, the zebrafish embryos exhibited defects in axis and neuroectoderm formation, suggesting its critical role in early embryogenesis. The embryo-developmental marker gene staining at different stages, phenotype analysis and rescue assays revealed that foxo3b acted its role through negatively regulating both maternal and zygotic Wnt/β-catenin signaling. Moreover, we found that foxo3b could interact with zebrafish β-catenin1 and β-catenin2 to suppress their transactivation in vitro and in vivo, further confirming its role relevant to the inhibition of Wnt/β-catenin signaling. Taken together, we revealed that foxo3b played a very important role in embryogenesis and negatively regulated maternal and zygotic Wnt/β-catenin signaling by directly interacting with both β-catenin1 and β-catenin2. Our studies provide an in vivo model for illustrating function of FOXO transcription factors in embryogenesis.

  6. Somatic embryogenesis and plant regeneration in embryo cultures of Euterpe edulis mart. (palmae).

    Science.gov (United States)

    Guerra, M P; Handro, W

    1988-12-01

    The induction of somatic embryogenesis in embryo cultures of Euterpe edulis is described. The basal medium was composed of LS salts and Morel & Wetmore vitamins. Activated charcoal was added to prevent explant oxidation. 2,4-D higher than 50 mg/l was necessary for inducing embryogenesis which occurs 45-180 days after the start of cultures. Embryos arise directly from surface proliferating tissues on the matrix structure , without callus formation. The transfer of tissues with embryo clusters to medium with NAA plus 2iP, or without growth regulators, induces embryo development into plantlets.

  7. Induction by thidiazuron of somatic embryogenesis in intact seedlings of peanut.

    Science.gov (United States)

    Saxena, P K; Malik, K A; Gill, R

    1992-06-01

    In planta differentiation of somatic embryos was induced in seedlings of peanut (Arachis hypogaea L.) obtained from mature seeds germinated on a medium supplemented with thidiazuron (TDZ: N-phenyl-N(1)- (1,2,3 thiadiazol-yl)urea). At optimum levels of TDZ (10 μM), all germinating seeds produced embryogenic seedlings, and somatic embryos developed in the apical region and on the surface of cotyledons and hypocotyls. These somatic embryos matured, germinated, and formed shoots which eventually developed into whole plants. Thidiazuron-induced direct embryogenesis from morphologically intact seedlings may provide an excellent experimental system for investigating somatic embryogenesis and the morphoregulatory role of TDZ.

  8. Estimating abundance: Chapter 27

    Science.gov (United States)

    Royle, J. Andrew

    2016-01-01

    This chapter provides a non-technical overview of ‘closed population capture–recapture’ models, a class of well-established models that are widely applied in ecology, such as removal sampling, covariate models, and distance sampling. These methods are regularly adopted for studies of reptiles, in order to estimate abundance from counts of marked individuals while accounting for imperfect detection. Thus, the chapter describes some classic closed population models for estimating abundance, with considerations for some recent extensions that provide a spatial context for the estimation of abundance, and therefore density. Finally, the chapter suggests some software for use in data analysis, such as the Windows-based program MARK, and provides an example of estimating abundance and density of reptiles using an artificial cover object survey of Slow Worms (Anguis fragilis).

  9. Stable Agrobacterium tumefaciens-mediated transformation related ...

    African Journals Online (AJOL)

    laddiya

    2012-07-17

    Jul 17, 2012 ... (2009) transformed the wheat late embryogenesis abundant (LEA) gene using ..... Effect of infection time on callus growth rate and transformation frequency according to various infection times (60, 30, 25, 20, 15 and 10 min) using 0.8 ..... This work was supported by Nutraceutical Bio Brain. Korea 21 Project ...

  10. Arabidopsis CDS blastp result: AK241134 [KOME

    Lifescience Database Archive (English)

    Full Text Available ontains Pfam PF05512: AWPM-19-like family; similar to late embryogenesis abundant protein, (Lea) with hydrophobic domain, high pI val...ue (11.6); 15kD protein; putative (GI:310570) {Glycine max} 3e-23 ...

  11. Genome-wide data (ChIP-seq) enabled identification of cell wall-related and aquaporin genes as targets of tomato ASR1, a drought stress-responsive transcription factor

    Science.gov (United States)

    Here we report efforts to take advantage of previous knowledge on well characterized proteins that extensively accumulate in dehydration, for example those belonging to the LEA (late embryogenesis abundant) superfamily. ASR proteins, a subgroup exclusive to the plant kingdom (albeit absent in Arabid...

  12. Book review: Working with academic literacies: case studies towards transformative practice. Lillis, T., Harrington, K., Lea, M.R. & Mitchell, S. eds.

    Directory of Open Access Journals (Sweden)

    Mastin Prinsloo

    2016-12-01

    Full Text Available This is a review of Lillis, T., Harrington, K., Lea, M.R. & Mitchell, S. (eds. 2015. Working with academic literacies: case studies towards transformative practice. Fort Collins, Colorado: WAC Clearinghouse.

  13. OXYGEN ABUNDANCES IN CEPHEIDS

    Energy Technology Data Exchange (ETDEWEB)

    Luck, R. E.; Andrievsky, S. M. [Department of Astronomy, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106-7215 (United States); Korotin, S. N.; Kovtyukh, V. V., E-mail: luck@fafnir.astr.cwru.edu, E-mail: serkor@skyline.od.ua, E-mail: val@deneb1.odessa.ua, E-mail: scan@deneb1.odessa.ua [Department of Astronomy and Astronomical Observatory, Odessa National University, Isaac Newton Institute of Chile, Odessa Branch, Shevchenko Park, 65014 Odessa (Ukraine)

    2013-07-01

    Oxygen abundances in later-type stars, and intermediate-mass stars in particular, are usually determined from the [O I] line at 630.0 nm, and to a lesser extent, from the O I triplet at 615.7 nm. The near-IR triplets at 777.4 nm and 844.6 nm are strong in these stars and generally do not suffer from severe blending with other species. However, these latter two triplets suffer from strong non-local thermodynamic equilibrium (NLTE) effects and thus see limited use in abundance analyses. In this paper, we derive oxygen abundances in a large sample of Cepheids using the near-IR triplets from an NLTE analysis, and compare those abundances to values derived from a local thermodynamic equilibrium (LTE) analysis of the [O I] 630.0 nm line and the O I 615.7 nm triplet as well as LTE abundances for the 777.4 nm triplet. All of these lines suffer from line strength problems making them sensitive to either measurement complications (weak lines) or to line saturation difficulties (strong lines). Upon this realization, the LTE results for the [O I] lines and the O I 615.7 nm triplet are in adequate agreement with the abundance from the NLTE analysis of the near-IR triplets.

  14. Somatic embryogenesis and plant regeneration of northern red oak (Quercus rubra L.)

    Science.gov (United States)

    G. Vengadesan; Paula M. Pijut

    2009-01-01

    A somatic embryogenesis protocol for plant regeneration of northern red oak (Quercus rubra) was established from immature cotyledon explants. Embryogenic callus cultures were induced on Murashige and Skoog medium (MS) containing 3% sucrose, 0.24% Phytagel™, and various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) after 4 weeks of...

  15. Ontogenic appearance of MHC class I (B-F) antigens during chicken embryogenesis

    DEFF Research Database (Denmark)

    Dunon, D; Salomonsen, J; Skjødt, K

    1990-01-01

    Expression of chicken MHC class I (B-F) antigens during ontogeny was determined by binding of anticlass I antibody and appearance of B-F transcripts by Northern blotting in chicken organs during embryogenesis until 2 weeks after hatching. MHC class I transcripts first become detectable in day 6...

  16. Embryogenesis from isolated microspores of tulip : towards developing F1 hybrid varieties

    NARCIS (Netherlands)

    Custers, J.B.M.; Ennik, E.; Eikelboom, W.; Dons, J.J.M.; Lookeren Campagne, van M.M.

    1997-01-01

    This report describes advances in the technique of embryogenesis from tulip microspores in culture. High temperature pretreatment (32°C) of bulbs, which contain fully developed inflorescences, had a positive effect on the production of microspore embryos. The pretreatment shortened the time for

  17. High-frequency plant regeneration through cyclic secondary somatic embryogenesis in black pepper (Piper nigrum L.).

    Science.gov (United States)

    Nair, R Ramakrishnan; Dutta Gupta, S

    2006-01-01

    A high-frequency plantlet regeneration protocol was developed for black pepper (Piper nigrum L.) through cyclic secondary somatic embryogenesis. Secondary embryos formed from the radicular end of the primary somatic embryos which were originally derived from micropylar tissues of germinating seeds on growth regulator-free SH medium in the absence of light. The process of secondary embryogenesis continued in a cyclic manner from the root pole of newly formed embryos resulting in clumps of somatic embryos. Strength of the medium and sucrose concentration influenced the process of secondary embryogenesis and fresh weight of somatic embryo clumps. Full-strength SH medium supplemented with 1.5% sucrose produced significantly higher fresh weight and numbers of secondary somatic embryos while 3.0 and 4.5% sucrose in the medium favored further development of proliferated embryos into plantlets. Ontogeny of secondary embryos was established by histological analysis. Secondary embryogenic potential was influenced by the developmental stage of the explanted somatic embryo and stages up to "torpedo" were more suitable. A single-flask system was standardized for proliferation, maturation, germination and conversion of secondary somatic embryos in suspension cultures. The system of cyclic secondary somatic embryogenesis in black pepper described here represents a permanent source of embryogenic material that can be used for genetic manipulations of this crop species.

  18. Cloning of B-1,3-glucanases expressed during Cichorium somatic embryogenesis

    NARCIS (Netherlands)

    Helleboid, S.; Chapman, A.; Hendriks, T.; Inze, D.; Vasseur, J.; Hilbert, J.L.

    2000-01-01

    Three different β-1,3-glucanase cDNA fragments, CG1, CG2 and CG3, were obtained by RT-PCR from RNA isolated from Cichorium hybrid `474' leaf fragments cultured for 11 days under somatic embryogenesis-inducing conditions. When expressed in Escherichia coli the proteins encoded by the three cDNAs were

  19. Glycogen Synthase Kinase-3 is involved in glycogen metabolism control and embryogenesis of Rhodnius prolixus.

    Science.gov (United States)

    Mury, Flávia B; Lugon, Magda D; DA Fonseca, Rodrigo Nunes; Silva, Jose R; Berni, Mateus; Araujo, Helena M; Fontenele, Marcio Ribeiro; Abreu, Leonardo Araujo DE; Dansa, Marílvia; Braz, Glória; Masuda, Hatisaburo; Logullo, Carlos

    2016-10-01

    Rhodnius prolixus is a blood-feeding insect that transmits Trypanosoma cruzi and Trypanosoma rangeli to vertebrate hosts. Rhodnius prolixus is also a classical model in insect physiology, and the recent availability of R. prolixus genome has opened new avenues on triatomine research. Glycogen synthase kinase 3 (GSK-3) is classically described as a key enzyme involved in glycogen metabolism, also acting as a downstream component of the Wnt pathway during embryogenesis. GSK-3 has been shown to be highly conserved among several organisms, mainly in the catalytic domain region. Meanwhile, the role of GSK-3 during R. prolixus embryogenesis or glycogen metabolism has not been investigated. Here we show that chemical inhibition of GSK-3 by alsterpaullone, an ATP-competitive inhibitor of GSK3, does not affect adult survival rate, though it alters oviposition and egg hatching. Specific GSK-3 gene silencing by dsRNA injection in adult females showed a similar phenotype. Furthermore, bright field and 4'-6-diamidino-2-phenylindole (DAPI) staining analysis revealed that ovaries and eggs from dsGSK-3 injected females exhibited specific morphological defects. We also demonstrate that glycogen content was inversely related to activity and transcription levels of GSK-3 during embryogenesis. Lastly, after GSK-3 knockdown, we observed changes in the expression of the Wingless (Wnt) downstream target β-catenin as well as in members of other pathways such as the receptor Notch. Taken together, our results show that GSK-3 regulation is essential for R. prolixus oogenesis and embryogenesis.

  20. The Pesticide Malathion Disrupts "Xenopus" and Zebrafish Embryogenesis: An Investigative Laboratory Exercise in Developmental Toxicology

    Science.gov (United States)

    Chemotti, Diana C.; Davis, Sarah N.; Cook, Leslie W.; Willoughby, Ian R.; Paradise, Christopher J.; Lom, Barbara

    2006-01-01

    Malathion is an organophosphorus insecticide, which is often sprayed to control mosquitoes. When applied to aquatic habitats, malathion can also influence the embryogenesis of non-target organisms such as frogs and fish. We modified the frog embryo teratogen assay in "Xenopus" (FETAX), a standard toxicological assay, into an investigative…

  1. Somatic embryogenesis as a tool to capture genetic gain from tree ...

    African Journals Online (AJOL)

    By virtue of its amenability to cryopreservation, it offers the opportunity to overcome the constraints associated with conventional vegetative propagation ... these restrictions are discussed together with the associated long-term potential of somatic embryogenesis, including genetic engineering and synthetic seed production.

  2. Congenital malformations in offspring of diabetic women treated with oral hypoglycaemic agents during embryogenesis

    DEFF Research Database (Denmark)

    Hellmuth, E; Damm, P; Mølsted-Pedersen, L

    1994-01-01

    in the offspring, the previously reported association was not confirmed. Thus we find no obvious indication for therapeutic abortions in patients who have accidentally been treated with oral hypoglycaemic agents during embryogenesis. On the contrary it seems reasonable to reassure these women with respect...

  3. The Arabidopsis kinase-associated protein phosphatase controls internalization of the somatic embryogenesis receptor kinase 1

    NARCIS (Netherlands)

    Shah, K.; Russinova, E.; Gadella, T.W.J.; Willemse, J.; Vries, de S.C.

    2002-01-01

    The AtSERK1 protein is a plasma membrane-located LRR receptor-like serine threonine kinase that is transiently expressed during plant embryogenesis. Our results show that AtSERK1 interacts with the kinase-associated protein phosphatase (KAPP) in vitro. The kinase interaction (KI) domain of KAPP does

  4. Imaging of polarity during zygotic and somatic embryogenesis of carrot (Daucus carota L.)

    NARCIS (Netherlands)

    Timmers, A.C.J.

    1993-01-01

    In this thesis a study of the regulation of coordinated growth and the development of polarity during embryogenesis of carrot, Daucus carota L., is described. To this end, several microscopical techniques were used, such as light microscopy, fluorescence microscopy,

  5. Microspore embryogenesis in barley: anther pre-treatment stimulates plant defence gene expression

    NARCIS (Netherlands)

    Jacquard, C.; Mazeyrat-Gourbeyre, F.; Devaux, P.; Boutilier, K.A.

    2009-01-01

    Microspore embryogenesis (ME) is a process in which the gametophytic pollen programme of the microspore is reorientated towards a new embryo sporophytic programme. This process requires a stress treatment, usually performed in the anther or isolated microspores for several days. Despite the

  6. Towards industrial production of tree varieties through somatic embryogenesis and other vegetative propagation technologies

    DEFF Research Database (Denmark)

    Find, Jens Iver

    2016-01-01

    The main focus of the research on somatic embryogenesis in nordmanns fir has until recently been on improving the basic protocols in each step of the process. However, with recent developments, one single set of methods has shown to be effective for production of plants from more than 500 differe...

  7. The Arabidopsis SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE1 protein complex includes BRASSINOSTEROID-INSENSITIVE1.

    NARCIS (Netherlands)

    Karlova, R.B.; Boeren, J.A.; Russinova, E.T.; Aker, J.C.M.; Vervoort, J.J.M.; Vries, de S.C.

    2006-01-01

    Arabidopsis thaliana SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE1 (SERK1) is a leucine-rich repeat receptor-like kinase (LRR-RLK) involved in the acquisition of embryogenic competence and in male sporogenesis. To determine the composition of the SERK1 signaling complex in vivo, we generated plants

  8. Polyamine and ethylene biosynthesis in relation to somatic embryogenesis in carrot (Daucus carota L.) cell cultures

    Science.gov (United States)

    Subhash C. Minocha; Cheryl A. Robie; Akhtar J. Khan; Nancy S. Papa; Andrew I. Samuelsen; Rakesh. Minocha

    1990-01-01

    Carrot cell cultures provide a model experimental system for the analysis of biochemical and molecular events associated with morphogenesis in plants (3, 4, 5, 14). Among the biochemical changes accompanying somatic embryogenesis in this tissue is an increased biosynthesis ofpolyamines (1, 2, 7, 10, 11, 13). A variety of inhibitors of polyamine biosynthetic enzymes...

  9. Germ band retraction as a landmark in glucose metabolism during Aedes aegypti embryogenesis

    Directory of Open Access Journals (Sweden)

    Logullo Carlos

    2010-02-01

    Full Text Available Abstract Background The mosquito A. aegypti is vector of dengue and other viruses. New methods of vector control are needed and can be achieved by a better understanding of the life cycle of this insect. Embryogenesis is a part of A. aegypty life cycle that is poorly understood. In insects in general and in mosquitoes in particular energetic metabolism is well studied during oogenesis, when the oocyte exhibits fast growth, accumulating carbohydrates, lipids and proteins that will meet the regulatory and metabolic needs of the developing embryo. On the other hand, events related with energetic metabolism during A. aegypti embryogenesis are unknown. Results Glucose metabolism was investigated throughout Aedes aegypti (Diptera embryonic development. Both cellular blastoderm formation (CBf, 5 h after egg laying - HAE and germ band retraction (GBr, 24 HAE may be considered landmarks regarding glucose 6-phosphate (G6P destination. We observed high levels of glucose 6-phosphate dehydrogenase (G6PDH activity at the very beginning of embryogenesis, which nevertheless decreased up to 5 HAE. This activity is correlated with the need for nucleotide precursors generated by the pentose phosphate pathway (PPP, of which G6PDH is the key enzyme. We suggest the synchronism of egg metabolism with carbohydrate distribution based on the decreasing levels of phosphoenolpyruvate carboxykinase (PEPCK activity and on the elevation observed in protein content up to 24 HAE. Concomitantly, increasing levels of hexokinase (HK and pyruvate kinase (PK activity were observed, and PEPCK reached a peak around 48 HAE. Glycogen synthase kinase (GSK3 activity was also monitored and shown to be inversely correlated with glycogen distribution during embryogenesis. Conclusions The results herein support the hypothesis that glucose metabolic fate changes according to developmental embryonic stages. Germ band retraction is a moment that was characterized as a landmark in glucose

  10. Loss of CMD2-mediated resistance to cassava mosaic disease in plants regenerated through somatic embryogenesis.

    Science.gov (United States)

    Beyene, Getu; Chauhan, Raj Deepika; Wagaba, Henry; Moll, Theodore; Alicai, Titus; Miano, Douglas; Carrington, James C; Taylor, Nigel J

    2016-09-01

    Cassava mosaic disease (CMD) and cassava brown streak disease (CBSD) are the two most important viral diseases affecting cassava production in Africa. Three sources of resistance are employed to combat CMD: polygenic recessive resistance, termed CMD1, the dominant monogenic type, named CMD2, and the recently characterized CMD3. The farmer-preferred cultivar TME 204 carries inherent resistance to CMD mediated by CMD2, but is highly susceptible to CBSD. Selected plants of TME 204 produced for RNA interference (RNAi)-mediated resistance to CBSD were regenerated via somatic embryogenesis and tested in confined field trials in East Africa. Although micropropagated, wild-type TME 204 plants exhibited the expected levels of resistance, all plants regenerated via somatic embryogenesis were found to be highly susceptible to CMD. Glasshouse studies using infectious clones of East African cassava mosaic virus conclusively demonstrated that the process of somatic embryogenesis used to regenerate cassava caused the resulting plants to become susceptible to CMD. This phenomenon could be replicated in the two additional CMD2-type varieties TME 3 and TME 7, but the CMD1-type cultivar TMS 30572 and the CMD3-type cultivar TMS 98/0505 maintained resistance to CMD after passage through somatic embryogenesis. Data are presented to define the specific tissue culture step at which the loss of CMD resistance occurs and to show that the loss of CMD2-mediated resistance is maintained across vegetative generations. These findings reveal new aspects of the widely used technique of somatic embryogenesis, and the stability of field-level resistance in CMD2-type cultivars presently grown by farmers in East Africa, where CMD pressure is high. © 2015 The Authors Molecular Plant Pathology Published by British Society for Plant Pathology and John Wiley & Sons Ltd.

  11. Acetylsalicylic acid enhances and synchronizes thidiazuron-induced somatic embryogenesis in geranium (Pelargonium x hortorum Bailey) tissue cultures.

    Science.gov (United States)

    Hutchinson, M J; Saxena, P K

    1996-03-01

    Thidiazuron (TDZ) effectively induced somatic embryogenesis in cultured hypocotyl explants of geranium (Pelargonium x hortorum Bailey) during only a 3-day period of induction. The presence of acetylsalicylic acid (ASA) during this period caused a two-fold increase in the number of somatic embryos and enhanced synchronization of embryo development compared to the TDZ treatment alone. Salicylic acid was ineffective in modulating similar embryogenic responses as ASA. The ASA-induced enhancement and synchronization of somatic embryogenesis could possibly be used as an experimental system to study the interplay of growth regulators in somatic embryogenesis.

  12. Salt deposits in Los Medanos area, Eddy and Lea counties, New Mexico

    Science.gov (United States)

    Jones, C.L.; with sections on Ground water hydrology, Cooley; and Surficial Geology, Bachman

    1973-01-01

    The salt deposits of Los Medanos area, in Eddy and Lea Counties, southeastern New Mexico, are being considered for possible use as a receptacle for radioactive wastes in a pilot-plant repository. The salt deposits of the area. are in three evaporite formations: the Castile, Salado, and Rustler Formations, in ascending order. The three formations are dominantly anhydrite and rock salt, but some gypsum, potassium ores, carbonate rock, and fine-grained clastic rocks are present. They have combined thicknesses of slightly more than 4,000 feet, of which roughly one-half belongs to the Salado. Both the Castile and the Rustler are-richer in anhydrite-and poorer in rock salt-than the Salado, and they provide this salt-rich formation with considerable Protection from any fluids which might be present in underlying or overlying rocks. The Salado Formation contains many thick seams of rock salt at moderate depths below the surface. The rock salt has a substantial cover of well-consolidated rocks, and it is very little deformed structurally. Certain geological details essential for Waste-storage purposes are unknown or poorly known, and additional study involving drilling is required to identify seams of rock salt suitable for storage purposes and to establish critical details of their chemistry, stratigraphy, and structure.

  13. Magellanic Cloud Cepheids - Abundances

    Science.gov (United States)

    Luck, R. Earle; Moffett, Thomas J.; Barnes, Thomas G., III; Gieren, Wolfgang P.

    1998-02-01

    We have determined parameters and abundances for three SMC and nine LMC Cepheids that have not been previously studied spectroscopically for metallicity, as well as three SMC and one LMC Cepheids observed previously by Luck and Lambert. This work nearly doubles the number of Cepheids with spectroscopic metallicities in the SMC and triples the number in the LMC. For Galactic reference stars we have determined the abundances of 11 supergiants of spectral type F0 to K1. These abundances are used herein to examine the dispersion in cosmic (Fe/H) values found in LMC Cepheids by Luck and Lambert. The Magellanic Cloud Cepheids tend to be longer-period Cepheids, and thus have physical gravities that are 1.0 dex and lower. Spectroscopic gravities are even lower. Fortunately, the (Fe/H) content as determined from Fe I is not strongly dependent on gravity and is thus well determined. We combine our abundances with other published high-resolution spectroscopic results, deriving a mean (Fe/H) for the SMC of -0.68. The total range seen in the (Fe/H) ratios derived herein is -0.84 to -0.65. This result is in agreement with previous studies showing a small intrinsic range in the metallicities in the SMC. For the LMC, from the Cepheids studied here we obtain -0.30, with a range of -0.55 to -0.19. Combining all available data, we obtain a mean (Fe/H) of -0.34. From this analysis, there is little evidence for a significant metallicity dispersion in the LMC.

  14. Effect of the combination of NAA, kinetin and sucrose on the induction of somatic embryogenesis in lulo (Solanum quitoense Lam.)

    OpenAIRE

    Hernando Criollo; Margarita Perea; Mariano Toribio; Johanna Muñoz

    2014-01-01

    Lulo is a species of great importance to the fruticulture of Colombia, but has significant phytosanitary problems that require an aggressive breeding program oriented toward the production of genotypes with tolerance to phytopathogens. These programs need to establish highly efficient mass plant propagation protocols, such as somatic embryogenesis. This study focused on research on the somatic embryogenesis of lulo using kinetin, naphthalene acetic acid-NAA (Plant Growth Regulators, PGRs), an...

  15. Somatic embryogenesis in Nicotiana tabacum L.: induction by thidiazuron of direct embryo differentiation from cultured leaf discs.

    Science.gov (United States)

    Gill, R; Saxena, P K

    1993-01-01

    Induction of somatic embryogenesis by different growth regulators was examined in leaf disc cultures of Nicotiana tabacum L. Direct differentiation of somatic embryos occurred on media supplemented with naphthaleneacetic acid (NAA) and N(6)-benzylaminopurine (BAP). Thidiazuron (N-phenyl-N'-1, 2,3,-thiadiazol-5-ylurea; TDZ) not only substituted for the most effective NAA-BAP combination but also induced a higher frequency of somatic embryogenesis. Regenerated somatic embryos were capable of developing into plants.

  16. Synchronization of Somatic Embryogenesis in Date Palm Suspension Culture Using Abscisic Acid.

    Science.gov (United States)

    Alwael, Hussain A; Naik, Poornananda M; Al-Khayri, Jameel M

    2017-01-01

    Somatic embryogenesis is considered the most effective method for commercial propagation of date palm. However, the limitation of obtaining synchronized development of somatic embryos remains an impediment. The synchronization of somatic embryo development is ideal for the applications to produce artificial seeds. Abscisic acid (ABA) is associated with stress response and influences in vitro growth and development. This chapter describes an effective method to achieve synchronized development of somatic embryos in date palm cell suspension culture. Among the ABA concentrations tested (0, 1, 10, 50, 100 μM), the best synchronized growth was obtained in response to 50-100 μM. Here we provide a comprehensive protocol for in vitro plant regeneration of date palm starting with shoot-tip explant, callus initiation and growth, cell suspension establishment, embryogenesis synchronization with ABA treatment, somatic embryo germination, and rooting as well as acclimatized plantlet establishment.

  17. Conservation of proteo-lipid nuclear membrane fusion machinery during early embryogenesis.

    Science.gov (United States)

    Byrne, Richard D; Veeriah, Selvaraju; Applebee, Christopher J; Larijani, Banafshé

    2014-01-01

    The fusogenic lipid diacylglycerol is essential for remodeling gamete and zygote nuclear envelopes (NE) during early embryogenesis. It is unclear whether upstream signaling molecules are likewise conserved. Here we demonstrate PLCγ and its activator SFK1, which co-operate during male pronuclear envelope formation, also promote the subsequent male and female pronuclear fusion. PLCγ and SFK1 interact directly at the fusion site leading to PLCγ activation. This is accompanied by a spatially restricted reduction of PtdIns(4,5)P2. Consequently, pronuclear fusion is blocked by PLCγ or SFK1 inhibition. These findings identify new regulators of events in the early embryo and suggest a conserved "toolkit" of fusion machinery drives successive NE fusion events during embryogenesis.

  18. Study of elemental variations during somatic embryogenesis in sugarcane using photon induced X-ray probe

    Science.gov (United States)

    Desai, N. S.; Joseph, D.; Suprasanna, P.; Bapat, V. A.

    2006-11-01

    Energy-dispersive X-ray fluorescence technique (EDXRF) has been extensively used to characterize trace element profiles during plant growth under stress and development. In this study, elemental accumulation was analyzed using EDXRF technique during somatic embryogenesis, from de-differentiated callus (S1) to proembryogenic callus (S2), embryogenic callus with developing embryos (S3) and embryo converted plantlets (S4, S5). There was much variation in Mg, K, Ca, Mn, Fe, Cu and Zn. Higher Mg (4.6%) K (1068 ppm) and Fe accumulation was observed in proembryogenic callus (S2) stage compared to other stages suggesting specific elemental accumulation in embryogenic callus. The results suggest that the information on the accumulation of elements during developmental stages in vitro could be useful for formulating a media for induction of high frequency of embryogenesis in sugarcane.

  19. Study of elemental variations during somatic embryogenesis in sugarcane using photon induced X-ray probe

    Energy Technology Data Exchange (ETDEWEB)

    Desai, N.S. [Dr. DY Patil Institute of Biotechnology and Bioinformatics, Sector 15, CBD Belapur, Navi Mumbai 400 614 (India); Joseph, D. [Nuclear Physics Division, Bhabha Atomic Research Center, Mumbai, Trombay 400 085 (India); Suprasanna, P. [Plant Cell Culture Technology Section, Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Center, Mumbai, Trombay 400 085 (India)]. E-mail: prasanna@barc.gov.in; Bapat, V.A. [Plant Cell Culture Technology Section, Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Center, Mumbai, Trombay 400 085 (India)

    2006-11-15

    Energy-dispersive X-ray fluorescence technique (EDXRF) has been extensively used to characterize trace element profiles during plant growth under stress and development. In this study, elemental accumulation was analyzed using EDXRF technique during somatic embryogenesis, from de-differentiated callus (S1) to proembryogenic callus (S2), embryogenic callus with developing embryos (S3) and embryo converted plantlets (S4, S5). There was much variation in Mg, K, Ca, Mn, Fe, Cu and Zn. Higher Mg (4.6%) K (1068 ppm) and Fe accumulation was observed in proembryogenic callus (S2) stage compared to other stages suggesting specific elemental accumulation in embryogenic callus. The results suggest that the information on the accumulation of elements during developmental stages in vitro could be useful for formulating a media for induction of high frequency of embryogenesis in sugarcane.

  20. Gene expression profiling of Atlantic cod (Gadus morhua) embryogenesis using microarray.

    Science.gov (United States)

    Drivenes, Øyvind; Taranger, Geir Lasse; Edvardsen, Rolf B

    2012-04-01

    Atlantic cod (Gadus morhua) is a fish species of high importance, as a key species in a range of Northern ecosystems, in fisheries, and as an emerging species in aquaculture. So far, little is known about the transcriptional activity during early developmental stages of Atlantic cod. Hence, we decided to use a cDNA microarray covering 7,000 genes to analyze the temporal activity of the transcriptome during cod embryogenesis. Twelve different embryonic time points were selected, covering major developmental stages and processes such as maternally derived mRNA, blastula, gastrula, segmentation, hatching, and first-feeding larval stage. The microarray analysis revealed a highly dynamic transcriptional profile, showing for the first time the differential expression of thousands of known and unknown genes during Atlantic cod embryogenesis. These initial findings will serve as an important baseline for future in-depth studies of candidate genes involved in development, reproductive control, disease resistance, growth, nutrient digestion, and metabolism.

  1. The δ-cyclin expression at early stages of embryogenesis of Brassica rapa L. under clinorotation

    Science.gov (United States)

    Artemenko, O. A.; Popova, A. F.

    We present some results of comparison studying of Brassica embryo development and the δ-cyclin genes expression under slow horizontal clinorotation and in the laboratory control. Some backlog of the δ1-cyclin genes expression at early stages of embryogenesis under clinorotation was revealed in comparison with the laboratory control. The similar level of the δ3-cyclin expression at all stages of embryo formation (from one to nine days) in both variants is shown. Some delays in the rate of Brassica rapa embryo development under clinorotation in comparison with the laboratory control can be a result of decrease of a level and some backlog of the δ1-cyclin expression at early stages of embryogenesis.

  2. Enhanced somatic embryogenesis in Theobroma cacao using the homologous BABY BOOM transcription factor.

    Science.gov (United States)

    Florez, Sergio L; Erwin, Rachel L; Maximova, Siela N; Guiltinan, Mark J; Curtis, Wayne R

    2015-05-16

    Theobroma cacao, the chocolate tree, is an important economic crop in East Africa, South East Asia, and South and Central America. Propagation of elite varieties has been achieved through somatic embryogenesis (SE) but low efficiencies and genotype dependence still presents a significant limitation for its propagation at commercial scales. Manipulation of transcription factors has been used to enhance the formation of SEs in several other plant species. This work describes the use of the transcription factor Baby Boom (BBM) to promote the transition of somatic cacao cells from the vegetative to embryonic state. An ortholog of the Arabidopsis thaliana BBM gene (AtBBM) was characterized in T. cacao (TcBBM). TcBBM expression was observed throughout embryo development and was expressed at higher levels during SE as compared to zygotic embryogenesis (ZE). TcBBM overexpression in A. thaliana and T. cacao led to phenotypes associated with SE that did not require exogenous hormones. While transient ectopic expression of TcBBM provided only moderate enhancements in embryogenic potential, constitutive overexpression dramatically increased SE proliferation but also appeared to inhibit subsequent development. Our work provides validation that TcBBM is an ortholog to AtBBM and has a specific role in both somatic and zygotic embryogenesis. Furthermore, our studies revealed that TcBBM transcript levels could serve as a biomarker for embryogenesis in cacao tissue. Results from transient expression of TcBBM provide confirmation that transcription factors can be used to enhance SE without compromising plant development and avoiding GMO plant production. This strategy could compliment a hormone-based method of reprogramming somatic cells and lead to more precise manipulation of SE at the regulatory level of transcription factors. The technology would benefit the propagation of elite varieties with low regeneration potential as well as the production of transgenic plants, which

  3. LONO1 encoding a nucleoporin is required for embryogenesis and seed viability in Arabidopsis.

    Science.gov (United States)

    Braud, Christopher; Zheng, Wenguang; Xiao, Wenyan

    2012-10-01

    Early embryogenesis in Arabidopsis (Arabidopsis thaliana) is distinguished by a predictable pattern of cell divisions and is a good system for investigating mechanisms of developmental pattern formation. Here, we identified a gene called LONO1 (LNO1) in Arabidopsis in which mutations can abolish the first asymmetrical cell division of the zygote, alter planes and number of cell divisions in early embryogenesis, and eventually arrest embryo development. LNO1 is highly expressed in anthers of flower buds, stigma papilla of open flowers, and embryo and endosperm during early embryogenesis, which is correlated with its functions in reproductive development. The homozygous lno1-1 seed is not viable. LNO1, a homolog of the nucleoporin NUP214 in human (Homo sapiens) and Nup159 in yeast (Saccharomyces cerevisiae), encodes a nucleoporin protein containing phenylalanine-glycine repeats in Arabidopsis. We demonstrate that LNO1 can functionally complement the defect in the yeast temperature-sensitive nucleoporin mutant nup159. We show that LNO1 specifically interacts with the Arabidopsis DEAD-box helicase/ATPase LOS4 in the yeast two-hybrid assay. Furthermore, mutations in AtGLE1, an Arabidopsis homolog of the yeast Gle1 involved in the same poly(A) mRNA export pathway as Nup159, also result in seed abortion. Our results suggest that LNO1 is a component of the nuclear pore complex required for mature mRNA export from the nucleus to the cytoplasm, which makes LNO1 essential for embryogenesis and seed viability in Arabidopsis.

  4. Differential proteome analysis during early somatic embryogenesis in Musa spp. AAA cv. Grand Naine.

    Science.gov (United States)

    Kumaravel, Marimuthu; Uma, Subbaraya; Backiyarani, Suthanthiram; Saraswathi, Marimuthu Somasundaram; Vaganan, Muthu Mayil; Muthusamy, Muthusamy; Sajith, Kallu Purayil

    2017-01-01

    Endogenous hormone secretion proteins along with stress and defense proteins play predominant role in banana embryogenesis. This study reveals the underlying molecular mechanism during transition from vegetative to embryogenic state. Banana (Musa spp.) is well known globally as a food fruit crop for millions. The requirement of quality planting material of banana is enormous. Although mass multiplication through tissue culture is in vogue, high-throughput techniques like somatic embryogenesis (SE) as a mass multiplication tool needs to be improved. Apart from clonal propagation, SE has extensive applications in genetic improvement and mutation. SE in banana is completely genome-dependent and most of the commercial cultivars exhibit recalcitrance. Thus, understanding the molecular basis of embryogenesis in Musa will help to develop strategies for mass production of quality planting material. In this study, differentially expressed proteins between embryogenic calli (EC) and non-embryogenic calli (NEC) with respect to the explant, immature male flower buds (IMFB), of cv. Grand Naine (AAA) were determined using two-dimensional gel electrophoresis (2DE). The 2DE results were validated through qRT-PCR. In total, 65 proteins were identified: 42 were highly expressed and 23 were less expressed in EC compared to NEC and IMFB. qRT-PCR analysis of five candidate proteins, upregulated in EC, were well correlated with expression at transcript level. Further analysis of proteins showed that embryogenesis in banana is associated with the control of oxidative stress. The regulation of ROS scavenging system and protection of protein structure occurred in the presence of heat shock proteins. Alongside, high accumulation of stress-related cationic peroxidase and plant growth hormone-related proteins like indole-3-pyruvate monooxygenase and adenylate isopentenyltransferase in EC revealed the association with the induction of SE.

  5. The role of γ -aminobutyric acid (Gaba) in somatic embryogenesis of Acca sellowiana Berg. (Myrtaceae)

    OpenAIRE

    Booz, Maristela Raitz; Gilberto B. Kerbauy; Guerra, Miguel Pedro; Pescador, Rosete

    2009-01-01

    The γ-aminobutyric acid (Gaba) is a non-protein amino acid found in prokaryotes and eukaryotes. Its role in plant development has not been fully established. This study reports a quantification of the levels of endogenous Gaba, as well as investigation of its role in different stages of somatic embryogenesis in Acca sellowiana Berg. (Myrtaceae). Zygotic embryos were used as explants and they were inoculated into the culture medium contained different concentrations of Gaba (0,2, 4, 6, 8 ...

  6. Morphological Characterization and Identification of Coffea liberica Callus of Somatic Embryogenesis Propagation.

    OpenAIRE

    Fitria Ardiyani

    2015-01-01

    Compared with other types of coffee, Liberica coffee is more difficult to be propagates using clonal methods. Meanwhile, demand for planting materials and consumption of this type of coffee is increasing lately. The objective of this paper is to present results of the work on morpological characterization of Liberica coffee (Coffea liberica) callus produced by somatic embryogenesis propagation. This research used C. liberica Arruminensis clone. This clone was one of Liberica coffee clones whi...

  7. Somatic embryogenesis in immature cotyledons of Manchurian ash (Fraxinus mandshurica Rupr.)

    Science.gov (United States)

    Somatic embryogenesis was obtained from immature cotyledon explants that were cultured on half-strength Murashige and Skoog (MS) salts and vitamins with 5.4 uM naphthaleneacetic acid (NAA) and 0.2 uM thidiazuron (TDZ) plus a 4x4 factorial combination of 0,9.8, 34.6, or 49.2 uM indole-3-butyric acid ...

  8. Plant Regeneration and Somatic Embryogenesis from Immature Embryos Derived through Interspecific Hybridization among Different Carica Species

    OpenAIRE

    Azad, Md. Abul; Rabbani, Md. Golam; Amin, Latifah

    2012-01-01

    Plant regeneration and somatic embryogenesis through interspecific hybridization among different Carica species were studied for the development of a papaya ringspot virus-resistant variety. The maximum fruit sets were recorded from the cross of the native variety C. papaya cv. Shahi with the wild species C. cauliflora. The highest hybrid embryos were recorded at 90 days after pollination and the embryos were aborted at 150 days after pollination. The immature hybrid embryos were used for pla...

  9. Phosphoenolpyruvate carboxykinase in bovine tick Rhipicephalus (Boophilus) micro plus embryogenesis and starvation larvae

    Energy Technology Data Exchange (ETDEWEB)

    Andrade, J.G. de; Mentizingen, L.G.; Logullo, C. [Universidade Estadual do Norte Fluminense Darcy Ribeiro (UENF), Campos dos Goytacazes, RJ (Brazil). Centro de Biociencias e Biotecnologia. Lab.de Quimica e Funcao de Proteinas e Peptideos (LQFPP); Andrade, C.P. de; Vaz Junior, Itabajara [Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, RS (Brazil). Centro de Biotecnologia; Daffre, S.; Esteves, E. [Universidade de Sao Paulo (USP), SP (Brazil). Inst. de Ciencias Biomedicas

    2008-07-01

    Full text: Phosphoenolpyruvate carboxykinase (PEPCK) is considered a key rate controlling enzyme in gluconeogenesis pathway. Gluconeogenesis is a highly regulated process, catalyzed by several enzymes subject to regulation by insulin. Normally, insulin rapidly and substantially inhibits PEPCK gene transcription and the PEPCK activity is proportional to the rate of gene transcription. The transcriptional regulation of the PEPCK gene has been extensively studied. CREM is the transcription factor that bind efficiently to the putative cyclic AMP response element (CRE) in PEPCK gene. Several other transcription factors can bind to this element and activate transcription. In oviparous animals, such as bovine tick R. microplus, the embryonic development occurs outside the maternal organism, implying that all the nutrients necessary for embryogenesis must be present in the oocytes. We observed the relationship between the main energy sources and the morphogenetic changes that occur during R. microplus tick embryogenesis. Energy homeostasis is maintained by glycogen mobilization in the beginning of embryogenesis, as its content is drastically decreased during the first five days of development. Afterwards, the activity of the gluconeogenesis enzyme PEPCK increases enormously, as indicated by a concomitant increase in glucose content (Moraes et al., 2007). Here, we analyzed PEPCK gene transcription by qPCR during the embryogenesis and starvation larvae. The PEPCK transcription was higher at first and 15th day eggs of the development. In larvae the levels of PEPCK transcripts is increased at fifth day after hatch. However, the activity is continuous increased in larvae the form first up to 15th day. Now we are investigating the involvement of CREM in the PEPCK gene transcription in these cells. In this sense, we obtained CREM sequence from TIGR ESTs R. microplus bank and designed the specific primers to qPCR. Taken together our results suggest the involvement of PEPCK to the

  10. Morphoregulatory role of thidiazuron : substitution of auxin and cytokinin requirement for the induction of somatic embryogenesis in geranium hypocotyl cultures.

    Science.gov (United States)

    Visser, C; Qureshi, J A; Gill, R; Saxena, P K

    1992-08-01

    Somatic embryogenesis was induced in hypocotyl explants of geranium (Pelargonium x hortorum) cultured on media supplemented with various concentrations of N-phenyl-N'-1,2,3-thiadiazol-5-ylurea (thidiazuron). In less than 2 weeks, somatic embryos were observed in treatments containing levels of thidiazuron (TDZ) ranging from 0.2 to 1.0 micromolar. The use of N(6)-benzylaminopurine in combination with indole-3-acetic acid also evoked embryogenesis, but the efficiency of somatic embryo production was significantly lower than that obtained with TDZ. Hypocotyl culture for only 2 days on TDZ-supplemented medium before transfer to a basal medium was sufficient for inducing somatic embryogenesis. This distinction between the induction and expression of embryogenesis may provide an experimental system for studying the developmental biology of somatic embryogenesis. Substitution of the auxin-cytokinin requirement for the induction of somatic embryogenesis by TDZ suggests the possibility of a novel mode of its action by modulation of endogenous growth regulators.

  11. Annual Reproductive Cycle and Unusual Embryogenesis of a Temperate Coral in the Mediterranean Sea.

    Directory of Open Access Journals (Sweden)

    Chiara Marchini

    Full Text Available The variety of reproductive processes and modes among coral species reflects their extraordinary regeneration ability. Scleractinians are an established example of clonal animals that can exhibit a mixed strategy of sexual and asexual reproduction to maintain their populations. This study provides the first description of the annual reproductive cycle and embryogenesis of the temperate species Caryophyllia inornata. Cytometric analyses were used to define the annual development of germ cells and embryogenesis. The species was gonochoric with three times more male polyps than female. Polyps were sexually mature from 6 to 8 mm length. Not only females, but also sexually inactive individuals (without germ cells and males were found to brood their embryos. Spermaries required 12 months to reach maturity, while oogenesis seemed to occur more rapidly (5-6 months. Female polyps were found only during spring and summer. Furthermore, the rate of gamete development in both females and males increased significantly from March to May and fertilization was estimated to occur from April to July, when mature germ cells disappeared. Gametogenesis showed a strong seasonal influence, while embryos were found throughout the year in males and in sexually inactive individuals without a defined trend. This unusual embryogenesis suggests the possibility of agamic reproduction, which combined with sexual reproduction results in high fertility. This mechanism is uncommon and only four other scleractinians (Pocillopora damicornis, Tubastraea diaphana, T. coccinea and Oulastrea crispata have been shown to generate their broods asexually. The precise nature of this process is still unknown.

  12. Tobacco arabinogalactan protein NtEPc can promote banana (Musa AAA) somatic embryogenesis.

    Science.gov (United States)

    Shu, H; Xu, L; Li, Z; Li, J; Jin, Z; Chang, S

    2014-12-01

    Banana is an important tropical fruit worldwide. Parthenocarpy and female sterility made it impossible to improve banana varieties through common hybridization. Genetic transformation for banana improvement is imperative. But the low rate that banana embryogenic callus was induced made the transformation cannot be performed in many laboratories. Finding ways to promote banana somatic embryogenesis is critical for banana genetic transformation. After tobacco arabinogalactan protein gene NtEPc was transformed into Escherichia coli (DE3), the recombinant protein was purified and filter-sterilized. A series of the sterilized protein was added into tissue culture medium. It was found that the number of banana immature male flowers developing embryogenic calli increased significantly in the presence of NtEPc protein compared with the effect of the control medium. Among the treatments, explants cultured on medium containing 10 mg/l of NtEPc protein had the highest chance to develop embryogenic calli. The percentage of lines that developed embryogenic calli on this medium was about 12.5 %. These demonstrated that NtEPc protein can be used to promote banana embryogenesis. This is the first paper that reported that foreign arabinogalactan protein (AGP) could be used to improve banana somatic embryogenesis.

  13. High Frequency of Plant Regeneration through Cyclic Secondary Somatic Embryogenesis in Panax ginseng.

    Science.gov (United States)

    Kim, Yu-Jin; Lee, Ok Ran; Kim, Kyung-Tack; Yang, Deok-Chun

    2012-10-01

    Somatic embryogenesis is one of good examples of the basic research for plant embryo development as well as an important technique for plant biotechnology such as medicinally important plants. Single embryos develop into normal plantlets with shoots and roots. Therefore, direct single embryogenesis derived from single cells is highly important for normal plant regeneration. Here we demonstrate that the cyclic secondary somatic embryogenesis in Panax ginseng Meyer is a permanent source of embryogenic material that can be used for genetic manipulations. Secondary somatic embryos were originated directly from the primary somatic embryos on hormone-free Murashige and Skoog medium, and proliferated further in a cyclic manner. EM medium (one third of modified MS medium [MS medium containing half amount of NH4NO3 and KNO3] with 2% to 3% sucrose) favored further development of proliferated secondary somatic embryos into plantlets with root system. The plantlets developed into plants with well-developed taproots in half-strength Schenk and Hildebrandt basal medium supplemented with 0.5% activated charcoal.

  14. Effects of p-chlorophenoxyisobutyric acid, arabinogalactan, and activated charcoal on microspore embryogenesis in kale.

    Science.gov (United States)

    Niu, R Q; Zhang, Y; Tong, Y; Liu, Z Y; Wang, Y H; Feng, H

    2015-04-27

    To improve embryogenesis in microspore cultures of kale (Brassica oleracea L. var. acephala DC.), 6-benzylaminopurine (6-BA), naphthaleneacetic acid (NAA), arabinogalactan (AG), p-chlorophenoxyisobutyric acid (PCIB), and activated charcoal (AC) were added to the medium using four varieties of kale. The results showed that the addition of AG (0.1-0.2 g/L), AC (0.1-0.2 g/L) or a combination of 6-BA (0.1-0.2 mg/L) and NAA (0.1-0.2 mg/L) promoted embryo-genesis. Adding 40 μM PCIB or a combination of 40 μM PCIB and 0.2 g/L AC to NLN-13 medium at pH 5.8 effectively enhanced embryogenesis. Treatment with a combination of 40 μM PCIB and 10 mg/L AG gave the highest rate of embryonic induction, especially in genotype "Y007," which showed a twelve-fold increase in yield.

  15. Genetic chimerism of Vitis vinifera cv. Chardonnay 96 is maintained through organogenesis but not somatic embryogenesis

    Directory of Open Access Journals (Sweden)

    Butterlin Gisèle

    2005-09-01

    Full Text Available Abstract Background Grapevine can be a periclinal chimera plant which is composed at least of two distinct cell layers (L1, L2. When the cell layers of this plant are separated by passage through somatic embryogenesis, regenerated plants could show distinct DNA profiles and a novel phenotype which proved different from that of the parent plant. Results Genetically Chardonnay clone 96 is a periclinal chimera plant in which is L1 and L2 cell layers are distinct. Plants obtained via organogenesis through meristematic bulks are shown to be composed of both cell layers. However, plants regenerated through somatic embryogenesis starting from anthers or nodal explants are composed only of L1 cells. These somaclones do not show phenotypic differences to the parental clone up to three years after regeneration. Interestingly, the only somaclone showing an atypical phenotype (asymmetric leave shows a genotypic modification. Conclusion These results suggest that the phenotype of Chardonnay 96 does not result from an interaction between the two distinct cell layers L1 and L2. If phenotype conformity is further confirmed, somatic embryogenesis will result in true-to-type somaclones of Chardonnay 96 and would be well suitable for gene transfer.

  16. Morphological Characterization and Identification of Coffea liberica Callus of Somatic Embryogenesis Propagation.

    Directory of Open Access Journals (Sweden)

    Fitria Ardiyani

    2015-08-01

    Full Text Available Compared with other types of coffee, Liberica coffee is more difficult to be propagates using clonal methods. Meanwhile, demand for planting materials and consumption of this type of coffee is increasing lately. The objective of this paper is to present results of the work on morpological characterization of Liberica coffee (Coffea liberica callus produced by somatic embryogenesis propagation. This research used C. liberica Arruminensis clone. This clone was one of Liberica coffee clones which had superior taste. Main activitis carried out in this experiment were explant sterilization, explant induction and histological analysis on the callus produced. The result of this research showed that non embryogenic callus was higher (72% than embryogenic callus (28%. The callus description can be used to identify type and characteristic of the callus. Therefore, it can be a parameter to choose type of callus for propagation material. This is important because choosing the right callus is determine of the succesfully process of Liberica somatic embryogenesis. Keywords: somatic embryogenesis, Liberica, embryogenic, non-embryogenic

  17. Embryo lipoproteins and yolk lipovitellin consumption during embryogenesis in Macrobrachium borellii (Crustacea: Palaemonidae).

    Science.gov (United States)

    García, F; Cunningham, M L; Garda, H; Heras, H

    2008-11-01

    The prawn Macrobrachium borellii has lecithotrophic eggs with highly-abbreviated development. The major yolk component is lipovitellin (LV), a lipoprotein with 30% lipids (by weight). LV consumption during embryogenesis was followed by ELISA and Western blot analysis using an anti-LV polyclonal antibody. No cross-reacting proteins were observed and LV-like lipoproteins were strongly recognized by the antibody in hemolymph (vitellogenin), yolk (LV) and embryos (LVe), as determined by Western Blot analysis. LV decreased significantly along development from 9.4 to 1.1 microg/mg egg. Consumption rate of LV was slow in early embryogenesis, followed by a rapid utilization in late embryonic stages. Significant LVe amounts were still present at hatching. LV apolipoproteins were selectively degraded during embryo development, being the highest molecular weight subunit the most affected. Comparison among in vitro, in vivo and theoretical proteolysis suggested that trypsin may be involved in LV degradation during late embryogenesis. Embryo lipoprotein (HDLe) synthesis was first detected at stage 6. HDLe shared the same density, MW and subunit composition as adult hemolymph HDL(1) and did not cross-react with LV-like lipoproteins. Though expressed at low concentration, it fulfilled embryo needs for lipid transport among organs.

  18. JUVENILE-MATURE GENETIC CORRELATIONS IN Pinus taeda CLONES PROPAGATED VIA SOMATIC EMBRYOGENESIS

    Directory of Open Access Journals (Sweden)

    Poliana Coqueiro Dias

    2016-04-01

    Full Text Available ABSTRACT This study aimed to estimate the genetic correlation among selection ages (juvenile - adult and efficiency of early selection for the height, diameter, and volume traits of individuals from Pinus taeda families propagated via somatic embryogenesis. This study was carried out by genetic-statistical analysis, estimation procedure of variance (Reml, and prediction components of breeding values (Blup, using the Selegen-Reml/Blup software. Genetic correlations among juvenile ages and rotation age were performed by applying the linear model developed by Lambeth (1980. In accordance with results of the established model, the early selection can be performed in clones of Pinus taeda with high selection efficiency. Ages from 4 to 6 years old are enough to select Pinus taeda clones propagated via somatic embryogenesis for harvesting at 8 and 12 years old; and 6 to 10 years old are enough to select them for harvesting at 20 years old. On the basis of the genetic correlations estimates from the environments, the clones' selection of Pinus taeda propagated via somatic embryogenesis should be developed specifically for each environment. The clones' selection can be performed considering the diameter due to the high correlation between volume and diameter.

  19. Nuclear lamins and peripheral nuclear antigens during fertilization and embryogenesis in mice and sea urchins

    Energy Technology Data Exchange (ETDEWEB)

    Schatten, G.; Schatten, H.; Simerly, C.; Maul, G.G.; Chaly, N.

    1985-07-01

    Nuclear structural changes during fertilization and embryogenesis in mice and sea urchins are traced using four antibodies. The oocytes from virgin female mice, morulae and blastocytes from mated females, and gametes from the sea urchin Lytechnius variegatis are studied using mouse monoclonal antibodies to nuclear lamin A/C, monoclonal antibody to P1, human autoimmune antibodies to lamin A/C, and to lamin B. The mouse fertilization data reveal no lamins on the oocyte; however, lamins are present on the pronuclei, and chromosomes are found on the oocytes and pronuclei. It is detected that on the sea urchin sperm the lamins are reduced to acrosomal and centriolar fossae and peripheral antigens are around the sperm nucleus. The mouse sperm bind lamin antibodies regionally and do not contain antigens. Lamins and antigens are observed on both pronuclei and chromosomes during sea urchin fertilization. Mouse embryogenesis reveals that lamin A/C is not recognized at morula and blastocyst stages; however, lamin B stains are retained. In sea urchin embryogenesis lamin recognition is lost at the blastrula, gastrula, and plutei stages. It is noted that nuclear lamins lost during spermatogenesis are restored at fertilization and peripheral antigens are associated with the surface of chromosomes during meiosis and mitosis and with the periphery of the pronuclei and nuclei during interphase. 32 references.

  20. Characterization of leafy cotyledon1-like during embryogenesis in Theobroma cacao L.

    Science.gov (United States)

    Alemanno, Laurence; Devic, Martine; Niemenak, Nicolas; Sanier, Christine; Guilleminot, Jocelyne; Rio, Mariannick; Verdeil, Jean-Luc; Montoro, Pascal

    2008-03-01

    Theobroma cacao L., an economically important crop for developing countries, can be experimentally propagated by somatic embryogenesis. Because of their potential roles in embryogenesis, a gene candidate strategy was initiated to find gene homologues of the members of the leafy cotyledon family of transcription factors. A homologue of the leafy cotyledon1-like gene, that encodes the HAP 3 subunit of the CCAAT box-binding factor, was found in the cocoa genome (TcL1L). The translated peptide shared a high amino acid sequence identity with the homologous genes of Arabidopsis thaliana, Phaseolus coccineus and Helianthus annuus. TcL1L transcripts mainly accumulated in young and immature zygotic embryos, and, to a lesser extent, in young and immature somatic embryos. In situ hybridization specified the localization of the transcripts as being mainly in embryonic cells of young embryos, the meristematic cells of the shoot and root apex of immature embryos, and in the protoderm and epidermis of young and immature embryos, either zygotic or somatic. Non-embryogenic explants did not show TcL1L expression. Ectopic expression of the TcL1L gene could partially rescue the Arabidopsis lec1 mutant phenotype, suggesting a similarity of function in zygotic embryogenesis.

  1. Potash resources in part of Los Medanos area of Eddy and Lea Counties, New Mexico

    Science.gov (United States)

    Jones, C.L.

    1975-01-01

    Los Medanos area of eastern Eddy and western Lea Counties, New Mexico, is being considered for possible siting of a repository Facility for experimental studies of nuclear-waste emplacement in salt beds of the Salado Formation of Permian age. The potential repository site encompasses about 29 square miles (75 square kilometres) of sandy terrain near the center of Los Medanos. The site is underlain by evaporite and red bed formations having an aggregate thickness of 4,462 feet (1,360 metres). The Salado Formation, which is the main salt-bearing unit of the area, lies at a depth of 1,000 feet (305 metres). The formation is almost 1,970 feet (600 metres) of rock salt with prominent interbeds of anhydrite, polyhalite, glauberite, some widely spaced seams of fine-grained clastic rocks, and a few potash deposits containing sylvite and langbeinite. The potash deposits occur in salt beds 517-871 feet (158-265 metres) below the top of the formation. The lower half of the formation includes some salt beds which may be used for the experimental emplacement of nuclear wastes. Most, if not nearly all, of the potential repository site is underlain by potash deposits that contain sylvite and langbeinite. The deposits grade 12-13 percent K2O as sylvite and 3-11 percent K2O as langbeinite, and must be regarded as having economic potential for potassium minerals. There is considerable uncertainty concerning the full extent and total range in quality of the ores in the deposit, and subsurface investigation will be required to assess their potential.

  2. Efficacy of Zingiber officinale ethanol extract on the viability, embryogenesis and infectivity of Toxocara canis eggs.

    Science.gov (United States)

    El-Sayed, Nagwa Mostafa

    2017-12-01

    This study evaluated the effect of Zingiber officinale (Z. officinale) ethanol extract on the viability, embryogenesis and infectivity Toxocara canis (T. canis) eggs. It was carried out both in vitro and in vivo. In the in vitro experiment, unembryonated T. canis eggs were incubated with 25, 50 and 100 mg/mL Z. officinale extract at 25 °C for 6, 12, and 24 h to assess the effect of Z. officinale on their viability and for two weeks to assess the effect of Z. officinale on their embryogenesis. In vivo experiment was performed to assess the effect of Z. officinale on infectivity of T. canis eggs. Treated embryonated eggs by Z. officinale extract at concentrations of 25, 50 and 100 mg/mL for 24 h were inoculated into mice and their livers were examined for the presence of T. canis larvae on the 7th day after infection and for histopathological evaluation at 14th day post-infection. Z. officinale showed a significant ovicidal activity on T. canis eggs. The best effect was observed with 100 mg/mL concentration after 24 h with an efficacy of 98.2%. However, the treated eggs by 25, 50 mg/mL of Z. officinale extract after 24 h showed ovicidal activity by 59.22 and 82.5% respectively. Moreover, this extract effectively inhibited T. canis eggs embryogenesis by 99.64% and caused their degeneration at the concentration of 100 mg/mL after 2 weeks of treatment. However, the lower concentrations, 25 and 50 mg/mL inhibited embryogenesis by 51.19 and 78.57% respectively. The effect of Z. officinale on the infectivity T. canis eggs was proven by the reduction of larvae recovery in the livers by 35.9, 62.8 and 89.5% in mice groups inoculated by Z. officinale treated eggs at concentrations of 25, 50 and 100 mg/mL respectively. Histopathologically, the liver tissues of mice infected with Z. officinale treated eggs at the concentration of 100 mg/mL appeared healthy with slight degenerative changes of hepatocytes, opposite to that recorded in the infected mice with

  3. HENRY CHARLES LEA Y SU RED DE COLABORADORES LATINOAMERICANOS: RAZONES PARA ESTUDIAR EL SANTO OFICIO EN LA SEGUNDA MITAD DEL SIGLO XIX

    Directory of Open Access Journals (Sweden)

    Doris Moreno

    2013-12-01

    Full Text Available Se aborda en este artículo la vida y pensamiento de Henry Charles Lea en relación a sus estudios sobre la historia de la Inquisición española. Se esboza brevemente la red de colaboradores que Lea estableció en toda Latinoamérica así como su perfil político e intelectual, subrayando su importancia para la escritura de la obra The Inquisition in the Spanish Dependencies (1908. Finalmente, se plantean los motivos que llevaron a estos intelectuales a colaborar intensamente con el historiador norteamericano en el marco de los procesos de construcción de identidades sociales y políticas en Latinoamérica en la segunda mitad del siglo XIX. Se toma como eje conductor de este planteamiento la correspondencia de Juan María Gutiérrez con Henry Charles Lea.

  4. High food abundance permits the evolution of placentotrophy: evidence from a placental lizard, Pseudemoia entrecasteauxii.

    Science.gov (United States)

    Van Dyke, James U; Griffith, Oliver W; Thompson, Michael B

    2014-08-01

    Mechanisms of reproductive allocation are major determinants of fitness because embryos cannot complete development without receiving sufficient nutrition from their parents. The nourishment of offspring via placentas (placentotrophy) has evolved repeatedly in vertebrates, including multiple times in squamate reptiles (lizards and snakes). Placentotrophy has been suggested to evolve only if food is sufficiently abundant throughout gestation to allow successful embryogenesis. If scarcity of food prevents successful embryogenesis, females should recoup nutrients allocated to embryos via abortion, reabsorption, and/or cannibalism. We tested these hypotheses in the placentotrophic southern grass skink Pseudemoia entrecasteauxii. We fed females one of four diets (high constant, high variable, low constant, and low variable) during gestation and tested the effects of both food amount and schedule of feeding on developmental success, cannibalism rate, placental nutrient transport, offspring size, and maternal growth and body condition. Low food availability reduced developmental success, placental nutrient transport, offspring size, and maternal growth and body condition. Cannibalism of offspring also increased when food was scarce. Schedule of feeding did not affect offspring or mothers. We suggest that high food abundance and ability to abort and cannibalize poor-quality offspring are permissive factors necessary for placentotrophy to be a viable strategy of reproductive allocation.

  5. Ultrastructural changes and the distribution of arabinogalactan proteins during somatic embryogenesis of banana (Musa spp. AAA cv. 'Yueyoukang 1').

    Science.gov (United States)

    Pan, Xiao; Yang, Xiao; Lin, Guimei; Zou, Ru; Chen, Houbin; Samaj, Jozef; Xu, Chunxiang

    2011-08-01

    A better understanding of somatic embryogenesis in banana (Musa spp.) may provide a practical way to improve regeneration of banana plants. In this study, we applied scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to visualize the ultrastructural changes during somatic embryogenesis of banana (Musa AAA cv. 'Yueyoukang 1'). We also used histological and immunohistochemical techniques with 16 monoclonal antibodies to study the spatial distribution and cellular/subcellular localization of different arabinogalactan protein (AGP) components of the cell wall during somatic embryogenesis. Histological study with periodic acid-Schiff staining documented diverse embryogenic stages from embryogenic cells (ECs) to the late embryos. SEM revealed a mesh-like structure on the surface of proembryos which represented an early structural marker of somatic embryogenesis. TEM showed that ECs were rich in juvenile mitochondria, endoplasmic reticulum and Golgi stacks. Cells in proembryos and early globular embryos resembled ECs, but they were more vacuolated, showed more regular nuclei and slightly more developed organelles. Immunocytochemical study revealed that the signal of most AGP epitopes was stronger in starch-rich cells when compared with typical ECs. The main AGP component in the extracellular matrix surface network of banana proembryos was the MAC204 epitope. Later, AGP immunolabelling patterns varied with the developmental stages of the embryos. These results about developmental regulation of AGP epitopes along with developmental changes in the ultrastructure of cells are providing new insights into the somatic embryogenesis of banana. Copyright © Physiologia Plantarum 2011.

  6. Proteome Analysis Unravels Mechanism Underling the Embryogenesis of the Honeybee Drone and Its Divergence with the Worker (Apis mellifera lingustica).

    Science.gov (United States)

    Fang, Yu; Feng, Mao; Han, Bin; Qi, Yuping; Hu, Han; Fan, Pei; Huo, Xinmei; Meng, Lifeng; Li, Jianke

    2015-09-04

    The worker and drone bees each contain a separate diploid and haploid genetic makeup, respectively. Mechanisms regulating the embryogenesis of the drone and its mechanistic difference with the worker are still poorly understood. The proteomes of the two embryos at three time-points throughout development were analyzed by applying mass spectrometry-based proteomics. We identified 2788 and 2840 proteins in the worker and drone embryos, respectively. The age-dependent proteome driving the drone embryogenesis generally follows the worker's. The two embryos however evolve a distinct proteome setting to prime their respective embryogenesis. The strongly expressed proteins and pathways related to transcriptional-translational machinery and morphogenesis at 24 h drone embryo relative to the worker, illustrating the earlier occurrence of morphogenesis in the drone than worker. These morphogenesis differences remain through to the middle-late stage in the two embryos. The two embryos employ distinct antioxidant mechanisms coinciding with the temporal-difference organogenesis. The drone embryo's strongly expressed cytoskeletal proteins signify key roles to match its large body size. The RNAi induced knockdown of the ribosomal protein offers evidence for the functional investigation of gene regulating of honeybee embryogenesis. The data significantly expand novel regulatory mechanisms governing the embryogenesis, which is potentially important for honeybee and other insects.

  7. Spatial expression of a sunflower SERK gene during induction of somatic embryogenesis and shoot organogenesis.

    Science.gov (United States)

    Thomas, Clément; Meyer, Denise; Himber, Christophe; Steinmetz, André

    2004-01-01

    Organogenesis or somatic embryogenesis can be induced on immature zygotic embryos (IZE) of sunflower depending on the culture conditions. Both morphogenic processes originate from the same group of cells and show identical kinetics. Using real-time PCR and in situ hybridisation, we showed that somatic embryogenesis receptor-like kinase (SERK) transcripts accumulate early after the beginning of the culture in the morphogenic zone of IZE explants whatever the induction conditions used, i.e. organogenic, embryogenic or highly embryogenic conditions. Quantitative analyses failed to show any correlation between the SERK expression level during the period decisive for the orientation of the morphogenic pathway, i.e. the first 2 days of culture, and the type of morphogenesis induced. However, after 2 days of culture on the organogenic medium, the SERK gene expression level was severely down-regulated in the IZE explants. At 4 days of culture, SERK transcripts were no longer detectable by in situ hybridisation in the developing shoot structures whereas they still continued to accumulate in the embryonic structures induced on both embryogenic and highly embryogenic culture media. The significance of these expression analyses was addressed by transfer medium experiments. Results revealed that IZE cultured on the organogenic medium were able to form somatic embryos when transferred on the highly embryogenic medium as long as the SERK transcripts accumulated at a high level in their morphogenic zone, i.e. first 2 days of culture. Passt this delay, explants rapidly lost their embryogenic competence. Indeed, after 4 days of culture on the organogenic medium, IZE were definitely oriented towards shoot organogenesis. Taken together, these data suggest that reactive cells of IZE develop the competence to somatic embryogenesis during the first day of culture whatever the morphogenic induction conditions used.

  8. The genomic distribution and function of histone variant HTZ-1 during C. elegans embryogenesis.

    Directory of Open Access Journals (Sweden)

    Christina M Whittle

    2008-09-01

    Full Text Available In all eukaryotes, histone variants are incorporated into a subset of nucleosomes to create functionally specialized regions of chromatin. One such variant, H2A.Z, replaces histone H2A and is required for development and viability in all animals tested to date. However, the function of H2A.Z in development remains unclear. Here, we use ChIP-chip, genetic mutation, RNAi, and immunofluorescence microscopy to interrogate the function of H2A.Z (HTZ-1 during embryogenesis in Caenorhabditis elegans, a key model of metazoan development. We find that HTZ-1 is expressed in every cell of the developing embryo and is essential for normal development. The sites of HTZ-1 incorporation during embryogenesis reveal a genome wrought by developmental processes. HTZ-1 is incorporated upstream of 23% of C. elegans genes. While these genes tend to be required for development and occupied by RNA polymerase II, HTZ-1 incorporation does not specify a stereotypic transcription program. The data also provide evidence for unexpectedly widespread independent regulation of genes within operons during development; in 37% of operons, HTZ-1 is incorporated upstream of internally encoded genes. Fewer sites of HTZ-1 incorporation occur on the X chromosome relative to autosomes, which our data suggest is due to a paucity of developmentally important genes on X, rather than a direct function for HTZ-1 in dosage compensation. Our experiments indicate that HTZ-1 functions in establishing or maintaining an essential chromatin state at promoters regulated dynamically during C. elegans embryogenesis.

  9. Plant regeneration via somatic embryogenesis and shoot organogenesis from immature cotyledons of Camellia nitidissima Chi.

    Science.gov (United States)

    Lü, Jinfeng; Chen, Rong; Zhang, Muhan; da Silva, Jaime A Teixeira; Ma, Guohua

    2013-09-01

    Camellia nitidissima Chi (Theaceae) is a world-famous economic and ornamental plant with golden-yellow flowers. It has been classified as one of the rarest and most endangered plants in China. Our objective was to induce somatic embryogenesis, shoot organogenesis and plant regeneration for C. nitidissima. Three types of callus (whitish, reddish and yellowish) were induced from immature cotyledons on improved woody plant medium (WPM) with different plant growth regulators (PGRs). Among the callus, whitish callus was induced by 4.5 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and reddish and yellowish callus were induced by strongly active cytokinins, thidiazuron (TDZ) or 6-benzylaminopurine (BAP), singly or combined with weakly active auxin, α-naphthaleneacetic acid (NAA). The embryogenic callus could differentiate into somatic embryos, nodular embryogenic structures (large embryo-like structures) or adventitious shoots depending on the PGR used in WPM. BAP was best for adventitious buds and zeatin was best for somatic embryogenesis while kinetin (Kt) was best for the formation of nodular embryogenic structures. The three regeneration pathways often occurred in the same embryogenic callus clumps. Most shoots (80.0%) developed roots in WPM supplemented with 24.6 μM IBA and 0.3 μM NAA while 47.5% of somatic embryos could germinate directly and develop into plantlets on induction medium supplemented with 0.9 μM BAP and 0.1 μM NAA. The nodular embryogenic structures could be sub-cultured and cyclically developed in one of two differentiation pathways: shoot organogenesis or somatic embryogenesis. Plantlets derived from shoot buds rooted and somatic embryos germinated when transplanted into soil in a greenhouse; 66.7% of plantlets from shoot culture and 78.6% of plantlets from somatic embryos survived after 8 weeks' acclimatization. Copyright © 2013 Elsevier GmbH. All rights reserved.

  10. Functional analysis of Bombyx Wnt1 during embryogenesis using the CRISPR/Cas9 system.

    Science.gov (United States)

    Zhang, Zhongjie; Aslam, Abu F M; Liu, Xiaojing; Li, Muwang; Huang, Yongping; Tan, Anjiang

    2015-08-01

    Recently established, custom-designed nuclease technologies such as the clustered regularly interspaced short palindromic repeat (CRISPR)-associated system provide attractive genome editing tools. Targeted gene mutagenesis using the CRISPR/Cas9 system has been achieved in several orders of insects. However, outside of studies on Drosophila melanogaster and the lepidopteron model insect Bombyx mori, little success has been reported, which is largely due to a lack of effective genetic manipulation tools that can be used in other insect orders. To create a simple and effective method of gene knockout analysis, especially for dissecting gene functioning during insect embryogenesis, we performed a functional analysis of the Bombyx Wnt1 (BmWnt1) gene using Cas9/sgRNA-mediated gene mutagenesis. The Wnt1 gene is required for embryonic patterning in various organisms, and its crucial roles during embryogenesis have been demonstrated in several insect orders. Direct injection of Cas9 mRNA and BmWnt1-specific sgRNA into Bombyx embryos induced a typical Wnt-deficient phenotype: injected embryos could not hatch and exhibited severe defects in body segmentation and pigmentation in a dose-dependent manner. Quantitative real-time PCR (qRT-PCR) analysis revealed that Hox genes were down-regulated after BmWnt1 depletion. Furthermore, large deletion, up to 18Kb, ware generated. The current study demonstrates that using the CRISPR/Cas9 system is a promising approach to achieve targeted gene mutagenesis during insect embryogenesis. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Somatic embryogenesis and plant regeneration in Wedelia calendulacea Less. an endangered medicinal plant

    OpenAIRE

    Shamima Akhtar Sharmin; Md. Jahangir Alam; Md. Mominul Islam Sheikh; Kanak Kanti Sarker; Muhammad Khalekuzzaman; Md. Anwarul Haque; Mohammad Firoz Alam; Iftekhar Alam

    2014-01-01

    In this work, plant regeneration via somatic embryogenesis was achieved from leaf and internode derived callus of Wedelia calendulacea, an endangered medicinal plant. Primary callus was induced by culturing leaf disc and internode explant on Murashige and Skoog medium supplemented with 2.0 mg L-1 of 2,4-D under light condition. Transfer of embryogenic callus on a reduced concentration of 2,4-D facilitated somatic embryo development while calluses remained unorganized at the same 2,4-D level. ...

  12. Packaging of Post Acclimatized Somatic Embryogenesis Cocoa Plantlet (Theobroma cacao L.

    Directory of Open Access Journals (Sweden)

    Soedarsianto Soedarsianto

    2009-05-01

    Full Text Available Clonal plants that produced by somatic embryogenesis technique is one of the best choice to produce supperior clonal cacao (Theobroma cacao L. planting materials. The somatic embryogenesis technique is a possible way for massive propagation, the outcome is true to type plants, the architecture similarity that the seedlings but there is not segregation like seedlings plants. At present mass production started of plantlets production until post-acclimatized plantlets of somatic embryogenesis cocoa was done at Indonesian Coffee and Cocoa Research Institute. Distribution system of the planting materials to whole areas in form of as up-rooted post-acclimatized plantlet. Some problems identified to reduce probability of decreasing viability of up-rooted post-acclimatized plantlets and one of them is extreme internal water deficit. This research investigate of the influece storage condition (air tight and non-air tight and box storage (mica plastic and cardboardbox. The first experiment result show, there is no significant different between mica plastic and cardboard box usage for storage of post-acclimatized cocoa pantlet. Viability of up-rooted post acclimatized cocoa plantlet influenced exactly by air tight and non-air tight storage condition. Air tight storage condition have better viability of up-rooted post acclimatised (81,58% than non-air tight storage condition (65,00%. Leaf sanasence on air tight storage condition (10,33% lower than non-air tight storage (32,58%. There is not significantly on volume storage per plantlet between 4.416 cm3 and 12.600 cm3. Relationship between fallen leaves and cocoa planlet viability follow negative linear correlation y = -1,4719x + 104,88 (R2 = 0,9772. The second experiment treatment showed that maximal storage periode of post cclimatized cocoa plantlet just until 6 days stored (97% and not significant with 3 days one. Viability of post acclimatized cocoa plantlet decreased after 6 days storage period

  13. La traducción de la voz. Una lectura de Maldición eterna a quien lea estas páginas de Manuel Puig

    Directory of Open Access Journals (Sweden)

    Delfina Cabrera

    2015-09-01

    Full Text Available This article aims to analyze the ways in which language and translation function in Puig’s Maldición eterna a quien lea estas páginas, paying special attention to the novel’s multilingual basis as well as to its questioning of the monolingual paradigm.

  14. Analysis of wheat microspore embryogenesis induction by transcriptome and small RNA sequencing using the highly responsive cultivar "Svilena".

    Science.gov (United States)

    Seifert, Felix; Bössow, Sandra; Kumlehn, Jochen; Gnad, Heike; Scholten, Stefan

    2016-04-21

    Microspore embryogenesis describes a stress-induced reprogramming of immature male plant gametophytes to develop into embryo-like structures, which can be regenerated into doubled haploid plants after whole genome reduplication. This mechanism is of high interest for both research as well as plant breeding. The objective of this study was to characterize transcriptional changes and regulatory relationships in early stages of cold stress-induced wheat microspore embryogenesis by transcriptome and small RNA sequencing using a highly responsive cultivar. Transcriptome and small RNA sequencing was performed in a staged time-course to analyze wheat microspore embryogenesis induction. The analyzed stages were freshly harvested, untreated uninucleate microspores and the two following stages from in vitro anther culture: directly after induction by cold-stress treatment and microspores undergoing the first nuclear divisions. A de novo transcriptome assembly resulted in 29,388 contigs distributing to 20,224 putative transcripts of which 9,305 are not covered by public wheat cDNAs. Differentially expressed transcripts and small RNAs were identified for the stage transitions highlighting various processes as well as specific genes to be involved in microspore embryogenesis induction. This study establishes a comprehensive functional genomics resource for wheat microspore embryogenesis induction and initial understanding of molecular mechanisms involved. A large set of putative transcripts presumably specific for microspore embryogenesis induction as well as contributing processes and specific genes were identified. The results allow for a first insight in regulatory roles of small RNAs in the reprogramming of microspores towards an embryogenic cell fate.

  15. Effects of Osmolytic Agents on Somatic Embryogenesis of Saffron (Crocus sativus L.

    Directory of Open Access Journals (Sweden)

    Maryam VAHEDI

    2015-03-01

    Full Text Available A protocol for callus induction from meristem tissues and subsequent somatic embryo formation were established in this study. Explants were taken from apical and lateral meristems of saffron and these explants were cultured on MS medium supplemented with combinations of 2.4-dichlorophenoxyacetic acid (2.4-D and Kinetin (Kn. The effects of osmotic agents such as abscisic acid (ABA, polyethylene glycol (PEG and Gelrite on somatic embryogenesis were also investigated. After 45 and 60 days of culture, calli were induced from apical and lateral meristems, respectively. The apical meristems yielded higher quality calli when compared to the lateral meristems. The highest frequency of callogenesis and the growth rate of callus were achieved from apical meristems on Murashige and Skoogs (MS medium supplemented with 2.4-D (2 mg/l and Kinetin (0.5 mg/l. After 45 days of subculture, the segments of nodular calli were transferred to plant growth regulator (PGR- free media for induction of pre-embryogenesis embryo formation. Pre-matured embryos were cultured on MS medium supplemented with different osmotic agents such as Gelrite, ABA and PEG to study their effects on embryo maturation. Both PEG and ABA proved more effective for somatic embryo maturation as compared to Gelrite.

  16. Effects of Osmolytic Agents on Somatic Embryogenesis of Saffron (Crocus sativus L.

    Directory of Open Access Journals (Sweden)

    Maryam VAHEDI

    2015-03-01

    Full Text Available A protocol for callus induction from meristem tissues and subsequent somatic embryo formation were established in this study. Explants were taken from apical and lateral meristems of saffron and these explants were cultured on MS medium supplemented with combinations of 2.4-dichlorophenoxyacetic acid (2.4-D and Kinetin (Kn. The effects of osmotic agents such as abscisic acid (ABA, polyethylene glycol (PEG and Gelrite on somatic embryogenesis were also investigated. After 45 and 60 days of culture, calli were induced from apical and lateral meristems, respectively. The apical meristems yielded higher quality calli when compared to the lateral meristems. The highest frequency of callogenesis and the growth rate of callus were achieved from apical meristems on Murashige and Skoogs (MS medium supplemented with 2.4-D (2 mg/l and Kinetin (0.5 mg/l. After 45 days of subculture, the segments of nodular calli were transferred to plant growth regulator (PGR- free media for induction of pre-embryogenesis embryo formation. Pre-matured embryos were cultured on MS medium supplemented with different osmotic agents such as Gelrite, ABA and PEG to study their effects on embryo maturation. Both PEG and ABA proved more effective for somatic embryo maturation as compared to Gelrite.

  17. Somatic embryogenesis and plant regeneration in Wedelia calendulacea Less. an endangered medicinal plant

    Directory of Open Access Journals (Sweden)

    Shamima Akhtar Sharmin

    2014-06-01

    Full Text Available In this work, plant regeneration via somatic embryogenesis was achieved from leaf and internode derived callus of Wedelia calendulacea, an endangered medicinal plant. Primary callus was induced by culturing leaf disc and internode explant on Murashige and Skoog medium supplemented with 2.0 mg L-1 of 2,4-D under light condition. Transfer of embryogenic callus on a reduced concentration of 2,4-D facilitated somatic embryo development while calluses remained unorganized at the same 2,4-D level. A histological analysis confirmed somatic embryo by revealing the presence of a closed vascular system in the developing embryos and lack of a vascularconnection with surrounding callus tissues. Somatic embryos germinated into plantlets upon transfer on MS medium containing 1.0 mg L-1 BAP plus 0.5 mg L-1 GA3. Plantlets were acclimatized successfully and survived under soil condition. This is the first on somatic embryogenesis of W.calendulacea. This result could facilitate genetic transformation of this important medicinal plant.

  18. Locus-Specific DNA Methylation Reprogramming During Early Porcine Embryogenesis1

    Science.gov (United States)

    Zhao, Ming-Tao; Rivera, Rocio M.; Prather, Randall S.

    2013-01-01

    ABSTRACT During early mammalian embryogenesis, there is a wave of DNA demethylation postfertilization and de novo methylation around implantation. The paternal genome undergoes active DNA demethylation, whereas the maternal genome is passively demethylated after fertilization in most mammals except for sheep and rabbits. However, the emerging genome-wide DNA methylation landscape has revealed a regulatory and locus-specific DNA methylation reprogramming pattern in mammalian preimplantation embryos. Here we optimized a bisulfite sequencing protocol to draw base-resolution DNA methylation profiles of several selected genes in gametes, early embryos, and somatic tissue. We observed locus-specific DNA methylation reprogramming in early porcine embryos. First, some pluripotency genes (POU5F1 and NANOG) followed a typical wave of DNA demethylation and remethylation, whereas CpG-rich regions of SOX2 and CDX2 loci were hypomethylated throughout development. Second, a differentially methylated region of an imprint control region in the IGF2/H19 locus exhibited differential DNA methylation which was maintained in porcine early embryos. Third, a centromeric repeat element retained a moderate DNA methylation level in gametes, early embryos, and somatic tissue. The diverse DNA methylation reprogramming during early embryogenesis is thought to be possibly associated with the multiple functions of DNA methylation in transcriptional regulation, genome stability and genomic imprinting. The latest technology such as oxidative bisulfite sequencing to identify 5-hydroxymethylcytosine will further clarify the DNA methylation reprogramming during porcine embryonic development. PMID:23303676

  19. Gametic embryogenesis and haploid technology as valuable support to plant breeding.

    Science.gov (United States)

    Germanà, Maria Antonietta

    2011-05-01

    Plant breeding is focused on continuously increasing crop production to meet the needs of an ever-growing world population, improving food quality to ensure a long and healthy life and address the problems of global warming and environment pollution, together with the challenges of developing novel sources of biofuels. The breeders' search for novel genetic combinations, with which to select plants with improved traits to satisfy both farmers and consumers, is endless. About half of the dramatic increase in crop yield obtained in the second half of the last century has been achieved thanks to the results of genetic improvement, while the residual advance has been due to the enhanced management techniques (pest and disease control, fertilization, and irrigation). Biotechnologies provide powerful tools for plant breeding, and among these ones, tissue culture, particularly haploid and doubled haploid technology, can effectively help to select superior plants. In fact, haploids (Hs), which are plants with gametophytic chromosome number, and doubled haploids (DHs), which are haploids that have undergone chromosome duplication, represent a particularly attractive biotechnological method to accelerate plant breeding. Currently, haploid technology, making possible through gametic embryogenesis the single-step development of complete homozygous lines from heterozygous parents, has already had a huge impact on agricultural systems of many agronomically important crops, representing an integral part in their improvement programmes. The aim of this review was to provide some background, recent advances, and future prospective on the employment of haploid technology through gametic embryogenesis as a powerful tool to support plant breeding.

  20. Cytomorphological studies on somatic embryogenesis of Gentiana tibetica (King and G. cruciata (L.

    Directory of Open Access Journals (Sweden)

    Anna Mikuła

    2014-01-01

    Full Text Available The process of plant regeneration via somatic embryogenesis of two gentianas, Gentiana tibetica and G. cruciata was described. For this purpose seedling explants were cultured on agar medium and later maintained in cell suspension. For callus initiation seedling explants like: cotyledons, hypocotyl and root were plated on a callus induction medium (CIM composed of MS (1962, supplemented with 0.5 mg/l 2,4-D and 1.0 mg/l Kin. For the formation of cell suspension culture, embryogenic callus was transferred into liquid maintained medium (MM composed of MS (1962, supplemented with 1.0 mg/l Dic + 0.1 mg/l NAA + 2.0 mg/l BAP + 80.0 mg/l SA. The conversion of somatic embryo into plantlets required a new medium (ECM based on MS (1962 mineral salts, supplemented with 0.5 mg/l GA3 + 1.0 mg/l Kin + 0.5 mg/l NAA. For cytomorphological studies of particular stages of embryogenesis, specimens were stained with dyes and reagents: 1. PAS reaction with leukofucsin, 2. Safranin + fast green, 3. Erlich's hematoxylin.

  1. Effect of microgravity and hypergravity on embryo axis alignment during postencystment embryogenesis in Artemia franciscana (Anostraca)

    Science.gov (United States)

    Rosowski, J. R.; Gouthro, M. A.; Schmidt, K. K.; Klement, B. J.; Spooner, B. S.

    1995-01-01

    Cysts of brine shrimp attached with a liquid adhesive to 12-mm diameter glass coverslips in a syringe-type fluid processing apparatus were flown aboard the NASA space shuttle Discovery, flight STS-60, from 3-11 February 1994, and were allowed to undergo postencystment embryogenesis and to hatch in microgravity. The shuttle flight and the ground-based control coverslips with attached cysts were parallel to the earth's surface during incubation in salt water. Based on the position of the cyst shell crack in the attached cyst population, the ground-control nauplii emerged mostly upward. On the shuttle in microgravity, although our method of detection of orientation would not reveal emergence toward the coverslip, the ratio of the position of the cyst shell crack in the population after hatching best fit the predicted values of a random direction for nauplii emergence. Centrifugation on earth was then used to create hypergravity forces of up to 73 g during postencystment embryogenesis and hatching. The upward orientation of emerging nauplii showed a high degree of correlation (r(2) =98.8%) with a linear relationship to the log of g, with 78.2% of the total hatching upward at 1 g and 91.0% hatching upward at 73 g.

  2. Functional redundancy of EGF-CFC genes in epiblast and extraembryonic patterning during early mouse embryogenesis.

    Science.gov (United States)

    Chu, Jianhua; Shen, Michael M

    2010-06-01

    During early mouse embryogenesis, multiple patterning and differentiation events require the activity of Nodal, a ligand of the transforming growth factor-beta (TGFbeta) family. Although Nodal signaling is known to require activity of EGF-CFC co-receptors in many contexts, it has been unclear whether all Nodal signaling in the early mouse embryo is EGF-CFC dependent. We have investigated the double null mutant phenotypes for the EGF-CFC genes Cripto and Cryptic, which encode co-receptors for Nodal, and have found that they have partially redundant functions in early mouse development. Expression of Cripto and Cryptic is non-overlapping prior to gastrulation, since Cripto is expressed solely in the epiblast whereas Cryptic is expressed in the primitive endoderm of the late blastocyst and the visceral endoderm after implantation. Despite these non-overlapping expression patterns, Cripto; Cryptic double mutants display severe defects in epiblast, extraembryonic ectoderm, and anterior visceral endoderm (AVE), resulting in phenotypes that are highly similar to those of Nodal null mutants. Our results indicate that both Cripto and Cryptic function non-cell-autonomously during normal development, and that most if not all Nodal activity in early mouse embryogenesis is EGF-CFC-dependent. Copyright 2010 Elsevier Inc. All rights reserved.

  3. Changes in ecdysteroids during embryogenesis of the blue crab, callinectes sapidus rathbun

    Energy Technology Data Exchange (ETDEWEB)

    McCarthy, J.F.; Skinner, D.M.

    1979-01-01

    Total ecdysteroid titers (estimated by radioimmunoassay (RIA)) in embryos of the blue crab increased from approx. 6 ng 20-hydroxyecdysone equivalents/g in the immature embryo to a maximum of approx. 500 ng 20-hydroxyecdysone equivalents/g in maturing embryos; titers dropped to approx. 300 ng 20-hydroxyecydsone equivalents/g in prehatch embryos. High-pressure reverse-phase chromatography of the embryo extracts resolved five RIA-active components, ..cap alpha..-Ecdysone and the polar conjugate of 20-hydroxyecdysone were present in low quantities. The concentration of 20-hydroxyecdysone increased during embryogenesis to a maximum of approx. 160 ng/g in maturing embryos and decreased only slightly in the prehatch embryos. Two unidentified components were also detected and the changes in their concentrations were estimated. One, an apolar component (peak III), accounted for as much as 63% of the total RIA activity as the embryos matured. The estimated concentration of this component increased from 85 ng/g in early embryos to 475 ng/g in maturing embryos, then decreased by 50% in the prehatch embryos. The level of the other, more polar component (peak II) increased from 7.5 to 75 ng/g as the embryos developed. The increase in the concentration of ecdysteroids during embryogenesis indicates that crab embryos have the capacity to synthesize ecdysteroids and suggests that these hormones may have a physiological role in the embryonic development of crustaceans.

  4. Rape embryogenesis. IV. Appearance and disappearance of starch during embryo development

    Directory of Open Access Journals (Sweden)

    Teresa Tykarska

    2014-01-01

    Full Text Available Starch appears first in the suspensor of the proembryo with two-cell apical part. It is observed in the embryo proper from the octant stage. At first it is visible in all the embryo cells in the form of minute transient grains which disappear during cell divisions. But the columella mother cells and their derivatives have persistent large grains. When the embryo turns green in the heart stage a gradual accumulation of storage starch begins and lasts to the end of embryogenesis. Storage starch grains appear first in the auter cortex layers of the hypocotyl where the largest grains are to be found later, and afterwards in all the other tissues. Starch is usually absent in the frequently dividing cells, but even there it appears in the form of minute grains after the end of cell divisions. Disappearance of starch starts when the intensive green colour of the seed coat begins to fade. The first to disappear are the smallest granules in the regions where they were noted latest. In the embryo axis the starch grains remain deposited longest in dermatogen and cortex cells in the lower hypocotyl part. They are visible there, still when the seed turns brown. In black seeds starch may be only found in the columella the cells of which throughout embryogenesis contain amyloplasts filled with starch. These grains disappear completely at the time when the seeds become dry.

  5. Somatic embryogenesis and enhanced shoot organogenesis in Metabriggsia ovalifolia W. T. Wang

    Science.gov (United States)

    Ouyang, Yao; Chen, Yulu; Lü, Jinfeng; Teixeira da Silva, Jaime A.; Zhang, Xinhua; Ma, Guohua

    2016-01-01

    An efficient protocol providing a dual regeneration pathway via direct shoot organogenesis and somatic embryogenesis for an endangered species, Metabriggsia ovalifolia W. T. Wang, was established from leaf explants. When applied at 2.5 μM, the cytokinins 6-benzyladenine (BA) or thidiazuron (TDZ) and the auxins indole-3-butyric acid (IBA), α-naphthaleneacetic acid (NAA) and indole-3-acetic acid (IAA) could induce shoots when on basal Murashige and Skoog (MS) medium. BA and TDZ could induce more adventitious shoots (19.1 and 31.2/explant, respectively) than NAA (4.6/explant), IBA (5.7/explant) or IAA (6.4/explant). BA and TDZ at 5–10 μM could induce both shoots and somatic embryos. A higher concentration of TDZ (25 μM) induced only somatic embryos (39.8/explant). The same concentration of BA induced both adventitious shoots (23.6/explant) and somatic embryos (9.7/explant). Thus, somatic embryogenesis in this plant needs a high cytokinin concentration (BA; TDZ), as evidenced by histology. Somatic embryos germinated easily when left on the same media, but formed adventitious roots in two weeks on MS supplemented with 0.5 μM NAA, 0.5 μM IBA and 0.1% activated charcoal. Over 93% of plantlets survived following acclimatization and transfer to a mixture of sand and vermiculite (1:1, v/v) in trays. PMID:27090564

  6. Decoding regulatory landscape of somatic embryogenesis reveals differential regulatory networks between japonica and indica rice subspecies

    Science.gov (United States)

    Indoliya, Yuvraj; Tiwari, Poonam; Chauhan, Abhisekh Singh; Goel, Ridhi; Shri, Manju; Bag, Sumit Kumar; Chakrabarty, Debasis

    2016-01-01

    Somatic embryogenesis is a unique process in plants and has considerable interest for biotechnological application. Compare to japonica, indica rice has been less responsive to in vitro culture. We used Illumina Hiseq 2000 sequencing platform for comparative transcriptome analysis between two rice subspecies at six different developmental stages combined with a tag-based digital gene expression profiling. Global gene expression among different samples showed greater complexity in japonica rice compared to indica which may be due to polyphyletic origin of two rice subspecies. Expression pattern in initial stage indicate major differences in proembryogenic callus induction phase that may serve as key regulator to observe differences between both subspecies. Our data suggests that phytohormone signaling pathways consist of elaborate networks with frequent crosstalk, thereby allowing plants to regulate somatic embryogenesis pathway. However, this crosstalk varies between the two rice subspecies. Down regulation of positive regulators of meristem development (i.e. KNOX, OsARF5) and up regulation of its counterparts (OsRRs, MYB, GA20ox1/GA3ox2) in japonica may be responsible for its better regeneration and differentiation of somatic embryos. Comprehensive gene expression information in the present experiment may also facilitate to understand the monocot specific meristem regulation for dedifferentiation of somatic cell to embryogenic cells. PMID:26973288

  7. Use of genomic DNA control features and predicted operon structure in microarray data analysis: ArrayLeaRNA – a Bayesian approach

    Directory of Open Access Journals (Sweden)

    Pin Carmen

    2007-11-01

    Full Text Available Abstract Background Microarrays are widely used for the study of gene expression; however deciding on whether observed differences in expression are significant remains a challenge. Results A computing tool (ArrayLeaRNA has been developed for gene expression analysis. It implements a Bayesian approach which is based on the Gumbel distribution and uses printed genomic DNA control features for normalization and for estimation of the parameters of the Bayesian model and prior knowledge from predicted operon structure. The method is compared with two other approaches: the classical LOWESS normalization followed by a two fold cut-off criterion and the OpWise method (Price, et al. 2006. BMC Bioinformatics. 7, 19, a published Bayesian approach also using predicted operon structure. The three methods were compared on experimental datasets with prior knowledge of gene expression. With ArrayLeaRNA, data normalization is carried out according to the genomic features which reflect the results of equally transcribed genes; also the statistical significance of the difference in expression is based on the variability of the equally transcribed genes. The operon information helps the classification of genes with low confidence measurements. ArrayLeaRNA is implemented in Visual Basic and freely available as an Excel add-in at http://www.ifr.ac.uk/safety/ArrayLeaRNA/ Conclusion We have introduced a novel Bayesian model and demonstrated that it is a robust method for analysing microarray expression profiles. ArrayLeaRNA showed a considerable improvement in data normalization, in the estimation of the experimental variability intrinsic to each hybridization and in the establishment of a clear boundary between non-changing and differentially expressed genes. The method is applicable to data derived from hybridizations of labelled cDNA samples as well as from hybridizations of labelled cDNA with genomic DNA and can be used for the analysis of datasets where

  8. EVALUATION OF THE FLOOD POTENTIAL OF THE SOUTH HOUSE (BLINEBRY) FIELD, LEA COUNTY, NEW MEXICO

    Energy Technology Data Exchange (ETDEWEB)

    L. Stephen Melzer

    2000-12-01

    The Blinebry (Permian) formation of eastern Lea County, NM has a long history of exploitation for petroleum and continues even today to be a strong target horizon for new drilling in the Permian Basin. Because of this long-standing interest it should be classified of strategic interest to domestic oil production; however, the formation has gained a reputation as a primary production target with limited to no flooding potential. In late May of 1999, a project to examine the feasibility of waterflooding the Blinebry formation was proposed to the U.S. Department of Energy's National Petroleum Technology Office (Tulsa, OK). A new well was proposed in one region (the South House area) to examine the reputation by acquiring core and borehole logging data for the collection of formation property data in order to conduct the waterflood evaluation. Notice of the DOE award was received on August 19, 1999 and the preparations for drilling, coring and logging were immediately made for a drilling start on 9/9/99. The Blinebry formation at 6000 feet, foot depth was reached on 9/16/99 and the coring of two 60 foot intervals of the Blinebry was completed on 9/19/99 with more than 98% core recovery. The well was drilled to a total depth of 7800 feet and the Blinebry interval was logged with spectral gamma ray, photoelectric cross section, porosity, resistivity, and borehole image logs on 8/24/99. The well was determined to be likely productive from the Blinebry interval and five & 1/2 inch casing was cemented in the hole on 9/25/99. Detailed core descriptions including environment of deposition have been accomplished. Whole core (a 4-inch diameter) and plug (1.5 inch diameter) testing for formation properties has been completed and are reported. Acquisition and analysis of the borehole logging results have been completed and are reported. Perforation of the Blinebry intervals was accomplished on November 8, 1999. The intervals were acidized and hydrofraced on 11/9 and 11

  9. EFFECT OF GLUTAMATE MANGANESE ON THE REPRODUCTIVE FUNCTION AND EMBRYOGENESIS OF FEMALE RATS

    Directory of Open Access Journals (Sweden)

    O. V. Shtapenko

    2015-10-01

    Full Text Available Pregnancy is associated with increased nutritional needs due to the physiologic changes of the female and the metabolic demands of the embryo/fetus. The use of chelating compounds with high biological activity increased the fertility of female rats by stimulation metabolism and functional activity of the reproductive system. Manganese is an essential element utilized by antioxidants, including superoxide dismutase (MnSOD, and others metalloenzymes that take part in reduction reactions, in multiple physiological processes including reproductive system. The aim of the research was to determine the effect of subcutaneous injections of Mn glutamate in liposomal forms on the reproductive system and the process of embryogenesis of experimental female rat. The study was conducted on female rats aged 2.5-3 months with body weight of 180-200 g. Rats with dated gestation were divided into three groups: two experimental and control. Female rabbits of the 1th experimental group were subcutaneous injected of 2 mg/ml for Mn glutamate in liposomal form one week before fertilization and the animals of the 2th group were obtained the same preparation during fertilization. Rats were euthanized on the twentieth day, ovariens were singled out of fiber, visual inspection and counting the number of yellow bodies were performed, absolute and relative mass indexes were determined, Mn glutamate influence index was calculated. Experimental results showed that the administration of Mn glutamate 7 days before fertilization and during fertilization significant increased the number of corpora lutea of pregnancy (p<0,001, number of live fetuses (p<0,001 due to the decrease in general and pre-implantation embryonic mortality compared with the control group. The experimental results showed improvement in key indicators of embryonic development. We observed significant increase in the number of implanted embryos on 1 female at 10,7±0,26 and 11,3±0,21 (p<0,001 (9,4±0,16 vs

  10. The function of nuclear receptors in regulation of female reproduction and embryogenesis in the red flour beetle, Tribolium castaneum.

    Science.gov (United States)

    Xu, Jingjing; Tan, Anjiang; Palli, Subba R

    2010-10-01

    Nineteen canonical and two Knirps-like family nuclear receptors (NRs) were identified in the genome of Tribolium castaneum. The current study was conducted to determine the function of these NRs in regulation of female reproduction and embryogenesis. RNA interference (RNAi)-aided knock-down in the expression of genes coding for 21 NRs showed that seven NRs E75, hormone receptor 3 (HR3), ecdysone receptor (EcR), ultraspiracle (USP), seven-up (SVP), FTZ transcription factor 1 (FTZ-F1) and hormone receptor 4 (HR4) are required for successful vitellogenesis and oogenesis. Knocking down the expression of genes coding for these seven NRs affected egg production by reducing the levels of vitellogenin mRNAs as well as by affecting the oocyte maturation. Expression of seven additional NRs hormone receptor 96 (HR96), hormone receptor 51 (HR51), hormone receptor 38 (HR38), hormone receptor 39 (HR39), Tailless (Tll), Dissatisfaction (Dsf) and Knirps-like is required for successful embryogenesis. The knock-down in the expression of genes coding for three other NRs (E78, hepatocyte nuclear factor 4, HNF4 and Eagle) partially blocked embryogenesis. This study showed that at least 17 out of the 21 NRs identified in T. castaneum play key roles in female reproduction and embryogenesis. Copyright 2010 Elsevier Ltd. All rights reserved.

  11. A relationship between seed development, Arabinogalactan-proteins (AGPs) and the AGP mediated promotion of somatic embryogenesis

    NARCIS (Netherlands)

    Hengel, van A.J.; Kammen, van A.; Vries, de S.C.

    2002-01-01

    Arabinogalactan-protein (AGP) epitopes are known to display developmentally regulated patterns of expression in several plant tissues. Therefore, AGPs have been suggested to play a role in plant development. Somatic embryogenesis is regulated by AGPs as well as by EP3 endochitinases. Using four

  12. DNA methyltransferase expressions in Japanese rice fish (Oryzias latipes) embryogenesis is developmentally regulated and modulated by ethanol and 5-azacytidine

    Science.gov (United States)

    We aimed to investigate the impact of the epigenome in inducting fetal alcohol spectrum disorder (FASD) phenotypes in Japanese rice fish embryogenesis. One of the significant events in epigenome is DNA methylation which is catalyzed by DNA methyl transferase (DNMT) enzymes. We analyzed DNMT enzyme m...

  13. Developmental regulation of neuroligin genes in Japanese rice fish (oryzias latipes) embryogenesis maintains the rhythym during ethanol-in

    Science.gov (United States)

    Although prenatal alcohol exposure is the potential cause of fetal alcohol spectrum disorder (FASD) in humans, the molecular mechanism(s) of FASD is yet unknown. We have used Japanese rice fish (Oryzias latipes) embryogenesis as an animal model of FASD and reported that this model has effectively ge...

  14. Bottlenecks in bog pine multiplication by somatic embryogenesis and their visualization with the environmental scanning electron microscope

    Czech Academy of Sciences Publication Activity Database

    Vlašínová, H.; Neděla, Vilém; Dordevic, B.; Havel, J.

    2017-01-01

    Roč. 254, č. 4 (2017), s. 1487-1497 ISSN 0033-183X R&D Projects: GA ČR(CZ) GA14-22777S Institutional support: RVO:68081731 Keywords : somatic embryogenesis * pinus uncinata subsp uliginosa * abnormalities * environmental scanning electron microscope Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 2.870, year: 2016

  15. Somatic embryogenesis from corolla tubes of interspecific amphiploids between cultivated sunflower (Helianthus annuus L.) and its wild species

    Science.gov (United States)

    Somatic embryogenesis in vitro provides an efficient means of plant multiplication, facilitating sunflower improvement and germplasm innovation. In the present study, using interspecific amphiploids (2n=4x=68) between cultivated sunflower and wild perennial Helianthus species as explant donors, soma...

  16. Loss of CMD2‐mediated resistance to cassava mosaic disease in plants regenerated through somatic embryogenesis

    Science.gov (United States)

    Chauhan, Raj Deepika; Wagaba, Henry; Moll, Theodore; Alicai, Titus; Miano, Douglas; Carrington, James C.; Taylor, Nigel J.

    2016-01-01

    Summary Cassava mosaic disease (CMD) and cassava brown streak disease (CBSD) are the two most important viral diseases affecting cassava production in Africa. Three sources of resistance are employed to combat CMD: polygenic recessive resistance, termed CMD1, the dominant monogenic type, named CMD2, and the recently characterized CMD3. The farmer‐preferred cultivar TME 204 carries inherent resistance to CMD mediated by CMD2, but is highly susceptible to CBSD. Selected plants of TME 204 produced for RNA interference (RNAi)‐mediated resistance to CBSD were regenerated via somatic embryogenesis and tested in confined field trials in East Africa. Although micropropagated, wild‐type TME 204 plants exhibited the expected levels of resistance, all plants regenerated via somatic embryogenesis were found to be highly susceptible to CMD. Glasshouse studies using infectious clones of East African cassava mosaic virus conclusively demonstrated that the process of somatic embryogenesis used to regenerate cassava caused the resulting plants to become susceptible to CMD. This phenomenon could be replicated in the two additional CMD2‐type varieties TME 3 and TME 7, but the CMD1‐type cultivar TMS 30572 and the CMD3‐type cultivar TMS 98/0505 maintained resistance to CMD after passage through somatic embryogenesis. Data are presented to define the specific tissue culture step at which the loss of CMD resistance occurs and to show that the loss of CMD2‐mediated resistance is maintained across vegetative generations. These findings reveal new aspects of the widely used technique of somatic embryogenesis, and the stability of field‐level resistance in CMD2‐type cultivars presently grown by farmers in East Africa, where CMD pressure is high. PMID:26662210

  17. Proteomic identification of differentially expressed proteins during the acquisition of somatic embryogenesis in oil palm (Elaeis guineensis Jacq.).

    Science.gov (United States)

    Silva, Rafael de Carvalho; Carmo, Lilian Silveira Travassos; Luis, Zanderluce Gomes; Silva, Luciano Paulino; Scherwinski-Pereira, Jonny Everson; Mehta, Angela

    2014-06-02

    In the present study we have identified and characterized the proteins expressed during different developmental stages of Elaeis guineensis calli obtained from zygotic embryos. We were interested in the possible proteomic changes that would occur during the acquisition of somatic embryogenesis and therefore samples were collected from zygotic embryos (E1), swollen explants 14days (E2) in induction medium, primary callus (E3), and pro-embryogenic callus (E4). The samples were grinded in liquid nitrogen, followed by total protein extraction using phenol and extraction buffer. Proteins were analyzed by two-dimensional electrophoresis (2-DE) and the differentially expressed protein spots were analyzed by MALDI-TOF mass spectrometry (MS and MS/MS). Interestingly, we have identified proteins, which can be used as potential candidates for future studies aiming at the development of biomarkers for embryogenesis acquisition and for the different stages leading to pro-embryogenic callus formation such as type IIIa membrane protein cp-wap13, fructokinase and PR proteins. The results obtained shed some light on the biochemical events involved in the process of somatic embryogenesis of E. guineensis obtained from zygotic embryos. The use of stage-specific protein markers can help monitor cell differentiation and contribute to improve the protocols for successfully cloning the species. Understanding the fate and dynamics of cells and tissues during callus formation is essential to understand totipotency and the mechanisms involved during acquisition of somatic embryogenesis (SE). In this study we have investigated the early stages of somatic embryogenesis induction in oil palm and have identified potential markers as well as proteins potentially involved in embryogenic competence acquisition. The use of these proteins can help improve tissue culture protocols in order to increase regeneration rates. This article is part of a Special Issue entitled: Environmental and structural

  18. 5-azacytidine promotes microspore embryogenesis initiation by decreasing global DNA methylation, but prevents subsequent embryo development in rapeseed and barley

    Directory of Open Access Journals (Sweden)

    María-Teresa eSolís

    2015-06-01

    Full Text Available Microspores are reprogrammed by stress in vitro towards embryogenesis. This process is an important tool in breeding to obtain double-haploid plants. DNA methylation is a major epigenetic modification that changes in differentiation and proliferation. We have shown changes in global DNA methylation during microspore reprogramming. 5-Azacytidine (AzaC cannot be methylated and leads to DNA hypomethylation. AzaC is a useful demethylating agent to study DNA dynamics, with a potential application in microspore embryogenesis. This work analyzes the effects of short and long AzaC treatments on microspore embryogenesis initiation and progression in two species, the dicot Brassica napus and the monocot Hordeum vulgare. This involved the quantitative analyses of proembryo and embryo production, the quantification of DNA methylation, 5mdC immunofluorescence and confocal microscopy, and the analysis of chromatin organization (condensation/ decondensation by light and electron microscopy. Four days of AzaC treatments (2.5 µM increased embryo induction, response associated with a decrease of DNA methylation, modified 5mdC and heterochromatin patterns compared to untreated embryos. By contrast, longer AzaC treatments diminished embryo production. Similar effects were found in both species, indicating that DNA demethylation promotes microspore reprogramming, totipotency acquisition and embryogenesis initiation, while embryo differentiation requires de novo DNA methylation and is prevented by AzaC. This suggests a role for DNA methylation in the repression of microspore reprogramming and possibly totipotency acquisition.Results provide new insights into the role of epigenetic modifications in microspore embryogenesis and suggest a potential benefit of inhibitors, such as AzaC, to improve the process efficiency in biotechnology and breeding programs.

  19. Enhanced Indirect Somatic Embryogenesis of Date Palm Using Low Levels of Seawater.

    Science.gov (United States)

    Taha, Rania A

    2017-01-01

    Date palm tolerates salinity, drought, and high temperatures. Arid and semiarid zones, especially the Middle East region, need a huge number of date palms for cultivation. To meet this demand, tissue culture techniques have great potential for mass production of plantlets, especially using the indirect embryogenesis technique; any improvement of these techniques is a worthy objective. Low levels of salinity can enhance growth and development of tolerant plants. A low level of seawater, a natural source of salinity, reduces the time required for micropropagation processes of date palm cv. Malkaby when added to MS medium. Medium containing seawater at 500 ppm total dissolved solid (TDS) (12.2 mL/L) improves callus proliferation, whereas 1500 ppm (36.59 mL/L) enhances plant regeneration including multiplication of secondary embryos, embryo germination, and rooting.

  20. Microspore Embryogenesis Through Anther Culture in Citrus clementina Hort. ex Tan.

    Science.gov (United States)

    Chiancone, Benedetta; Germanà, Maria Antonietta

    2016-01-01

    Anther culture is a biotechnological method that allows to obtain, in one step, homozygous plants, very important to plant breeding, due to their numerous applications in mutation research, selection, genome sequencing, genetic analysis, and transformation. To induce the microspores, i.e., the immature male gametes, to switch from the normal gametophytic pathway to the sporophytic one, it is necessary to submit them to a type of stress, such as high or low temperature, starvation, or magnetic field. Stress can be applied to the donor plants and/or the floral buds or the anthers or the isolated microspores, before or during the culture. In this chapter, the protocol to induce gametic embryogenesis from anther culture of several cultivars of Citrus clementina Hort. ex Tan. is reported.

  1. Reduction in chromosome mobility accompanies nuclear organization during early embryogenesis in Caenorhabditis elegans.

    Science.gov (United States)

    Arai, Ritsuko; Sugawara, Takeshi; Sato, Yuko; Minakuchi, Yohei; Toyoda, Atsushi; Nabeshima, Kentaro; Kimura, Hiroshi; Kimura, Akatsuki

    2017-06-16

    In differentiated cells, chromosomes are packed inside the cell nucleus in an organised fashion. In contrast, little is known about how chromosomes are packed in undifferentiated cells and how nuclear organization changes during development. To assess changes in nuclear organization during the earliest stages of development, we quantified the mobility of a pair of homologous chromosomal loci in the interphase nuclei of Caenorhabditis elegans embryos. The distribution of distances between homologous loci was consistent with a random distribution up to the 8-cell stage but not at later stages. The mobility of the loci was significantly reduced from the 2-cell to the 48-cell stage. Nuclear foci corresponding to epigenetic marks as well as heterochromatin and the nucleolus also appeared around the 8-cell stage. We propose that the earliest global transformation in nuclear organization occurs at the 8-cell stage during C. elegans embryogenesis.

  2. Dual lineage-specific expression of Sox17 during mouse embryogenesis

    DEFF Research Database (Denmark)

    Choi, Eunyoung; Kraus, Marine R C; Lemaire, Laurence A

    2012-01-01

    Sox17 is essential for both endoderm development and fetal hematopoietic stem cell (HSC) maintenance. While endoderm-derived organs are well known to originate from Sox17-expressing cells, it is less certain whether fetal HSCs also originate from Sox17-expressing cells. By generating a Sox17(GFPCre......) allele and using it to assess the fate of Sox17-expressing cells during embryogenesis, we confirmed that both endodermal and a part of definitive hematopoietic cells are derived from Sox17-positive cells. Prior to E9.5, the expression of Sox17 is restricted to the endoderm lineage. However, at E9.5 Sox17...... is expressed in the endothelial cells (ECs) at the para-aortic splanchnopleural region that contribute to the formation of HSCs at a later stage. The identification of two distinct progenitor cell populations that express Sox17 at E9.5 was confirmed using fluorescence-activated cell sorting together with RNA...

  3. THE EFFECT OF PICLORAM AND LIGHT ON SOMATIC EMBRYOGENESIS REGENERATION OF PINEAPPLE

    Directory of Open Access Journals (Sweden)

    Ika Roostika

    2012-10-01

    Full Text Available Smooth Cayenne is the largest pineapple type cultivated in Indonesia, but its vegetative planting materials for mass propagation are limited. Somatic embryogenesis is a potential method to be applied. The aim of this study was to investigate the somatic embryogenesis regeneration under the effect of picloram and light. Callus formation was induced by picloram (21, 41 and 62 μM added with 9 μM thidiazuron. The calli were transferred onto MS or Bac medium  enriched with N-organic compounds with or without addition of 21 μM picloram under dark or light condition. The compact calli were subcultured onto MS medium supplemented with 4.65 μM kinetin, while the friable calli were  transferred onto BIG medium (modified MS + 1.1 μM benzyl adenine + 0.9 μM indole butyric acid + 0.09 μM giberelic acid or B medium (MS + 0.018 mM benzyl adenine. The results showed that the events of somatic embryogenesis were started from cell polarization, asymmetrical division, proembryo formation as  embryogenic tissues and friable embryogenic tissues, and embryo development. The best treatment for callus induction was 21 μM picloram. The addition of 21 μM picloram on N-organic enriched medium and the use of light condition  proliferated embryogenic calli. The N-organic enriched Bac medium and light condition yielded the highest number of mature somatic embryos (17 embryos perexplant in 2 months. The B medium was better than BIG medium to develop  somatic embryos from friable embryogenic tissues. The somatic embryogenesis method presented is potential for pineapple mass propagation and artificial seedproduction.Abstrak Bahasa IndonesiaSmooth Cayenne merupakan kultivar nenas yang banyak dibudidayakan di  Indonesia, namun ketersediaan benih untuk perbanyakan massal masih terbatas. Embriogenesis somatikadalah metode yang potensial untuk produksi bibit secara massal. Tujuan penelitian adalah untuk mempelajari pengaruh pikloram dan pencahayaan terhadap regenerasi

  4. Histological Evidence of Indirect Somatic Embryogenesis from Immature Female Date Palm Inflorescences.

    Science.gov (United States)

    Zayed, Eman M M; Abdelbar, Ola H

    2017-01-01

    Rapid production of somatic embryogenesis and date palm regeneration is achieved by culturing immature female inflorescence explants. Inflorescence explants are soft, creamy in color, average 6-7 cm in length, and cultured on Murashige and Skoog (MS) medium containing 1 mg/L thidiazuron (TDZ). Callus induction occurs after 4-5 weeks of culture on the callus induction medium. Subsequently, callus develops embryogenic calli on MS medium supplemented with 0.1 mg/L naphthalene acetic acid (NAA). Histological samples were collected successively at the culturing time and during morphogenetic changes throughout the developmental stages of somatic embryos. Initiation of callus and different successive developmental stages for somatic embryos including two-celled, four-celled, globular, bipolar, and fully developed cotyledonary somatic embryos were observed. Mature somatic embryos develop within 10-12 weeks after culture establishment.

  5. Establishment and maintenance of heritable chromatin structure during early Drosophila embryogenesis.

    Science.gov (United States)

    Blythe, Shelby A; Wieschaus, Eric F

    2016-11-23

    During embryogenesis, the initial chromatin state is established during a period of rapid proliferative activity. We have measured with 3-min time resolution how heritable patterns of chromatin structure are initially established and maintained during the midblastula transition (MBT). We find that regions of accessibility are established sequentially, where enhancers are opened in advance of promoters and insulators. These open states are stably maintained in highly condensed mitotic chromatin to ensure faithful inheritance of prior accessibility status across cell divisions. The temporal progression of establishment is controlled by the biological timers that control the onset of the MBT. In general, acquisition of promoter accessibility is controlled by the biological timer that measures the nucleo-cytoplasmic (N:C) ratio, whereas timing of enhancer accessibility is regulated independently of the N:C ratio. These different timing classes each associate with binding sites for two transcription factors, GAGA-factor and Zelda, previously implicated in controlling chromatin accessibility at ZGA.

  6. The polarity protein Par6 is coupled to the microtubule network during molluscan early embryogenesis

    Energy Technology Data Exchange (ETDEWEB)

    Homma, Taihei [Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan); Shimizu, Miho [Kuroda Chiromorphology Team, ERATO-SORST, JST, Komaba, Meguro-ku, Tokyo 153-8902 (Japan); Kuroda, Reiko, E-mail: ckuroda@mail.ecc.u-tokyo.ac.jp [Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan); Kuroda Chiromorphology Team, ERATO-SORST, JST, Komaba, Meguro-ku, Tokyo 153-8902 (Japan); Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Komaba, Meguro-ku, Tokyo 153-8902 (Japan)

    2011-01-07

    Research highlights: {yields} The cDNAs encoding Par6 and aPKC homologues were cloned from the snail Lymnaea stagnalis. {yields} L. stagnalis Par6 directly interacts with tubulin and microtubules and localizes to the microtubule cytoskeleton during the early embryogenesis. {yields} Identical sequence and localization of LsPar6 for the dextral and the sinistral snails exclude the possibility of the gene being the primary determinant of body handedness. -- Abstract: Cell polarity, which directs the orientation of asymmetric cell division and segregation of fate determinants, is a fundamental feature of development and differentiation. Regulators of polarity have been extensively studied, and the critical importance of the Par (partitioning-defective) complex as the polarity machinery is now recognized in a wide range of eukaryotic systems. The Par polarity module is evolutionarily conserved, but its mechanism and cooperating factors vary among different systems. Here we describe the cloning and characterization of a pond snail Lymnaea stagnalis homologue of partitioning-defective 6 (Lspar6). The protein product LsPar6 shows high affinity for microtubules and localizes to the mitotic apparatus during embryonic cell division. In vitro assays revealed direct binding of LsPar6 to tubulin and microtubules, which is the first evidence of the direct interaction between the two proteins. The interaction is mediated by two distinct regions of LsPar6 both located in the N-terminal half. Atypical PKC, a functional partner of Par6, was also found to localize to the mitotic spindle. These results suggest that the L. stagnalis Par complex employs the microtubule network in cell polarity processes during the early embryogenesis. Identical sequence and localization of LsPar6 for the dextral and the sinistral snails exclude the possibility of the gene being the primary determinant of handedness.

  7. Effects of cadmium on chick embryogenesis and some comparisons with lead

    Energy Technology Data Exchange (ETDEWEB)

    King, D.W.; Chen, D.C.C.; Hsu, J.L.

    1978-07-01

    During the last ten years because of the severity of the problem of pollution and the part that heavy metals play in it we have been doing research on the effects of some heavy metals on chick embryogenesis in order to get a comparative study and to elucidate their mechanisms of action. Experiments were performed using 431 fertilized white Leghorn eggs to study the effect of cadmium on chick embryogenesis. Cadmium acetate at 0.015, 0.030, 0.045, 0.060, 0.12 or 0.24 mg/egg and lead acetate at 0.02, 0.04 or 0.075 mg/egg was injected in ovo on the fourth day of incubation. The embryos were taken out on the 19th day and examined for gross defects. Electrocardiograms were recorded on some embryos. Hemoglobin determinations were done on others. The changes in plasma delta-aminolevulinic acid dehydrase (ALAD) of the embryos due to cadmium and lead acetate were also determined. It was found that the LD50 of cadmium acetate was close to 0.045 mg. The highest incidence of abnormality, 30.9% of the surviving embryos, appeared in the 0.030 mg group although malformed embryos were also found in the 0.015, 0.045 and 0.060 mg groups. The most common malformations occurred in the liver (58%) and the cardiovascular system, with edema totalling over 90%. Lesser abnormalities were observed in the limbs. Lead acetate affected ALAD more than cadmium acetate. There was no significant difference on hemoglobin concentration or EKG between the distilled water control and either the cadmium or lead treated groups. Thus, embryolethality, embryotoxicity, congenital abnormalities and changes in ALAD were all observed in the cadmium-treated chick embryos although lead acetate seemed to inhibit the ALAD activity more effectively than cadmium acetate.

  8. Cefotaxime prevents microbial contamination and improves microspore embryogenesis in wheat and triticale.

    Science.gov (United States)

    Asif, Muhammad; Eudes, François; Randhawa, Harpinder; Amundsen, Eric; Yanke, Jay; Spaner, Dean

    2013-10-01

    Cefotaxime (100 mg/l) mitigate occasional gram negative bacterial contamination in wheat and triticale microspore culture and most importantly it increases cell growth and green plant production. Isolated microspore culture is a promising option to rapidly fix the product of meiotic recombination of F1 hybrids, in the process of varietal development. Clean culture and high embryogenesis rate are essential to commercial triticale and wheat microspore cultures. So, this study investigated (1) contaminants from isolated microspores cultures, (2) two antibiotics to control bacterial growth, and (3) the contribution of antibiotics to increased microspore-derived embryo-like structures (ELS), green and albino plants. Five species of bacteria were identified in contaminated cultures (Erwinia aphidicola, Pantoea agglomerans, Pseudomonas sp., Staphylococcus epidermis and Staphylococcus warneri) using fatty acid analysis and 16S ribosomal RNA sequences analysis, and yeast. Antibacterial susceptibility test using Cefotaxime and Vancomycin resulted in strong inhibition of 24 bacterial isolates, using Cefotaxime at 100 mg/l, but not Pseudomonas sp. Other antibiotic treatments inhibited bacterial growth at least partially. Microspore induction medium supplemented with the same antibiotics treatments resulted in successful microspore embryogenesis and green plant production. Antibiotic treatments were first tested in triticale and then validated in wheat cultivars AC Carberry and AC Andrew. Induction medium supplemented with Cefotaxime at 50 and 100 mg/l substantially increased the formation of ELS and green plants in triticale and wheat, respectively. Incidentally, it also affected the occurrence of albinism in all genotypes. Our results demonstrated dual purpose of Cefotaxime for isolated microspore culture, most importantly it increases cell growth and success of microspore cultures in triticale and wheat genotypes, but would also prevent accidental loss of cultures with

  9. Differential expression of ribosome-inactivating protein genes during somatic embryogenesis in spinach (Spinacia oleracea).

    Science.gov (United States)

    Kawade, Kensuke; Ishizaki, Takuma; Masuda, Kiyoshi

    2008-10-01

    Root segments from spinach (Spinacia oleracea L. cv. Jiromaru) seedlings form embryogenic callus (EC) that responded to exogenous GA(3) by accumulating a 31-kDa glycoprotein [BP31 or S. oleracea ribosome-inactivating protein (EC 3.2.2.22) (SoRIP1)] in association with the expression of embryogenic potential. Microsequencing of this protein revealed significant similarity with type 1 RIPs. We identified cDNAs for SoRIP1 and S. oleracea RIP2 (SoRIP2), a novel RIP having a consensus shiga/ricin toxic domain and performed a comparative analysis of the expression of SoRIPs during somatic embryogenesis. Western blotting and quantitative polymerase chain reaction analyses revealed that the expression of SoRIP1 in calli increased remarkably in association with the acquisition of embryogenic potential, although the expression in somatic embryos decreased moderately with their development. However, the expression of SoRIP2 in calli remained low and constant but increased markedly with the development of somatic embryos. Treatment of callus with GA(3) and/or ABA for 24 h, or with ABA for a longer period, failed to stimulate the expression of either gene. Immunohistochemistry showed that SoRIP1 preferentially accumulated in the proembryos and peripheral meristem of somatic embryos early in development. Appreciable expression of SoRIP2 was not detected in the callus, but intense expression was found in the epidermis of somatic embryos. These results suggest that the expression of spinach RIP genes is differentially regulated in a development-dependent fashion during somatic embryogenesis in spinach.

  10. Exploring nervous system transcriptomes during embryogenesis and metamorphosis in Xenopus tropicalis using EST analysis

    Directory of Open Access Journals (Sweden)

    Wegnez Maurice

    2007-05-01

    Full Text Available Abstract Background The western African clawed frog Xenopus tropicalis is an anuran amphibian species now used as model in vertebrate comparative genomics. It provides the same advantages as Xenopus laevis but is diploid and has a smaller genome of 1.7 Gbp. Therefore X. tropicalis is more amenable to systematic transcriptome surveys. We initiated a large-scale partial cDNA sequencing project to provide a functional genomics resource on genes expressed in the nervous system during early embryogenesis and metamorphosis in X. tropicalis. Results A gene index was defined and analysed after the collection of over 48,785 high quality sequences. These partial cDNA sequences were obtained from an embryonic head and retina library (30,272 sequences and from a metamorphic brain and spinal cord library (27,602 sequences. These ESTs are estimated to represent 9,693 transcripts derived from an estimated 6,000 genes. Comparison of these cDNA sequences with protein databases indicates that 46% contain their start codon. Further annotation included Gene Ontology functional classification, InterPro domain analysis, alternative splicing and non-coding RNA identification. Gene expression profiles were derived from EST counts and used to define transcripts specific to metamorphic stages of development. Moreover, these ESTs allowed identification of a set of 225 polymorphic microsatellites that can be used as genetic markers. Conclusion These cDNA sequences permit in silico cloning of numerous genes and will facilitate studies aimed at deciphering the roles of cognate genes expressed in the nervous system during neural development and metamorphosis. The genomic resources developed to study X. tropicalis biology will accelerate exploration of amphibian physiology and genetics. In particular, the model will facilitate analysis of key questions related to anuran embryogenesis and metamorphosis and its associated regulatory processes.

  11. Equine zona protein synthesis and ZP structure during folliculogenesis, oocyte maturation, and embryogenesis.

    Science.gov (United States)

    Kölle, Sabine; Dubois, Clotilde S; Caillaud, Maud; Lahuec, Cécile; Sinowatz, Fred; Goudet, Ghylène

    2007-07-01

    In the equine, the zona pellucida (ZP) is the major barrier to successful in vitro fertilization. Therefore the aim of our studies was to analyze species-specific features of the equine ZP in regard to structure and glycoprotein ZPB and ZPC expression sites during oocyte development and embryogenesis. The equine ZP revealed high immunological cross-reactivity to porcine ZPB and ZPC. In the ovary, the distribution of ZPB and ZPC was co-localized and correlated with the developmental stage of the follicle. ZPB and ZPC expression started in the oocyte of the late primordial and primary follicle. In the secondary follicle, both the oocyte and the cumulus cells contributed to ZPB and ZPC synthesis. After in vivo maturation the oocyte stopped ZPB and ZPC production whereas the cumulus cells continued synthesis. Contrary, in vitro matured (IVM) cumulus-oocyte-complexes (COCs) revealed a reverse expression pattern. This was correlated to alterations in the distribution, number, and size of pores in the ZP. In the zona, N-acetylglucosamine residues were co-localized with ZPC. The acellular glycoprotein capsule surrounding early equine embryos was negative for ZPB and ZPC. Our results imply that in the horse ZPB and ZPC glycoprotein expression is differentially regulated during folliculogenesis, oocyte maturation, and embryogenesis. Contrary to the bovine and porcine, zona protein synthesis during in vivo maturation is completely overtaken by the cumulus cells implying that in the horse these cells are crucial for zona integrity. During IVM, the cumulus cells lose their ability to synthesize glycoproteins leading to alterations in the zona structure.

  12. Plant Regeneration and Somatic Embryogenesis from Immature Embryos Derived through Interspecific Hybridization among Different Carica Species

    Directory of Open Access Journals (Sweden)

    Latifah Amin

    2012-12-01

    Full Text Available Plant regeneration and somatic embryogenesis through interspecific hybridization among different Carica species were studied for the development of a papaya ringspot virus-resistant variety. The maximum fruit sets were recorded from the cross of the native variety C. papaya cv. Shahi with the wild species C. cauliflora. The highest hybrid embryos were recorded at 90 days after pollination and the embryos were aborted at 150 days after pollination. The immature hybrid embryos were used for plant regeneration and somatic embryogenesis. The 90-day-old hybrid embryos from the cross of C. papaya cv. Shahi × C. cauliflora showed the highest percentage of germination, as well as plant regeneration on growth regulators free culture medium after 7 days pre-incubation on half-strength MS medium supplemented with 0.2 mg/L BAP, 0.5 mg/L NAA and 60 g/L sucrose. The 90-day-old hybrid embryos from the cross of C. papaya cv. Shahi × C. cauliflora produced maximum callus, as well as somatic embryos when cultured on half-strength MS medium containing 5 mg/L 2,4-D, 100 mg/L glutamine, 100 mg/L casein hydrolysate and 60 g/L sucrose. The somatic embryos were transferred into half-strength MS medium containing 0.5 mg/L BAP and 0.2 mg/L NAA and 60 g/L sucrose for maturation. The highest number of regenerated plants per hybrid embryo (10.33 was recorded from the cross of C. papaya cv. Shahi × C. cauliflora. Isoenzyme and dendrogram cluster analysis using UPGMA of the regenerated F1 plantlets confirmed the presence of the hybrid plantlets.

  13. Plant regeneration and somatic embryogenesis from immature embryos derived through interspecific hybridization among different Carica species.

    Science.gov (United States)

    Azad, Md Abul Kalam; Rabbani, Md Golam; Amin, Latifah

    2012-12-12

    Plant regeneration and somatic embryogenesis through interspecific hybridization among different Carica species were studied for the development of a papaya ringspot virus-resistant variety. The maximum fruit sets were recorded from the cross of the native variety C. papaya cv. Shahi with the wild species C. cauliflora. The highest hybrid embryos were recorded at 90 days after pollination and the embryos were aborted at 150 days after pollination. The immature hybrid embryos were used for plant regeneration and somatic embryogenesis. The 90-day-old hybrid embryos from the cross of C. papaya cv. Shahi × C. cauliflora showed the highest percentage of germination, as well as plant regeneration on growth regulators free culture medium after 7 days pre-incubation on half-strength MS medium supplemented with 0.2 mg/L BAP, 0.5 mg/L NAA and 60 g/L sucrose. The 90-day-old hybrid embryos from the cross of C. papaya cv. Shahi × C. cauliflora produced maximum callus, as well as somatic embryos when cultured on half-strength MS medium containing 5 mg/L 2,4-D, 100 mg/L glutamine, 100 mg/L casein hydrolysate and 60 g/L sucrose. The somatic embryos were transferred into half-strength MS medium containing 0.5 mg/L BAP and 0.2 mg/L NAA and 60 g/L sucrose for maturation. The highest number of regenerated plants per hybrid embryo (10.33) was recorded from the cross of C. papaya cv. Shahi × C. cauliflora. Isoenzyme and dendrogram cluster analysis using UPGMA of the regenerated F(1) plantlets confirmed the presence of the hybrid plantlets.

  14. Developmental toxicity of endocrine disruptors in early life stages of zebrafish, a genetic and embryogenesis study.

    Science.gov (United States)

    Santos, Dércia; Matos, Manuela; Coimbra, Ana M

    2014-01-01

    Endocrine disrupting compounds (EDCs) are capable of interfering with the endocrine system and are increasingly widespread in the aquatic environments. In the present study, zebrafish (Danio rerio) embryos and larvae were used to assess how EDCs may interfere with embryogenesis. Therefore, zebrafish embryos were exposed to 17α-ethinylestradiol (EE2: 0.4, 2, 4 and 20 ng/L), genistein (Gen: 2, 20, 200 and 2000 ng/L) and fadrozole (Fad: 2, 10, 50 and 250 μg/L), between 2 and 144 h post-fertilization (hpf). Somite development, heartbeat, malformations, mortality and hatching rates were evaluated. In parallel, the expression patterns of hormone receptors (esr1, esr2a, esr2b and ar) and apoptotic pathways related genes (p53 and c-jun) were determined using quantitative real-time PCR. Results showed that EE2, Gen and Fad caused a higher mortality and also malformations in larvae compared with control. A significant toxic effect was observed in the heartbeat rate, at 144 hpf, in larvae exposed to EE2 and Fad. QPCR revealed alterations in the expression levels of all the evaluated genes, at different time points. esr1 and c-jun genes were upregulated by EE2 and Gen exposure while the expression of esr2a, esr2b and ar genes was downregulated. Fad exposure decreased esr1, p53 and c-jun expression levels. This study shows a toxic effect of EE2, Gen and Fad to vertebrate embryogenesis and a relation between hormones action and apoptosis pathways. Copyright © 2014 Elsevier Inc. All rights reserved.

  15. Thermal manipulation during embryogenesis affects myoblast proliferation and skeletal muscle growth in meat-type chickens.

    Science.gov (United States)

    Piestun, Yogev; Yahav, Shlomo; Halevy, Orna

    2015-10-01

    Thermal manipulation (TM) of 39.5°C applied during mid-embryogenesis (embryonic d 7 to 16) has been proven to promote muscle development and enhance muscle growth and meat production in meat-type chickens. This study aimed to elucidate the cellular basis for this effect. Continuous TM or intermittent TM (for 12 h/d) increased myoblast proliferation manifested by higher (25 to 48%) myoblast number in the pectoral muscles during embryonic development but also during the first week posthatch. Proliferation ability of the pectoral-muscle-derived myoblasts in vitro was significantly higher in the TM treatments until embryonic d 15 (intermittent TM) or 13 (continuous TM) compared to that of controls, suggesting increased myogenic progeny reservoir in the muscle. However, the proliferation ability of myoblasts was lower in the TM treatments vs. control during the last days of incubation. This coincided with higher levels of myogenin expression in the muscle, indicating enhanced cell differentiation in the TM muscle. A similar pattern was observed posthatch: Myoblast proliferation was significantly higher in the TM chicks relative to controls during the peak of posthatch cell proliferation until d 6, followed by lower cell number 2 wk posthatch as myoblast number sharply decreases. Higher myogenin expression was observed in the TM chicks on d 6. This resulted in increased muscle growth, manifested by significantly higher relative weight of breast muscle in the embryo and posthatch. It can be concluded that temperature elevation during mid-term embryogenesis promotes myoblast proliferation, thus increasing myogenic progeny reservoir in the muscle, resulting in enhanced muscle growth in the embryo and posthatch. © 2015 Poultry Science Association Inc.

  16. Effect of pyriproxyfen on developing stages and embryogenesis of California red scale (CRS), Aonidiella aurantii.

    Science.gov (United States)

    Eliahu, Miriam; Blumberg, Daniel; Horowitz, A Rami; Ishaaya, Isaac

    2007-08-01

    During the past decade, the insect growth regulator pyriproxyfen has been the most used insecticide for controlling the California red scale (CRS), Aonidiella aurantii (Maskell), an important pest in citrus groves worldwide. The aim of the present research was to develop bioassays for determining the effect of pyriproxyfen on developing stages and embryogenesis of CRS that can serve as baselines for monitoring resistance. Pyriproxyfen was very effective against first- and second-instar nymphs of a susceptible strain of CRS; 0.1 and 0.2 mg AI L(-1), resulting in 80 and 100% mortality of first-instar nymphs, and in 62.5 and 91.7% mortality of second-instar nymphs respectively. According to probit analysis, the LC(50) and LC(90) values of pyriproxyfen for first-instar nymphs were 0.03 and 0.15 mg AI L(-1), and for second-instar nymphs they were 0.05 and 0.25 mg AI L(-1) respectively. Pyriproxyfen was quite effective on first-instar nymphs of a CRS field strain originating from a citrus grove where two successive applications of the compound had been applied. Mortality rates were 86 and 100% at concentrations of 0.2 and 0.4 mg AI L(-1) respectively. Pyriproxyfen suppressed embryogenesis when CRS females were treated with pyriproxyfen. Young CRS females treated with 1.0 and 5.0 mg AI L(-1) pyriproxyfen produced 6.6 N(1)/F and 2.4 N(1)/F as compared with 10.9 N(1)/F in untreated control. Copyright (c) 2007 Society of Chemical Industry

  17. Ethylene is integrated into the nitric oxide regulation of Arabidopsis somatic embryogenesis

    Directory of Open Access Journals (Sweden)

    Mohamed M. Mira

    2015-06-01

    Full Text Available The study confirms the role of the two Arabidopsis hemoglobin genes (Glb1 and Glb2 during somatic embryogenesis and proposes the involvement of ethylene in the regulation of embryo development. Suppression of both Glb1 and Glb2 results in accumulation of nitric oxide (NO and a different embryogenic response. Compared to WT tissue, down-regulation of Glb1 (Glb1 RNAi line compromises the embryogenic process, while repression of Glb2 (Glb2−/− line increases the number of embryos. These differences were ascribed to the differential accumulation of NO in the two lines, as Glb1 is a more effective NO scavenger compared to Glb2. A high elevation of NO level [achieved pharmacologically using the NO donor sodium nitroprusside (SNP, or genetically using the Glb1 suppressing line], activated the two ethylene biosynthetic genes 1-aminocyclopropane-1-carboxylate synthase (ACC synthase and 1-aminocyclopropane-1-carboxylate oxidase (ACC oxidase. Ethylene accumulation repressed embryogenesis, as shown by the decreased embryo number observed in tissue treated with the ethylene releasing agent Ethephon (ETH, as well as by the increased embryo production obtained with the two ethylene insensitive mutant lines (ein2-1 and ein3-1. A repression in ethylene level increased the expression of many auxin biosynthetic genes and favored the accumulation of the auxin indole-acetic acid (IAA at the sites of the explants where embryogenic tissue will form. Collectively these data reveal that high levels of NO, generated by the Glb1 suppressing line, but not by the Glb2 suppressing line, might increase the level of ethylene, which represses the production of auxin. Auxin is the inductive signal required for the formation of the embryogenic tissue.

  18. Ground-water investigations of the Project Gnome area, Eddy and Lea Counties, New Mexico

    Science.gov (United States)

    Cooper, J.B.

    1962-01-01

    The U.S. Atomic Energy Commission, through the Office of Test Operations, Albuquerque Operations Office, plans to detonate a nuclear device in a massive salt bed 1,200 feet beneath the land surface. The project, known as Project Gnome, is an element of the Plowshare program--a study of peacetime applications of nuclear fission. The location of the proposed underground shot is in a sparsely-populated area in southeastern Eddy County, N. Mex., east of the Pecos River and about 25 miles southeast of the city of Carlsbad. The area is arid to Semiarid and ground water is a vital factor in the economic utilization of the land, which is primarily used for stock raising. An investigation of the Project Gnome site and surrounding area for the purposes of evaluating the ground-water resources and the possible effect upon them from the detonation of the nuclear shot was desired by the Commission. This report describes work done by the U.S. Geological Survey on behalf of the Commission and presents results of the investigation of the ground-water resources and geology of the area. The most intensive investigations were made within a 15-mile radius of the site of Project Gnome and mainly on the east side of the Pecos River. The total area of study of over 1,200 square miles includes parts of Eddy and Lea Counties, N. Mex. The Project Gnome site is in the sedimentary Delaware Basin. It is underlain by about 18,000 feet of sedimentary rocks ranging in age from Ordovician to Recent. Upper Permian evaporitic rocks, which contain the principal source of potash available in the United States, are worked in nearby mines. The potash minerals are found in a massive salt bed about 1,400 feet thick in the Salado Formation of Permian age. The land surface of the area is covered mostly by a wind-blown sand and caliche; however, rocks of the Rustler Formation of Permian age and younger rocks of Permian, Triassic, Pleistocene(?) and Recent age crop out at several localities. Solution by

  19. NEFSC Survey Indices of Abundance

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Northeast Fisheries Survey Bottom trawl survey indices of abundance such as stratified mean number per tow or mean weight per tow by species stock. Includes indices...

  20. Line differences in Cor/Lea and fructan biosynthesis-related gene transcript accumulation are related to distinct freezing tolerance levels in synthetic wheat hexaploids.

    Science.gov (United States)

    Yokota, Hirokazu; Iehisa, Julio C M; Shimosaka, Etsuo; Takumi, Shigeo

    2015-03-15

    In common wheat, cultivar differences in freezing tolerance are considered to be mainly due to allelic differences at two major loci controlling freezing tolerance. One of the two loci, Fr-2, is coincident with a cluster of genes encoding C-repeat binding factors (CBFs), which induce downstream Cor/Lea genes during cold acclimation. Here, we conducted microarray analysis to study comprehensive changes in gene expression profile under long-term low-temperature (LT) treatment and to identify other LT-responsive genes related to cold acclimation in leaves of seedlings and crown tissues of a synthetic hexaploid wheat line. The microarray analysis revealed marked up-regulation of a number of Cor/Lea genes and fructan biosynthesis-related genes under the long-term LT treatment. For validation of the microarray data, we selected four synthetic wheat lines that contain the A and B genomes from the tetraploid wheat cultivar Langdon and the diverse D genomes originating from different Aegilops tauschii accessions with distinct levels of freezing tolerance after cold acclimation. Quantitative RT-PCR showed increased transcript levels of the Cor/Lea, CBF, and fructan biosynthesis-related genes in more freezing-tolerant lines than in sensitive lines. After a 14-day LT treatment, a significant difference in fructan accumulation was observed among the four lines. Therefore, the fructan biosynthetic pathway is associated with cold acclimation in development of wheat freezing tolerance and is another pathway related to diversity in freezing tolerance, in addition to the CBF-mediated Cor/Lea expression pathway. Copyright © 2014 Elsevier GmbH. All rights reserved.

  1. Comparative analysis of the heat stable proteome of radicles of Medicago truncatula seeds during germination identifies late embryogenesis abundant proteins associated with desiccation tolerance

    NARCIS (Netherlands)

    Boudet, J.; Buitink, J.; Hoekstra, F.A.; Rogniaux, H.; Larré, C.; Satour, P.; Leprince, O.

    2006-01-01

    A proteomic analysis was performed on the heat stable protein fraction of imbibed radicles of Medicago truncatula seeds to investigate whether proteins can be identified that are specifically linked to desiccation tolerance (DT). Radicles were compared before and after emergence (2.8 mm long) in

  2. Solar and stellar photospheric abundances

    Directory of Open Access Journals (Sweden)

    Carlos Allende Prieto

    2016-07-01

    Full Text Available Abstract The determination of photospheric abundances in late-type stars from spectroscopic observations is a well-established field, built on solid theoretical foundations. Improving those foundations to refine the accuracy of the inferred abundances has proven challenging, but progress has been made. In parallel, developments on instrumentation, chiefly regarding multi-object spectroscopy, have been spectacular, and a number of projects are collecting large numbers of observations for stars across the Milky Way and nearby galaxies, promising important advances in our understanding of galaxy formation and evolution. After providing a brief description of the basic physics and input data involved in the analysis of stellar spectra, a review is made of the analysis steps, and the available tools to cope with large observational efforts. The paper closes with a quick overview of relevant ongoing and planned spectroscopic surveys, and highlights of recent research on photospheric abundances.

  3. Identification of novel genes potentially involved in somatic embryogenesis in chicory (Cichorium intybus L.

    Directory of Open Access Journals (Sweden)

    Quillet Marie-Christine

    2010-06-01

    Full Text Available Abstract Background In our laboratory we use cultured chicory (Cichorium intybus explants as a model to investigate cell reactivation and somatic embryogenesis and have produced 2 chicory genotypes (K59, C15 sharing a similar genetic background. K59 is a responsive genotype (embryogenic capable of undergoing complete cell reactivation i.e. cell de- and re-differentiation leading to somatic embryogenesis (SE, whereas C15 is a non-responsive genotype (non-embryogenic and is unable to undergo SE. Previous studies 1 showed that the use of the β-D-glucosyl Yariv reagent (β-GlcY that specifically binds arabinogalactan-proteins (AGPs blocked somatic embryo production in chicory root explants. This observation indicates that β-GlcY is a useful tool for investigating somatic embryogenesis (SE in chicory. In addition, a putative AGP (DT212818 encoding gene was previously found to be significantly up-regulated in the embryogenic K59 chicory genotype as compared to the non-embryogenic C15 genotype suggesting that this AGP could be involved in chicory re-differentiation 2. In order to improve our understanding of the molecular and cellular regulation underlying SE in chicory, we undertook a detailed cytological study of cell reactivation events in K59 and C15 genotypes, and used microarray profiling to compare gene expression in these 2 genotypes. In addition we also used β-GlcY to block SE in order to identify genes potentially involved in this process. Results Microscopy confirmed that only the K59, but not the C15 genotype underwent complete cell reactivation leading to SE formation. β-GlcY-treatment of explants blocked in vitro SE induction, but not cell reactivation, and induced cell wall modifications. Microarray analyses revealed that 78 genes were differentially expressed between induced K59 and C15 genotypes. The expression profiles of 19 genes were modified by β-GlcY-treatment. Eight genes were both differentially expressed between K59 and C

  4. Estimating species abundance from occurrence

    OpenAIRE

    He, F.; Gaston, K.J.

    2000-01-01

    The number of individuals, or the abundance, of a species\\ud in an area is a fundamental ecological parameter and a\\ud critical consideration when making management and conservation decisions (Andrewartha and Birch 1954; Krebs\\ud 1978; Gaston 1994; Caughley and Gunn 1996). However,\\ud unless the scale is very fine or localized (e.g., in a measurable habitat or a forest stand), abundance is not readily determined. At coarse or regional scales for many species, information on commonness and rar...

  5. Comparison of the Lea Symbols and HOTV charts for preschool vision screening from age 3 to 4.5 years old

    Directory of Open Access Journals (Sweden)

    Ya-Hui Zhang

    2014-12-01

    Full Text Available AIM: To evaluate the applicability and the development of the normal visual acuity from age 3 to 3.5 years using Lea Symbols and HOTV charts.METHODS: It was a survey research study. Totally, 133 preschoolers(266 eyesbetween 3 to 4.5 years old recruited from two kid-gardens in Guangzhou were tested with both the Lea Symbols chart and the HOTV chart. Outcome measures were monocular logarithm of the minimum angle of resolution(logMARvisual acuity and inter-eye acuity difference in logMAR units for each test. RESULTS: The testability rates of the two charts were high(96.24% vs 92.48%, respectively, but difference was not statistically significant(P>0.05. The difference between the two kind of monocular vision was not statistically significant(the right eye: t=0.517, P=0.606; the left eye: t=-0.618, P=0.538. There was no significant difference between different eye(Lea Symbols chart: t=0.638, P=0.525; HOTV chart: t=-0.897, P=0.372. The visual acuities of the boys were better than those of the girls, but the difference was not statistically significant(P>0.05. The results which came from visual acuities with the two charts for the corresponding age groups(3-year-old group, 3.5-year-old group, 4-year-old group, 4.5-year-old groupindicated that the visual acuities of the preschoolers were improving with increasing age, but the difference among the visual acuities with the Lea Symbols chart was not statistically significant(the right eye: F=2.662, P=0.052; the left eye: F=1.850, P=0.143. However the difference among the visual acuities with the HOTV chart was statistically significant(the right eye: F=4.518, P=0.005; the left eye: F=3.893, P=0.011.CONCLUSION: Both Lea Symbols and HOTV chars are accepted and appropriate for preschool vision screening from 3 to 4.5 years old. The monocular visual acuity of preschoolers from age 3 to 4.5 years could be assessed was similar using the two charts. There is no correlation between visual acuity and different eye

  6. 3D early embryogenesis image filtering by nonlinear partial differential equations.

    Science.gov (United States)

    Krivá, Z; Mikula, K; Peyriéras, N; Rizzi, B; Sarti, A; Stasová, O

    2010-08-01

    We present nonlinear diffusion equations, numerical schemes to solve them and their application for filtering 3D images obtained from laser scanning microscopy (LSM) of living zebrafish embryos, with a goal to identify the optimal filtering method and its parameters. In the large scale applications dealing with analysis of 3D+time embryogenesis images, an important objective is a correct detection of the number and position of cell nuclei yielding the spatio-temporal cell lineage tree of embryogenesis. The filtering is the first and necessary step of the image analysis chain and must lead to correct results, removing the noise, sharpening the nuclei edges and correcting the acquisition errors related to spuriously connected subregions. In this paper we study such properties for the regularized Perona-Malik model and for the generalized mean curvature flow equations in the level-set formulation. A comparison with other nonlinear diffusion filters, like tensor anisotropic diffusion and Beltrami flow, is also included. All numerical schemes are based on the same discretization principles, i.e. finite volume method in space and semi-implicit scheme in time, for solving nonlinear partial differential equations. These numerical schemes are unconditionally stable, fast and naturally parallelizable. The filtering results are evaluated and compared first using the Mean Hausdorff distance between a gold standard and different isosurfaces of original and filtered data. Then, the number of isosurface connected components in a region of interest (ROI) detected in original and after the filtering is compared with the corresponding correct number of nuclei in the gold standard. Such analysis proves the robustness and reliability of the edge preserving nonlinear diffusion filtering for this type of data and lead to finding the optimal filtering parameters for the studied models and numerical schemes. Further comparisons consist in ability of splitting the very close objects which

  7. EFFECT OF GLUTAMATE MANGANESE ON THE REPRODUCTIVE FUNCTION AND EMBRYOGENESIS OF FEMALE RATS

    Directory of Open Access Journals (Sweden)

    Shtapenko O. V.

    2015-12-01

    Full Text Available Pregnancy is associated with increased nutritional needs due to the physiologic changes of the female and the metabolic demands of the embryo/fetus. The use of chelating compounds with high biological activity increased the fertility of female rats by stimulation metabolism and functional activity of the reproductive system. Manganese is an essential element utilized by antioxidants, including superoxide dismutase (MnSOD, and others metalloenzymes that take part in reduction reactions, in multiple physiological processes including reproductive system. The aim of the research was to determine the effect of subcutaneous injections of Mn glutamate in liposomal forms on the reproductive system and the process of embryogenesis of experimental female rat. The study was conducted on female rats aged 2.5-3 months with body weight of 180-200 g. Rats with dated gestation were divided into three groups: two experimental and control. Female rabbits of the 1th experimental group were subcutaneous injected of 2 mg/ml for Mn glutamate in liposomal form one week before fertilization and the animals of the 2th group were obtained the same preparation during fertilization. Rats were euthanized on the twentieth day, ovariens were singled out of fiber, visual inspection and counting the number of yellow bodies were performed, absolute and relative mass indexes were determined, Mn glutamate influence index was calculated. Experimental results showed that the administration of Mn glutamate 7 days before fertilization and during fertilization significant increased the number of corpora lutea of pregnancy (p<0,001, number of live fetuses (p<0,001 due to the decrease in general and pre-implantation embryonic mortality compared with the control group. The experimental results showed improvement in key indicators of embryonic development. We observed significant increase in the number of implanted embryos on 1 female at 10,7±0,26 and 11,3±0,21 (p<0,001 (9,4±0,16 vs

  8. Induction of cassava somatic embryogenesis in liquid medium associated to floating membrane rafts

    Directory of Open Access Journals (Sweden)

    Elizabete Keiko Takahashi

    2000-01-01

    Full Text Available The objective of this study was to examine the effect of two culture systems, liquid medium associated to floating membranes and solid medium, both supplemented with different concentrations of 2,4-D, in the induction of somatic embryogenesis of cassava (Manihot esculenta Crantz. Only 28% of the young leaf lobes (with 9 µM 2,4-D were induced to form organized embryogenic structures (OES with membrane rafts, compared to 50% of the explants presenting this type of tissue in solid medium with 36 µM of 2,4-D. Despite the lower response observed in liquid medium with membrane, the amount of OES/explant in all 2,4-D concentrations was higher than solid medium. Based on the results and considering the high cost of the membrane rafts, this system was not distinctly superior than solid medium for inducing somatic embryogenesis in cassava.O objetivo deste estudo foi comparar a indução de embriogênese somática em mandioca (Manihot esculenta Crantz utilizando o sistema de cultivo em meio líquido associado com membranas flutuantes com meio sólido, ambos suplementados com diferentes doses de 2,4-D. Utilizando membranas flutuantes, o melhor resultado foi obtido na concentração de 9 µM 2,4-D, onde apenas 28% dos explantes foliares apresentaram estruturas embriogênicas organizadas (OES. Por outro lado, em explantes cultivados em meio sólido suplementado com 36 µM de 2,4-D a frequência de OES foi de 50%. Embora a frequência de indução embriogênica tenha sido inferior em meio líquido associado com membranas flutuantes, a quantidade de OES por explante foi igual ou superior ao do meio sólido em todas as concentrações de 2,4-D testadas. Baseado nestes resultados, e considerando o elevado custo das membranas, este sistema de cultura não apresentou vantagens significativas para indução de embriogênese somática em mandioca em relação ao meio sólido.

  9. Characterization of expressed sequence tags obtained by SSH during somatic embryogenesis in Cichorium intybus L.

    Science.gov (United States)

    Legrand, Sylvain; Hendriks, Theo; Hilbert, Jean-Louis; Quillet, Marie-Christine

    2007-06-06

    Somatic embryogenesis (SE) is an asexual propagation pathway requiring a somatic-to-embryonic transition of differentiated somatic cells toward embryogenic cells capable of producing embryos in a process resembling zygotic embryogenesis. In chicory, genetic variability with respect to the formation of somatic embryos was detected between plants from a population of Cichorium intybus L. landrace Koospol. Though all plants from this population were self incompatible, we managed by repeated selfing to obtain a few seeds from one highly embryogenic (E) plant, K59. Among the plants grown from these seeds, one plant, C15, was found to be non-embryogenic (NE) under our SE-inducing conditions. Being closely related, we decided to exploit the difference in SE capacity between K59 and its descendant C15 to study gene expression during the early stages of SE in chicory. Cytological analysis indicated that in K59 leaf explants the first cell divisions leading to SE were observed at day 4 of culture. In contrast, in C15 explants no cell divisions were observed and SE development seemed arrested before cell reactivation. Using mRNAs isolated from leaf explants from both genotypes after 4 days of culture under SE-inducing conditions, an E and a NE cDNA-library were generated by SSH. A total of 3,348 ESTs from both libraries turned out to represent a maximum of 2,077 genes. In silico subtraction analysis sorted only 33 genes as differentially expressed in the E or NE genotype, indicating that SSH had resulted in an effective normalisation. Real-time RT-PCR was used to verify the expression levels of 48 genes represented by ESTs from either library. The results showed preferential expression of genes related to protein synthesis and cell division in the E genotype, and related to defence in the NE genotype. In accordance with the cytological observations, mRNA levels in explants from K59 and C15 collected at day 4 of SE culture reflected differential gene expression that presumably

  10. Characterization of expressed sequence tags obtained by SSH during somatic embryogenesis in Cichorium intybus L

    Directory of Open Access Journals (Sweden)

    Quillet Marie-Christine

    2007-06-01

    Full Text Available Abstract Background Somatic embryogenesis (SE is an asexual propagation pathway requiring a somatic-to-embryonic transition of differentiated somatic cells toward embryogenic cells capable of producing embryos in a process resembling zygotic embryogenesis. In chicory, genetic variability with respect to the formation of somatic embryos was detected between plants from a population of Cichorium intybus L. landrace Koospol. Though all plants from this population were self incompatible, we managed by repeated selfing to obtain a few seeds from one highly embryogenic (E plant, K59. Among the plants grown from these seeds, one plant, C15, was found to be non-embryogenic (NE under our SE-inducing conditions. Being closely related, we decided to exploit the difference in SE capacity between K59 and its descendant C15 to study gene expression during the early stages of SE in chicory. Results Cytological analysis indicated that in K59 leaf explants the first cell divisions leading to SE were observed at day 4 of culture. In contrast, in C15 explants no cell divisions were observed and SE development seemed arrested before cell reactivation. Using mRNAs isolated from leaf explants from both genotypes after 4 days of culture under SE-inducing conditions, an E and a NE cDNA-library were generated by SSH. A total of 3,348 ESTs from both libraries turned out to represent a maximum of 2,077 genes. In silico subtraction analysis sorted only 33 genes as differentially expressed in the E or NE genotype, indicating that SSH had resulted in an effective normalisation. Real-time RT-PCR was used to verify the expression levels of 48 genes represented by ESTs from either library. The results showed preferential expression of genes related to protein synthesis and cell division in the E genotype, and related to defence in the NE genotype. Conclusion In accordance with the cytological observations, mRNA levels in explants from K59 and C15 collected at day 4 of SE

  11. The Roles of the Wnt-Antagonists Axin and Lrp4 during Embryogenesis of the Red Flour Beetle Tribolium castaneum

    Directory of Open Access Journals (Sweden)

    Romy Prühs

    2017-10-01

    Full Text Available In both vertebrates and invertebrates, the Wnt-signaling pathway is essential for numerous processes in embryogenesis and during adult life. Wnt activity is fine-tuned at various levels by the interplay of a number of Wnt-agonists (Wnt ligands, Frizzled-receptors, Lrp5/6 coreceptors and Wnt-antagonists (among them Axin, Secreted frizzled and Lrp4 to define anterior–posterior polarity of the early embryo and specify cell fate in organogenesis. So far, the functional analysis of Wnt-pathway components in insects has concentrated on the roles of Wnt-agonists and on the Wnt-antagonist Axin. We depict here additional features of the Wnt-antagonist Axin in the flour beetle Tribolium castaneum. We show that Tc-axin is dynamically expressed throughout embryogenesis and confirm its essential role in head development. In addition, we describe an as yet undetected, more extreme Tc-axin RNAi-phenotype, the ectopic formation of posterior abdominal segments in reverse polarity and a second hindgut at the anterior. For the first time, we describe here that an lrp4 ortholog is involved in axis formation in an insect. The Tribolium Lrp4 ortholog is ubiquitously expressed throughout embryogenesis. Its downregulation via maternal RNAi results in the reduction of head structures but not in axis polarity reversal. Furthermore, segmentation is impaired and larvae develop with a severe gap-phenotype. We conclude that, as in vertebrates, Tc-lrp4 functions as a Wnt-inhibitor in Tribolium during various stages of embryogenesis. We discuss the role of both components as negative modulators of Wnt signaling in respect to axis formation and segmentation in Tribolium.

  12. Jasmonic acid is a downstream component in the modulation of somatic embryogenesis by Arabidopsis Class 2 phytoglobin

    Science.gov (United States)

    Mira, Mohamed M.; Wally, Owen S. D.; Elhiti, Mohamed; El-Shanshory, Adel; Reddy, Dhadi S.; Hill, Robert D.; Stasolla, Claudio

    2016-01-01

    Previous studies have shown that the beneficial effect of suppression of the Arabidopsis phytoglobin 2 gene, PGB2, on somatic embryogenesis occurs through the accumulation of nitric oxide (NO) within the embryogenic cells originating from the cultured explant. NO activates the expression of Allene oxide synthase (AOS) and Lipoxygenase 2 (LOX2), genes encoding two key enzymes of the jasmonic acid (JA) biosynthetic pathway, elevating JA content within the embryogenic tissue. The number of embryos in the single aos1-1 mutant and pgb2-aos1-1 double mutant declined, and was not rescued by increasing levels of NO stimulating embryogenesis in wild-type tissue. NO also influenced JA responses by up-regulating PLANT DEFENSIN 1 (PDF1) and JASMONATE-ZIM-PROTEIN (JAZ1), as well as down-regulating MYC2. The NO and JA modulation of MYC2 and JAZ1 controlled embryogenesis. Ectopic expression of JAZ1 or suppression of MYC2 promoted the formation of somatic embryos, while repression of JAZ1 and up-regulation of MYC2 reduced the embryogenic performance. Sustained expression of JAZ1 induced the transcription of several indole acetic acid (IAA) biosynthetic genes, resulting in higher IAA levels in the embryogenic cells. Collectively these data fit a model integrating JA in the PGB2 regulation of Arabidopsis embryogenesis. Suppression of PGB2 increases JA through NO. Elevated levels of JA repress MYC2 and induce JAZ1, favoring the accumulation of IAA in the explants and the subsequent production of somatic embryos. PMID:26962208

  13. Alternative evolution of a spheroidal colony in volvocine algae: developmental analysis of embryogenesis in Astrephomene (Volvocales, Chlorophyta).

    Science.gov (United States)

    Yamashita, Shota; Arakaki, Yoko; Kawai-Toyooka, Hiroko; Noga, Akira; Hirono, Masafumi; Nozaki, Hisayoshi

    2016-11-09

    Volvocine algae, which range from the unicellular Chlamydomonas to the multicellular Volvox with a germ-soma division of labor, are a model for the evolution of multicellularity. Within this group, the spheroidal colony might have evolved in two independent lineages: Volvocaceae and the goniacean Astrephomene. Astrephomene produces spheroidal colonies with posterior somatic cells. The feature that distinguishes Astrephomene from the volvocacean algae is lack of inversion during embryogenesis; the volvocacean embryo undergoes inversion after successive divisions to orient flagella toward the outside. The mechanisms of inversion at the molecular and cellular levels in volvocacean algae have been assessed in detail, particularly in Volvox carteri. However, embryogenesis in Astrephomene has not been subjected to such investigations. This study relied on light microscopy time-lapse imaging using an actively growing culture of a newly established strain to conduct a developmental analysis of Astrephomene as well as to perform a comparison with the similar spheroidal volvocacean Eudorina. During the successive divisions involved in Astrephomene embryogenesis, gradual rotation of daughter protoplasts resulted in movement of their apical portions toward the embryonic posterior, forming a convex-to-spheroidal cell sheet with the apical ends of protoplasts on the outside. Differentiation of the posterior somatic cells from the embryo periphery was traced based on cell lineages during embryogenesis. In contrast, in Eudorina, the rotation of daughter protoplasts did not occur during successive cell divisions; however, inversion occurred after such divisions, and a spheroidal embryo was formed. Indirect immunofluorescence microscopy of basal bodies and nuclei verified this difference between Astrephomene and Eudorina in the movement of embryonic protoplasts. These results suggest different tactics for spheroidal colony formation between the two lineages: rotation of daughter

  14. Are genetics and epigenetic instabilities of plant embryogenic cells a fatality? The experience of coffee somatic embryogenesis

    OpenAIRE

    Etienne, Hervé; Bertrand, Benoît; Dechamp, Eveline; Maurel, Patrick; Georget, Frederic; Guyot, Romain; Breitler, Jean-Christophe

    2016-01-01

    In plants, undifferentiated or totally differentiated cells can be easily in vitro cultured to generate undifferentiated embryogenic cells that can regenerate complete plants. This is the most spectacular expression of totipotency. Embryogenic cells represent a key material in plant biotechnologies as they are used in many processes aiming at asexual reproduction by somatic embryogenesis (SE), genetic transformation, protoplast regeneration and cryopreservation. Nevertheless, tissue culture s...

  15. Regeneration of soapnut tree through somatic embryogenesis and assessment of genetic fidelity through ISSR and RAPD markers.

    Science.gov (United States)

    Singh, Reetika; Kashyap, Sarvesh Pratap; Kumari, Nishi; Singh, Major

    2016-07-01

    Somatic embryogenic system was developed in Sapindus mukorossi Gaertn. using rachis as explants from a mature tree. Explants showed callus initiation on Murashige and Skoog medium supplemented with TDZ (1-Phenyl-3-(1, 2, 3-thiadiazol-5-yl) urea), zeatin or 6-benzylaminopurine. Induction of somatic embryogenesis was achieved on both MS basal medium and MS medium supplemented with 8.88 µM 6-benzylaminopurine. Hundred percent embryogenesis was observed on MS medium supplemented with 8.88 µM 6-benzylaminopurine with maximum intensity of embryogenesis (51.92 ± 0.40 a). Maximum maturation of somatic embryos (92.86 ± 0.34 a) was observed on induction medium supplemented with 0.0378 µM abscisic and treated for 21 days. Germination of somatic embryos was maximum (77.33 ± 0.58 a) on MS medium supplemented with 8.88 µM 6-benzylaminopurine. In vitro raised plantlets were hardened, acclimatized and transferred to the field. Survival frequency of plantlets was 80 % in field conditions. The genetic fidelity of in vitro regenerated plants was also evaluated and compared with mother plant using random amplified polymorphic DNA and inter simple sequence repeat. Both markers showed similarity in molecular profile of mother plant and in vitro regenerated plants.

  16. Somatic Embryogenesis and Plant Regeneration in Sapindus mukorossi Gaertn. from Leaf-Derived Callus Induced with 6-Benzylaminopurine.

    Science.gov (United States)

    Singh, Reetika; Rai, Manoj Kumar; Kumari, Nishi

    2015-09-01

    A somatic embryogenesis system was developed for Sapindus mukorossi Gaertn. from leaf explants obtained from fresh flushes of a mature tree. Callus was induced from the midrib region of leaf explants on Murashige and Skoog (MS) medium containing different concentrations of 2,4-dichlorophenoxyacetic acid or 6-benzylaminopurine. Callus induction and somatic embryogenesis was significantly influenced by the size, physiological age, and orientation of leaf explants on the culture medium and plant growth regulators. Adaxial-side-up orientation of leaf explants significantly promoted embryogenesis in comparison with abaxial-side-up orientation. Maximum number of somatic embryos was induced on MS medium supplemented with 8.88 μM 6-benzylaminopurine. Scanning electron microscopy of embryogenic callus revealed somatic embryo origin and the development of globular-, heart-, and cotyledonary-stage somatic embryos. The frequency of maturation as well as germination of somatic embryos was higher on MS medium containing 8.88 μM 6-benzylaminopurine than on medium without 6-benzylaminopurine. Plantlets which developed from somatic embryos were acclimatized successfully with 90 % survival.

  17. Annotation of Differentially Expressed Genes in the Somatic Embryogenesis of Musa and Their Location in the Banana Genome

    Directory of Open Access Journals (Sweden)

    Josefina Ines Maldonado-Borges

    2013-01-01

    Full Text Available Analysis of cDNA-AFLP was used to study the genes expressed in zygotic and somatic embryogenesis of Musa acuminata Colla ssp. malaccensis, and a comparison was made between their differential transcribed fragments (TDFs and the sequenced genome of the double haploid- (DH- Pahang of the malaccensis subspecies that is available in the network. A total of 253 transcript-derived fragments (TDFs were detected with apparent size of 100–4000 bp using 5 pairs of AFLP primers, of which 21 were differentially expressed during the different stages of banana embryogenesis; 15 of the sequences have matched DH-Pahang chromosomes, with 7 of them being homologous to gene sequences encoding either known or putative protein domains of higher plants. Four TDF sequences were located in all Musa chromosomes, while the rest were located in one or two chromosomes. Their putative individual function is briefly reviewed based on published information, and the potential roles of these genes in embryo development are discussed. Thus the availability of the genome of Musa and the information of TDFs sequences presented here opens new possibilities for an in-depth study of the molecular and biochemical research of zygotic and somatic embryogenesis of Musa.

  18. Annotation of differentially expressed genes in the somatic embryogenesis of musa and their location in the banana genome.

    Science.gov (United States)

    Maldonado-Borges, Josefina Ines; Ku-Cauich, José Roberto; Escobedo-Graciamedrano, Rosa Maria

    2013-01-01

    Analysis of cDNA-AFLP was used to study the genes expressed in zygotic and somatic embryogenesis of Musa acuminata Colla ssp. malaccensis, and a comparison was made between their differential transcribed fragments (TDFs) and the sequenced genome of the double haploid- (DH-) Pahang of the malaccensis subspecies that is available in the network. A total of 253 transcript-derived fragments (TDFs) were detected with apparent size of 100-4000 bp using 5 pairs of AFLP primers, of which 21 were differentially expressed during the different stages of banana embryogenesis; 15 of the sequences have matched DH-Pahang chromosomes, with 7 of them being homologous to gene sequences encoding either known or putative protein domains of higher plants. Four TDF sequences were located in all Musa chromosomes, while the rest were located in one or two chromosomes. Their putative individual function is briefly reviewed based on published information, and the potential roles of these genes in embryo development are discussed. Thus the availability of the genome of Musa and the information of TDFs sequences presented here opens new possibilities for an in-depth study of the molecular and biochemical research of zygotic and somatic embryogenesis of Musa.

  19. Embryogenesis induction, callogenesis, and plant regeneration by in vitro culture of tomato isolated microspores and whole anthers.

    Science.gov (United States)

    Seguí-Simarro, José M; Nuez, Fernando

    2007-01-01

    In this work, some of the different in vitro developmental pathways into which tomato microspores or microsporocytes can be deviated experimentally were explored. The two principal ones are direct embryogenesis from isolated microspores and callus formation from meiocyte-containing anthers. By means of light and electron microscopy, the process of early embryogenesis from isolated microspores and the disruption of normal meiotic development and change of developmental fate towards callus proliferation, morphogenesis, and plant regeneration have been shown. From microspores isolated at the vacuolate stage, embryos can be directly induced, thus avoiding non-androgenic products. In contrast, several different morphogenic events can be triggered in cultures of microsporocyte-containing anthers under adequate conditions, including indirect embryogenesis, adventitious organogenesis, and plant regeneration. Both callus and regenerated plants may be haploid, diploid, and mostly mixoploid. The results demonstrate that both gametophytic and sporophytic calli occur in cultured tomato anthers, and point to an in vitro-induced disturbance of cytokinesis and subsequent fusion of daughter nuclei as a putative cause for mixoploidy and genome doubling during both tetrad compartmentalization and callus proliferation. The potential implications of the different alternative pathways are discussed in the context of their application to the production of doubled-haploid plants in tomato, which is still very poorly developed.

  20. Global transcriptome analysis identifies regulated transcripts and pathways activated during oogenesis and early embryogenesis in Atlantic cod.

    Science.gov (United States)

    Kleppe, Lene; Edvardsen, Rolf Brudvik; Furmanek, Tomasz; Taranger, Geir Lasse; Wargelius, Anna

    2014-07-01

    The molecular mechanisms underlying oogenesis and maternally controlled embryogenesis in fish are not fully understood, especially in marine species. Our aim was to study the egg and embryo transcriptome during oogenesis and early embryogenesis in Atlantic cod. Follicles from oogenesis stages (pre-, early-, and late-vitellogenic), ovulated eggs, and two embryonic stages (blastula, gastrula) were collected from broodstock fish and fertilized eggs. Gene expression profiles were measured in a 44 K oligo microarray consisting of 23,000 cod genes. Hundreds of differentially expressed genes (DEGs) were identified in the follicle stages investigated, implicating a continuous accumulation and degradation of polyadenylated transcripts throughout oogenesis. Very few DEGs were identified from ovulated egg to blastula, showing a more stable maternal RNA pool in early embryonic stages. The highest induction of expression was observed between blastula and gastrula, signifying the onset of zygotic transcription. During early vitellogenesis, several of the most upregulated genes are linked to nervous system signaling, suggesting increasing requirements for ovarian synaptic signaling to stimulate the rapid growth of oocytes. Highly upregulated genes during late vitellogenesis are linked to protein processing, fat metabolism, osmoregulation, and arrested meiosis. One of the genes with the highest upregulation in the ovulated egg is involved in oxidative phosphorylation, reflecting increased energy requirements during fertilization and the first rapid cell divisions of early embryogenesis. In conclusion, this study provides a large-scale presentation of the Atlantic cod's maternally controlled transcriptome in ovarian follicles through oogenesis, ovulated eggs, and early embryos. Published 2014 Wiley Periodicals, Inc.

  1. Oxygen changes drive non-uniform scaling in Drosophila melanogaster embryogenesis [version 1; referees: 2 approved, 1 approved with reservations

    Directory of Open Access Journals (Sweden)

    Steven G. Kuntz

    2015-10-01

    Full Text Available We previously demonstrated that, while changes in temperature produce dramatic shifts in the time elapsed during Drosophila melanogaster embryogenesis, the relative timing of events within embryogenesis does not change. However, it was unclear if this uniform scaling is an intrinsic property of developing embryos, or if it is specific to thermal fluctuations. To investigate this, here we characterize the embryonic response to changes in oxygen concentration, which also impact developmental rate, using time-lapse imaging, and find it fundamentally different from the temperature response. Most notably, changes in oxygen levels drive developmental heterochrony, with the timing of several morphological processes showing distinct scaling behaviors. Gut formation is severely slowed by decreases in oxygen, while head involution and syncytial development are less impacted than the rest of development, and the order of several developmental landmarks is inverted at different oxygen levels. These data reveal that the uniform scaling seen with changes in temperature is not a trivial consequence of adjusting developmental rate. The developmental rate changes produced by changing oxygen concentrations dwarf those induced by temperature, and greatly impact survival. While extreme temperatures increase early embryo mortality, mild hypoxia increases arrest and death during mid-embryogenesis and mild hyperoxia increases survival over normoxia.

  2. Coho Abundance - Linear Features [ds183

    Data.gov (United States)

    California Department of Resources — The CalFish Abundance Database contains a comprehensive collection of anadromous fisheries abundance information. Beginning in 1998, the Pacific States Marine...

  3. Chinook Abundance - Point Features [ds180

    Data.gov (United States)

    California Department of Resources — The CalFish Abundance Database contains a comprehensive collection of anadromous fisheries abundance information. Beginning in 1998, the Pacific States Marine...

  4. Steelhead Abundance - Linear Features [ds185

    Data.gov (United States)

    California Department of Resources — The CalFish Abundance Database contains a comprehensive collection of anadromous fisheries abundance information. Beginning in 1998, the Pacific States Marine...

  5. Steelhead Abundance - Point Features [ds184

    Data.gov (United States)

    California Department of Resources — The CalFish Abundance Database contains a comprehensive collection of anadromous fisheries abundance information. Beginning in 1998, the Pacific States Marine...

  6. Coho Abundance - Point Features [ds182

    Data.gov (United States)

    California Department of Resources — The CalFish Abundance Database contains a comprehensive collection of anadromous fisheries abundance information. Beginning in 1998, the Pacific States Marine...

  7. Abundance estimation and Conservation Biology

    Directory of Open Access Journals (Sweden)

    Nichols, J. D.

    2004-06-01

    Full Text Available Abundance is the state variable of interest in most population–level ecological research and in most programs involving management and conservation of animal populations. Abundance is the single parameter of interest in capture–recapture models for closed populations (e.g., Darroch, 1958; Otis et al., 1978; Chao, 2001. The initial capture–recapture models developed for partially (Darroch, 1959 and completely (Jolly, 1965; Seber, 1965 open populations represented efforts to relax the restrictive assumption of population closure for the purpose of estimating abundance. Subsequent emphases in capture–recapture work were on survival rate estimation in the 1970’s and 1980’s (e.g., Burnham et al., 1987; Lebreton et al.,1992, and on movement estimation in the 1990’s (Brownie et al., 1993; Schwarz et al., 1993. However, from the mid–1990’s until the present time, capture–recapture investigators have expressed a renewed interest in abundance and related parameters (Pradel, 1996; Schwarz & Arnason, 1996; Schwarz, 2001. The focus of this session was abundance, and presentations covered topics ranging from estimation of abundance and rate of change in abundance, to inferences about the demographic processes underlying changes in abundance, to occupancy as a surrogate of abundance. The plenary paper by Link & Barker (2004 is provocative and very interesting, and it contains a number of important messages and suggestions. Link & Barker (2004 emphasize that the increasing complexity of capture–recapture models has resulted in large numbers of parameters and that a challenge to ecologists is to extract ecological signals from this complexity. They offer hierarchical models as a natural approach to inference in which traditional parameters are viewed as realizations of stochastic processes. These processes are governed by hyperparameters, and the inferential approach focuses on these hyperparameters. Link & Barker (2004 also suggest that

  8. Abundance estimation and conservation biology

    Science.gov (United States)

    Nichols, J.D.; MacKenzie, D.I.

    2004-01-01

    Abundance is the state variable of interest in most population–level ecological research and in most programs involving management and conservation of animal populations. Abundance is the single parameter of interest in capture–recapture models for closed populations (e.g., Darroch, 1958; Otis et al., 1978; Chao, 2001). The initial capture–recapture models developed for partially (Darroch, 1959) and completely (Jolly, 1965; Seber, 1965) open populations represented efforts to relax the restrictive assumption of population closure for the purpose of estimating abundance. Subsequent emphases in capture–recapture work were on survival rate estimation in the 1970’s and 1980’s (e.g., Burnham et al., 1987; Lebreton et al.,1992), and on movement estimation in the 1990’s (Brownie et al., 1993; Schwarz et al., 1993). However, from the mid–1990’s until the present time, capture–recapture investigators have expressed a renewed interest in abundance and related parameters (Pradel, 1996; Schwarz & Arnason, 1996; Schwarz, 2001). The focus of this session was abundance, and presentations covered topics ranging from estimation of abundance and rate of change in abundance, to inferences about the demographic processes underlying changes in abundance, to occupancy as a surrogate of abundance. The plenary paper by Link & Barker (2004) is provocative and very interesting, and it contains a number of important messages and suggestions. Link & Barker (2004) emphasize that the increasing complexity of capture–recapture models has resulted in large numbers of parameters and that a challenge to ecologists is to extract ecological signals from this complexity. They offer hierarchical models as a natural approach to inference in which traditional parameters are viewed as realizations of stochastic processes. These processes are governed by hyperparameters, and the inferential approach focuses on these hyperparameters. Link & Barker (2004) also suggest that our attention

  9. Stress induced acquisition of somatic embryogenesis in common bean Phaseolus vulgaris L.

    Science.gov (United States)

    Cabrera-Ponce, José Luis; López, Liliana; León-Ramírez, Claudia G; Jofre-Garfias, Alba E; Verver-y-Vargas, Aurora

    2015-03-01

    Common bean Phaseolus vulgaris L. has been shown to be a recalcitrant plant to induce somatic embryogenesis (SE) under in vitro conditions. We used an alternative strategy to induce SE in common bean based upon the use of a cytokinin (BAP) coupled with osmotic stress adaptation instead of SE response that is induced by auxins. Explants derived from zygotic embryos of common bean were subjected to osmotic stress (sucrose 12 % w/v, 0.5 M) in the presence of BAP 10 mg/L and adenine free base 40 mg/L to induce somatic embryos from specific competent cells of the apical meristem and cotyledonary node. Somatic embryos were obtained from the competent cells in a direct response (direct SE). In a secondary response (secondary SE), those somatic embryos formed proembryogenic masses (PEM) that originated/developed into secondary somatic embryos and showed the SE ontogeny. Maturation of somatic embryos was achieved by using different osmolality media and converted to plants. Full-visible light spectrum was necessary to achieve efficient plant regeneration. Long-term recurrent SE was demonstrated by propagation of PEM at early stages of SE. This protocol is currently being applied for stable genetic transformation by means of Agrobacterium tumefaciens and bioballistics as well as for basic biochemical and molecular biology experiments.

  10. Improved protocol for somatic embryogenesis and calcium alginate encapsulation in Anethum graveolens L.: a medicinal herb.

    Science.gov (United States)

    Dhir, Richa; Shekhawat, G S; Alam, Afroz

    2014-08-01

    An improved procedure has been developed for efficient somatic embryogenesis in Anethum graveolens. Green friable embryogenic callus was obtained from hypocotyl segments on medium augmented with 2,4-dichlorophenoxyacetic acid (2,4-D). The highest embryogenic callus induction frequency of 87 % was obtained on Murashige and Skoog (MS) medium containing 1.13 μM 2,4-D. At lower concentration of 2,4-D (0.34 μM) callus turned dark in color and slow growing. Embryogenic cultures (76 %) responded with a mean number of 43 globular and 18 heart stage embryos. Somatic embryo maturation and subsequent conversion into plantlets took place on MS lacking growth regulators. Maximum number of somatic embryos developed on MS medium was 128.3 (per flask) and a plantlet conversion of 82 % was observed. Calcium alginate beads were produced by encapsulating somatic embryos. Highest percent germination (83 %) was observed on 0.8 % agar solidified MS medium with the plantlets acquiring an average length of 2.1 cm. Encapsulated somatic embryos could be stored at 4 °C up to 60 days with a conversion frequency of 49.3 %. Highest protein and proline content has been observed in embryogenic callus with small globular embryos. During morphological differentiation of the somatic embryos, changes in the antioxidant enzymatic system were observed. Superoxide dismutase (SOD) activity increased during initial stages and decreased catalase (CAT), peroxidase (POD), and ascorbate peroxidase (APX) activities were detected.

  11. Expression of phosphatidylcholine biosynthetic enzymes during early embryogenesis in the amphibian Bufo arenarum.

    Science.gov (United States)

    Fernández-Bussy, Rodrigo; Mouguelar, Valeria; Banchio, Claudia; Coux, Gabriela

    2015-04-01

    In the principal route of phosphatidylcholine (PC) synthesis the regulatory steps are catalysed by CTP:phosphocholine cytidylyltransferase (CCT) and choline kinase (CK). Knock-out mice in Pcyt1a (CCT gene) and Chka1 (CK gene) resulted in preimplantation embryonic lethality, demonstrating the essential role of this pathway. However, there is still a lack of detailed CCT and CK expression analysis during development. The aim of the current work was to study the expression during early development of both enzymes in the external-fertilization vertebrate Bufo arenarum. Reverse transcription polymerase chain reaction (RT-PCR) and western blot confirmed their presence in unfertilized eggs. Analysis performed in total extracts from staged embryos showed constant protein levels of both enzymes until the 32-cell stage: then they decreased, reaching a minimum in the gastrula before starting to recover. CTP:phosphocholine cytidylyltransferase is an amphitropic enzyme that inter-converts between cytosolic inactive and membrane-bound active forms. Immunoblot analysis demonstrated that the cytosolic:total CCT protein ratio does not change throughout embryogenesis, suggesting a progressive decline of CCT activity in early development. However, PC (and phosphatidylethanolamine) content per egg/embryo remained constant throughout the stages analysed. In conclusion, the current data for B. arenarum suggest that net synthesis of PC mediated by CCT and CK is not required in early development and that supplies for membrane biosynthesis are fulfilled by lipids already present in the egg/embryo reservoirs.

  12. Regulation of embryogenesis by light and its ecological significance in the Asian Tadpole shrimp Triops granarius.

    Science.gov (United States)

    Horiguchi, Tsutomu; Ito, Chihiro; Numata, Hideharu

    2009-07-01

    Triops granarius (Lucas) (Notostraca: Triopsidae) lives In paddy fields from the Kanto district to northern Kyushu, Japan. Changes in the size distribution of this species were examined in the paddy fields and then the effect of light on hatching was examined under quasi-natural and laboratory conditions. Adult tadpole shrimps were found about one week after irrigation and plowing in two paddy fields in Sakai, Japan. They developed rapidly and disappeared altogether about one month later. Under conditions of natural daylength and temperature, eggs laid in the soil did not hatch without being removed from the soil. Under constant light at 25 degrees C, the lower the light intensity was, the longer the eggs took to hatch. Moreover, most eggs kept in constant darkness did not hatch, but many of them hatched within a short period after being transferred to constant light with an intensity of 0.3 W/m(2) or more. Because a 1-h light pulse was found to induce hatching, light is considered necessary for the resumption of embryonic development. These results suggest that eggs of T. granarius laid in the soil do not hatch without exposure to light; consequently, this species has a univoltine life cycle in the paddy fields. Histological observations revealed that under constant darkness, embryonic development was arrested at an early stage of organogenesis, in which the nauplius eye had not yet formed. We discuss the role of light in the regulation of embryogenesis in T. granarius.

  13. An Observation-Driven Agent-Based Modeling and Analysis Framework for C. elegans Embryogenesis.

    Science.gov (United States)

    Wang, Zi; Ramsey, Benjamin J; Wang, Dali; Wong, Kwai; Li, Husheng; Wang, Eric; Bao, Zhirong

    2016-01-01

    With cutting-edge live microscopy and image analysis, biologists can now systematically track individual cells in complex tissues and quantify cellular behavior over extended time windows. Computational approaches that utilize the systematic and quantitative data are needed to understand how cells interact in vivo to give rise to the different cell types and 3D morphology of tissues. An agent-based, minimum descriptive modeling and analysis framework is presented in this paper to study C. elegans embryogenesis. The framework is designed to incorporate the large amounts of experimental observations on cellular behavior and reserve data structures/interfaces that allow regulatory mechanisms to be added as more insights are gained. Observed cellular behaviors are organized into lineage identity, timing and direction of cell division, and path of cell movement. The framework also includes global parameters such as the eggshell and a clock. Division and movement behaviors are driven by statistical models of the observations. Data structures/interfaces are reserved for gene list, cell-cell interaction, cell fate and landscape, and other global parameters until the descriptive model is replaced by a regulatory mechanism. This approach provides a framework to handle the ongoing experiments of single-cell analysis of complex tissues where mechanistic insights lag data collection and need to be validated on complex observations.

  14. A chimeric arabinogalactan protein promotes somatic embryogenesis in cotton cell culture.

    Science.gov (United States)

    Poon, Simon; Heath, Robyn Louise; Clarke, Adrienne Elizabeth

    2012-10-01

    Arabinogalactan proteins (AGPs) are a family of extracellular plant proteoglycans implicated in many aspects of plant growth and development, including in vitro somatic embryogenesis (SE). We found that specific AGPs were produced by cotton (Gossypium hirsutum) calli undergoing SE and that when these AGPs were isolated and incorporated into tissue culture medium, cotton SE was promoted. When the AGPs were partly or fully deglycosylated, SE-promoting activity was not diminished. Testing of AGPs separated by reverse-phase high-performance liquid chromatography revealed that the SE-promoting activity resided in a hydrophobic fraction. We cloned a full-length complementary DNA (cotton PHYTOCYANIN-LIKE ARABINOGALACTAN-PROTEIN1 [GhPLA1]) that encoded the protein backbone of an AGP in the active fraction. It has a chimeric structure comprising an amino-terminal signal sequence, a phytocyanin-like domain, an AGP-like domain, and a hydrophobic carboxyl-terminal domain. Recombinant production of GhPLA1 in tobacco (Nicotiana tabacum) cells enabled us to purify and analyze a single glycosylated AGP and to demonstrate that this chimeric AGP promotes cotton SE. Furthermore, the nonglycosylated phytocyanin-like domain from GhPLA1, which was bacterially produced, also promoted SE, indicating that the glycosylated AGP domain was unnecessary for in vitro activity.

  15. Evaluation of Somatic Embryogenesis Ability in Robusta Coffee (Coffea canephora Pierre

    Directory of Open Access Journals (Sweden)

    Priyono Priyono

    2010-08-01

    Full Text Available Embriogenesis somatik diharapkan sebagai metode perbanyakan tanaman yang sangat efektif pada kopi. Evaluasi dua jenis proses embriogenesis somatik, yaitu proses langsung dan tidak langsung akan bermanfaat untuk menggambarkan kemampuan proliferasi sel. Penelitian untuk mengevaluasi embriogenesis somatik kopi Robusta (Coffea canephora yang mempunyai tingkat keragaman genetik tinggi telah dilakukan di Nestlé R&D Centre Tours, Perancis. Bahan tanam menggunakan kopi Robusta koleksi Nestle Perancis dan tiga klon koleksi Pusat Penelitian Kopi dan Kakao Indonesia (Puslitkoka. Tiga aspek, yaitu proses embriogenesis, keragaman embriogenesis dan kemantapan embriogenesis dievaluasi dalam penelitian ini. Hasil penelitian menunjukkan bahwa baik embriogenesis somatik langsung maupun tidak langsung dapat diamati. Penelitian ini menunjukkan bahwa kedua proses embriogenesis somatik tersebut merupakan dua mekanisme yang berbeda. Dalam penelitian ini ditunjukkan bahwa kemampuan embriognesis somatik tergantung pada genotipe, baik antar maupun di dalam kelompok genetik kopi Robusta, yaitu Congolese,Guinean dan Conillon. Lebih lanjut diketahui bahwa kedua proses embriogenesis somatik tersebut stabil terhadap indukan sebagai sumber eksplan. Kemampuan embriogenesis somatik tidak langsung ketiga klon Puslitkoka (BP409, BP961 dan Q121 sangat beragam, sehingga memberikan harapan adanya pola segregasi yang baik berdasarkan kemampuan embriogenesis somatik tidak langsung pada populasi yang dibuat dari silangan klon tersebut.Key words: Coffea canephora, somatic embryogenesis, variability, stability, genotype.

  16. Somatic embryogenesis and vegetative cutting capacity are under distinct genetic control in Coffea canephora Pierre.

    Science.gov (United States)

    Priyono; Florin, Bruno; Rigoreau, Michel; Ducos, Jean-Paul; Sumirat, Ucu; Mawardi, Surip; Lambot, Charles; Broun, Pierre; Pétiard, Vincent; Wahyudi, Teguh; Crouzillat, Dominique

    2010-04-01

    The purpose of the study was to evaluate the possible genetic effect on vegetative propagation of Coffea canephora. Diversity for somatic embryogenesis (SE) ability was observed not only among two groups of C. canephora Pierre (Congolese and Guinean), but also within these different genetic groups. The results therefore showed that, under given experimental conditions, SE ability is depending on genotype. Furthermore the detection of quantitative trait loci (QTLs) controlling the SE and cutting abilities of C. canephora was performed on a large number of clones including accessions from a core collection, three parental clones and their segregating progenies. On the one hand we detected eight QTLs determining SE. Six positive QTLs for SE ability, whatever the criteria used to quantify this ability, were localized on one single chromosome region of the consensus genetic map. Two negative QTLs for SE ability (frequency of micro calli without somatic embryo) were detected on another linkage group. Deep analysis of the six QTLs detected for SE ability came to the conclusion that they can be assimilated to one single QTL explaining 8.6-12.2% of the observed variation. On the other hand, two QTLs for average length of roots and length of the longest sprouts of cuttings were detected in two linkage groups. These QTLs detected for cutting ability are explaining 12-27% of the observed variation. These observations led to conclude that SE and cutting abilities of C. canephora Pierre appeared to be genetic dependent but through independent mechanisms.

  17. Embryogenesis of the Uropygial Glands in the Laysan Albatross (Phoebastria immutabilis (Rothschild, 1893): Procellariiformes).

    Science.gov (United States)

    Rehorek, S J; Wu, J L; Smith, T D; Beeching, S C

    2017-08-01

    An avian uropygial gland is located on the mid-dorsum of the tail, and is the only external gland found in birds. Most studies have focused on the function, gross anatomy and chemical nature of this gland, with little research on its ontogeny. The purpose of this study was to examine the development of this gland in a series of Laysan Albatross (Phoebastria immutabilis) embryos. Specimens were examined anatomically and histologically. It was found that grooves preceded glandular development by many stages. The embryogenesis of the uropygial gland was divided into 6 phases: preinception, groove inception, mesodermal separation, migrating mesodermal cells, oval shaped "depressions", constriction and finally glandular inception. No other gland is known to develop similarly, though there may be parallels with femoral gland development. In comparison to other bird species, the length of the development period in the Albatross, as well as other compounding factors, make it difficult to determine the significance of these observations. The development of a mesodermal band, soon to be a connective tissue capsule, is more complex than originally described in ducks. Thus, the unique nature of this gland is established, but the significance of the observations required further studies into uropygial gland development. Anat Rec, 300:1420-1428, 2017. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  18. Somatic embryogenesis in cv. Navolean using shoots apexes from axillary buds

    Directory of Open Access Journals (Sweden)

    Jorge López

    2005-04-01

    Full Text Available In order to develop embryogenic cultures in AAB Musa genotypes without persistent male inflorescence, the process has had greater success from proliferating meristems for callus formation with embryogenic structures. Based on the previous information, other alternative explant sources for somatic embryogenesis development in cv. Navolean. Meristematic apexes were cultured in p5 culture medium supplemented with thidiazuron and ancymidol (0.2, 0.4, 0.6, mg.l-1 to obtain axillary buds. Later, axillary buds and proliferated meristems were tested for callus induction with embryogenic structures combinations with different 2,4-D concentrations. The best growth regulator for obtaining axillary buds was ancymidol (0.2 mg.l-1. For callus formation with embryogenic structures, axillary buds at 1.0 mg.l-1 2,4-D provided a higher percentage (13.6%. These results permitted the development of embryogenic cell suspensions from somatic embryos. Key words: Ancymidol, embryogenic cell suspension, plantain

  19. Neptune is involved in posterior axis and tail formation in Xenopus embryogenesis.

    Science.gov (United States)

    Takeda, Masatoshi; Kurauchi, Takayuki; Yamazaki, Takeshi; Izutsu, Yumi; Maéno, Mitsugu

    2005-09-01

    In order to elucidate the molecular mechanisms underlying the posterior axis and tail formation in embryogenesis, the function of Neptune, a zinc-finger transcription factor, in Xenopus laevis embryos was investigated. Injection of neptune mRNA into the animal pole area of embryos resulted in the formation of an additional tail structure that included a neural tube and muscle tissue. This activity required FGF signaling since coinjection of a dominant-negative FGF receptor RNA (XFD) completely blocked the formation of a tail structure. A loss-of-function experiment using a fusion construct of neptune and Drosophila engrailed (en-neptune) RNA showed that endogenous Neptune is necessary for formation of the posterior trunk and tail. Furthermore, activity of Neptune was necessary for the endogenous expression of brachyury and fgf-8 at the late gastrula stage. These findings demonstrate a novel function of Neptune in the process of anterior-posterior axis formation through the FGF and brachyury signaling cascades. An experiment using a combination explant with ventral and dorsal marginal tissues showed that cooperation of these two distinct tissues is important for the tail formation and that expression of Neptune in prospective ventral cells may be involved in the activation of the process of tail formation. Copyright 2005 Wiley-Liss, Inc.

  20. SWATH-MS data of Drosophila melanogaster proteome dynamics during embryogenesis

    Directory of Open Access Journals (Sweden)

    Bertrand Fabre

    2016-12-01

    Full Text Available Embryogenesis is one of the most important processes in the life of an animal. During this dynamic process, progressive cell division and cellular differentiation are accompanied by significant changes in protein expression at the level of the proteome. However, very few studies to date have described the dynamics of the proteome during the early development of an embryo in any organism. In this dataset, we monitor changes in protein expression across a timecourse of more than 20 h of Drosophila melanogaster embryonic development. Mass-spectrometry data were produced using a SWATH acquisition mode on a Sciex Triple-TOF 6600. A spectral library built in-house was used to analyse these data and more than 1950 proteins were quantified at each embryonic timepoint. The files presented here are a permanent digital map and can be reanalysed to test against new hypotheses. The data have been deposited with the ProteomeXchange Consortium with the dataset identifier PRIDE: PXD0031078.

  1. Light sheet-based imaging and analysis of early embryogenesis in the fruit fly.

    Science.gov (United States)

    Khairy, Khaled; Lemon, William C; Amat, Fernando; Keller, Philipp J

    2015-01-01

    The fruit fly is an excellent model system for investigating the sequence of epithelial tissue invaginations constituting the process of gastrulation. By combining recent advancements in light sheet fluorescence microscopy (LSFM) and image processing, the three-dimensional fly embryo morphology and relevant gene expression patterns can be accurately recorded throughout the entire process of embryogenesis. LSFM provides exceptionally high imaging speed, high signal-to-noise ratio, low level of photoinduced damage, and good optical penetration depth. This powerful combination of capabilities makes LSFM particularly suitable for live imaging of the fly embryo.The resulting high-information-content image data are subsequently processed to obtain the outlines of cells and cell nuclei, as well as the geometry of the whole embryo tissue by image segmentation. Furthermore, morphodynamics information is extracted by computationally tracking objects in the image. Towards that goal we describe the successful implementation of a fast fitting strategy of Gaussian mixture models.The data obtained by image processing is well-suited for hypothesis testing of the detailed biomechanics of the gastrulating embryo. Typically this involves constructing computational mechanics models that consist of an objective function providing an estimate of strain energy for a given morphological configuration of the tissue, and a numerical minimization mechanism of this energy, achieved by varying morphological parameters.In this chapter, we provide an overview of in vivo imaging of fruit fly embryos using LSFM, computational tools suitable for processing the resulting images, and examples of computational biomechanical simulations of fly embryo gastrulation.

  2. The α-tocopherol transfer protein is essential for vertebrate embryogenesis.

    Directory of Open Access Journals (Sweden)

    Galen W Miller

    Full Text Available The hepatic α-tocopherol transfer protein (TTP is required for optimal α-tocopherol bioavailability in humans; mutations in the human TTPA gene result in the heritable disorder ataxia with vitamin E deficiency (AVED, OMIM #277460. TTP is also expressed in mammalian uterine and placental cells and in the human embryonic yolk-sac, underscoring TTP's significance during fetal development. TTP and vitamin E are essential for productive pregnancy in rodents, but their precise physiological role in embryogenesis is unknown. We hypothesize that TTP is required to regulate delivery of α-tocopherol to critical target sites in the developing embryo. We tested to find if TTP is essential for proper vertebrate development, utilizing the zebrafish as a non-placental model. We verify that TTP is expressed in the adult zebrafish and its amino acid sequence is homologous to the human ortholog. We show that embryonic transcription of TTP mRNA increases >7-fold during the first 24 hours following fertilization. In situ hybridization demonstrates that Ttpa transcripts are localized in the developing brain, eyes and tail bud at 1-day post fertilization. Inhibiting TTP expression using oligonucleotide morpholinos results in severe malformations of the head and eyes in nearly all morpholino-injected embryos (88% compared with 5.6% in those injected with control morpholinos or 1.7% in non-injected embryos. We conclude that TTP is essential for early development of the vertebrate central nervous system.

  3. Effects of gravity on meiosis, fertilization and early embryogenesis in Caenorhabditis elegans

    Science.gov (United States)

    Sasagawa, Y.; Saito, Y.; Shimizu, M.; Ishioka, N.; Yamashita, M.; Takahashi, H.; Higashitani, A.

    The embryonic development of the nematode Caenorhabditis elegans was examined under different gravitational conditions. The first cleavage plane in the 1-cell embryo was slid to some extent by re-orientation of liquid culture vessel, but the pattern and timing of cleavages were not affected. Under 100G of hypergravity condition with swing-centrifuge, the number of eggs laid from an adult hermaphrodite decreased and their hatching rate was drastically reduced. On the other hand, the embryonic development after fertilization normally occurred and grew to adulthood at more than 100G of hypergravity. When the adult hermaphrodites cultured under 100G of hypergravity transferred to a ground condition (1G), the newly fertilized embryos normally developed and their hatching rate was fully recovered. These results indicated that the reproductive process except spermatogenesis, oogenesis and embryogenesis after fertilization is impaired under 100G of hypergravity condition, and the effect is transient. Namely, the fertilization process including meiotic divisions I and II is sensitive to hypergravity in the nematode C. elegans.

  4. Genome editing reveals a role for OCT4 in human embryogenesis.

    Science.gov (United States)

    Fogarty, Norah M E; McCarthy, Afshan; Snijders, Kirsten E; Powell, Benjamin E; Kubikova, Nada; Blakeley, Paul; Lea, Rebecca; Elder, Kay; Wamaitha, Sissy E; Kim, Daesik; Maciulyte, Valdone; Kleinjung, Jens; Kim, Jin-Soo; Wells, Dagan; Vallier, Ludovic; Bertero, Alessandro; Turner, James M A; Niakan, Kathy K

    2017-10-05

    Despite their fundamental biological and clinical importance, the molecular mechanisms that regulate the first cell fate decisions in the human embryo are not well understood. Here we use CRISPR-Cas9-mediated genome editing to investigate the function of the pluripotency transcription factor OCT4 during human embryogenesis. We identified an efficient OCT4-targeting guide RNA using an inducible human embryonic stem cell-based system and microinjection of mouse zygotes. Using these refined methods, we efficiently and specifically targeted the gene encoding OCT4 (POU5F1) in diploid human zygotes and found that blastocyst development was compromised. Transcriptomics analysis revealed that, in POU5F1-null cells, gene expression was downregulated not only for extra-embryonic trophectoderm genes, such as CDX2, but also for regulators of the pluripotent epiblast, including NANOG. By contrast, Pou5f1-null mouse embryos maintained the expression of orthologous genes, and blastocyst development was established, but maintenance was compromised. We conclude that CRISPR-Cas9-mediated genome editing is a powerful method for investigating gene function in the context of human development.

  5. ROLUL NEGUSTORILOR ALOGENI ÎN PROCESUL DE CONSTITUIRE A BURGHEZIEI COMERCIALE DIN PRINCIPATUL MOLDOVA (sfârşitul sec. al XVIII-lea – începutul sec. al XIX-lea

    Directory of Open Access Journals (Sweden)

    Irina CEREŞ

    2015-11-01

    Full Text Available În articol este a investigat rolul comercianţilor străini în crearea burgheziei comerciale în Principatul Moldova (de la sfârşitul secolului al XVIII – începutul secolului al XIX-lea. La sfârşitul secolul al XVIII-lea – începutul secolului al XIX-lea burghezia comercială a Principatului Moldova a fost o categorie ce se afla într-o stare de formare. Astfel, se poate considera că negustorii străini au fost un element im-portant în formarea lentă a burgheziei din zona carpatică. Negustorii străini stabiliţi în Moldova s-au dovedit a fi un factor de progres în ceea ce priveşte evoluţia economiei locale, au contribuit la diversificarea activităţilor economice, la dezvoltarea comerţului interior şi exterior, precum şi la consolidarea rolului burgheziei în formarea categoriilor profesionale productive şi dinamice – de meşteşugari şi meseriaşi.LE RÔLE DES MARCHANDS ALLOGÉNIQUES DANS LE PROCESSUS DE FORMATION DE LA BOURGEOISIE COMMERCIALE DE LA PRINCIPAUTÉ DE MOLDAVIE (À LA FIN DU XVIIIÈME SIÈCLE – DÉBUT DU XIX ÈME SIÈCLE  La Bourgeoisie commerciale de la Principauté de Moldavie existante à la fin du XVIIIème siècle – début du XIXème siècle constitue une catégorie sociale en phase embryonnaire. Ses représentants sont généralement des éléments allogènes, avec peu d’autochtones. On sait qu’en effet toute bourgeoisie a commencé son développement à l’aide des étrangers. La séparation de ceux-ci s’est produite uniquement après une certaine période de temps, après avoir fait leur éducation économique dans le pays d’origine. On rencontre le même processus au cas des Principautés Roumains. Les commerçants étrangers ont constitué l’un des éléments les plus importants dans le processus lent de cristallisation de la bourgeoisie autochtone. Par leurs activités commerciales et par leurs propres intérêts ils on contribué à faire évoluer les activités de commerce de

  6. Hematite Abundance on Martian Surface

    Science.gov (United States)

    1999-01-01

    This figure shows the concentration of hematite measured by the Mars Global Surveyor Thermal Emission Spectrometer (TES) instrument. The abundance of hematite is shown in red, with increasing brightness indicating increasing hematite abundance. The location and size of the individual TES observations on the surface are indicated by the individual squares. Black squares indicate observations with no detectable hematite. Data from 11 separate orbits acquired between Nov. 22, 1997 and April 25, 1998 are shown in this image. The TES data are superimposed on a Viking photomosaic for context. The image extends from 10 S to 10 N latitude and 350 W to 15 W longitude, covering an area 1500 km (940 miles) in longitude by 1200 km (750 miles) in latitude.The TES instrument was built by Santa Barbara Remote Sensing and is operated by Philip R. Christensen, of Arizona State University, Tempe, AZ. The MGS mission is managed for NASA by the Jet Propulsion Laboratory, Pasadena CA.

  7. Chlorine Abundances in Martian Meteorites

    Science.gov (United States)

    Bogard, D.D.; Garrison, D.H.; Park, J.

    2009-01-01

    Chlorine measurements made in martian surface rocks by robotic spacecraft typically give Chlorine (Cl) abundances of approximately 0.1-0.8%. In contrast, Cl abundances in martian meteorites appear lower, although data is limited, and martian nakhlites were also subjected to Cl contamination by Mars surface brines. Chlorine abundances reported by one lab for whole rock (WR) samples of Shergotty, ALH77005, and EET79001 range 108-14 ppm, whereas Cl in nakhlites range 73-1900 ppm. Measurements of Cl in various martian weathering phases of nakhlites varied 0.04-4.7% and reveal significant concentration of Cl by martian brines Martian meteorites contain much lower Chlorine than those measured in martian surface rocks and give further confirmation that Cl in these surface rocks was introduced by brines and weathering. It has been argued that Cl is twice as effective as water in lowering the melting point and promoting melting at shallower martian depths, and that significant Cl in the shergottite source region would negate any need for significant water. However, this conclusion was based on experiments that utilized Cl concentrations more analogous to martian surface rocks than to shergottite meteorites, and may not be applicable to shergottites.

  8. In vitro somatic embryogenesis and plantlet regeneration from immature male inflorescence of adult dura and tenera palms of Elaeis guineensis (Jacq.).

    Science.gov (United States)

    Jayanthi, Madhavan; Susanthi, Bollarapu; Murali Mohan, Nandiganti; Mandal, Pranab Kumar

    2015-01-01

    We report here a method for plant regeneration through somatic embryogenesis from explants collected from immature male inflorescence of adult oil palm cultivated in India. Callus induction was successful from tissues of immature male inflorescence collected from both dura and tenera varieties of oil palm. A modified Y3 (Eeuwens) media supplemented with several additives and activated charcoal (3%) were used for the experiments. Out of four different auxin treatments, 4-amino-3,5,6-trichloro-2-pyridinecarboxylic acid (picloram) produced maximum callus induction (82%) and it was not significantly different from 2,4-dichlorophenoxyacetic acid (2,4-D) and a combination of 2,4-D + picloram. The callus induction obtained with auxin α-naphthalene acetic acid was only 54% and it was significantly low as compared to the other treatments. Highest embryogenesis was obtained with a combination of 2,4-D + picloram (4.9%) followed by picloram (3.4%). Genotypic variation in response to the same auxins was observed both for callus induction and embryogenesis. Callus induction and embryogenesis ranged from 42 to 72% and 6.8 to 9.35%, respectively in tenera. The formation of embryogenic calli was marked by the appearance of white to yellowish globular or nodular structures which subsequently formed clear somatic embryos. Somatic embryogenesis was asynchronous and at one time we could find different stages of embryogenesis like the globular, torpedo and the cotyledonary stages. The somatic embryos when exposed to light in the same basal media along with 6-benzyladenine (18 µM), abscisic acid (3.78 µM) and gibberellic acid (5.78 µM) regenerated into plantlets. To the best of our knowledge this is the first report o f callus induction and somatic embryogenesis from immature male inflorescence of oil palm.

  9. IN VITRO REGENERATION OF THREE CHRYSANTHEMUM (Dendrathema grandiflora VARIETIES “VIA” ORGANOGENESIS AND SOMATIC EMBRYOGENESIS

    Directory of Open Access Journals (Sweden)

    Elizabeth Hodson de Jaramillo

    2008-09-01

    Full Text Available Chrysanthemum (Dendrathema grandiflora has a high demand in the Colombian and international cut flower markets.Since commercial production of this ornamental species is strongly affected by fungal diseases such as chrysanthemumwhite rust (Puccinia horiana, high doses of fungicides are being used posing increased environmental and commercialcosts. Assessment of in vitro regeneration systems from leaf discs was a first step in developing a plant genetic transformationprotocol to obtain fungi-resistant plants. Leaf discs of White Albatross, Yellow Albatross, and Escapade varieties wereestablished in vitro on MS medium supplemented with NAA (0 - 4.83 μM and BAP (0 - 13.32 μM alone and incombination. Leaf discs were also cultured on MumB medium containing 2,4-D (0 - 4.52 μM for 7, 14, and 21 days priorto their transferral to a 2,4-D free MumB medium. Regenerated shoots were individualized, rooted, and hardened. Resultsshow that MS with 4.83 μM NAA + 4.44 μM BAP and 4.83 μM NAA + 13.32 μM BAP induce organogenesis, and MumBwith 2.26 μM 2,4-D induces somatic embryogenesis on all three varieties, with exposition periods to 2,4-D of 14 days forWhite Albatross and 21 days for Yellow Albatross and Escapade. Shoot development from somatic embryos was observedin the three varieties when cultured on a 2,4-D free MumB medium. Spontaneous rooting was recorded in 85% of the shootsthus facilitating hardening and successful transfer to soil.

  10. Label-Free Quantitative Proteomics of Embryogenic and Non-Embryogenic Callus during Sugarcane Somatic Embryogenesis.

    Directory of Open Access Journals (Sweden)

    Angelo Schuabb Heringer

    Full Text Available The development of somatic cells in to embryogenic cells occurs in several stages and ends in somatic embryo formation, though most of these biochemical and molecular changes have yet to be elucidated. Somatic embryogenesis coupled with genetic transformation could be a biotechnological tool to improve potential crop yields potential in sugarcane cultivars. The objective of this study was to observe somatic embryo development and to identify differentially expressed proteins in embryogenic (E and non-embryogenic (NE callus during maturation treatment. E and NE callus were cultured on maturation culture medium supplemented with different concentrations (0.0, 0.75, 1.5 and 2.0 g L(-1 of activated charcoal (AC. Somatic embryo formation and differential protein expression were evaluated at days 0 and 21 using shotgun proteomic analyses. Treatment with 1.5 g L(-1 AC resulted in higher somatic embryo maturation rates (158 somatic embryos in 14 days in E callus but has no effect in NE callus. A total of 752 co-expressed proteins were identified through the SUCEST (The Sugarcane EST Project, including many housekeeping proteins. E callus showed 65 exclusive proteins on day 0, including dehydrogenase, desiccation-related protein, callose synthase 1 and nitric oxide synthase. After 21 days on maturation treatment, 14 exclusive proteins were identified in E callus, including catalase and secreted protein. NE callus showed 23 exclusive proteins on day 0 and 10 exclusive proteins after 21 days on maturation treatment, including many proteins related to protein degradation. The induction of maturation leads to somatic embryo development, which likely depends on the expression of specific proteins throughout the process, as seen in E callus under maturation treatment. On the other hand, some exclusive proteins can also specifically prevent of somatic embryos development, as seen in the NE callus.

  11. Zebrafish Noxa promotes mitosis in early embryonic development and regulates apoptosis in subsequent embryogenesis.

    Science.gov (United States)

    Zhong, J-X; Zhou, L; Li, Z; Wang, Y; Gui, J-F

    2014-06-01

    Noxa functions in apoptosis and immune system of vertebrates, but its activities in embryo development remain unclear. In this study, we have studied the role of zebrafish Noxa (zNoxa) by using zNoxa-specifc morpholino knockdown and overexpression approaches in developing zebrafish embryos. Expression pattern analysis indicates that zNoxa transcript is of maternal origin, which displays a uniform distribution in early embryonic development until shield stage, and the zygote zNoxa transcription is initiated from this stage and mainly localized in YSL of the embryos. The zNoxa expression alterations result in strong embryonic development defects, demonstrating that zNoxa regulates apoptosis from 75% epiboly stage of development onward, in which zNoxa firstly induces the expression of zBik, and then cooperates with zBik to regulate apoptosis. Moreover, zNoxa knockdown also causes a reduction in number of mitotic cells before 8 h.p.f., suggesting that zNoxa also promotes mitosis before 75% epiboly stage. The effect of zNoxa on mitosis is mediated by zWnt4b in early embryos, whereas zMcl1a and zMcl1b suppress the ability of zNoxa to regulate mitosis and apoptosis at different developmental stages. In addition, mammalian mouse Noxa (mNoxa) mRNA was demonstrated to rescue the arrest of mitosis when zNoxa was knocked down, suggesting that mouse and zebrafish Noxa might have similar dual functions. Therefore, the current findings indicate that Noxa is a novel regulator of early mitosis before 75% epiboly stage when it translates into a key mediator of apoptosis in subsequent embryogenesis.

  12. Drosophila MOF controls Checkpoint protein2 and regulates genomic stability during early embryogenesis.

    Science.gov (United States)

    Pushpavalli, Sreerangam N C V L; Sarkar, Arpita; Ramaiah, M Janaki; Chowdhury, Debabani Roy; Bhadra, Utpal; Pal-Bhadra, Manika

    2013-01-24

    In Drosophila embryos, checkpoints maintain genome stability by delaying cell cycle progression that allows time for damage repair or to complete DNA synthesis. Drosophila MOF, a member of MYST histone acetyl transferase is an essential component of male X hyperactivation process. Until recently its involvement in G2/M cell cycle arrest and defects in ionizing radiation induced DNA damage pathways was not well established. Drosophila MOF is highly expressed during early embryogenesis. In the present study we show that haplo-insufficiency of maternal MOF leads to spontaneous mitotic defects like mitotic asynchrony, mitotic catastrophe and chromatid bridges in the syncytial embryos. Such abnormal nuclei are eliminated and digested in the yolk tissues by nuclear fall out mechanism. MOF negatively regulates Drosophila checkpoint kinase 2 tumor suppressor homologue. In response to DNA damage the checkpoint gene Chk2 (Drosophila mnk) is activated in the mof mutants, there by causing centrosomal inactivation suggesting its role in response to genotoxic stress. A drastic decrease in the fall out nuclei in the syncytial embryos derived from mof¹/+; mnkp⁶/+ females further confirms the role of DNA damage response gene Chk2 to ensure the removal of abnormal nuclei from the embryonic precursor pool and maintain genome stability. The fact that mof mutants undergo DNA damage has been further elucidated by the increased number of single and double stranded DNA breaks. mof mutants exhibited genomic instability as evidenced by the occurance of frequent mitotic bridges in anaphase, asynchronous nuclear divisions, disruption of cytoskeleton, inactivation of centrosomes finally leading to DNA damage. Our findings are consistent to what has been reported earlier in mammals that; reduced levels of MOF resulted in increased genomic instability while total loss resulted in lethality. The study can be further extended using Drosophila as model system and carry out the interaction of MOF

  13. Mcm10 is required for oogenesis and early embryogenesis in Drosophila.

    Science.gov (United States)

    Reubens, Michael C; Biller, Megan D; Bedsole, Sidney E; Hopkins, Lucas T; Ables, Elizabeth T; Christensen, Tim W

    2015-11-01

    Efficient replication of the genome and the establishment of endogenous chromatin states are processes that are essential to eukaryotic life. It is well documented that Mcm10 is intimately linked to both of these important biological processes; therefore, it is not surprising that Mcm10 is commonly misregulated in many human cancers. Most of the research regarding the biological roles of Mcm10 has been performed in single-cell or cell-free in-vitro systems. Though these systems are informative, they are unable to provide information on the cell-specific function of Mcm10 in the context of the tissue and organ systems that comprise multicellular eukaryotes. We therefore sought to identify the potential biological functions of Mcm10 in the context of a complex multicellular organism by continuing our analysis in Drosophila using three novel hypomorphic alleles. Observation of embryonic nuclear morphology and quantification of embryo hatch rates reveal that maternal loading of Mcm10 is required for embryonic nuclear stability, and suggest a role for Mcm10 post zygotic transition. Contrary to the essential nature of Mcm10 depicted in the literature, it does not appear to be required for adult viability in Drosophila if embryonic requirements are met. Although not required for adult somatic viability, analysis of fecundity and ovarian morphology in mutant females suggest that Mcm10 plays a role in maintenance of the female germline. Taken together, our results demonstrate critical roles for Mcm10 during early embryogenesis, and mark the first data linking Mcm10 to female specific reproduction in multicellular eukaryotes. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Mitotic waves in the early embryogenesis ofDrosophila: Bistability traded for speed.

    Science.gov (United States)

    Vergassola, Massimo; Deneke, Victoria E; Di Talia, Stefano

    2018-02-15

    Early embryogenesis of most metazoans is characterized by rapid and synchronous cleavage divisions. Chemical waves of Cdk1 activity were previously shown to spread across Drosophila embryos, and the underlying molecular processes were dissected. Here, we present the theory of the physical mechanisms that control Cdk1 waves in Drosophila The in vivo dynamics of Cdk1 are captured by a transiently bistable reaction-diffusion model, where time-dependent reaction terms account for the growing level of cyclins and Cdk1 activation across the cell cycle. We identify two distinct regimes. The first one is observed in mutants of the mitotic switch. There, waves are triggered by the classical mechanism of a stable state invading a metastable one. Conversely, waves in wild type reflect a transient phase that preserves the Cdk1 spatial gradients while the overall level of Cdk1 activity is swept upward by the time-dependent reaction terms. This unique mechanism generates a wave-like spreading that differs from bistable waves for its dependence on dynamic parameters and its faster speed. Namely, the speed of "sweep" waves strikingly decreases as the strength of the reaction terms increases and scales as the powers 3/4, -1/2, and 7/12 of Cdk1 molecular diffusivity, noise amplitude, and rate of increase of Cdk1 activity in the cell-cycle S phase, respectively. Theoretical predictions are supported by numerical simulations and experiments that couple quantitative measurements of Cdk1 activity and genetic perturbations of the accumulation rate of cyclins. Finally, our analysis bears upon the inhibition required to suppress Cdk1 waves at the cell-cycle pause for the maternal-to-zygotic transition.

  15. Nuclear glycogen synthase kinase-3 {beta} (GSK-3) in Rhipicephalus (Boophilus) microplus tick embryogenesis

    Energy Technology Data Exchange (ETDEWEB)

    Mentzingen, Leticia; Andrade, Josiana G. de; Logullo, Carlos [Universidade Estadual do Norte Fluminense Darcy Ribeiro (UENF), Campos dos Goytacazes, RJ (Brazil). Centro de Biociencias e Biotecnologia. Lab. de Quimica e Funcao de Proteinas e Peptideos (LQFPP); Andrade, Caroline P. de; Vaz Junior, Itabajara [Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, RS (Brazil). Centro de Biotecnologia

    2008-07-01

    Full text: Glycogen synthase kinase-3 (GSK3) is recognized as a key component of a large number of cellular processes and diseases. Several mechanisms play a part in controlling the actions of GSK3, including phosphorylation, protein complex formation, and subcellular distribution. Recent observations point to functions for phosphorylases several transcription factors in the nucleus. Also, GSK3b participate of the canonical W nt signalling pathway, which has been studied intensively in embryonic and cancer cells. Like in many other signaling pathways, most components in W nt signal transduction were highly conserved during the evolution. More than 40 proteins have been reported to be phosphorylated by GSK3, including over a dozen transcription factors. Although the mechanisms regulating GSK3 are not fully understood, precise control appears to be achieved by a combination of phosphorylation, localization, and interactions with GSK3-binding proteins. Although GSK3 is traditionally considered a cytosolic protein, it is also present in nuclei. Nuclear GSK3 is particularly interesting because of the many transcription factors that it regulates enabling GSK3 to influence many signaling pathways that converge on these transcription factors, thereby regulating the expression of many genes. Our group identified that GSK-3 {beta} could be detected in different stage eggs of R. micro plus. In this work we detected the GSK-3 in isolated nuclear fraction from the egg homogenates of R. micro plus by western-blot analysis, using anti-GSK- 3 {beta} antibodies. The enzyme activity was also detected radiochemically throughout embryogenesis in same fraction. The GSK-3 activity was inhibiting by using SB 216763 (selective molecule inhibitors of GSK-3). Taken together our results suggest that GSK-3 {beta} isoform probably is involved in gene transcription factors during R. micro plus embryo development.

  16. Current (2004-07) Conditions and Changes in Ground-Water Levels from Predevelopment to 2007, Southern High Plains Aquifer, Southeast New Mexico-Lea County Underground Water Basin

    Science.gov (United States)

    Tillery, Anne

    2008-01-01

    The Southern High Plains aquifer is the principal aquifer and primary source of water in southeastern New Mexico. The Lea County portion of the aquifer covers approximately the northern two thirds of the 4,393-square-mile county. Successful water-supply planning for New Mexico's Southern High Plains requires knowledge of the current aquifer conditions and a context from which to estimate future trends given current aquifer-management policy. Maps representing water-level declines, current (2007) water levels, aquifer saturated thickness, and depth to water accompanied by hydrographs from representative wells for the Southern High Plains aquifer in the Lea County Underground Water Basin were prepared in cooperation with the New Mexico Office of the State Engineer. Results of this mapping effort show the water level has declined as much as 97 feet in the Lea County Underground Water Basin from predevelopment (1914-54) to 2007 with rates as high as 0.88 feet per year.

  17. Codo de tenis (tendinosis epicondílea externa): tratamiento esclerosante ecoguiado con polidocanol. A propósito de dos casos

    OpenAIRE

    Álvarez Rey, Guillermo; Álvarez Rey, Ignacio; Álvarez Bustos, Guillermo

    2006-01-01

    Dos pacientes (2 varones de 42 y 55 años) con tendinosis epicondílea externa dolorosa fueron tratados mediante un novedoso tratamiento que se basa en la esclerosis con Polidocanol, un agente esclerosante, ecoguiada con Power Color (PC) mediante Doppler color de polidocanol, un agente esclerosante. Tras el tratamiento se produjo una marcada reducción del dolor recogida mediante una escala analógica visual (VAS) y desaparición y disminución de la vascularización en cada caso, respectivamente. E...

  18. Estudio de concordancia de mediciones radiológicas en fracturas supracondíleas de húmero en niños

    OpenAIRE

    Abril Aguilar, Andrés Camilo; Buitrago Buitrago, Luis Guillermo; Vergara Amador, Enrique Manuel

    2010-01-01

    Antecedente. En la valoración radiológica de las fracturas supracondíleas de codo en niños existen algunas mediciones utilizadas como el ángulo de Baumann, el ángulo de inclinación diafisario-condíleo, la línea humeral anterior y la valoración rotacional. No se dispone de evidencia acerca de la concordancia de estas mediciones y los parámetros para determinar un tratamiento o los resultados del mismo, bajo esos criterios. Objetivo. Determinar la concordancia de esas mediciones y su relaci...

  19. Abundance measurements in stellar environments

    Science.gov (United States)

    Leone, F.

    2014-05-01

    Most of what we know about stars, and systems of stars, is derived from the analysis of their electromagnetic radiation. This lesson is an attempt to describe to Physicists, without any Astrophysical background, the framework to understand the present status of abundance determination in stellar environments and its limit. These notes are dedicated to the recently passed, November 21, 2013, Prof. Dimitri Mihalas who spent his life confuting the 19th century positivist philosopher Auguste Comte who stated that we shall not at all be able to determine the chemical composition of stars.

  20. Abundance measurements in stellar environments

    Energy Technology Data Exchange (ETDEWEB)

    Leone, F. [Università di Catania, Dipartimento di Fisica e Astronomia, Sezione Astrofisica, Via S. Sofia 78, 95123 Catania (Italy)

    2014-05-09

    Most of what we know about stars, and systems of stars, is derived from the analysis of their electromagnetic radiation. This lesson is an attempt to describe to Physicists, without any Astrophysical background, the framework to understand the present status of abundance determination in stellar environments and its limit. These notes are dedicated to the recently passed, November 21, 2013, Prof. Dimitri Mihalas who spent his life confuting the 19th century positivist philosopher Auguste Comte who stated that we shall not at all be able to determine the chemical composition of stars.

  1. Recurrent somatic embryogenesis in long-term cultures of Gentiana lutea L. as a source for synthetic seed production for medium-term preservation

    Directory of Open Access Journals (Sweden)

    Holobiuc Irina

    2012-01-01

    Full Text Available Our aim was to establish an efficient and reproducible system for producing synthetic seeds from recurrent somatic embryogenesis in long-term cultures of Gentiana lutea L. This species is a vulnerable medicinal plant, protected both at the national and international levels, and is included in different Red Lists and Books. In vitro culture, as an alternative to classical methods of preservation, allows for the cyclic multiplication of plant material and short-, medium- and long-term preservation of tissue collections. Biotechnological approaches allow for maintenance of the plant material in a confined space and protection against biotic and abiotic factors. Somatic embryogenesis (SE is the most efficient way to regenerate plants, ensuring material for preservation and fundamental research. In our experiment, recurrent somatic embryogenesis was developed in long-term cultures in the presence of sugar alcohols (mannitol, sorbitol and in the absence of growth factors. This process proceeded at a high rate, with adventive somatic embryos being generated in a continuous process, followed by maturation, germination and development into plants. To follow the somatic embryogenesis process, histological samples were made. We used these embryogenic cultures for synthetic seed production and medium-term conservation. The viability of somatic embryos after moderate osmotic stress treatment was tested using TTC. Our methodology relied on the induction of somatic embryogenesis in the presence of auxins in the first cycle of in vitro cultures, long-term high embryogenic culture maintenance in the presence of sugar alcohols and synthetic seed production.

  2. Doubled haploid production from Spanish onion (Allium cepa L.) germplasm: embryogenesis induction, plant regeneration and chromosome doubling

    Science.gov (United States)

    Fayos, Oreto; Vallés, María P.; Garcés-Claver, Ana; Mallor, Cristina; Castillo, Ana M.

    2015-01-01

    The use of doubled haploids in onion breeding is limited due to the low gynogenesis efficiency of this species. Gynogenesis capacity from Spanish germplasm, including the sweet cultivar Fuentes de Ebro, the highly pungent landrace BGHZ1354 and the two Valenciana type commercial varieties Recas and Rita, was evaluated and optimized in this study. The OH-1 population, characterized by a high gynogenesis induction, was used as control. Growing conditions of the donor plants were tested with a one-step protocol and field plants produced a slightly higher percentage of embryogenesis induction than growth chamber plants. A one-step protocol was compared with a two-step protocol for embryogenesis induction. Spanish germplasm produced a 2–3 times higher percentage of embryogenesis with the two-step protocol, Recas showing the highest percentage (2.09%) and Fuentes de Ebro the lowest (0.53%). These percentages were significantly lower than those from the OH-1 population, with an average of 15% independently of the protocol used. The effect of different containers on plant regeneration was tested using both protocols. The highest percentage of acclimated plants was obtained with the two-step protocol in combination with Eco2box (70%), whereas the lowest percentage was observed with glass tubes in the two protocols (20–23%). Different amiprofos-methyl (APM) treatments were applied to embryos for chromosome doubling. A similar number of doubled haploid plants were recovered with 25 or 50 μM APM in liquid medium. However, the application of 25 μM in solid medium for 24 h produced the highest number of doubled haploid plants. Somatic regeneration from flower buds of haploid and mixoploid plants proved to be a successful approach for chromosome doubling, since diploid plants were obtained from the four regenerated lines. In this study, doubled haploid plants were produced from the four Spanish cultivars, however further improvements are needed to increase their gynogenesis

  3. Confirmation of Transgenic Robusta Coffee (Coffea canephora Transformed by Chitinase-encoding Gene and Its Propagation Through Somatic Embryogenesis

    Directory of Open Access Journals (Sweden)

    Priyono .

    2005-08-01

    Full Text Available Genetic engineering of Robusta coffee resistant to fungal diseases might be done by introducing a chitinase-encoding gene into genome of this plant. This research was aimed to confirm transgenic plant of BP 308 clone Robusta coffee transformed by chi gene and to evaluate its ability for the somatic embryogenesis. Confirmation of transgenic was carried out by analysis the presence of NPTII gene as a selectable marker for Canamysin resistant using PCR technique. The somatic embryo initiation and reproduction were evaluated in 11 plant accessions. Three kinds of sucrose concentration, 20%, 30% and 40% were applied in initiation stage of somatic embryo germination. The suitability of 4 medium, namely M1 (without addition by liquid medium, M2 (addition by liquid medium contained 0.25 mg/l kinetin, M3 (addition by liquid medium contained 0.25 mg/l IAA and M4 (addition by liquid medium contained 0.25 mg/l GA3 was evaluated for somatic embryo maturation. The result showed that 8 out of 10 plant accessions tested were transgenic and they could be propagated through somatic embryogenesis. The ability of transgenic plant for somatic embryo initiation, reproduction and regeneration were similar with that of nontransgenic one. Germination of somatic embryo could be improved by using 40% sucrose. Maturation of somatic embryo could be improved by addition of fresh liquid medium on the ancient gelled medium that used for somatic embryos reproduction. The best result was obtained on addition of fresh medium contained 0.25 mg/l GA 3 in which 65% of the somatic embryos developed to pre-germinate somatic embryo. Key words: Coffea canephora, transgenic plant, somatic embryogenesis.

  4. Desiccation Treatment and Endogenous IAA Levels Are Key Factors Influencing High Frequency Somatic Embryogenesis in Cunninghamia lanceolata (Lamb. Hook

    Directory of Open Access Journals (Sweden)

    Xiaohong Zhou

    2017-12-01

    Full Text Available Cunninghamia lanceolata (Lamb. Hook (Chinese fir is an important tree, commercially and ecologically, in southern China. The traditional regenerating methods are based on organogenesis and cutting propagation. Here, we report the development of a high-frequency somatic embryogenesis (SE regeneration system synchronized via a liquid culture from immature zygotic embryos. Following synchronization, PEM II cell aggregates were developmentally equivalent in appearance to cleaved zygotic embryos. Embryo and suspensor growth and subsequent occurrence of the apical and then the cotyledonary meristems were similar for zygotic and SE embryo development. However, SE proembryos exhibited a more reddish coloration than zygotic proembryos, and SE embryos were smaller than zygotic embryos. Mature somatic embryos gave rise to plantlets on hormone-free medium. For juvenile explants, low concentrations of endogenous indole-3-acetic acid in initial explants correlated with improved proembryogenic mass formation, and high SE competency. Analysis of karyotypes and microsatellites detected no major genetic variation in the plants regenerated via SE, and suggest a potential in the further development of this system as a reliable methodology for true-to-type seedling production. Treatment with polyethylene glycol (PEG and abscisic acid (ABA were of great importance to proembryo formation and complemented each other. ABA assisted the growth of embryonal masses, whereas PEG facilitated the organization of the proembryo-like structures. SOMATIC EMBRYOGENESIS RECEPTOR KINASE SERK and the WUSCHEL homeobox (WOX transcription factor served as molecular markers during early embryogenesis. Our results show that ClSERKs are conserved and redundantly expressed during SE. SERK and WOX transcript levels were highest during development of the proembryos and lowest in developed embryos. ClWOX13 expression correlates with the critical transition from proembryogenic masses to

  5. A SIMPLE PROTOCOL FOR SOMATIC EMBRYOGENESIS INDUCTION OF IN VITRO SUGARCANE ( Saccharum officinarum. L BY 2,4-D AND BAP

    Directory of Open Access Journals (Sweden)

    Laily Ilman Widuri

    2016-05-01

    Full Text Available Induction of in vitro sugarcane through somatic embryogenesis technique influenced by addition of plant growth regulator. The objective of this research was to determine appropriate formulation medium for indirect somatic embryogenesis induction on two potential sugarcane SUT Event 02 and PS 881. This research carried out in three steps, callus induction, callus proliferation, and shoot regeneration. Explants taken from basal of in vitro plantlet one month SUT Event 02 and PS 881 resulted from shoot regeneration previously. Five different medium formulas applied for callus induction and one formula for proliferation and shoot regeneration. Research using completely randomized design (CRD factorial with five different formulation induction mediums. The result showed that the best respond of indirect somatic embryogenesis on SUT Event 02 and PS 881 was medium containing  3 mgL-1of  2,4-D.

  6. Thidiazuron induces shoot organogenesis at low concentrations and somatic embryogenesis at high concentrations on leaf and petiole explants of African violet (Saintpaulia ionantha Wendl).

    Science.gov (United States)

    Mithila, J; Hall, J C; Victor, J M R; Saxena, P K

    2003-01-01

    Regeneration via shoot organogenesis and somatic embryogenesis was observed from thidiazuron (TDZ)-treated leaf and petiole explants of greenhouse- and in vitro-grown African violet plants. The response of cultures to other growth regulators over a range of 0.5 microM to 10 microM was 50% less than that observed with TDZ. A comparative study among several cultivars of African violet indicated that "Benjamin" and "William" had the highest regeneration potential. In "Benjamin", higher frequencies of shoot organogenesis (twofold) and somatic embryogenesis (a 50% increase) were observed from in vitro- and greenhouse-grown plants, respectively. At concentrations lower than 2.5 microM, TDZ induced shoot organogenesis, whereas at higher doses (5-10 microM) somatic embryos were formed. These findings provide the first report of simultaneous shoot organogenesis and somatic embryogenesis of African violet explants in response to TDZ.

  7. 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.

  8. Macondo crude oil from the Deepwater Horizon oil spill disrupts specific developmental processes during zebrafish embryogenesis

    Directory of Open Access Journals (Sweden)

    de Soysa T Yvanka

    2012-05-01

    Full Text Available Abstract Background The Deepwater Horizon disaster was the largest marine oil spill in history, and total vertical exposure of oil to the water column suggests it could impact an enormous diversity of ecosystems. The most vulnerable organisms are those encountering these pollutants during their early life stages. Water-soluble components of crude oil and specific polycyclic aromatic hydrocarbons have been shown to cause defects in cardiovascular and craniofacial development in a variety of teleost species, but the developmental origins of these defects have yet to be determined. We have adopted zebrafish, Danio rerio, as a model to test whether water accumulated fractions (WAF of the Deepwater Horizon oil could impact specific embryonic developmental processes. While not a native species to the Gulf waters, the developmental biology of zebrafish has been well characterized and makes it a powerful model system to reveal the cellular and molecular mechanisms behind Macondo crude toxicity. Results WAF of Macondo crude oil sampled during the oil spill was used to treat zebrafish throughout embryonic and larval development. Our results indicate that the Macondo crude oil causes a variety of significant defects in zebrafish embryogenesis, but these defects have specific developmental origins. WAF treatments caused defects in craniofacial development and circulatory function similar to previous reports, but we extend these results to show they are likely derived from an earlier defect in neural crest cell development. Moreover, we demonstrate that exposure to WAFs causes a variety of novel deformations in specific developmental processes, including programmed cell death, locomotor behavior, sensory and motor axon pathfinding, somitogenesis and muscle patterning. Interestingly, the severity of cell death and muscle phenotypes decreased over several months of repeated analysis, which was correlated with a rapid drop-off in the aromatic and alkane

  9. Plant regeneration through somatic embryogenesis and genome size analysis of Coriandrum sativum L.

    Science.gov (United States)

    Ali, Muzamil; Mujib, A; Tonk, Dipti; Zafar, Nadia

    2017-01-01

    In the present study, an improved plant regeneration protocol via primary and secondary somatic embryogenesis was established in two Co-1 and Rajendra Swathi (RS) varieties of Coriandrum sativum L. Callus was induced from root explants on 2, 4-D (0.5-2.0 mg/l) supplemented MS. The addition of BA (0.2 mg/l) improved callus induction and proliferation response significantly. The maximum callus induction frequency was on 1.0 mg/l 2, 4-D and 0.2 mg/l BA added MS medium (77.5 % in Co-1 and 72.3 % in RS). The callus transformed into embryogenic callus on 2, 4-D added MS with maximum embryogenic frequency was on 1.0 mg/l. The granular embryogenic callus differentiated into globular embryos on induction medium, which later progressed to heart-, torpedo- and cotyledonary embryos on medium amended with 0.5 mg/l NAA and 0.2 mg/l BA. On an average, 2-3 secondary somatic embryos (SEs) were developed on mature primary SEs, which increased the total embryo numbers in culture. Histology and scanning electron microscopy (SEM) studies are presented for the origin, development of primary and secondary embryos in coriander. Later, these induced embryos converted into plantlets on 1.0 mg/l BA and 0.2 mg/l NAA-amended medium. The regenerated plantlets were cultured on 0.5 mg/l IBA added ½ MS for promotion of roots. The well-rooted plantlets were acclimatized and transferred to soil. The genetic stability of embryo-regenerated plant was analyzed by flow cytometry with optimized Pongamia pinnata as standard. The 2C DNA content of RS coriander variety was estimated to 5.1 pg; the primary and secondary somatic embryo-derived plants had 5.26 and 5.44 pg 2C DNA content, respectively. The regenerated plants were genetically stable, genome size similar to seed-germinated coriander plants.

  10. Macondo crude oil from the Deepwater Horizon oil spill disrupts specific developmental processes during zebrafish embryogenesis.

    Science.gov (United States)

    de Soysa, T Yvanka; Ulrich, Allison; Friedrich, Timo; Pite, Danielle; Compton, Shannon L; Ok, Deborah; Bernardos, Rebecca L; Downes, Gerald B; Hsieh, Shizuka; Stein, Rachael; Lagdameo, M Caterina; Halvorsen, Katherine; Kesich, Lydia-Rose; Barresi, Michael J F

    2012-05-04

    The Deepwater Horizon disaster was the largest marine oil spill in history, and total vertical exposure of oil to the water column suggests it could impact an enormous diversity of ecosystems. The most vulnerable organisms are those encountering these pollutants during their early life stages. Water-soluble components of crude oil and specific polycyclic aromatic hydrocarbons have been shown to cause defects in cardiovascular and craniofacial development in a variety of teleost species, but the developmental origins of these defects have yet to be determined. We have adopted zebrafish, Danio rerio, as a model to test whether water accumulated fractions (WAF) of the Deepwater Horizon oil could impact specific embryonic developmental processes. While not a native species to the Gulf waters, the developmental biology of zebrafish has been well characterized and makes it a powerful model system to reveal the cellular and molecular mechanisms behind Macondo crude toxicity. WAF of Macondo crude oil sampled during the oil spill was used to treat zebrafish throughout embryonic and larval development. Our results indicate that the Macondo crude oil causes a variety of significant defects in zebrafish embryogenesis, but these defects have specific developmental origins. WAF treatments caused defects in craniofacial development and circulatory function similar to previous reports, but we extend these results to show they are likely derived from an earlier defect in neural crest cell development. Moreover, we demonstrate that exposure to WAFs causes a variety of novel deformations in specific developmental processes, including programmed cell death, locomotor behavior, sensory and motor axon pathfinding, somitogenesis and muscle patterning. Interestingly, the severity of cell death and muscle phenotypes decreased over several months of repeated analysis, which was correlated with a rapid drop-off in the aromatic and alkane hydrocarbon components of the oil. Whether these

  11. Glucose 6-phosphate dehydrogenase deficiency enhances germ cell apoptosis and causes defective embryogenesis in Caenorhabditis elegans

    Science.gov (United States)

    Yang, H-C; Chen, T-L; Wu, Y-H; Cheng, K-P; Lin, Y-H; Cheng, M-L; Ho, H-Y; Lo, S J; Chiu, D T-Y

    2013-01-01

    Glucose 6-phosphate dehydrogenase (G6PD) deficiency, known as favism, is classically manifested by hemolytic anemia in human. More recently, it has been shown that mild G6PD deficiency moderately affects cardiac function, whereas severe G6PD deficiency leads to embryonic lethality in mice. How G6PD deficiency affects organisms has not been fully elucidated due to the lack of a suitable animal model. In this study, G6PD-deficient Caenorhabditis elegans was established by RNA interference (RNAi) knockdown to delineate the role of G6PD in animal physiology. Upon G6PD RNAi knockdown, G6PD activity was significantly hampered in C. elegans in parallel with increased oxidative stress and DNA oxidative damage. Phenotypically, G6PD-knockdown enhanced germ cell apoptosis (2-fold increase), reduced egg production (65% of mock), and hatching (10% of mock). To determine whether oxidative stress is associated with G6PD knockdown-induced reproduction defects, C. elegans was challenged with a short-term hydrogen peroxide (H2O2). The early phase egg production of both mock and G6PD-knockdown C. elegans were significantly affected by H2O2. However, H2O2-induced germ cell apoptosis was more dramatic in mock than that in G6PD-deficient C. elegans. To investigate the signaling pathways involved in defective oogenesis and embryogenesis caused by G6PD knockdown, mutants of p53 and mitogen-activated protein kinase (MAPK) pathways were examined. Despite the upregulation of CEP-1 (p53), cep-1 mutation did not affect egg production and hatching in G6PD-deficient C. elegans. Neither pmk-1 nor mek-1 mutation significantly affected egg production, whereas sek-1 mutation further decreased egg production in G6PD-deficient C. elegans. Intriguingly, loss of function of sek-1 or mek-1 dramatically rescued defective hatching (8.3- and 9.6-fold increase, respectively) induced by G6PD knockdown. Taken together, these findings show that G6PD knockdown reduces egg production and hatching in C. elegans

  12. Hematite Abundance Map at Echo

    Science.gov (United States)

    2004-01-01

    This image shows the hematite abundance map for a portion of the Meridiani Planum rock outcrop near where the Mars Exploration Rover Opportunity landed. It was acquired by the rover's miniature thermal emission spectrometer instrument from a spot called 'Echo.' Portions of the inner crater wall in this region appear rich in hematite (red). The sharp boundary from hematite-rich to hematite-poor (yellow and green) surfaces corresponds to a change in the surface texture and color. The hematite-rich surfaces have ripple-like forms suggesting wind transported hematite to these surfaces. The bounce marks produced during landing at the base of the slope on the left are low in hematite (blue). The hematite grains that originally covered the surface were pushed below the surface by the lander, exposing a soil that has less hematite.

  13. Immuno-cytochemical localization of indole-3-acetic acid during induction of somatic embryogenesis in cultured sunflower embryos.

    Science.gov (United States)

    Thomas, Clément; Bronner, Roberte; Molinier, Jean; Prinsen, Els; van Onckelen, Harry; Hahne, Günther

    2002-08-01

    Immature zygotic embryos of sunflower (Helianthus annuus L.) produce somatic embryos when cultured on medium supplemented with a cytokinin as the sole source of exogenous growth regulators. The timing of the induction phase and subsequent morphogenic events have been well characterized in previous work. We address here the question of the role of endogenous indole-3-acetic acid (IAA), since auxins are known to have a crucial role in the induction of somatic embryogenesis in many other culture and regeneration systems. The fact that in the sunflower system no exogenous auxin is required for the induction of somatic embryos makes this system very suitable for the study of the internal dynamics of IAA. We used an immuno-cytochemical approach to visualize IAA distribution within the explants before, during and after the induction phase. IAA accumulated transiently throughout cultured embryos during the induction phase. The detected signal was not uniform but certain tissues, such as the root cap and the root meristem, accumulated IAA in a more pronounced manner. IAA accumulation was not restricted to the reactive zone but the kinetics of endogenous variations strikingly mimic the pulse of IAA that is usually provoked by exogenous IAA application. The direct evidence presented here indicates that an endogenous auxin pulse is indeed among the first signals leading to the induction of somatic embryogenesis.

  14. Effect of the combination of NAA, kinetin and sucrose on the induction of somatic embryogenesis in lulo (Solanum quitoense Lam.

    Directory of Open Access Journals (Sweden)

    Hernando Criollo

    2014-08-01

    Full Text Available Lulo is a species of great importance to the fruticulture of Colombia, but has significant phytosanitary problems that require an aggressive breeding program oriented toward the production of genotypes with tolerance to phytopathogens. These programs need to establish highly efficient mass plant propagation protocols, such as somatic embryogenesis. This study focused on research on the somatic embryogenesis of lulo using kinetin, naphthalene acetic acid-NAA (Plant Growth Regulators, PGRs, and different sucrose concentrations in a MS medium. Two lulo varieties, Solanum quitoense var. septentrionale and S. quitoense var. quitoense, and two explant types (hypocotyl and cotyledon were used, incubated in dark conditions at 25±2°C. The highest production percentage of the embryos was obtained when 50 muM of NAA were added to the medium with sucrose (50.0 and 263.1 muM for the two explant types used. In lulo with spines, the highest percentage of embryonic structures (50% was observed with cotyledonary leaf explants and 50 muM of NAA ; while in the spineless lulo, the embryonic structures were observed in the same type of explant with 50 muM of NAA + 263.1 muM of sucrose (32%.

  15. Bottlenecks in bog pine multiplication by somatic embryogenesis and their visualization with the environmental scanning electron microscope.

    Science.gov (United States)

    Vlašínová, Helena; Neděla, Vilem; Đorđević, Biljana; Havel, Ladislav

    2017-07-01

    Somatic embryogenesis (SE) is an important biotechnological technique used for the propagation of many pine species in vitro. However, in bog pine, one of the most endangered tree species in the Czech Republic, limitations were observed, which negatively influenced the development and further germination of somatic embryos. Although initiation frequency was very low-0.95 %, all obtained cell lines were subjected to maturation. The best responding cell line (BC1) was used and subjected to six different variants of the maturation media. The media on which the highest number of early-precotyledonary/cotyledonary somatic embryos was formed was supplemented with 121 μM abscisic acid (ABA) and with 6 % maltose. In the end of maturation experiments, different abnormalities in formation of somatic embryos were observed. For visualization and identification of abnormalities in meristem development during proliferation and maturation processes, the environmental scanning electron microscope was used. In comparison to the classical light microscope, the non-commercial environmental scanning electron microscope AQUASEM II has been found as a very useful tool for the quick recognition of apical meristem disruption and abnormal development. To our knowledge, this is the first report discussing somatic embryogenesis in bog pine. Based on this observation, the cultivation procedure could be enhanced and the method for SE of bog pine optimized.

  16. Somatic Embryogenesis in Coffee: The Evolution of Biotechnology and the Integration of Omics Technologies Offer Great Opportunities.

    Science.gov (United States)

    Campos, Nádia A; Panis, Bart; Carpentier, Sebastien C

    2017-01-01

    One of the most important crops cultivated around the world is coffee. There are two main cultivated species, Coffea arabica and C. canephora. Both species are difficult to improve through conventional breeding, taking at least 20 years to produce a new cultivar. Biotechnological tools such as genetic transformation, micropropagation and somatic embryogenesis (SE) have been extensively studied in order to provide practical results for coffee improvement. While genetic transformation got many attention in the past and is booming with the CRISPR technology, micropropagation and SE are still the major bottle neck and urgently need more attention. The methodologies to induce SE and the further development of the embryos are genotype-dependent, what leads to an almost empirical development of specific protocols for each cultivar or clone. This is a serious limitation and excludes a general comprehensive understanding of the process as a whole. The aim of this review is to provide an overview of which achievements and molecular insights have been gained in (coffee) somatic embryogenesis and encourage researchers to invest further in the in vitro technology and combine it with the latest omics techniques (genomics, transcriptomics, proteomics, metabolomics, and phenomics). We conclude that the evolution of biotechnology and the integration of omics technologies offer great opportunities to (i) optimize the production process of SE and the subsequent conversion into rooted plantlets and (ii) to screen for possible somaclonal variation. However, currently the usage of the latest biotechnology did not pass the stage beyond proof of potential and needs to further improve.

  17. Efficient somatic embryogenesis and molecular marker based analysis as effective tools for conservation of red-listed plant Commiphora wightii

    Directory of Open Access Journals (Sweden)

    ASHOK KUMAR PARMAR

    2014-08-01

    Full Text Available A refined and high efficiency protocol for in vitro regeneration of Commiphora wightii, a red-listed medicinal plant of medicinal importance, has been developed through optimized somatic embryogenesis pathway. Cultures from immature fruits were induced and proliferated on B5 medium supplemented with 2.26 µM 2,4-D. Embryogenic calli were obtained and then maintained for extended periods by alternately subculturing on modified MS medium supplemented with 1.11 µM BAP, 0.57 µM IBA and with 0.5% activated charcoal or without PGR every 3-4 weeks. Cyclic embryogenesis was obtained. Late torpedo and early cotyledonary stages somatic embryos were regularly harvested from PGR-free modified MS medium. It was found that percent moisture available in culture containers play a critical role in maturation and subsequent germination of somatic embryos of C. wighti. Maximum germination of more than 80% was achieved through media recycling. Somatic embryo derived plants (emblings were acclimatized. After 5 months, acclimatized plants were out-planted in experimental field. These morphologically normal plants have been surviving for over 3 years. Molecular polymorphism was clearly evident when these plants were tested using RAPD primers, making the plants suitable for use in its species restoration program.

  18. Somatic Embryogenesis in Coffee: The Evolution of Biotechnology and the Integration of Omics Technologies Offer Great Opportunities

    Directory of Open Access Journals (Sweden)

    Nádia A. Campos

    2017-08-01

    Full Text Available One of the most important crops cultivated around the world is coffee. There are two main cultivated species, Coffea arabica and C. canephora. Both species are difficult to improve through conventional breeding, taking at least 20 years to produce a new cultivar. Biotechnological tools such as genetic transformation, micropropagation and somatic embryogenesis (SE have been extensively studied in order to provide practical results for coffee improvement. While genetic transformation got many attention in the past and is booming with the CRISPR technology, micropropagation and SE are still the major bottle neck and urgently need more attention. The methodologies to induce SE and the further development of the embryos are genotype-dependent, what leads to an almost empirical development of specific protocols for each cultivar or clone. This is a serious limitation and excludes a general comprehensive understanding of the process as a whole. The aim of this review is to provide an overview of which achievements and molecular insights have been gained in (coffee somatic embryogenesis and encourage researchers to invest further in the in vitro technology and combine it with the latest omics techniques (genomics, transcriptomics, proteomics, metabolomics, and phenomics. We conclude that the evolution of biotechnology and the integration of omics technologies offer great opportunities to (i optimize the production process of SE and the subsequent conversion into rooted plantlets and (ii to screen for possible somaclonal variation. However, currently the usage of the latest biotechnology did not pass the stage beyond proof of potential and needs to further improve.

  19. Identification of QTLs Associated with Callogenesis and Embryogenesis in Oil Palm Using Genetic Linkage Maps Improved with SSR Markers

    Science.gov (United States)

    Ting, Ngoot-Chin; Jansen, Johannes; Nagappan, Jayanthi; Ishak, Zamzuri; Chin, Cheuk-Weng; Tan, Soon-Guan; Cheah, Suan-Choo; Singh, Rajinder

    2013-01-01

    Clonal reproduction of oil palm by means of tissue culture is a very inefficient process. Tissue culturability is known to be genotype dependent with some genotypes being more amenable to tissue culture than others. In this study, genetic linkage maps enriched with simple sequence repeat (SSR) markers were developed for dura (ENL48) and pisifera (ML161), the two fruit forms of oil palm, Elaeis guineensis. The SSR markers were mapped onto earlier reported parental maps based on amplified fragment length polymorphism (AFLP) and restriction fragment length polymorphism (RFLP) markers. The new linkage map of ENL48 contains 148 markers (33 AFLPs, 38 RFLPs and 77 SSRs) in 23 linkage groups (LGs), covering a total map length of 798.0 cM. The ML161 map contains 240 markers (50 AFLPs, 71 RFLPs and 119 SSRs) in 24 LGs covering a total of 1,328.1 cM. Using the improved maps, two quantitative trait loci (QTLs) associated with tissue culturability were identified each for callusing rate and embryogenesis rate. A QTL for callogenesis was identified in LGD4b of ENL48 and explained 17.5% of the phenotypic variation. For embryogenesis rate, a QTL was detected on LGP16b in ML161 and explained 20.1% of the variation. This study is the first attempt to identify QTL associated with tissue culture amenity in oil palm which is an important step towards understanding the molecular processes underlying clonal regeneration of oil palm. PMID:23382832

  20. Establishment of embryonic shoot–root axis is involved in auxin and cytokinin response during Arabidopsis somatic embryogenesis

    Directory of Open Access Journals (Sweden)

    Ying Hua eSu

    2015-01-01

    Full Text Available Auxin and cytokinin signaling participates in regulating a large spectrum of developmental and physiological processes in plants. The shoots and roots of plants have specific and sometimes even contrary responses to these hormones. Recent studies have clearly shown that establishing the spatiotemporal distribution of auxin and cytokinin response signals is central for the control of shoot apical meristem (SAM induction in cultured tissues. However, little is known about the role of these hormones in root apical meristem (RAM initiation. Here, we found that the expression patterns of several regulatory genes critical for RAM formation were correlated with the establishment of the embryonic root meristem during somatic embryogenesis in Arabidopsis. Interestingly, the early expression of the WUS-RELATED HOMEOBOX 5 (WOX5 and WUSCHEL (WUS genes was induced and was nearly overlapped within the embryonic callus when somatic embryos (SEs could not be identified morphologically. Their correct expression was essential for RAM and SAM initiation and embryonic shoot–root axis establishment. Furthermore, we analyzed the auxin and cytokinin response during SE initiation. Notably, cytokinin response signals were detected in specific regions that were correlated with induced WOX5 expression and subsequent SE formation. Overexpression of the ARABIDOPSIS RESPONSE REGULATOR genes ARR7 and ARR15 (feedback repressors of cytokinin signaling, disturbed RAM initiation and SE induction. These results provide new information on auxin and cytokinin-regulated apical–basal polarity formation of shoot–root axis during somatic embryogenesis.

  1. Identification of QTLs associated with callogenesis and embryogenesis in oil palm using genetic linkage maps improved with SSR markers.

    Directory of Open Access Journals (Sweden)

    Ngoot-Chin Ting

    Full Text Available Clonal reproduction of oil palm by means of tissue culture is a very inefficient process. Tissue culturability is known to be genotype dependent with some genotypes being more amenable to tissue culture than others. In this study, genetic linkage maps enriched with simple sequence repeat (SSR markers were developed for dura (ENL48 and pisifera (ML161, the two fruit forms of oil palm, Elaeis guineensis. The SSR markers were mapped onto earlier reported parental maps based on amplified fragment length polymorphism (AFLP and restriction fragment length polymorphism (RFLP markers. The new linkage map of ENL48 contains 148 markers (33 AFLPs, 38 RFLPs and 77 SSRs in 23 linkage groups (LGs, covering a total map length of 798.0 cM. The ML161 map contains 240 markers (50 AFLPs, 71 RFLPs and 119 SSRs in 24 LGs covering a total of 1,328.1 cM. Using the improved maps, two quantitative trait loci (QTLs associated with tissue culturability were identified each for callusing rate and embryogenesis rate. A QTL for callogenesis was identified in LGD4b of ENL48 and explained 17.5% of the phenotypic variation. For embryogenesis rate, a QTL was detected on LGP16b in ML161 and explained 20.1% of the variation. This study is the first attempt to identify QTL associated with tissue culture amenity in oil palm which is an important step towards understanding the molecular processes underlying clonal regeneration of oil palm.

  2. The Fate of Integrated Ri T-DNA rol Genes during Regeneration via Somatic Embryogenesis in Tylophora indica

    Directory of Open Access Journals (Sweden)

    Dipasree Roychowdhury

    2015-01-01

    Full Text Available The fate of integrated Ri T-DNA rol genes during regeneration via indirect somatic embryogenesis and stability of its effect on morphology and tylophorine content of Ri-transformed plants have been studied in Tylophora indica. Integration and expression of Ri T-DNA genes in transformed embryogenic callus lines derived from transformed root lines, 300 Ri-transformed somatic embryos, and 23 Ri-transformed plant lines were analysed. Fifty root lines studied showed integration and expression of four rol genes of TL-DNA. Spontaneous regeneration via indirect somatic embryogenesis was obtained from root lines that were TL+/TR−. Stable integration and expression of rol genes were observed in root lines, embryogenic callus lines, and the spontaneously induced somatic embryos. Nineteen out of the 23 Ri-transformed plant lines and their clones showed phenotypic and genetic stability over the period of 3 years. Four Ri-transformed plants were morphologically similar to nontransformed plants but showed variation with the integration and expression of the rolA gene and absence of other rol genes. Variant Ri-transformed plant line A428#1-V showed highest tylophorine content (2.93±0.03 mg gDW−1 among plant lines studied. The effects of T-DNA genes on growth, morphology, and tylophorine content of the Ri-transformed plants were stable in the long term culture.

  3. The influence of α-Naphtaleneacetic Acid (NAA on somatic embryogenesis moon orchid Phalaenopsis amabilis (L. Bl.

    Directory of Open Access Journals (Sweden)

    EDY SETITI WIDA UTAMI

    2007-10-01

    Full Text Available An experiment to analyze the effect of plant growth regulator α-Naphtaleneacetic Acid (NAA on somatic embryogenesis moon orchid Ph amabilis (L. Bl. was carried out. One year old of plantlets were used as explants sources. Basal leaf of these explants were cultured in medium New Phalaenopsis (NP added with 0,1 mg/L, 1 mg/L, 2 mg/L, 3 mg/L, and 4 mg/L NAA. The explants were cultured in medium NP without NAA were used as control. The formation of embryogenic callus was observed every day, while the formation of somatic embryo was observed every week for 6 week using a dissecting microscope. The result showed that somatic embryogenesis Ph amabilis (L. Bl were influenced by plant growth regulator NAA. Explants were cultured in medium NP without NAA didn’t embryo formed, while explants were cultured in medium NP added with 2 mg/L, 3 mg/L, and 4 mg/L NAA embryo formed at embryogenic callus appear. These somatic embryos formed as an indirect via callus phase.

  4. G-protein-coupled estrogen receptor 1 is involved in brain development during zebrafish (Danio rerio) embryogenesis

    Energy Technology Data Exchange (ETDEWEB)

    Shi, Yanan; Liu, Xiaochun [State Key Laboratory of Biocontrol, Institute of Aquatic Economic Animals and Guangdong Provincial Key Laboratory for Aquatic Economic Animals, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275 (China); Zhu, Pei; Li, Jianzhen; Sham, Kathy W.Y. [School of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong (China); Cheng, Shuk Han [Department of Biology and Chemistry, City University of Hong Kong, Kowloon, Hong Kong (China); Li, Shuisheng; Zhang, Yong [State Key Laboratory of Biocontrol, Institute of Aquatic Economic Animals and Guangdong Provincial Key Laboratory for Aquatic Economic Animals, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275 (China); Cheng, Christopher H.K., E-mail: chkcheng@cuhk.edu.hk [School of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong (China); Lin, Haoran, E-mail: lsslhr@mail.sysu.edu.cn [State Key Laboratory of Biocontrol, Institute of Aquatic Economic Animals and Guangdong Provincial Key Laboratory for Aquatic Economic Animals, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275 (China); College of Ocean, Hainan University, Haikou 570228, Hainan (China)

    2013-05-24

    Highlights: •The Gper expression was detected in the developing brain of zebrafish. •Gper morpholino knockdown induced apoptosis of brain cells. •Gper morpholino knockdown reduced expression in neuron markers. •Zebrafish Gper may be involved in neuronal development. -- Abstract: G-protein-coupled estrogen receptor 1 (Gper, formerly known as GPR30) is found to be a trophic and protective factor in mediating action of estrogen in adult brain, while its role in developing brain remains to be elucidated. Here we present the expression pattern of Gper and its functions during embryogenesis in zebrafish. Both the mRNA and protein of Gper were detected throughout embryogenesis. Whole mount in situ hybridization (WISH) revealed a wide distribution of gper mRNAs in various regions of the developing brain. Gper knockdown by specific morpholinos resulted in growth retardation in embryos and morphological defects in the developing brain. In addition, induced apoptosis, decreased proliferation of the brain cells and maldevelopment of sensory and motor neurons were also found in the morphants. Our results provide novel insights into Gper functions in the developing brain, revealing that Gper can maintain the survival of the brain cells, and formation and/or differentiation of the sensory and motor neurons.

  5. OsMPK6 plays a critical role in cell differentiation during early embryogenesis in Oryza sativa.

    Science.gov (United States)

    Yi, Jakyung; Lee, Yang-Seok; Lee, Dong-Yeon; Cho, Man-Ho; Jeon, Jong-Seong; An, Gynheung

    2016-04-01

    The formation of body axes is the basis of morphogenesis during plant embryogenesis. We identified embryo-lethal mutants of rice (Oryza sativa) in which T-DNAs were inserted in OsMPK6 Embryonic organs were absent because their development was arrested at the globular stage. Similar to observations made with gle4, shootless, and organless, the osmpk6 mutations affected the initial step of cell differentiation. Expression of an apical-basal axis marker gene, OSH1, was reduced in the mutant embryos while that of the radial axes marker genes OsSCR and OsPNH1 was not detected. The signal for ROC1, a protodermal cell marker, was weak at the globular stage and gradually disappeared. Transcript levels of auxin and gibberellin biosynthesis genes were diminished in osmpk6 embryos. In addition, phytoalexin biosynthesis genes were down-regulated in osmpk6 and a major diterpene phytoalexin, momilactone A, did not accumulate in the mutant embryos. These results indicate that OsMPK6 begins to play a critical role during early embryogenesis, especially when the L1 radial axis is being formed. © The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  6. Influence of growth regulators on somatic embryogenesis, plantlet regeneration, and post-transplant survival of Echinochloa frumentacea.

    Science.gov (United States)

    Sankhla, A; Davis, T D; Sankhla, D; Sankhla, N; Upadhyaya, A; Joshi, S

    1992-07-01

    After placement on Murashige and Skoog's basal medium supplemented with 3-5 mg/l 2,4-D, immature inflorescence expiants of Echinochloa frumentacea gave rise to three distinct types of callus: a) loosely arranged and soft; b) compact and translucent; c) compact, sticky and mucilaginous. Somatic embryo formation occurred in type 'b' callus in about 18-24 d. Callus types 'a' and 'c' did not produce somatic embryos. The highest percentage of cultures exhibiting somatic embryogenesis occurred on the medium containing 5 mg/l 2,4-D and 0.5 mg/l kinetin. Somatic embryos also formed directly on the inflorescence (without intervening callus formation) in about 15% of the expiants placed on this medium. The addition of paclobutrazol or uniconazole (0.25 or 1 mg/l) to the medium had no influence on the percentage of cultures exhibiting direct somatic embryogenesis, but paclobutrazol slightly increased the mean number of somatic embryos per culture. Many of the callus-derived somatic embryos germinated when subcultured on basal MS medium supplemented with kinetin. Addition of paclobutrazol or uniconazole to the culture medium at 0.25 or 1 mg/l decreased somatic embryo germination and shoot elongation but increased root length and leaf width. Both paclobutrazol and uniconazole increased survival of the plantlets following transplanting to soil. Increased post-transplant survival was accompanied by reduced water loss from plantlets produced on culture media containing triazoles.

  7. A Modified Murine Embryonic Stem Cell Test for Evaluating the Teratogenic Effects of Drugs on Early Embryogenesis.

    Directory of Open Access Journals (Sweden)

    Ruoxing Yu

    Full Text Available Mammalian fetal development is easily disrupted by exogenous agents, making it essential to test new drug candidates for embryotoxicity and teratogenicity. To standardize the testing of drugs that might be used to treat pregnant women, the U.S. Food and Drug Administration (FDA formulated special grade categories, labeled A, B, C, D and X, that define the level of risk associated with the use of a specific drug during pregnancy. Drugs in categories (Cat. D and X are those with embryotoxic and/or teratogenic effects on humans and animals. However, which stages of pregnancy are affected by these agents and their molecular mechanisms are unknown. We describe here an embryonic stem cell test (EST that classifies FDA pregnancy Cat.D and Cat.X drugs into 4 classes based on their differing effects on primitive streak formation. We show that ~84% of Cat.D and Cat.X drugs target this period of embryogenesis. Our results demonstrate that our modified EST can identify how a drug affects early embryogenesis, when it acts, and its molecular mechanism. Our test may thus be a useful addition to the drug safety testing armamentarium.

  8. Embriogênese somática do caquizeiro Somatic embryogenesis of japanese persimmon

    Directory of Open Access Journals (Sweden)

    Dayse Cristina de Carvalho

    2004-08-01

    through somatic embryogenesis. Zygotic embryos excised from fruits collected from adult plants were tested at several developmental phases. They were collected during 22 weeks after 4 weeks blossoming. The basic medium tested was MS½NO3. Initial induction medium was supplemented with 20 µM 2,4-D + 2 µM kinetin. Dark calluses obtained were transferred to induction medium, with concentrations of 10 and 20 µM 2,4-D + 2 µM kinetin. The calli with pro-embryogenic masses were transferred to a maintenance and multiplication medium, with 2 µM kinetin and 2,4-D at the concentrations of 2.5, 5.0 and 10 µM. Embryogenic masses formed were transferred to a maturation medium supplemented with 0.5 µM IBA and 5, 10 and 20 µM 2iP. Embryos formed were isolated in two conversion media, one with 5 µM 2-iP + 5 µM GA3 and 0.5 µM IBA, and another medium with 0.5 µM GA3 and BAP at 0, 0.25, 0.5 and 1.0 µM. Indirect somatic embryogenesis were obtained from mature zygotic embryos collected after 22 weeks, when cultivated in culture medium with 10 µM 2,4D combined with 2 µM kinetin. Embryos maintenance and multiplication were more efficient with 5 µM 2,4-D, in which the pro-embryos advanced to the globular embryos phase. At the maturation phase the concentrations of 2-iP tested promoted globular embryos to more advanced stages of ontogeny, such as cordiform, torpedo and cotiledonary. Supplementation of the culture medium with 1 µM BAP generated better developed plants, with highest number of leaves and size.

  9. Effects of increasing use of trifluralin and glyphosate on the microbial activity of a lea soil; Efeito de doses crescentes dos herbicidas trifluralina e glifosato sobre a atividade microbiana de um solo de varzea

    Energy Technology Data Exchange (ETDEWEB)

    Barros, Edna Santos de; Monteiro, Regina Teresa Rosim [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil); Peixoto, Maria de Fatima da Silva Pinto [Sao Paulo Univ., Piracicaba, SP (Brazil). Escola Superior de Agricultura Luiz de Queiroz. Dept. de Quimica. E-mail: mfsppeix@carpa.ciagri.usp.br; Fay, Elizabeth Francisconi [Empresa Brasileira de Pesquisa Agropecuaria, Jaguariuna, SP (Brazil). Centro Nacional de Pesquisa de Defesa da Agricultura. E-mail: bethfay@cnpma.embrapa.com.br

    1997-07-01

    This work considers the importance of the glyphosate and trifluralin, which are the most used herbicides by the brazilian plantations, applying approximately fifteen and nine millions of liters by crop, respectively, for the evaluation of the increasing use of these herbicides effects on the microbial activity of a lea soil which are used for beans cultivation.

  10. Subjektiivinen teatterihistoria / Lea Tormis

    Index Scriptorium Estoniae

    Tormis, Lea, 1932-

    1996-01-01

    Eesti teatriajalugu, näitlejad, teatritegijad, teatrid. Lisaks "Yhteydet yli Suomenlahden", lk. 286; "Tanssitaide kotimaistuu", lk. 288; "Esimerkkejä Viron ja Suomen teatterisuhteiden kehityksestä Viron puolelta katsottuna" (1868-1985)

  11. Saatuslikud kohtumised / Lea Arme

    Index Scriptorium Estoniae

    Arme, Lea

    2001-01-01

    Laive Poskast, Eesti Eriolümpia rahvuslikust direktorist, vaimupuudega laste ja noorte treenerist. Tema õpilasest Andrei Kolovanist, kes 1995. a. võitis Connecticutis paraolümpiamängudel kahevõistluses tõstmises hõbemedali

  12. Progresso da ferrugem e da cercosporiose do cafeeiro consorciado com grevílea, com ingazeiro e a pleno sol em Lavras - MG Progress of rust and coffee plant cercosporiose mixed with grevílea, with ingazeiro and in the full sunshine in Lavras - MG

    Directory of Open Access Journals (Sweden)

    Bruno Grandi Salgado

    2007-08-01

    Full Text Available Com o presente trabalho, objetivou-se avaliar a incidência da ferrugem e da cercosporiose do cafeeiro em diferentes sistemas de cultivo, agroflorestais e a pleno sol, através da curva de progresso dessas doenças. Utilizou-se o delineamento inteiramente casualizado, com três tratamentos e sete repetições. Os tratamentos foram compostos por cafeeiros consorciados com ingazeiro, cafeeiros consorciados com grevílea e cafeeiros cultivados convencionalmente a pleno sol. As avaliações foram realizadas mensalmente no período de abril de 2001 a março de 2003. Realizou-se o cálculo da área abaixo da curva de progresso da doença (AACPD, a qual foi submetida à análise de variância. Para a incidência da ferrugem do cafeeiro o consórcio cafeeiro x ingazeiro mostrou-se com maiores índices da doença, sendo que os tratamentos consórcio cafeeiro x grevílea e cafeeiro a pleno sol não diferiram entre si e apresentaram menores incidências da doença. Diminuição na luz solar direta e maior umidade podem ter favorecido a ferrugem no sistema cafeeiro x ingazeiro. Para a incidência de cercosporiose, os cafeeiros a pleno sol obtiveram maiores valores de incidência da doença, seguidos por cafeeiros x grevílea, e menores taxas da doença foram observadas no consórcio cafeeiros x ingazeiro. A incidência de radiação solar direta pode ter favorecido a maior incidência de cercosporiose nos cafeeiros a pleno sol.The present work aimed to evaluate the incidence of rust and coffee plant cercosporiose in different systems of agroforest cultivation and in the full sunshine through the progress curve of those diseases. The completely randomized design with three treatments and seven replicates was utilized. The treatments were made up of coffee plants mixed with ingazeiro, coffee plants mixed with grevílea and coffee plants cultivated conventionally in the full sunshine. The evaluations were performed monthly during the period of April 2001 to

  13. Hydrocarbon Reserves: Abundance or Scarcity

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2005-07-01

    IFP and the OAPEC jointly organize a regular international seminar dealing with world oil-related problems appearing in the news. For the first time, this seminar has been opened to oil and gas company specialists, service companies, research centers and independents. This year's theme concerns oil and gas reserves: are they abundant or are we headed towards the shortages announced by some experts? This theme is especially topical in that: oil and gas currently meet two thirds of world energy needs and almost completely dominate the transport sector; the reserves declared by the OAPEC countries account for nearly half of world reserves; the price of a barrel of oil went through the roof in 2004; world energy demand is growing fast and alternative sources of energy are far from ready to take over from oil and gas in the next few decades. Since the reserves correspond to the volume it is technically and economically viable to produce, the seminar has, of course, dealt with the technical and economic questions that arise in connection with exploration and production, but it has also considered changes in the geopolitical context. Presentations by the leading companies of the OAPEC countries and by the IFP group were completed by presentation from the International Energy Agency (IEA), the United States Geological Survey (USGS), the IHS Energy Group, Total and Gaz de France. This document gathers the transparencies of the following presentations: Hydrocarbon reserves in OAPEC members countries: current and future (M. Al-Lababidi); Non OAPEC liquid reserves and production forecasts (Y. Mathieu); World oil and gas resources and production outlook (K. Chew); Global investments in the upstream (F. Birol); Total's policy in the oil and gas sector (C. de Margerie); Gaz de France's policy in the oil and gas sector (J. Abiteboul); NOC/IOC's opportunities in OPEC countries (I. Sandrea); Relationships between companies, countries and investors: How they may

  14. The Arabidopsis thaliana AAA protein CDC48A interacts in vivo with the somatic embryogenesis receptor-like kinase 1 receptor at the plasma membrane.

    NARCIS (Netherlands)

    Aker, J.C.M.; Borst, J.W.; Karlova, R.B.; Vries, de S.C.

    2006-01-01

    Fluorescent cell division cycle (CDC)48 proteins were studied in living plant protoplasts. CDC48A and somatic embryogenesis receptor like kinase 1 (SERK1) were found to co-localize in the endoplasmatic reticulum (ER) and at the plasma membrane (PM), but not in endosomal compartments. Fluorescent

  15. PLEXIN-D1, a novel plexin family member, is expressed in vascular endothelium and the central nervous system during mouse embryogenesis.

    NARCIS (Netherlands)

    Zwaag, A. van der; Hellemons, A.J.; Leenders, W.P.J.; Burbach, J.P.H.; Brunner, H.G.; Padberg, G.W.A.M.; Bokhoven, J.H.L.M. van

    2002-01-01

    The genetic defect in Mobius syndrome 2 (MBS2, MIM 601471), a dominantly inherited disorder characterised by paralysis of the facial nerve, is situated at chromosome 3q21-q22. We characterised the cDNA and predicted protein, and examined the expression pattern during mouse embryogenesis of a

  16. Expression of C4.4A, a structural uPAR homolog, reflects squamous epithelial differentiation in the adult mouse and during embryogenesis

    DEFF Research Database (Denmark)

    Kriegbaum, Mette Camilla; Jacobsen, Benedikte; Hald, Andreas

    2011-01-01

    -glandular portion of the rodent stomach, anus, vagina, cornea, and skin. This epithelial confinement was particularly evident from the abrupt termination of C4.4A expression at the squamo-columnar transition zones found at the ano-rectal and utero-vaginal junctions, for example. During mouse embryogenesis, C4.4A...

  17. Rosmarinic acid plays a protective role in the embryogenesis of zebrafish exposed to food colours through its influence on aurora kinase A level.

    Science.gov (United States)

    Swarnalatha, Y; Jerrine Joseph, I S; Jayakrishna, Tippabathani

    2017-05-01

    To evaluate the protective nature of the rosmarinic acid from Sphaeranthus amaranthoides during zebra fish embryogenesis. Rosmarinic acid was isolated from the S. amaranthoides. An accurate, sensitive and simple LC-MS analysis was performed to determine the rosmarinic acid from S. amaranthoides. In the present study, zebrafish embryos were exposed to crimson red and sunset yellow at a concentration of 0.1 and 0.5mg/l and the effect of these food colours on the levels of aurora kinase A was studied individually. Aurora kinase A levels are crucial for embryogenesis in zebrafish which is used as model in this study. The decrease of aurora kinase A levels in food colour treated embryos influences the embryogenesis, resulting in short and bent trunk leading to cell death and growth retardation. Elevated levels of aurora kinase A in rosmarinic acid treated groups can be attributed to the restoration of normal growth in zebra fish embryos with well developed brain and eyes. Further insilico docking studies were carried out and target was identified as rosmarinic acid. From the docking studies the docking poses and binding energy confirms that aurora kinase A is the target for rosmarinic acid. Rosmarinic acid was found to play a protective role in the embryogenesis of zebra fish exposed to food colours (crimson red and sunset yellow) through its influence on aurora kinase A levels. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  18. Induction of embryogenesis in Brassica napus microspores produces a callosic subintinal layer and abnormal cell walls with altered levels of callose and cellulose

    Directory of Open Access Journals (Sweden)

    Veronica eParra-Vega

    2015-11-01

    Full Text Available The induction of microspore embryogenesis produces dramatic changes in different aspects of the cell physiology and structure. Changes at the cell wall level are among the most intriguing and poorly understood. In this work, we used high pressure freezing and freeze substitution, immunolocalization, confocal and electron microscopy to analyze the structure and composition of the first cell walls formed during conventional Brassica napus microspore embryogenesis, and in cultures treated to alter the intracellular Ca2+ levels. Our results revealed that one of the first signs of embryogenic commitment is the formation of a callose-rich, cellulose-deficient layer beneath the intine (the subintinal layer, and of irregular, incomplete cell walls. In these events, Ca2+ may have a role. We propose that abnormal cell walls are due to a massive callose synthesis and deposition of excreted cytoplasmic material, and the parallel inhibition of cellulose synthesis. These features were absent in pollen-like structures and in microspore-derived embryos, few days after the end of the heat shock, where abnormal cell walls were no longer produced. Together, our results provide an explanation to a series of relevant aspects of microspore embryogenesis including the role of Ca2+ and the occurrence of abnormal cell walls. In addition, our discovery may be the explanation to why nuclear fusions take place during microspore embryogenesis.

  19. Arabidopsis transcription factor genes NF-YA1, 5, 6, and 9 play redundant roles in male gametogenesis, embryogenesis, and seed development.

    Science.gov (United States)

    Mu, Jinye; Tan, Helin; Hong, Sulei; Liang, Yan; Zuo, Jianru

    2013-01-01

    Nuclear factor Y (NF-Y) is a highly conserved transcription factor presented in all eukaryotic organisms, and is a heterotrimer consisting of three subunits: NF-YA, NF-YB, and NF-YC. In Arabidopsis, these three subunits are encoded by multigene families. The best-studied member of the NF-Y transcription factors is LEAFY COTYLEDON1 (LEC1), a NF-YB family member, which plays a critical role in embryogenesis and seed maturation. However, the function of most NF-Y genes remains elusive. Here, we report the characterization of four genes in the NF-YA family. We found that a gain-of-function mutant of NF-YA1 showed defects in male gametogenesis and embryogenesis. Consistently, overexpression of NF-YA1, 5, 6, and 9 affects male gametogenesis, embryogenesis, seed morphology, and seed germination, with a stronger phenotype when overexpressing NF-YA1 and NF-YA9. Moreover, overexpression of these NF-YA genes also causes hypersensitivity to abscisic acid (ABA) during seed germination, retarded seedling growth, and late flowering at different degrees. Intriguingly, overexpression of NF-YA1, 5, 6, and 9 is sufficient to induce the formation of somatic embryos from the vegetative tissues. However, single or double mutants of these NF-YA genes do not have detectable phenotype. Collectively, these results provide evidence that NF-YA1, 5, 6, and 9 play redundant roles in male gametophyte development, embryogenesis, seed development, and post-germinative growth.

  20. Changes in ecdysteroid levels and expression patterns of ecdysteroid-responsive factors and neuropeptide hormones during the embryogenesis of the blue crab, Callinectes sapidus.

    Science.gov (United States)

    Techa, Sirinart; Alvarez, Javier V; Sook Chung, J

    2015-04-01

    Embryogenesis requires the involvement and coordination of multiple networks of various genes, according to a timeline governing development. Crustacean embryogenesis usually includes the first molt, a process that is known to be positively controlled by ecdysteroids. We determined the amounts of ecdysteroids, as well as other related factors: the ecdysone receptor (CasEcR), the retinoid X receptor (CasRXR), the molt-inhibiting hormone (CasMIH), and crustacean hyperglycemic hormone (CasCHH) during the ovarian and embryonic developments of Callinectes sapidus. In summary, the ovaries at stages 1-4 have expression levels of maternal CasEcR and CasRXR 10-50 times higher than levels seen in embryos at the yolk stage. This large difference in the amount of the these factors in C. sapidus ovaries suggests that these maternal ecdysteroid-responsive factors may be utilized at the initiation of embryogenesis. During embryogenesis, the changes in total ecdysteroids and levels of CasEcR and CasRXR expression are similar to those observed in juvenile molts. The full-length cDNA sequence of the C. sapidus BTB domain protein (CasBTBDP) initially isolated from Y-organ cDNA, contains only Broad-Complex, Tramtrack, and Bric a brac (BTB) domains. The levels of CasBTBDP are kept constant throughout embryogenesis. The expression profiles of CasMIH and CasCHH are similar to the titers of ecdysteroids. However, the timing of their appearance is followed by increases in CasEcRs and CasRXRs, implying that the expressions of these neuropeptides may be influenced by ecdysteroids. Moreover, the ecdysteroid profile during embryogenesis may track directly with the timing of organogenesis of Y-organs and their activity. Our work reports, for first time, the observed expression and changes of ecdysteroid-responsive factors, along with CasCHH and CasMIH, during embryogenesis in the crustacean C. sapidus. Copyright © 2014 Elsevier Inc. All rights reserved.

  1. Stardust Abundance Variations among Interplanetary Dust Particles

    Science.gov (United States)

    Messenger, S.; Keller, L. P.; Nakamura-Messenger, K.; Nguyen, A. N.; Walker, Robert M.

    2009-01-01

    Presolar grain abundances reflect the degree of processing primitive materials have experienced. This is evidenced by the wide range of silicate stardust abundances among primitive meteorites (10 to 300 ppm) [1], attributable to parent body hydrothermal processing. Stardust abundance variations are also pronounced in anhydrous interplanetary dust particles (CPIDPs), that have not experienced parent body processing (300 to > 10,000 ppm) [2-4]. The large range in stardust abundances among CP IDPs thus reflect nebular processing. Here we present results of a systematic search for stardust among cluster CP IDPs. Our goals are to establish mineralogical trends among IDPs with different stardust abundances. This may shed light into the nature of isotopically normal presolar grains (GEMS grains?; 5) if their abundances vary similarly to that of isotopically exotic stardust grains.

  2. Messenger RNA during early embryogenesis in Artemia salina: altered translatability and sequence complexity.

    Science.gov (United States)

    James, T C; Tata, J R

    1980-02-01

    RNA from developing embryos of Artemia salina (5, 10, and 20 h after re-initiation of development) was translated 3-10 times more efficiently in a rabbit reticulocyte lysate cell-free protein synthesizing system than RNA from dormant gastrulae. The latter did not appear to contain any significant amount of translation inhibitor activity. Ninety percent of the translatable activity in dormant gastrulae was recovered as poly(A)--RNA, whereas 80% of that in post-gastrular developing embryos was present as poly(A)+-RNA. The size of most polypeptides coded for by dormant gastrular RNA was less than 130,000 daltons whereas the size of those coded for by developing embryonic RNA was up to 200,000 daltons, which correlated with a corresponding shift to poly A-containing RNA of higher molecular weight. Two major polypeptides of about 37,000 daltons coded for by dormant gastrular RNA disappeared at 20 h after resumption of development. Hybridization of complementary DNA (cDNA) to a 1000-fold excess of the homologous poly(A)+-RNA revealed the presence of three complexity classes of mRNA. Forty-five percent, 30%, and 25% of RNA in dormant gastrulae were present as high, middle, and low abundance classes comprising about 10, 80, and 9700 species, respectively whereas in the nauplii there were 10, 150, and 7900 species of high, middle, and low abundancy sequences, respectively. Heterologous hybridizations using cDNA complementary to highly abundant messenger population of nauplii (isolated by chromatography on hydroxyapatite) to poly(A)+-RNA from dormant cysts showed considerably divergence in this class of messengers from the two developmental stages. Re-initiation of development of dormant Artemia gastrulae is thus characterized by a "re-programming" seen as a simultaneous and rapid increase in the polyadenylation and translatability of poly(A)+-RNA accompanied by a qualitative change in its sequence complexity.

  3. Conférence de l'EAS "Biotechnologies for quality". Barcelone, Espagne, 20-23 octobre 2004 - Rapport de mission

    OpenAIRE

    Lacroix, Denis

    2004-01-01

    Le congrès annuel de l'EAS (European Aquaculture Society) a rassemblé pendant 4 jours 531 chercheurs, producteurs et responsables institutionnels de 47 nationalités. Il a donné lieu à la présentation de 105 communications orales et de 274 posters. La thématique générale du congrès était construite sur le thème général suivant: Biotechnologies for quality. Il s'agissait de montrer que les biotechnologies, qui font parfois peur au consommateur comme au citoyen, constituent un outil privilégié d...

  4. Correlation between some environmental variables and abundance ...

    African Journals Online (AJOL)

    Correlation between some environmental variables and abundance of Almophrya mediovacuolata (Ciliophora: Anoplophryidae) endocommensal ciliate of an anecic earthworms (Oligochaeta: Annelida) in Bambui (North-West Cameroon)

  5. Dynamics of the concentration of IAA and some of its conjugates during the induction of somatic embryogenesis in Coffea canephora.

    Science.gov (United States)

    Ayil-Gutiérrez, Benajmín; Galaz-Ávalos, Rosa; Peña-Cabrera, Eduardo; Loyola-Vargas, Victor

    2013-11-01

    Most of the somatic embryogenesis (SE) process requires the presence, either before or during the embryogenic process, of at least one exogenous auxin. This exogenous auxin induces the presence of endogenous auxins, which appears to be essential for SE induction. We found that during the preincubation period of SE in Coffea canephora, there is an important increase in both free and conjugated indole-3-acetic acid (IAA), as well as indole-3-butyric acid. This increase is accompanied by an increase in the expression of YUCCA (CcYUC), TRYPTOPHAN AMINOTRANSFERASE OF ARABIDOPSIS 1 (CcTAA1), and GRETCHEN HAGEN 3 (GH3) genes. On the other hand, most of the IAA compounds decreased during the induction of SE. The results presented in this research suggest that a balance between free IAA and its amide conjugates is necessary to allow the expression of SE-related genes.

  6. Genome-wide analysis of transcription factors during somatic embryogenesis in banana (Musa spp.) cv. Grand Naine.

    Science.gov (United States)

    Shivani; Awasthi, Praveen; Sharma, Vikrant; Kaur, Navjot; Kaur, Navneet; Pandey, Pankaj; Tiwari, Siddharth

    2017-01-01

    Transcription factors BABY BOOM (BBM), WUSCHEL (WUS), BSD, LEAFY COTYLEDON (LEC), LEAFY COTYLEDON LIKE (LIL), VIVIPAROUS1 (VP1), CUP SHAPED COTYLEDONS (CUC), BOLITA (BOL), and AGAMOUS LIKE (AGL) play a crucial role in somatic embryogenesis. In this study, we identified eighteen genes of these nine transcription factors families from the banana genome database. All genes were analyzed for their structural features, subcellular, and chromosomal localization. Protein sequence analysis indicated the presence of characteristic conserved domains in these transcription factors. Phylogenetic analysis revealed close evolutionary relationship among most transcription factors of various monocots. The expression patterns of eighteen genes in embryogenic callus containing somatic embryos (precisely isolated by Laser Capture Microdissection), non-embryogenic callus, and cell suspension cultures of banana cultivar Grand Naine were analyzed. The application of 2, 4-dichlorophenoxyacetic acid (2, 4-D) in the callus induction medium enhanced the expression of MaBBM1, MaBBM2, MaWUS2, and MaVP1 in the embryogenic callus. It suggested 2, 4-D acts as an inducer for the expression of these genes. The higher expression of MaBBM2 and MaWUS2 in embryogenic cell suspension (ECS) as compared to non-embryogenic cells suspension (NECS), suggested that these genes may play a crucial role in banana somatic embryogenesis. MaVP1 showed higher expression in both ECS and NECS, whereas MaLEC2 expression was significantly higher in NECS. It suggests that MaLEC2 has a role in the development of non-embryogenic cells. We postulate that MaBBM2 and MaWUS2 can be served as promising molecular markers for the embryogencity in banana.

  7. Aspects on gametogenesis, fertilization and embryogenesis of two deep-sea polychaetes from Eastern Atlantic cold seeps

    Science.gov (United States)

    Gaudron, S. M.; Hourdez, S.; Olu, K.

    2017-11-01

    We investigated two gonochoristic species of annelid polychaetes (one siboglinid and one polynoid) from cold seeps that ranged from 525 m to 3300 m in depth (Guiness, Worm Hole and Regab pockmarks) on the Gabon and Congo continental margins (Gulf of Guinea). Different aspects of gametogenesis (oocyte diameter, presence of ovisac, spermatozoa shape, and fecundity), fertilization (in vitro fertilization experiments: IVF) and embryogenesis (cleavage rate) were studied. The sampled siboglinid was a new species of Lamellibrachia and the second population of this genus in the Eastern Atlantic. Mean oocyte diameter was about 100 μm and fully-grown primary oocytes were stored in an ovisac, as in other studied siboglinids. The presence of a single spermatozoon was noted within an oviduct, indicating a possible internal fertilization. The rate of cell division at 6 °C was one cleavage every 20 h. Embryos developed normally to the blastula stage after 5-d post-fertilization at atmospheric pressure suggesting some pressure tolerance. The second polychaete was the scale-worm Branchipolynoe cf. seepensis that lives in commensalism in the mantle cavity of Bathymodiolus aff. boomerang. Anatomical reproductive features were similar to those described in B. seepensis from hydrothermal vents on Mid-Atlantic Ridge, with lecithotrophic larval development and continuous gametogenesis. We performed the first IVF carried out on gametes for any deep-sea polynoid species. Fertilization and development occurred but a number of abnormalities were observed demonstrating a limitation to embryogenesis at atmospheric pressure. The rate of cell division was three times faster at 8 °C than at 4 °C with a maximum stage of 8-cells reached after 72 h post-fertilization. We surprisingly observed some oocytes from the negative seawater control during IVF experiments cleaved to the 2-cell stage, demonstrating the possible occurrence of internal fertilization prior to IVF experiment or the accidental

  8. Wuschel overexpression promotes somatic embryogenesis and induces organogenesis in cotton (Gossypium hirsutum L.) tissues cultured in vitro.

    Science.gov (United States)

    Bouchabké-Coussa, O; Obellianne, M; Linderme, D; Montes, E; Maia-Grondard, A; Vilaine, F; Pannetier, C

    2013-05-01

    This work shows that overexpression of the WUS gene from Arabidopsis enhanced the expression of embryogenic competence and triggered organogenesis from some cells of the regenerated embryo-like structures. Agrobacterium-mediated genetic transformation of cotton was described in the late 1980s, but is still time consuming and largely genotype dependant due to poor regeneration. To help solve this bottleneck, we over-expressed the WUSCHEL (WUS) gene, a homeobox transcription factor cloned in Arabidopsis thaliana, known to stimulate organogenesis and/or somatic embryogenesis in Arabidopsis tissues cultured in vitro. The AtWUS gene alone, and AtWUS gene fused to the GFP marker were compared to the GFP gene alone and to an empty construct used as a control. Somatic embryogenesis was improved in WUS expressed calli, as the percentage of explants giving rise to embryogenic tissues was significantly higher (×3) when WUS gene was over-expressed than in the control. An interesting result was that WUS embryogenic lines evolved in green embryo-like structures giving rise to ectopic organogenesis never observed in any of our previous transformation experiments. Using our standard in vitro culture protocol, the overexpression of AtWUS in tissues of a recalcitrant variety did not result in the production of regenerated plants. This achievement will still require the optimization of other non-genetic factors, such as the balance of exogenous phytohormones. However, our results suggest that targeted expression of the WUS gene is a promising strategy to improve gene transfer in recalcitrant cotton cultivars.

  9. Impaired imprinting and social behaviors in chicks exposed to mifepristone, a glucocorticoid receptor antagonist, during the final week of embryogenesis.

    Science.gov (United States)

    Nishigori, Hideo; Kagami, Keisuke; Nishigori, Hidekazu

    2014-03-15

    The effects of glucocorticoid receptor dysfunction during embryogenesis on the imprinting abilities and social behaviors of hatchlings were examined using "fertile hen's egg-embryo-chick" system. Of embryos treated with mifepristone (0.4μmol/egg) on day 14, over 75% hatched a day later than the controls (day 22) without external anomalies. The mifepristone-treated hatchlings were assayed for imprinting ability on post-hatching day 2 and for social behaviors on day 3. The findings were as follows: imprinting ability (expressed as preference score) was significantly lower in mifepristone-treated hatchlings than in controls (0.65±0.06 vs. 0.92±0.02, P<0.005). Aggregation tests to evaluate the speed (seconds) required for four chicks, individually isolated with cardboard dividers in a box, to form a group after removal of the barriers showed that aggregation was significantly slower in mifepristone-treated hatchlings than in controls (8.7±1.1 vs. 2.6±0.3, P<0.001). In belongingness tests to evaluate the speed (seconds) for a chick isolated at a corner to join a group of three chicks placed at the opposite corner, mifepristone-treated hatchlings took significantly longer than controls (4.5±0.4/40 cm vs. 2.4±0.08/40 cm, P<0.001). In vocalization tests, using a decibel meter to measure average decibel level/30s (chick vocalization), mifepristone-treated hatchlings had significantly weaker vocalizations than controls (14.2±1.9/30s vs. 26.4±1.3/30s P<0.001). In conclusion, glucocorticoid receptor dysfunction during the last week embryogenesis altered the programming of brain development, resulting in impaired behavioral activities in late life. Copyright © 2013 Elsevier B.V. All rights reserved.

  10. Early embryogenesis-specific expression of the rice transposon Ping enhances amplification of the MITE mPing.

    Directory of Open Access Journals (Sweden)

    Shota Teramoto

    2014-06-01

    Full Text Available Miniature inverted-repeat transposable elements (MITEs are numerically predominant transposable elements in the rice genome, and their activities have influenced the evolution of genes. Very little is known about how MITEs can rapidly amplify to thousands in the genome. The rice MITE mPing is quiescent in most cultivars under natural growth conditions, although it is activated by various stresses, such as tissue culture, gamma-ray irradiation, and high hydrostatic pressure. Exceptionally in the temperate japonica rice strain EG4 (cultivar Gimbozu, mPing has reached over 1000 copies in the genome, and is amplifying owing to its active transposition even under natural growth conditions. Being the only active MITE, mPing in EG4 is an appropriate material to study how MITEs amplify in the genome. Here, we provide important findings regarding the transposition and amplification of mPing in EG4. Transposon display of mPing using various tissues of a single EG4 plant revealed that most de novo mPing insertions arise in embryogenesis during the period from 3 to 5 days after pollination (DAP, and a large majority of these insertions are transmissible to the next generation. Locus-specific PCR showed that mPing excisions and insertions arose at the same time (3 to 5 DAP. Moreover, expression analysis and in situ hybridization analysis revealed that Ping, an autonomous partner for mPing, was markedly up-regulated in the 3 DAP embryo of EG4, whereas such up-regulation of Ping was not observed in the mPing-inactive cultivar Nipponbare. These results demonstrate that the early embryogenesis-specific expression of Ping is responsible for the successful amplification of mPing in EG4. This study helps not only to elucidate the whole mechanism of mPing amplification but also to further understand the contribution of MITEs to genome evolution.

  11. Effects of Radiation From Contaminated Soil and Moss in Fukushima on Embryogenesis and Egg Hatching of the Aphid Prociphilus oriens.

    Science.gov (United States)

    Akimoto, Shin-Ichi; Li, Yang; Imanaka, Tetsuji; Sato, Hitoshi; Ishida, Ken

    2018-02-14

    Radiation-contaminated soils are widespread around the Fukushima Daiichi Nuclear Power Plant, and such soils raise concerns over its harmful effect on soil-dwelling organisms. We evaluated the effects of contaminated soil and moss sampled in Fukushima on the embryogenesis and hatching of aphid eggs, along with the measurement of the egg exposure dose. Cs-137 concentration in soil and moss from Fukushima ranged from 2200 to 3300 Bq/g and from 64 to 105 Bq/g, respectively. Eggs of the eriosomatine aphid Prociphilus oriens that were collected from a non-contaminated area were directly placed on the soil and moss for 4 or 3 months during diapause and then incubated until hatching. The total exposure dose to the eggs was estimated as ca. 100-200 mGy in the 4-month soil experiment and 4-10 mGy in the 4-month moss experiment. There was no significant difference in egg hatchability between the contaminated soil treatment and the control. No morphological abnormalities were detected in the first instars that hatched from the contaminated soil treatment. However, we found weak effects of radiation on egg hatching; eggs placed on the contaminated moss hatched earlier than did the control eggs. On the contaminated soil, the effects of radiation on egg hatching were not obvious because of uncontrolled environmental differences among containers. The effects of radiation on egg hatching were detected only in containers where high hatchability was recorded. Through the experiments, we concluded that the aphid eggs responded to ultra-low-dose radiation by advancing embryogenesis. © The American Genetic Association 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  12. Gene expression and metabolite profiling of gibberellin biosynthesis during induction of somatic embryogenesis in Medicago truncatula Gaertn.

    Science.gov (United States)

    Igielski, Rafał; Kępczyńska, Ewa

    2017-01-01

    Gibberellins (GAs) are involved in the regulation of numerous developmental processes in plants including zygotic embryogenesis, but their biosynthesis and role during somatic embryogenesis (SE) is mostly unknown. In this study we show that during three week- long induction phase, when cells of leaf explants from non-embryogenic genotype (M9) and embryogenic variant (M9-10a) were forming the callus, all the bioactive gibberellins from non-13-hydroxylation (GA4, GA7) and 13-hydroxylation (GA1, GA5, GA3, GA6) pathways were present, but the contents of only a few of them differed between the tested lines. The GA53 and GA19 substrates synthesized by the 13-hydroxylation pathway accumulated specifically in the M9-10a line after the first week of induction; subsequently, among the bioactive gibberellins detected, only the content of GA3 increased and appeared to be connected with acquisition of embryogenic competence. We fully annotated 20 Medicago truncatula orthologous genes coding the enzymes which catalyze all the known reactions of gibberellin biosynthesis. Our results indicate that, within all the genes tested, expression of only three: MtCPS, MtGA3ox1 and MtGA3ox2, was specific to embryogenic explants and reflected the changes observed in GA53, GA19 and GA3 contents. Moreover, by analyzing expression of MtBBM, SE marker gene, we confirmed the inhibitory effect of manipulation in GAs metabolism, applying exogenous GA3, which not only impaired the production of somatic embryos, but also significantly decreased expression of this gene.

  13. Taking species abundance distributions beyond individuals

    NARCIS (Netherlands)

    Morlon, Helene; White, Ethan P.; Etienne, Rampal S.; Green, Jessica L.; Ostling, Annette; Alonso, David; Enquist, Brian J.; He, Fangliang; Hurlbert, Allen; Magurran, Anne E.; Maurer, Brian A.; McGill, Brian J.; Olff, Han; Storch, David; Zillio, Tommaso; Chave, Jérôme

    The species abundance distribution (SAD) is one of the few universal patterns in ecology. Research on this fundamental distribution has primarily focused on the study of numerical counts, irrespective of the traits of individuals. Here we show that considering a set of Generalized Species Abundance

  14. Diversity and abundance of ammonia-oxidizing

    NARCIS (Netherlands)

    Cardoso, J.F.M.F.; van Bleijswijk, J.D.L.; Witte, H.; van Duyl, F.C.

    2013-01-01

    We analysed the diversity and abundance of ammonia-oxidizing Archaea (AOA) and Bacteria (AOB) in the shallow warm-water sponge Halisarca caerulea and the deep cold-water sponges Higginsia thielei and Nodastrella nodastrella. The abundance of AOA and AOB was analysed using catalyzed reporter

  15. On the solar abundance of indium

    NARCIS (Netherlands)

    Vitas, N.; Vince, I.; Lugaro, M.A.; Andriyenko, O.; Gosic, M.; Rutten, R.J.

    2008-01-01

    The generally adopted value for the solar abundance of indium is over six times higher than the meteoritic value. We address this discrepancy through numerical synthesis of the 451.13-nm line on which all indium abundance studies are based, both for the quiet Sun and the sunspot umbra spectrum,

  16. Seasonal variation in fish abundance and physicochemical ...

    African Journals Online (AJOL)

    There is a positive correlation between fish abundance and biomass for wet and dry seasons (r = 0.60 and 0.76 respectively). There was no significant difference between fish abundance in the two seasons. Variations occurred between physico-chemical parameters of water samples. Analysis of the lagoon waters showed ...

  17. On protein abundance distributions in complex mixtures.

    Science.gov (United States)

    Koziol, Ja; Griffin, Nm; Long, F; Li, Y; Latterich, M; Schnitzer, Je

    2013-01-29

    Mass spectrometry, an analytical technique that measures the mass-to-charge ratio of ionized atoms or molecules, dates back more than 100 years, and has both qualitative and quantitative uses for determining chemical and structural information. Quantitative proteomic mass spectrometry on biological samples focuses on identifying the proteins present in the samples, and establishing the relative abundances of those proteins. Such protein inventories create the opportunity to discover novel biomarkers and disease targets. We have previously introduced a normalized, label-free method for quantification of protein abundances under a shotgun proteomics platform (Griffin et al., 2010). The introduction of this method for quantifying and comparing protein levels leads naturally to the issue of modeling protein abundances in individual samples. We here report that protein abundance levels from two recent proteomics experiments conducted by the authors can be adequately represented by Sichel distributions. Mathematically, Sichel distributions are mixtures of Poisson distributions with a rather complex mixing distribution, and have been previously and successfully applied to linguistics and species abundance data. The Sichel model can provide a direct measure of the heterogeneity of protein abundances, and can reveal protein abundance differences that simpler models fail to show.

  18. Interstellar Abundances Toward X Per, Revisited

    Science.gov (United States)

    Valencic, Lynne A.; Smith, Randall K.

    2014-01-01

    The nearby X-ray binary X Per (HD 24534) provides a useful beacon with which to measure elemental abundances in the local ISM. We examine absorption features of 0, Mg, and Si along this line of sight using spectra from the Chandra Observatory's LETG/ ACIS-S and XMM-Newton's RGS instruments. In general, we find that the abundances and their ratios are similar to those of young F and G stars and the most recent solar values. We compare our results with abundances required by dust grain models.

  19. The solar system abundance of tin

    Energy Technology Data Exchange (ETDEWEB)

    Loss, R.D.; Rosman, K.K.R.; De Laeter, J.R. (Curtin Univ. of Technology, Perth (Australia))

    1989-04-01

    The elemental abundance of tin has been determined by the mass spectrometric isotope dilution technique for four carbonaceous chondrites, including the Cl chondrites Orgueil and Ivuna. A solar system abundance of 3.98 (normalized to Si = 10{sup 6} atoms) can be derived from these data, which is in excellent agreement with the presently accepted value. Recently, a theoretical value of 2.82 {plus minus} 0.21 (Si = 10{sup 6} atoms) has been proposed for the abundance of tin based on an s-process nucleosynthesis model. This value is 26% lower than the presently accepted solar system abundance. The present data confirm the higher value for tin and indicate that a reexamination of the assumptions on which the nucleosynthetic value is based is required.

  20. Chinook Abundance - Linear Features [ds181

    Data.gov (United States)

    California Department of Resources — The dataset 'ds181_Chinook_ln' is a product of the CalFish Adult Salmonid Abundance Database. Data in this shapefile are collected from stream sections or reaches...

  1. Abundance, composition and distribution of simple sequence ...

    Indian Academy of Sciences (India)

    Abundance, composition and distribution of simple sequence repeats and dinucleotide compositional bias within WSSV genomes. MALATHI SHEKAR, INDRANI KARUNASAGAR and IDDYA KARUNASAGAR*. Department of Fishery Microbiology, UNESCO Centre for Marine Biotechnology,. Karnataka Veterinary, Animal ...

  2. Estimating Squirrel Abundance From Live trapping Data

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — A reprint of an article from the Journal of Wildlife Management entitled "Estimating Squirrel Abundance from Live Trapping Data" by Nixon, Edwards and Eberhardt. The...

  3. Abundance estimation of spectrally similar minerals

    CSIR Research Space (South Africa)

    Debba, Pravesh

    2009-07-01

    Full Text Available This paper evaluates a spectral unmixing method for estimating the partial abundance of spectrally similar minerals in complex mixtures. The method requires formulation of a linear function of individual spectra of individual minerals. The first...

  4. Stochastic species abundance models involving special copulas

    Science.gov (United States)

    Huillet, Thierry E.

    2018-01-01

    Copulas offer a very general tool to describe the dependence structure of random variables supported by the hypercube. Inspired by problems of species abundances in Biology, we study three distinct toy models where copulas play a key role. In a first one, a Marshall-Olkin copula arises in a species extinction model with catastrophe. In a second one, a quasi-copula problem arises in a flagged species abundance model. In a third model, we study completely random species abundance models in the hypercube as those, not of product type, with uniform margins and singular. These can be understood from a singular copula supported by an inflated simplex. An exchangeable singular Dirichlet copula is also introduced, together with its induced completely random species abundance vector.

  5. SWFSC/MMTD: Vaquita Abundance Survey 1997

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — In 1997, the Southwest Fisheries Science Center (SWFSC) conducted a survey designed to estimate the abundance of vaquita, the Gulf of California harbor porpoise...

  6. Developmental localization and the role of hydroxyproline rich glycoproteins during somatic embryogenesis of banana (Musa spp. AAA).

    Science.gov (United States)

    Xu, Chunxiang; Takáč, Tomáš; Burbach, Christian; Menzel, Diedrik; Samaj, Jozef

    2011-02-24

    Hydroxyproline rich glycoproteins (HRGPs) are implicated to have a role in many aspects of plant growth and development but there is limited knowledge about their localization and function during somatic embryogenesis of higher plants. In this study, the localization and function of hydroxyproline rich glycoproteins in embryogenic cells (ECs) and somatic embryos of banana were investigated by using immunobloting and immunocytochemistry with monoclonal JIM11 and JIM20 antibodies as well as by treatment with 3,4-dehydro-L-proline (3,4-DHP, an inhibitor of extensin biosynthesis), and by immunomodulation with the JIM11 antibody. Immunofluorescence labelling of JIM11 and JIM20 hydroxyproline rich glycoprotein epitopes was relatively weak in non-embryogenic cells (NECs), mainly on the edge of small cell aggregates. On the other hand, hydroxyproline rich glycoprotein epitopes were found to be enriched in early embryogenic cells as well as in various developmental stages of somatic embryos. Embryogenic cells (ECs), proembryos and globular embryos showed strong labelling of hydroxyproline rich glycoprotein epitopes, especially in their cell walls and outer surface layer, so-called extracellular matrix (ECM). This hydroxyproline rich glycoprotein signal at embryo surfaces decreased and/or fully disappeared during later developmental stages (e.g. pear-shaped and cotyledonary stages) of embryos. In these later developmental embryogenic stages, however, new prominent hydroxyproline rich glycoprotein labelling appeared in tri-cellular junctions among parenchymatic cells inside these embryos. Overall immunofluorescence labelling of late stage embryos with JIM20 antibody was weaker than that of JIM11. Western blot analysis supported the above immunolocalization data. The treatment with 3,4-DHP inhibited the development of embryogenic cells and decreased the rate of embryo germination. Embryo-like structures, which developed after 3,4-DHP treatment showed aberrant non

  7. Thermal manipulation during embryogenesis has long-term effects on muscle and liver metabolism in fast-growing chickens.

    Directory of Open Access Journals (Sweden)

    Thomas Loyau

    Full Text Available Fast-growing chickens have a limited ability to tolerate high temperatures. Thermal manipulation during embryogenesis (TM has previously been shown to lower chicken body temperature (Tb at hatching and to improve thermotolerance until market age, possibly resulting from changes in metabolic regulation. The aim of this study was to evaluate the long-term effects of TM (12 h/d, 39.5°C, 65% RH from d 7 to 16 of embryogenesis vs. 37.8°C, 56% RH continuously and of a subsequent heat challenge (32°C for 5 h at 34 d on the mRNA expression of metabolic genes and cell signaling in the Pectoralis major muscle and the liver. Gene expression was analyzed by RT-qPCR in 8 chickens per treatment, characterized by low Tb in the TM groups and high Tb in the control groups. Data were analyzed using the general linear model of SAS considering TM and heat challenge within TM as main effects. TM had significant long-term effects on thyroid hormone metabolism by decreasing the muscle mRNA expression of deiodinase DIO3. Under standard rearing conditions, the expression of several genes involved in the regulation of energy metabolism, such as transcription factor PGC-1α, was affected by TM in the muscle, whereas for other genes regulating mitochondrial function and muscle growth, TM seemed to mitigate the decrease induced by the heat challenge. TM increased DIO2 mRNA expression in the liver (only at 21°C and reduced the citrate synthase activity involved in the Krebs cycle. The phosphorylation level of p38 Mitogen-activated-protein kinase regulating the cell stress response was higher in the muscle of TM groups compared to controls. In conclusion, markers of energy utilization and growth were either changed by TM in the Pectoralis major muscle and the liver by thermal manipulation during incubation as a possible long-term adaptation limiting energy metabolism, or mitigated during heat challenge.

  8. Developmental localization and the role of hydroxyproline rich glycoproteins during somatic embryogenesis of banana (Musa spp. AAA

    Directory of Open Access Journals (Sweden)

    Menzel Diedrik

    2011-02-01

    Full Text Available Abstract Background Hydroxyproline rich glycoproteins (HRGPs are implicated to have a role in many aspects of plant growth and development but there is limited knowledge about their localization and function during somatic embryogenesis of higher plants. In this study, the localization and function of hydroxyproline rich glycoproteins in embryogenic cells (ECs and somatic embryos of banana were investigated by using immunobloting and immunocytochemistry with monoclonal JIM11 and JIM20 antibodies as well as by treatment with 3,4-dehydro-L-proline (3,4-DHP, an inhibitor of extensin biosynthesis, and by immunomodulation with the JIM11 antibody. Results Immunofluorescence labelling of JIM11 and JIM20 hydroxyproline rich glycoprotein epitopes was relatively weak in non-embryogenic cells (NECs, mainly on the edge of small cell aggregates. On the other hand, hydroxyproline rich glycoprotein epitopes were found to be enriched in early embryogenic cells as well as in various developmental stages of somatic embryos. Embryogenic cells (ECs, proembryos and globular embryos showed strong labelling of hydroxyproline rich glycoprotein epitopes, especially in their cell walls and outer surface layer, so-called extracellular matrix (ECM. This hydroxyproline rich glycoprotein signal at embryo surfaces decreased and/or fully disappeared during later developmental stages (e.g. pear-shaped and cotyledonary stages of embryos. In these later developmental embryogenic stages, however, new prominent hydroxyproline rich glycoprotein labelling appeared in tri-cellular junctions among parenchymatic cells inside these embryos. Overall immunofluorescence labelling of late stage embryos with JIM20 antibody was weaker than that of JIM11. Western blot analysis supported the above immunolocalization data. The treatment with 3,4-DHP inhibited the development of embryogenic cells and decreased the rate of embryo germination. Embryo-like structures, which developed after 3,4-DHP

  9. Coronae of Stars with Supersolar Elemental Abundances

    Science.gov (United States)

    Peretz, Uria; Behar, Ehud; Drake, Stephen A.

    2015-01-01

    Coronal elemental abundances are known to deviate from the photospheric values of their parent star, with the degree of deviation depending on the first ionization potential (FIP). This study focuses on the coronal composition of stars with supersolar photospheric abundances. We present the coronal abundances of six such stars: 11 LMi, iota Hor, HR 7291, tau Boo, and alpha Cen A and B. These stars all have high-statistics X-ray spectra, three of which are presented for the first time. The abundances we measured were obtained using the line-resolved spectra of the Reflection Grating Spectrometer (RGS) in conjunction with the higher throughput EPIC-pn camera spectra onboard the XMM-Newton observatory. A collisionally ionized plasma model with two or three temperature components is found to represent the spectra well. All elements are found to be consistently depleted in the coronae compared to their respective photospheres. For 11 LMi and tau Boo no FIP effect is present, while iota Hor, HR 7291, and alpha Cen A and B show a clear FIP trend. These conclusions hold whether the comparison is made with solar abundances or the individual stellar abundances. Unlike the solar corona, where low-FIP elements are enriched, in these stars the FIP effect is consistently due to a depletion of high-FIP elements with respect to actual photospheric abundances. A comparison with solar (instead of stellar) abundances yields the same fractionation trend as on the Sun. In both cases, a similar FIP bias is inferred, but different fractionation mechanisms need to be invoked.

  10. TEA: A CODE CALCULATING THERMOCHEMICAL EQUILIBRIUM ABUNDANCES

    Energy Technology Data Exchange (ETDEWEB)

    Blecic, Jasmina; Harrington, Joseph; Bowman, M. Oliver, E-mail: jasmina@physics.ucf.edu [Planetary Sciences Group, Department of Physics, University of Central Florida, Orlando, FL 32816-2385 (United States)

    2016-07-01

    We present an open-source Thermochemical Equilibrium Abundances (TEA) code that calculates the abundances of gaseous molecular species. The code is based on the methodology of White et al. and Eriksson. It applies Gibbs free-energy minimization using an iterative, Lagrangian optimization scheme. Given elemental abundances, TEA calculates molecular abundances for a particular temperature and pressure or a list of temperature–pressure pairs. We tested the code against the method of Burrows and Sharp, the free thermochemical equilibrium code Chemical Equilibrium with Applications (CEA), and the example given by Burrows and Sharp. Using their thermodynamic data, TEA reproduces their final abundances, but with higher precision. We also applied the TEA abundance calculations to models of several hot-Jupiter exoplanets, producing expected results. TEA is written in Python in a modular format. There is a start guide, a user manual, and a code document in addition to this theory paper. TEA is available under a reproducible-research, open-source license via https://github.com/dzesmin/TEA.

  11. Factors influencing cucumber (Cucumis sativus L.) somatic embryogenesis. I. The crucial role of pH and nitrogen in suspension culture

    OpenAIRE

    Tadeusz Wróblewski; Marcin K. Filipecki; Stefan Malepszy

    2014-01-01

    A method of obtaining and the characteristics of an embryogenic stabilised cucumber (Cucumis sativus L.) suspension culture which has many similarities to the carrot model are presented. The Specific Type I cells and proembryogenic mass were present in such a suspension. The maintenance of the proembryogenic stage took place in medium containing 2,4-D as the sole growth regulator, subsequent stages of embryogenesis occurred in hormone-free medium. Embryonic structures were also observed in me...

  12. High genetic and epigenetic stability in Coffea arabica plants derived from embryogenic suspensions and secondary embryogenesis as revealed by AFLP, MSAP and the phenotypic variation rate.

    Directory of Open Access Journals (Sweden)

    Roberto Bobadilla Landey

    Full Text Available Embryogenic suspensions that involve extensive cell division are risky in respect to genome and epigenome instability. Elevated frequencies of somaclonal variation in embryogenic suspension-derived plants were reported in many species, including coffee. This problem could be overcome by using culture conditions that allow moderate cell proliferation. In view of true-to-type large-scale propagation of C. arabica hybrids, suspension protocols based on low 2,4-D concentrations and short proliferation periods were developed. As mechanisms leading to somaclonal variation are often complex, the phenotypic, genetic and epigenetic changes were jointly assessed so as to accurately evaluate the conformity of suspension-derived plants. The effects of embryogenic suspensions and secondary embryogenesis, used as proliferation systems, on the genetic conformity of somatic embryogenesis-derived plants (emblings were assessed in two hybrids. When applied over a 6 month period, both systems ensured very low somaclonal variation rates, as observed through massive phenotypic observations in field plots (0.74% from 200,000 plant. Molecular AFLP and MSAP analyses performed on 145 three year-old emblings showed that polymorphism between mother plants and emblings was extremely low, i.e. ranges of 0-0.003% and 0.07-0.18% respectively, with no significant difference between the proliferation systems for the two hybrids. No embling was found to cumulate more than three methylation polymorphisms. No relation was established between the variant phenotype (27 variants studied and a particular MSAP pattern. Chromosome counting showed that 7 of the 11 variant emblings analyzed were characterized by the loss of 1-3 chromosomes. This work showed that both embryogenic suspensions and secondary embryogenesis are reliable for true-to-type propagation of elite material. Molecular analyses revealed that genetic and epigenetic alterations are particularly limited during coffee

  13. Detailed expression profile of all six Glypicans and their modifying enzyme Notum during chick embryogenesis and their role in dorsal-ventral patterning of the neural tube.

    Science.gov (United States)

    Saad, Kawakeb; Otto, Anthony; Theis, Susanne; Kennerley, Niki; Munsterberg, Andrea; Luke, Graham; Patel, Ketan

    2017-04-20

    Vertebrate development is orchestrated by secreted signalling molecules that regulate cell behaviour and cell fate decisions during early embryogenesis. The activity of key signalling molecules including members of Hedgehog, Bone Morphogenetic Proteins and Wnt families are regulated by Glypicans, a family of GPI linked polypeptides. Glypicans either promote or inhibit the action of signalling molecules and add a layer of complexity that needs to be understood in order to fully decipher the processes that regulate early vertebrate development. Here we present a detailed expression profile of all six Glypicans and their modifying enzyme Notum during chick embryogenesis. Our results strongly suggest that these proteins have many as yet undiscovered roles to play during early embryogenesis. Finally, we have taken an experimental approach to investigate their role during the patterning of a key embryonic structure - the neural tube. In particular, we show that over-expression of Notum leads to the dorsalisation of this structure. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. VHA-19 is essential in Caenorhabditis elegans oocytes for embryogenesis and is involved in trafficking in oocytes.

    Directory of Open Access Journals (Sweden)

    Alison J Knight

    Full Text Available There is an urgent need to develop new drugs against parasitic nematodes, which are a significant burden on human health and agriculture. Information about the function of essential nematode-specific genes provides insight to key nematode-specific processes that could be targeted with drugs. We have characterized the function of a novel, nematode-specific Caenorhabditis elegans protein, VHA-19, and show that VHA-19 is essential in the germline and, specifically, the oocytes, for the completion of embryogenesis. VHA-19 is also involved in trafficking the oocyte receptor RME-2 to the oocyte plasma membrane and is essential for osmoregulation in the embryo, probably because VHA-19 is required for proper eggshell formation via exocytosis of cortical granules or other essential components of the eggshell. VHA-19 may also have a role in cytokinesis, either directly or as an indirect effect of its role in osmoregulation. Critically, VHA-19 is expressed in the excretory cell in both larvae and adults, suggesting that it may have a role in osmoregulation in C. elegans more generally, probably in trafficking or secretion pathways. This is the first time a role for VHA-19 has been described.

  15. WNT4-like protein is a cortical granule component in mouse oocytes and functions in regulating preimplantation embryogenesis.

    Science.gov (United States)

    Liu, Min; Yang, Huei-Ting

    2016-01-01

    Mammalian cortical granules (CG) are membrane-bound organelles located in the cortex of the unfertilized oocytes. Upon fertilization, CG undergo exocytosis to function in blocking polyspermy. While cortical granules are important in fertilization, their exact biochemical composition and reproductive function have not been fully defined. In the present study, a 66 kDa wingless-type MMTV integration site family, member 4 (WNT4)-like protein, with mouse CG origin was identified. Oocytes that were double labeled with lectin Lens culinaris agglutinin (LCA) and WNT4 antibody showed colocalization of the WNT4 molecules and cortical granules. The disappearance of WNT4 molecules in the artificially activated oocytes that were devoid of cortical granules confirmed their granule origin. Following fertilization, WNT4 remained associated with zygotes and blastomeres of 2-cell and 8-cell embryos; however the amount of protein present was reduced more than 2-fold as embryos developed. Prior to implantation, WNT4 appeared to be detectable only in the trophoblast cells. Our functional study revealed that WNT4 molecules were involved in regulating zygotic cleavage and early embryogenesis. To our knowledge, this is the first study demonstrating mammalian cortical granules contain signaling molecules that are involved in the regulation of the first phase of embryonic development.

  16. 3D Visualization of Developmental Toxicity of 2,4,6-Trinitrotoluene in Zebrafish Embryogenesis Using Light-Sheet Microscopy

    Directory of Open Access Journals (Sweden)

    Juneyong Eum

    2016-11-01

    Full Text Available Environmental contamination by trinitrotoluene is of global concern due to its widespread use in military ordnance and commercial explosives. Despite known long-term persistence in groundwater and soil, the toxicological profile of trinitrotoluene and other explosive wastes have not been systematically measured using in vivo biological assays. Zebrafish embryos are ideal model vertebrates for high-throughput toxicity screening and live in vivo imaging due to their small size and transparency during embryogenesis. Here, we used Single Plane Illumination Microscopy (SPIM/light sheet microscopy to assess the developmental toxicity of explosive-contaminated water in zebrafish embryos and report 2,4,6-trinitrotoluene-associated developmental abnormalities, including defects in heart formation and circulation, in 3D. Levels of apoptotic cell death were higher in the actively developing tissues of trinitrotoluene-treated embryos than controls. Live 3D imaging of heart tube development at cellular resolution by light-sheet microscopy revealed trinitrotoluene-associated cardiac toxicity, including hypoplastic heart chamber formation and cardiac looping defects, while the real time PCR (polymerase chain reaction quantitatively measured the molecular changes in the heart and blood development supporting the developmental defects at the molecular level. Identification of cellular toxicity in zebrafish using the state-of-the-art 3D imaging system could form the basis of a sensitive biosensor for environmental contaminants and be further valued by combining it with molecular analysis.

  17. Sensitivity of eastern oyster (Crassostrea virginica) spermatozoa and oocytes to dispersed oil: Cellular responses and impacts on fertilization and embryogenesis.

    Science.gov (United States)

    Vignier, J; Volety, A K; Rolton, A; Le Goïc, N; Chu, F-L E; Robert, R; Soudant, P

    2017-06-01

    The 2010 Deepwater Horizon (DWH) oil spill released millions of barrels of oil and dispersant into the Gulf of Mexico. The timing of the spill coincided with the spawning season of Crassostrea virginica. Consequently, gametes released in the water were likely exposed to oil and dispersant. This study aimed to (i) evaluate the cellular effects of acute exposure of spermatozoa and oocytes to surface slick oil, dispersed mechanically (HEWAF) and chemically (CEWAF), using flow-cytometric (FCM) analyses, and (ii) determine whether the observed cellular effects relate to impairments of fertilization and embryogenesis of gametes exposed to the same concentrations of CEWAF and HEWAF. Following a 30-min exposure, the number of spermatozoa and their viability were reduced due to a physical action of oil droplets (HEWAF) and a toxic action of CEWAF respectively. Additionally, reactive oxygen species (ROS) production in exposed oocytes tended to increase with increasing oil concentrations suggesting that exposure to dispersed oil resulted in an oxidative stress. The decrease in fertilization success (1-h), larval survival (24-h) and increase in abnormalities (6-h and 24-h) may be partly related to altered cellular characteristics. FCM assays are a good predictor of sublethal effects especially on fertilization success. These data suggest that oil/dispersant are cytotoxic to gametes, which may affect negatively the reproduction success and early development of oysters. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Melanoma. Shall we move away from the sun and focus more on embryogenesis, body weight and longevity?☆

    Science.gov (United States)

    Bataille, Veronique

    2013-01-01

    There are many observations regarding the behaviour of melanoma which points away from sunshine as the main cause of this tumour. Incidence data shows that the increase is mostly seen for thin melanomas which cannot be attributed to sun exposure but increasing screening over the last 20 years. Melanoma behaves in a similar fashion all over the world regarding age of onset, gender differences and histological subtypes. An excess of naevi is the strongest risk factor for melanoma and their appearance and involution throughout life, and the differences in naevus distribution according to gender is giving us a lot of clues about melanoma biology. Melanoma like all cancers is a complex disease with the involvement of many common and low penetrance genes many of them involved in pigmentation and naevogenesis but these only explain a very small portion of melanoma susceptibility. Genes involved in melanocyte differentiation early on in embryogenesis are also becoming relevant for melanoma initiation and progression. Reduced senescence and longevity as well as body weight and energy expenditure are also relevant for melanoma susceptibility. These observations with links between melanoma and non-sun related phenotypes as well as gene discoveries should help to assess the relative contribution of genetic and environmental factors in its causation. PMID:23796690

  19. Melanoma. Shall we move away from the sun and focus more on embryogenesis, body weight and longevity?

    Science.gov (United States)

    Bataille, Veronique

    2013-11-01

    There are many observations regarding the behaviour of melanoma which points away from sunshine as the main cause of this tumour. Incidence data shows that the increase is mostly seen for thin melanomas which cannot be attributed to sun exposure but increasing screening over the last 20 years. Melanoma behaves in a similar fashion all over the world regarding age of onset, gender differences and histological subtypes. An excess of naevi is the strongest risk factor for melanoma and their appearance and involution throughout life, and the differences in naevus distribution according to gender is giving us a lot of clues about melanoma biology. Melanoma like all cancers is a complex disease with the involvement of many common and low penetrance genes many of them involved in pigmentation and naevogenesis but these only explain a very small portion of melanoma susceptibility. Genes involved in melanocyte differentiation early on in embryogenesis are also becoming relevant for melanoma initiation and progression. Reduced senescence and longevity as well as body weight and energy expenditure are also relevant for melanoma susceptibility. These observations with links between melanoma and non-sun related phenotypes as well as gene discoveries should help to assess the relative contribution of genetic and environmental factors in its causation. Copyright © 2013 The Author. Published by Elsevier Ltd.. All rights reserved.

  20. A functional genome-wide in vivo screen identifies new regulators of signalling pathways during early Xenopus embryogenesis.

    Directory of Open Access Journals (Sweden)

    Siwei Zhang

    Full Text Available Embryonic development requires exquisite regulation of several essential processes, such as patterning of tissues and organs, cell fate decisions, and morphogenesis. Intriguingly, these diverse processes are controlled by only a handful of signalling pathways, and mis-regulation in one or more of these pathways may result in a variety of congenital defects and diseases. Consequently, investigating how these signalling pathways are regulated at the molecular level is essential to understanding the mechanisms underlying vertebrate embryogenesis, as well as developing treatments for human diseases. Here, we designed and performed a large-scale gain-of-function screen in Xenopus embryos aimed at identifying new regulators of MAPK/Erk, PI3K/Akt, BMP, and TGF-β/Nodal signalling pathways. Our gain-of-function screen is based on the identification of gene products that alter the phosphorylation state of key signalling molecules, which report the activation state of the pathways. In total, we have identified 20 new molecules that regulate the activity of one or more signalling pathways during early Xenopus development. This is the first time that such a functional screen has been performed, and the findings pave the way toward a more comprehensive understanding of the molecular mechanisms regulating the activity of important signalling pathways under normal and pathological conditions.

  1. 3D Visualization of Developmental Toxicity of 2,4,6-Trinitrotoluene in Zebrafish Embryogenesis Using Light-Sheet Microscopy.

    Science.gov (United States)

    Eum, Juneyong; Kwak, Jina; Kim, Hee Joung; Ki, Seoyoung; Lee, Kooyeon; Raslan, Ahmed A; Park, Ok Kyu; Chowdhury, Md Ashraf Uddin; Her, Song; Kee, Yun; Kwon, Seung-Hae; Hwang, Byung Joon

    2016-11-17

    Environmental contamination by trinitrotoluene is of global concern due to its widespread use in military ordnance and commercial explosives. Despite known long-term persistence in groundwater and soil, the toxicological profile of trinitrotoluene and other explosive wastes have not been systematically measured using in vivo biological assays. Zebrafish embryos are ideal model vertebrates for high-throughput toxicity screening and live in vivo imaging due to their small size and transparency during embryogenesis. Here, we used Single Plane Illumination Microscopy (SPIM)/light sheet microscopy to assess the developmental toxicity of explosive-contaminated water in zebrafish embryos and report 2,4,6-trinitrotoluene-associated developmental abnormalities, including defects in heart formation and circulation, in 3D. Levels of apoptotic cell death were higher in the actively developing tissues of trinitrotoluene-treated embryos than controls. Live 3D imaging of heart tube development at cellular resolution by light-sheet microscopy revealed trinitrotoluene-associated cardiac toxicity, including hypoplastic heart chamber formation and cardiac looping defects, while the real time PCR (polymerase chain reaction) quantitatively measured the molecular changes in the heart and blood development supporting the developmental defects at the molecular level. Identification of cellular toxicity in zebrafish using the state-of-the-art 3D imaging system could form the basis of a sensitive biosensor for environmental contaminants and be further valued by combining it with molecular analysis.

  2. Molecular cloning and in silico analysis of three somatic embryogenesis receptor kinase mRNA from date palm

    Directory of Open Access Journals (Sweden)

    Rekik Imen

    2013-01-01

    Full Text Available We report here the isolation and characterizations of three somatic embryogenesis receptor kinase (PhSERK genes from palm date by a rapid amplification of cDNA ends (RACE approach. PhSERKs belong to a small family of receptor kinase genes, share a conserved structure and extensive sequence homology with previously reported plant SERK genes. Sequence analysis of these genes revealed the sequence size of 11051 pb (PhSERK1, 7981 pb (PhSERK2 and 10510 pb (PhSERK3. The open reading frames of PhSERK1, PhSERK2 and PhSERK3 are 1914 pb, 1797 pb and 1719 pb respectively. PhSERKs belongs to the LRR-type cell surface RLKs, which possess a number of characteristic domains. These include an extracellular domain (EX containing a variable number of LRR units, signal pepetide (SP immediately followed by a single transmembrane domain (TM and an intracellular kinase domain. The phylogenetic tree shows that the protein PhSERK1, PhSERK2 and PhSERK3 clustered within monocots SERKs proteins groups. We also predicted the secondary and tertiary with ligand binding sites structure of the protein PhSERKs.

  3. Plant Explants Grown on Medium Supplemented with Fe3O4 Nanoparticles Have a Significant Increase in Embryogenesis

    Directory of Open Access Journals (Sweden)

    Inese Kokina

    2017-01-01

    Full Text Available Development of nanotechnology leads to the increasing release of nanoparticles in the environment that results in accumulation of different NPs in living organisms including plants. This can lead to serious changes in plant cultures which leads to genotoxicity. The aims of the present study were to detect if iron oxide NPs pass through the flax cell wall, to compare callus morphology, and to estimate the genotoxicity in Linum usitatissimum L. callus cultures induced by different concentrations of Fe3O4 nanoparticles. Two parallel experiments were performed: experiment A, where flax explants were grown on medium supplemented with 0.5 mg/l, 1 mg/l, and 1.5 mg/l Fe3O4 NPs for callus culture obtaining, and experiment B, where calluses obtained from basal MS medium were transported into medium supplemented with concentrations of NPs identical to experiment A. Obtained results demonstrate similarly in both experiments that 25 nm Fe3O4 NPs pass into callus cells and induce low toxicity level in the callus cultures. Nevertheless, calluses from experiment A showed 100% embryogenesis in comparison with experiment B where 100% rhizogenesis was noticed. It could be associated with different stress levels and adaptation time for explants and calluses that were transported into medium with Fe3O4 NPs supplementation.

  4. The Homeobox Genes of Caenorhabditis elegans and Insights into Their Spatio-Temporal Expression Dynamics during Embryogenesis

    Science.gov (United States)

    Abou-Zied, Akram M.; Lüppert, Martin; Dethlefsen, Johan; Mukherjee, Krishanu; Tong, Yong Guang; Tang, Lois; Gangishetti, Umesh; Baillie, David L.; Bürglin, Thomas R.

    2015-01-01

    Homeobox genes play crucial roles for the development of multicellular eukaryotes. We have generated a revised list of all homeobox genes for Caenorhabditis elegans and provide a nomenclature for the previously unnamed ones. We show that, out of 103 homeobox genes, 70 are co-orthologous to human homeobox genes. 14 are highly divergent, lacking an obvious ortholog even in other Caenorhabditis species. One of these homeobox genes encodes 12 homeodomains, while three other highly divergent homeobox genes encode a novel type of double homeodomain, termed HOCHOB. To understand how transcription factors regulate cell fate during development, precise spatio-temporal expression data need to be obtained. Using a new imaging framework that we developed, Endrov, we have generated spatio-temporal expression profiles during embryogenesis of over 60 homeobox genes, as well as a number of other developmental control genes using GFP reporters. We used dynamic feedback during recording to automatically adjust the camera exposure time in order to increase the dynamic range beyond the limitations of the camera. We have applied the new framework to examine homeobox gene expression patterns and provide an analysis of these patterns. The methods we developed to analyze and quantify expression data are not only suitable for C. elegans, but can be applied to other model systems or even to tissue culture systems. PMID:26024448

  5. Clathrin heavy chain is important for viability, oviposition, embryogenesis and, possibly, systemic RNAi response in the predatory mite Metaseiulus occidentalis.

    Directory of Open Access Journals (Sweden)

    Ke Wu

    Full Text Available Clathrin heavy chain has been shown to be important for viability, embryogenesis, and RNA interference (RNAi in arthropods such as Drosophila melanogaster. However, the functional roles of clathrin heavy chain in chelicerate arthropods, such as the predatory mite Metaseiulus occidentalis, remain unknown. We previously showed that dsRNA ingestion, followed by feeding on spider mites, induced systemic and robust RNAi in M. occidentalis females. In the current study, we performed a loss-of-function analysis of the clathrin heavy chain gene in M. occidentalis using RNAi. We showed that ingestion of clathrin heavy chain dsRNA by M. occidentalis females resulted in gene knockdown and reduced longevity. In addition, clathrin heavy chain dsRNA treatment almost completely abolished oviposition by M. occidentalis females and the few eggs produced did not hatch. Finally, we demonstrated that clathrin heavy chain gene knockdown in M. occidentalis females significantly reduced a subsequent RNAi response induced by ingestion of cathepsin L dsRNA. The last finding suggests that clathrin heavy chain may be involved in systemic RNAi responses mediated by orally delivered dsRNAs in M. occidentalis.

  6. Clustering in the stellar abundance space

    Science.gov (United States)

    Boesso, R.; Rocha-Pinto, H. J.

    2018-03-01

    We have studied the chemical enrichment history of the interstellar medium through an analysis of the n-dimensional stellar abundance space. This work is a non-parametric analysis of the stellar chemical abundance space. The main goal is to study the stars from their organization within this abundance space. Within this space, we seek to find clusters (in a statistical sense), that is, stars likely to share similar chemo-evolutionary history, using two methods: the hierarchical clustering and the principal component analysis. We analysed some selected abundance surveys available in the literature. For each sample, we labelled the group of stars according to its average abundance curve. In all samples, we identify the existence of a main enrichment pattern of the stars, which we call chemical enrichment flow. This flow is set by the structured and well-defined mean rate at which the abundances of the interstellar medium increase, resulting from the mixture of the material ejected from the stars and stellar mass-loss and interstellar medium gas. One of the main results of our analysis is the identification of subgroups of stars with peculiar chemistry. These stars are situated in regions outside of the enrichment flow in the abundance space. These peculiar stars show a mismatch in the enrichment rate of a few elements, such as Mg, Si, Sc and V, when compared to the mean enrichment rate of the other elements of the same stars. We believe that the existence of these groups of stars with peculiar chemistry may be related to the accretion of planetary material on to stellar surfaces or may be due to production of the same chemical element by different nucleosynthetic sites.

  7. Molecular characterization and expression analysis of SERK1 and SERK2 in Brassica napus L.: implication for microspore embryogenesis and plant regeneration.

    Science.gov (United States)

    Ahmadi, Behzad; Masoomi-Aladizgeh, Farhad; Shariatpanahi, Mehran E; Azadi, Pejman; Keshavarz-Alizadeh, Mehdi

    2016-01-01

    The BnSERK1 and BnSERK2 are involved in the process of microspore embryogenesis induction, development, and plantlet regeneration. Little is known about regulatory role of somatic embryogenesis-related kinase (SERK) genes family in the induction of microspore embryogenesis, development and plant regeneration. In this study, the expression of two SERK genes (SERK1 and SERK2) was assessed during the microspore embryogenesis and plantlet regeneration in Brassica napus L. The BnSERK1 was severely up-regulated 1-5 days following microspore culture and its expression drastically decreased in the globular-heart and also torpedo staged microspore-derived embryos (MDEs). In addition, high levels of BnSERK1 transcript were detected in the MDE maturation phase and in the roots and shoots of the regenerated plantlets which indicates a broader role(s) of BnSERK1 in the organ formation, rather than being specific to the embryogenesis. Results of partial sequencing indicated that the BnSERK1 shares a conserved serine-threonine kinase catalytic domain and exhibited 95 % similarity with AtSERK1, CsSERK1, BrSERK1, NaSERK1, and NbSERK1. A steady increase in the expression of BnSERK2 was observed during the MDE initiation and development so that, the highest expression was noted in the MDE maturation phase i.e., late cotyledonary MDEs. Our results also indicated low amounts of BnSERK2 transcript at the onset of rhyzogenesis but significantly higher expression in the developing roots. In contrast, the BnSERK2 strongly up-regulated during the both initially and developed shoots. The BnSERK2 shares highly conserved LRR-RLK domain when compared with different species tested so that, high homology (100 %) was noticed with BrSERK2. Based on our findings, MDE formation and plantlet regeneration seem to be correlated with both BnSERK1 and BnSERK2 expression.

  8. Bracken: estimating species abundance in metagenomics data

    Directory of Open Access Journals (Sweden)

    Jennifer Lu

    2017-01-01

    Full Text Available Metagenomic experiments attempt to characterize microbial communities using high-throughput DNA sequencing. Identification of the microorganisms in a sample provides information about the genetic profile, population structure, and role of microorganisms within an environment. Until recently, most metagenomics studies focused on high-level characterization at the level of phyla, or alternatively sequenced the 16S ribosomal RNA gene that is present in bacterial species. As the cost of sequencing has fallen, though, metagenomics experiments have increasingly used unbiased shotgun sequencing to capture all the organisms in a sample. This approach requires a method for estimating abundance directly from the raw read data. Here we describe a fast, accurate new method that computes the abundance at the species level using the reads collected in a metagenomics experiment. Bracken (Bayesian Reestimation of Abundance after Classification with KrakEN uses the taxonomic assignments made by Kraken, a very fast read-level classifier, along with information about the genomes themselves to estimate abundance at the species level, the genus level, or above. We demonstrate that Bracken can produce accurate species- and genus-level abundance estimates even when a sample contains multiple near-identical species.

  9. REVIEW: Can habitat selection predict abundance?

    Science.gov (United States)

    Boyce, Mark S; Johnson, Chris J; Merrill, Evelyn H; Nielsen, Scott E; Solberg, Erling J; van Moorter, Bram

    2016-01-01

    Habitats have substantial influence on the distribution and abundance of animals. Animals' selective movement yields their habitat use. Animals generally are more abundant in habitats that are selected most strongly. Models of habitat selection can be used to distribute animals on the landscape or their distribution can be modelled based on data of habitat use, occupancy, intensity of use or counts of animals. When the population is at carrying capacity or in an ideal-free distribution, habitat selection and related metrics of habitat use can be used to estimate abundance. If the population is not at equilibrium, models have the flexibility to incorporate density into models of habitat selection; but abundance might be influenced by factors influencing fitness that are not directly related to habitat thereby compromising the use of habitat-based models for predicting population size. Scale and domain of the sampling frame, both in time and space, are crucial considerations limiting application of these models. Ultimately, identifying reliable models for predicting abundance from habitat data requires an understanding of the mechanisms underlying population regulation and limitation. © 2015 The Authors. Journal of Animal Ecology published by John Wiley & Sons Ltd on behalf of British Ecological Society.

  10. Temporal Changes in Gametogenesis of the Invasive Chinese Pond Mussel Sinanodonta woodiana (Lea, 1834) (Bivalvia: Unionidae) from the Konin Lakes System (Central Poland).

    Science.gov (United States)

    Hliwa, Piotr; Zdanowski, Bogusław; Dietrich, Grzegorz J; Andronowska, Aneta; Król, Jarosław; Ciereszko, Andrzej

    2015-01-01

    Gametogenesis and the temporal changes occurring in the ovaries and testes throughout the reproductive cycle in the invasive alien bivalve, Chinese pond mussel Sinanodonta woodiana (Lea), from the heated Konin lakes system (central Poland) were studied using histological techniques. S. woodiana was confirmed to be a gonochoristic species with overall sex ratio of 1:1. The examined morphological parameters of Chinese pond mussel spermatozoa, i.e. 42 μm mean total length; 4.3 μm mean head length and the maximum size of previtellogenic (34-43 μm) and vitellogenic oocytes (75-83 μm) are consistent with values established for closely related members of the Unionidae family. Our results suggest that S. woodiana in the Konin lakes system are able to spawn throughout March to October, with a season of higher reproductive activity in females extending from March to April. This type of reproductive biology may contribute to the Chinese pond mussel's success in thriving in freshwater ecosystems.

  11. Chemical Abundances of Compact Planetary Nebulae

    Science.gov (United States)

    Lee, Ting-Hui; Shaw, Richard A.; Stanghellini, letizia; Riley, Ben

    2015-08-01

    We present preliminary results from an optical spectroscopic survey of compact planetary nebulae (PNe) in the Galactic disk. This is an ongoing optical+infrared spectral survey of 150 compact PNe to build a deep sample of PN chemical abundances. We obtained optical spectra of PNe with the Southern Astrophysical Research (SOAR) Telescope and Goodman High-Throughput Spectrograph between 2012 and 2015. These data were used to calculate the nebulae diagnostics such as electron temperature and density for each PN, and to derive the elemental abundances of He, N, O Ne, S and Ar. These abundances are vital to understanding the nature of the PNe, and their low- to intermediate-mass progenitor stars.

  12. Estimating the relationship between abundance and distribution

    DEFF Research Database (Denmark)

    Rindorf, Anna; Lewy, Peter

    2012-01-01

    Numerous studies investigate the relationship between abundance and distribution using indices reflecting one of the three aspects of distribution: proportion of area occupied, aggregation, and geographical range. Using simulations and analytical derivations, we examine whether these indices...... based on Euclidean distance to the centre of gravity of the spatial distribution. Only the proportion of structurally empty areas, Lloyds index, and indices of the distance to the centre of gravity of the spatial distribution are unbiased at all levels of abundance. The remaining indices generate...... relationships between abundance and distribution even in cases where no underlying relationships exists, although the problem decreases for measures derived from Lorenz curves when samples contain more than four individuals on average. To illustrate the problem, the indices are applied to juvenile North Sea cod...

  13. Age-abundance relationships for neutral communities

    Science.gov (United States)

    Danino, Matan; Shnerb, Nadav M.

    2015-10-01

    Neutral models for the dynamics of a system of competing species are often used to describe a wide variety of empirical communities. These models are used in many situations, ranging from population genetics and ecological biodiversity to macroevolution and cancer tumors. One of the main issues discussed within this framework is the relationships between the abundance of a species and its age. Here we provide a comprehensive analysis of the age-abundance relationships for fixed-size and growing communities. Explicit formulas for the average and the most likely age of a species with abundance n are given, together with the full probability distribution function. We further discuss the universality of these results and their applicability to the tropical forest community.

  14. INTERSTELLAR ABUNDANCES TOWARD X Per, REVISITED

    Energy Technology Data Exchange (ETDEWEB)

    Valencic, Lynne A. [Johns Hopkins University, Baltimore, MD 21218 (United States); Smith, Randall K. [Smithsonian Astrophysical Observatory, Cambridge, MA 02138 (United States)

    2013-06-10

    The nearby X-ray binary X Per (HD 24534) provides a useful beacon with which to examine dust grain types and measure elemental abundances in the local interstellar medium (ISM). The absorption features of O, Fe, Mg, and Si along this line of sight were measured using spectra from the Chandra X-Ray Observatory's LETG/ACIS-S and XMM-Newton's RGS instruments, and the Spex software package. The spectra were fit with dust analogs measured in the laboratory. The O, Mg, and Si abundances were compared to those from standard references, and the O abundance was compared to that along lines of sight toward other X-ray binaries. The results are as follows. First, it was found that a combination of MgSiO{sub 3} (enstatite) and Mg{sub 1.6}Fe{sub 0.4}SiO{sub 4} (olivine) provided the best fit to the O K edge, with N(MgSiO{sub 3})/N(Mg{sub 1.6}Fe{sub 0.4}SiO{sub 4}) = 3.4. Second, the Fe L edge could be fit with models that included metallic iron, but it was not well described by the laboratory spectra currently available. Third, the total abundances of O, Mg, and Si were in very good agreement with that of recently re-analyzed B stars, suggesting that they are good indicators of abundances in the local ISM, and the depletions were also in agreement with expected values for the diffuse ISM. Finally, the O abundances found from X-ray binary absorption spectra show a similar correlation with Galactocentric distances as seen in other objects.

  15. Abundances in planetary nebulae : NGC 6826

    NARCIS (Netherlands)

    Surendiranath, R.; Pottasch, S. R.

    Aims. We determine the chemical abundances and other parameters of the nebula NGC 6826 and its central star. Methods. We present new ISO spectra and combine them with archival IUE and optical spectra from the literature to get a complete, extinction-corrected, spectrum. The chemical composition of

  16. Abundances of planetary nebula NGC2392

    NARCIS (Netherlands)

    Pottasch, S. R.; Bernard-Salas, J.; Roellig, T. L.

    The spectra of the planetary nebula NGC2392 is reanalysed using spectral measurements made in the mid-infrared with the Spitzer Space Telescope. The aim is to determine the chemical composition of this object. We also make use of IUE and ground based spectra. Abundances determined from the

  17. Catch Composition, Abundance and Length- Weight Relationships ...

    African Journals Online (AJOL)

    loading Salter digital balance for small fish (<2.0 kg) or on a hanging Salter ... white- spotted grouper,. Epinephelus caeruleopunctatus, was numerically the most abundant (15.6%) in the catch followed by Cephalopholis boenak. (13%) and E. fasciatus ... Vanga, the commonly landed species were the white-spotted grouper,.

  18. A biogeographical perspective on species abundance distributions

    DEFF Research Database (Denmark)

    Matthews, Thomas J.; Borges, Paulo A. V.; de Azevedo, Eduardo Brito

    2017-01-01

    the fit of different SAD models to data from a single site is to collect abundance data from a variety of sites, and then build models to analyse how different SAD properties (e.g. form, skewness) vary with different predictor variables. Such a biogeographical approach to SAD research is potentially very...

  19. Will Abundant Natural Gas Solve Climate Change?

    Science.gov (United States)

    McJeon, H. C.; Edmonds, J.; Bauer, N.; Leon, C.; Fisher, B.; Flannery, B.; Hilaire, J.; Krey, V.; Marangoni, G.; Mi, R.; Riahi, K.; Rogner, H.; Tavoni, M.

    2015-12-01

    The rapid deployment of hydraulic fracturing and horizontal drilling technologies enabled the production of previously uneconomic shale gas resources in North America. Global deployment of these advanced gas production technologies could bring large influx of economically competitive unconventional gas resources to the energy system. It has been hoped that abundant natural gas substituting for coal could reduce carbon dioxide (CO2) emissions, which in turn could reduce climate forcing. Other researchers countered that the non-CO2 greenhouse gas (GHG) emissions associated with shale gas production make its lifecycle emissions higher than those of coal. In this study, we employ five state-of-the-art integrated assessment models (IAMs) of energy-economy-climate systems to assess the full impact of abundant gas on climate change. The models show large additional natural gas consumption up to +170% by 2050. The impact on CO2 emissions, however, is found to be much smaller (from -2% to +11%), and a majority of the models reported a small increase in climate forcing (from -0.3% to +7%) associated with the increased use of abundant gas. Our results show that while globally abundant gas may substantially change the future energy market equilibrium, it will not significantly mitigate climate change on its own in the absence of climate policies.

  20. SPECIES RICHNESS AND ABUNDANCE OF CLINID FISH ...

    African Journals Online (AJOL)

    Data analyses. Stepwise multiple regression analysis was used to determine which environmental factors. (independent variables) were significant predictors of number of clinid species, and clinid abundance and ... models to detect possible non-linear relations between dependent and independent variables. Sepkoski ...

  1. Palaeoceanographic implications of abundance and mean ...

    Indian Academy of Sciences (India)

    Abstract. Temporal variation in abundance and mean proloculus diameter of the benthic foraminiferal species Epistominella exigua has been reconstructed over the last ∼ 50,000 yr BP,from a core collected from the distal Bay of Bengal fan,to assess its potential application in palaeoceanographic reconstruction studies.

  2. Electron Density and Temperature Measurements, and Abundance ...

    Indian Academy of Sciences (India)

    Using spectra obtained from the SUMER (Solar Ultraviolet Measurements of Emitted Radiation) spectrograph on the spacecraft SOHO (Solar and Heliospheric Observatory), we investigate the height dependence of electron density, temperature and abundance anomalies in the solar atmosphere. In particular, we present ...

  3. Composition, abundance, distribution and seasonality of larval ...

    African Journals Online (AJOL)

    Composition, abundance, distribution and seasonality of larval fishes in the Sundays Estuary, South Africa. K. Sutherland, N.A. Strydom, T.H. Wooldridge. Abstract. The larval fish assemblage was studied in the permanently open Sundays Estuary on the southeast coast of South Africa. Seasonal samples were collected ...

  4. Composition, abundance, distribution and seasonality of ...

    African Journals Online (AJOL)

    Composition, abundance, distribution and seasonality of zooplankton in the Sundays Estuary, South Africa. K Sutherland, TH Wooldridge, NA Strydom. Abstract. The zooplankton assemblage in the permanently open Sundays Estuary, on the south-east coast of South Africa, was studied seasonally. Samples were collected ...

  5. In Abundance: Networked Participatory Practices as Scholarship

    Science.gov (United States)

    Stewart, Bonnie E.

    2015-01-01

    In an era of knowledge abundance, scholars have the capacity to distribute and share ideas and artifacts via digital networks, yet networked scholarship often remains unrecognized within institutional spheres of influence. Using ethnographic methods including participant observation, interviews, and document analysis, this study investigates…

  6. Occurrence, abundance and distribution of benthic ...

    African Journals Online (AJOL)

    A baseline study was conducted of the occurrence of macroinvertebrates at 26 sites in the Nyando River catchment in 2005–2006. A total of 13 orders and 16 families of Arthropoda, Mollusca, Platyhelminthes and Annelida were collected, with the order Ephemeroptera being most abundant in the up- and mid-stream ...

  7. Short communication: Abundance, distribution and population ...

    African Journals Online (AJOL)

    The Nile crocodile (Crocodylus niloticus) is an iconic or keystone species in many aquatic ecosystems. In order to understand the abundance, distribution, and population trends of Nile crocodiles in Gonarezhou National Park (GNP), southeastern Zimbabwe, we carried out 4 annual aerial surveys, using a Super Cub aircraft ...

  8. Benthic macrofauna community composition, abundance and ...

    African Journals Online (AJOL)

    Macroinvertebrate abundance was higher in the shallow nearshore waters than in the deep offshore waters. The communities of the shallow, intermediate and deep environments were composed mainly of molluscs, worms and insects. Gastropods (52.68%) and bivalves (40.18%) constituted the major part of the benthic ...

  9. Microarray analysis of siberian ginseng cyclic somatic embryogenesis culture systems provides insight into molecular mechanisms of embryogenic cell cluster generation.

    Directory of Open Access Journals (Sweden)

    Chenguang Zhou

    Full Text Available Four systems of cyclic somatic embryogenesis of Siberian ginseng (Eleutherococcus senticosus Maxim were used to study the mechanism of embryonic cell cluster generation. The first, direct somatic embryo induction (DSEI, generates secondary embryos directly from the primary somatic embryos; the second, direct embryogenic cell cluster induction (DEC, induces embryogenic cell clusters directly from somatic embryos in agar medium. Subsequently, we found that when DEC-derived somatic embryos are transferred to suspension culture or a bioreactor culture, only somatic embryos are induced, and embryogenic cell clusters cannot form. Therefore, these new lines were named DEC cultured by liquid medium (ECS and DEC cultured by bioreactor (ECB, respectively. Transmission electron microscopy showed that DEC epidermal cells contained a variety of inclusions, distinct from other lines. A cDNA library of DEC was constructed, and 1,948 gene clusters were obtained and used as probes. RNA was prepared from somatic embryos from each of the four lines and hybridized to a microarray. In DEC, 7 genes were specifically upregulated compared with the other three lines, and 4 genes were downregulated. EsXTH1 and EsPLT1, which were among the genes upregulated in DEC, were cloned using the rapid amplification of cDNA ends (RACE. Real-time quantitative PCR showed EsXTH1 was more highly expressed in DEC than in other lines throughout the culture cycle, and EsPLT1 expression in DEC increased as culture duration increased, but remained at a low expression level in other lines. These results suggest that EsXTH1 and EsPLT1 may be the essential genes that play important roles during the induction of embryogenic cell clusters.

  10. Transient Exposure to Ethanol during Zebrafish Embryogenesis Results in Defects in Neuronal Differentiation: An Alternative Model System to Study FASD

    Science.gov (United States)

    Joya, Xavier; Garcia-Algar, Oscar; Vall, Oriol; Pujades, Cristina

    2014-01-01

    Background The exposure of the human embryo to ethanol results in a spectrum of disorders involving multiple organ systems, including the impairment of the development of the central nervous system (CNS). In spite of the importance for human health, the molecular basis of prenatal ethanol exposure remains poorly understood, mainly to the difficulty of sample collection. Zebrafish is now emerging as a powerful organism for the modeling and the study of human diseases. In this work, we have assessed the sensitivity of specific subsets of neurons to ethanol exposure during embryogenesis and we have visualized the sensitive embryonic developmental periods for specific neuronal groups by the use of different transgenic zebrafish lines. Methodology/Principal Findings In order to evaluate the teratogenic effects of acute ethanol exposure, we exposed zebrafish embryos to ethanol in a given time window and analyzed the effects in neurogenesis, neuronal differentiation and brain patterning. Zebrafish larvae exposed to ethanol displayed small eyes and/or a reduction of the body length, phenotypical features similar to the observed in children with prenatal exposure to ethanol. When neuronal populations were analyzed, we observed a clear reduction in the number of differentiated neurons in the spinal cord upon ethanol exposure. There was a decrease in the population of sensory neurons mainly due to a decrease in cell proliferation and subsequent apoptosis during neuronal differentiation, with no effect in motoneuron specification. Conclusion Our investigation highlights that transient exposure to ethanol during early embryonic development affects neuronal differentiation although does not result in defects in early neurogenesis. These results establish the use of zebrafish embryos as an alternative research model to elucidate the molecular mechanism(s) of ethanol-induced developmental toxicity at very early stages of embryonic development. PMID:25383948

  11. Modulation of DNA methylation machineries in Japanese rice fish (Oryzias latipes) embryogenesis by ethanol and 5-azacytidine.

    Science.gov (United States)

    Dasmahapatra, Asok K; Khan, Ikhlas A

    2016-01-01

    As a sequel of our investigations on the impact of epigenome in inducing fetal alcohol spectrum disorder (FASD) phenotypes in Japanese rice fish, we have investigated on several DNA methylation machinery genes including DNA methyl transferase 3ba (dnmt3ba) and methyl binding proteins (MBPs), namely, mbd1b, mbd3a, mbd3b, and mecp2 at the transcription level. Studies were made during normal development, from 0day post fertilization (dpf) to hatching, and also exposing the fertilized eggs to ethanol or a DNMT inhibitor, 5-azacytidine (5-azaC). We observed that during development, all these genes followed distinct expression patterns, generally high mRNA copies in early phases (0-1dpf) and significantly low mRNA copies prior to or after hatching. Ethanol (100-500mM, 0-2dpf) was unable to alter any of these mRNAs in 2dpf; additional four day (2-6dpf) maintenance of these embryos in ethanol-free environment, on 6dpf, was also unable to establish any significant difference in these mRNA levels in comparison with the corresponding controls. However, continuous exposure of fertilized eggs in 300mM ethanol, 0-6dpf, showed significantly high mRNA copies only in MBPs (mbd1b, mbd3a, mbd3b, mecp2). 5-azaC (2mM) on 2dpf was able to enhance only mbd3b mRNA. Removal of 5-azaC and maintenance of these embryos in clean medium, 2-6dpf, showed significantly enhanced mbd3b and mecp2 mRNAs compared to corresponding controls on 6dpf. Our studies showed that in Japanese rice fish embryogenesis both ethanol and 5-azaC have the potential to specifically modulate the developmental rhythm of DNA methylation machineries. Published by Elsevier Inc.

  12. DNA methyltransferase expressions in Japanese rice fish (Oryzias latipes) embryogenesis is developmentally regulated and modulated by ethanol and 5-azacytidine.

    Science.gov (United States)

    Dasmahapatra, Asok K; Khan, Ikhlas A

    2015-01-01

    We aimed to investigate the impact of the epigenome in inducting fetal alcohol spectrum disorder (FASD) phenotypes in Japanese rice fish embryogenesis. One of the significant events in epigenome is DNA methylation which is catalyzed by DNA methyltransferase (DNMT) enzymes. We analyzed DNMT enzyme mRNA expressions in Japanese rice fish development starting from fertilized eggs to hatching and also in embryos exposed for first 48h of development either to ethanol (300mM) or to 5-azacytidine (5-azaC; 2mM), an inhibitor of DNMT enzyme activity. As observed in FASD phenotypes, 5-azaC exposure was able to induce microcephaly and craniofacial cartilage deformities in Japanese rice fish. Moreover, we have observed that expression of DNMTs (dnmt1, dnmt3aa, and dnmt3bb.1) are developmentally regulated; high mRNA copies were found in early stages (1-2day-post-fertilization, dpf), followed by gradual reduction until hatched. In ethanol-treated embryos, compared to controls, dnmt1 mRNA is in reduced level in 2dpf and in enhanced level in 6dpf embryos. While dnmt3aa and 3bb.1 remained unaltered. In contrast, embryos exposed to 5-azaC have an enhanced level of dnmt1 and dnmt3bb.1 mRNAs both in 2 and 6dpf embryos while dnmt3aa is enhanced only in 6dpf embryos. Moreover, endocannabinoid receptor 1a (cnr1a) mRNA which was found to be reduced by ethanol remained unaltered and cnr1b and cnr2 mRNAs, which were remained unaltered by ethanol, were increased significantly by 5-azaC in 6dpf embryos. This study indicates that the craniofacial defects observed in FASD phenotypes are the results of dysregulations in DNMT expressions. Published by Elsevier Inc.

  13. Carbon source dependent somatic embryogenesis and plant regeneration in cotton, Gossypium hirsutum L. cv. SVPR2 through suspension cultures.

    Science.gov (United States)

    Ganesan, M; Jayabalan, N

    2005-10-01

    Highly reproducible and simple protocol for cotton somatic embryogenesis is described here by using different concentrations of maltose, glucose, sucrose and fructose. Maltose (30 g/l) is the best carbon source for embryogenic callus induction and glucose (30 g/l) was suitable for induction, maturation of embryoids and plant regeneration. Creamy white embryogenic calli of hypocotyl explants were formed on medium containing MS basal salts, myo-inositol (100 mg/l), thiamine HCI (0.3 mg/l), picloram (0.3 mg/l), Kin (0.1 mg/l) and maltose (30 g/l). During embryo induction and maturation, accelerated growth was observed in liquid medium containing NH3NO4 (1 g/l), picloram (2.0 mg/l), 2 ip (0.2 mg/l), Kin (0.1 mg/l) and glucose (30 g/l). Before embryoid induction, large clumps of embryogenic tissue were formed. These tissues only produced viable embryoids. Completely matured somatic embryos were germinated successfully on the medium fortified with MS salts, myo-inositol (50 mg/l), thiamine HCl (0.2 mg/l), GA3 (0.2 mg/l), BA (1.0 mg/l) and glucose (30 g/l). Compared with earlier reports, 65% of somatic embryo germination was observed. The abnormal embryo formation was highly reduced by using glucose (30 g/l) compared to other carbon sources. The regenerated plantlets were fertile but smaller in height than the seed derived control plants.

  14. The effects of thermal manipulations during embryogenesis of broiler chicks on growth of embryo and skeletal traits

    Energy Technology Data Exchange (ETDEWEB)

    Aygün, Ali, E-mail: aaygun@selcuk.edu.tr [Selcuk University, Faculty of Agriculture, Department of Animal Science, Konya, 42075 (Turkey); Narinç, Doğan, E-mail: narincd@gmail.com [Namik Kemal University, Faculty of Veterinary Medicine, Department of Genetics, Tekirdag, 59100 (Turkey)

    2016-04-18

    Incubation temperature is one of the important environmental factors that can induce epigenetic thermal adaptation of different physiological control systems. Thus, post hatch thermo tolerance ability of birds may be gained using these manipulations during different incubation periods. The current study was carried out to reveal the effects of temperature manipulations during early and late embryogenesis on weight of embryo and size of skeletal bilateral traits (face, wings, metatarsus, tibia, and femur) in broiler chicken embryos. One thousand commercial broiler eggs from 46 week old breeder flock were used in study. Treatments consisted of eggs incubated at 37.8°C and 55% relative humidity throughout (control; DG1), heated to 36.9°C and supplied 60% relative humidity for 6 hours daily from day 0 to 8 (DG2), heated to 36.9°C and supplied 60% relative humidity for 6 hours daily from day 10 to 18 (DG3), heated to 41°C and supplied 65% relative humidity for 3 hours daily from day 8 to 10 (DG4), and heated to 41°C and supplied 65% relative humidity for 3 hours daily from day 16 to 18 (DG5). Measurements of embryo weight and bilateral traits were obtained at 20 day of incubation and at hatch (at day 21). It was determined that the live weights of embryo and chick were affected significantly by treatment; DG3 group has shown higher mean values than the other treatment groups (P<0.05). There were differences in lengths of femur, tibia and metatarsus among treatment groups at hatch. Particularly, the high incubator temperatures at the second half of incubation accelerated growth of body and bone in embryos. These consequences of the treatments performed at different temperatures and times indicate that the different metabolic shifts realized by the embryos.

  15. Comprehensive analysis of cystatin family genes suggests their putative functions in sexual reproduction, embryogenesis, and seed formation.

    Science.gov (United States)

    Zhao, Peng; Zhou, Xue-mei; Zou, Jie; Wang, Wei; Wang, Lu; Peng, Xiong-bo; Sun, Meng-xiang

    2014-09-01

    Cystatins are tightly bound and reversible inhibitors of cysteine proteases in C1A and C13 peptidase families, which have been identified in several species and shown to function in vegetative development and response to biotic/abiotic stresses in plants. Recent work revealed their critical role in regulating programmed cell death during embryogenesis in tobacco and suggested their more comprehensive roles in the process of sexual plant reproduction, although little is known about cystatin family genes in the processes. Here, 10 cystatin family genes in Nicotiana tabacum were identified using an expressed sequence tag (EST)-based gene clone strategy. Analysis of their biochemical properties showed that nine of them have the potency to inhibit the activities of both commercial cathepsin L-like proteases and extracted cysteine proteases from seeds, but with different K i values depending on the types of proteases and the developmental stages of the seed tested. This suggests that cystatin-dependent cathepsin L-like proteolytic pathways are probably important for early seed development. Comprehensive expression profile analysis revealed that cystatin family genes showed manifold variations in their transcription levels in different plant cell types, including the sperm, egg, and zygote, especially in the embryo and seed at different developmental stages. More interestingly, intracellular localization analysis of each cystatin revealed that most members of cystatin families are recognized as secretory proteins with signal peptides that direct them to the endoplasmic reticulum. These results suggest their widespread roles in cell fate determination and cell-cell communication in the process of sexual reproduction, especially in gamete and embryo development, as well as in seed formation. © The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  16. Comparative toxicities of selected rare earth elements: Sea urchin embryogenesis and fertilization damage with redox and cytogenetic effects.

    Science.gov (United States)

    Pagano, Giovanni; Guida, Marco; Siciliano, Antonietta; Oral, Rahime; Koçbaş, Fatma; Palumbo, Anna; Castellano, Immacolata; Migliaccio, Oriana; Thomas, Philippe J; Trifuoggi, Marco

    2016-05-01

    Broad-ranging adverse effects are known for rare earth elements (REE), yet only a few studies tested the toxicity of several REE, prompting studies focusing on multi-parameter REE toxicity. Trichloride salts of Y, La, Ce, Nd, Sm, Eu and Gd were tested in Paracentrotus lividus sea urchin embryos and sperm for: (1) developmental defects in either REE-exposed larvae or in the offspring of REE-exposed sperm; (2) fertilization success; (3) mitotic anomalies in REE-exposed embryos and in the offspring of REE-exposed sperm, and (4) reactive oxygen species (ROS) formation, and malondialdehyde (MDA) and nitric oxide (NO) levels. REEs affected P. lividus larvae with concentration-related increase in developmental defects, 10(-6) to 10(-4)M, ranking as: Gd(III)>Y(III)>La(III)>Nd(III)≅Eu(III)>Ce(III)≅Sm(III). Nominal concentrations of REE salts were confirmed by inductively coupled plasma mass spectrometry (ICP-MS). Significant increases in MDA levels, ROS formation, and NO levels were found in REE-exposed embryos. Sperm exposure to REEs (10(-5) to 10(-4)M) resulted in concentration-related decrease in fertilization success along with increase in offspring damage. Decreased mitotic activity and increased aberration rates were detected in REE-exposed embryos and in the offspring of REE-exposed sperm. REE-associated toxicity affecting embryogenesis, fertilization, cytogenetic and redox endpoints showed different activities of tested REEs. Damage to early life stages, along with redox and cytogenetic anomalies should be the focus of future REE toxicity studies. Copyright © 2016 Elsevier Inc. All rights reserved.

  17. Muscle formation during embryogenesis of the polychaete Ophryotrocha diadema (Dorvilleidae – new insights into annelid muscle patterns

    Directory of Open Access Journals (Sweden)

    Paululat Achim

    2008-01-01

    Full Text Available Abstract Background The standard textbook information that annelid musculature consists of oligochaete-like outer circular and inner longitudinal muscle-layers has recently been called into question by observations of a variety of complex muscle systems in numerous polychaete taxa. To clarify the ancestral muscle arrangement in this taxon, we compared myogenetic patterns during embryogenesis of Ophryotrocha diadema with available data on oligochaete and polychaete myogenesis. This work addresses the conflicting views on the ground pattern of annelids, and adds to our knowledge of the evolution of lophotrochozoan taxa. Results Somatic musculature in Ophryotrocha diadema can be classified into the trunk, prostomial/peristomial, and parapodial muscle complexes. The trunk muscles comprise strong bilateral pairs of distinct dorsal and ventral longitudinal strands. The latter are the first to differentiate during myogenesis. They originate within the peristomium and grow posteriorly through the continuous addition of myocytes. Later, the longitudinal muscles also expand anteriorly and form a complex arrangement of prostomial muscles. Four embryonic parapodia differentiate in an anterior-to-posterior progression, significantly contributing to the somatic musculature. Several diagonal and transverse muscles are present dorsally. Some of the latter are situated external to the longitudinal muscles, which implies they are homologous to the circular muscles of oligochaetes. These circular fibers are only weakly developed, and do not appear to form complete muscle circles. Conclusion Comparison of embryonic muscle patterns showed distinct similarities between myogenetic processes in Ophryotrocha diadema and those of oligochaete species, which allows us to relate the diverse adult muscle arrangements of these annelid taxa to each other. These findings provide significant clues for the interpretation of evolutionary changes in annelid musculature.

  18. Transient exposure to ethanol during zebrafish embryogenesis results in defects in neuronal differentiation: an alternative model system to study FASD.

    Directory of Open Access Journals (Sweden)

    Xavier Joya

    Full Text Available The exposure of the human embryo to ethanol results in a spectrum of disorders involving multiple organ systems, including the impairment of the development of the central nervous system (CNS. In spite of the importance for human health, the molecular basis of prenatal ethanol exposure remains poorly understood, mainly to the difficulty of sample collection. Zebrafish is now emerging as a powerful organism for the modeling and the study of human diseases. In this work, we have assessed the sensitivity of specific subsets of neurons to ethanol exposure during embryogenesis and we have visualized the sensitive embryonic developmental periods for specific neuronal groups by the use of different transgenic zebrafish lines.In order to evaluate the teratogenic effects of acute ethanol exposure, we exposed zebrafish embryos to ethanol in a given time window and analyzed the effects in neurogenesis, neuronal differentiation and brain patterning. Zebrafish larvae exposed to ethanol displayed small eyes and/or a reduction of the body length, phenotypical features similar to the observed in children with prenatal exposure to ethanol. When neuronal populations were analyzed, we observed a clear reduction in the number of differentiated neurons in the spinal cord upon ethanol exposure. There was a decrease in the population of sensory neurons mainly due to a decrease in cell proliferation and subsequent apoptosis during neuronal differentiation, with no effect in motoneuron specification.Our investigation highlights that transient exposure to ethanol during early embryonic development affects neuronal differentiation although does not result in defects in early neurogenesis. These results establish the use of zebrafish embryos as an alternative research model to elucidate the molecular mechanism(s of ethanol-induced developmental toxicity at very early stages of embryonic development.

  19. Non-Salmonid Abundance - Line Features [ds186

    Data.gov (United States)

    California Department of Resources — The CalFish Abundance Database contains a comprehensive collection of anadromous fisheries abundance information. The "Other Fish" category contains data collected...

  20. Chemical Cartography in the Milky Way with SDSS/APOGEE: Multi-element abundances and abundance ratio variations

    Science.gov (United States)

    Holtzman, Jon A.; Hasselquist, Sten; Johnson, Jennifer; Bird, Jonathan C.; Majewski, Steven R.; SDSS/APOGEE Team

    2017-01-01

    The SDSS/APOGEE project is measuring abundances of multiple elements for several hundred thousand stars across the Milky Way. These allow the mapping of abundances and abundance ratio variations. Results will be presented for multiple abundance ratios across of the Galactic disk. The interpretation of mean abundance maps is complicated by variations in star formation history across the disk and by changing abundance ratios that result from an overall metallicity gradient. Variations in chemical abundance sequences, however, show the potential for using abundance ratios to track the movement of stars through the disk, and provide key information for constraining Galaxy formation and chemical evolution models.

  1. Deuterium Abundance in Consciousness and Current Cosmology

    Science.gov (United States)

    Rauscher, Elizabeth A.

    We utilize the deuterium-hydrogen abundances and their role in setting limits on the mass and other conditions of cosmogenesis and cosmological evolution. We calculate the dependence of a set of physical variables such as density, temperature, energy mass, entropy and other physical variable parameters through the evolution of the universe under the Schwarzschild conditions as a function from early to present time. Reconciliation with the 3°K and missing mass is made. We first examine the Schwarzschild condition; second, the geometrical constraints of a multidimensional Cartesian space on closed cosmologies, and third we will consider the cosmogenesis and evolution of the universe in a multidimensional Cartesian space, obeying the Schwarzschild condition. Implications of this model for matter creation are made. We also examine experimental evidence for closed versus open cosmologies; x-ray detection of the "missing mass" density. Also the interstellar deuterium abundance, along with the value of the Hubble constant set a general criterion on the value of the curvature constant, k. Once the value of the Hubble constant, H is determined, the deuterium abundance sets stringent restrictions on the value of the curvature constant k by an detailed discussion is presented. The experimental evidences for the determination of H and the primary set of coupled equations to determine D abundance is given. 'The value of k for an open, closed, or flat universe will be discussed in terms of the D abundance which will affect the interpretation of the Schwarzschild, black hole universe. We determine cosmology solutions to Einstein's field obeying the Schwarzschild solutions condition. With this model, we can form a reconciliation of the black hole, from galactic to cosmological scale. Continuous creation occurs at the dynamic blackhole plasma field. We term this new model the multiple big bang or "little whimper model". We utilize the deuteriumhydrogen abundances and their role in

  2. Emigrare o restare in Germania? Tre differenti risposte di autrici ebree tedesche di seconda generazione (Barbara Honigmann, Lea Fleischmann, Esther Dischereit

    Directory of Open Access Journals (Sweden)

    Alessandro Costazza

    2014-04-01

    Full Text Available The purpose of this paper is to investigate, through the analysis of the writings of three German Jewish authors of second generation – Lea Fleischmann, Esther Dischereit and Barbara Honigmann – their different responses to the complex post-war situation of Jews in both West- and East-Germany. Despite the different origins and experiences of their parents under Nazism, all three make in their youth the painful experience of a Judaism lived mainly as absence, as silence about the past and as lack of Jewish tradition and religion. Trying to achieve their own identity, Fleischmann and Dischereit seek a possible integration in post-war West-Germany in the political commitment of the 70s, but must then recognize that anti-Semitism is still widespread in that country. This discovery compels Fleischmann to leave Germany and seek in Israel and eventually in the religion of her fathers a Jewish identity. Esther Dischereit decides on the contrary to perform in Germany her "exercises to be Jewish", assuming the role of critical conscience of German society. Different is the situation of Barbara Honigmann, born and raised in East-Germany, who reacts to the dominant materialism in that state with a need for spirituality that will take her to seek in religion – lived outside of Germany and so far from "negative symbiosis" between Jews and Germans, but also far from Israel - her own diasporic identity. Despite the three different outcomes resulting from the search for Jewish identity in post-war Germany, for all three authors writing plays a crucial function as memory and critical conscience and since this writing is a German one, it imposes eventually a tight and thorough confrontation with German culture.

  3. The effect of auxin 2,4-D and cytokinin 2-ip on direct somatic embryogenesis formation of Coffea arabica L. leaf explant

    Directory of Open Access Journals (Sweden)

    Rina Arimarsetiowati

    2011-08-01

    Full Text Available One of the propagation technique for coffee plant production is tissue culture. Tissue culture technique for Coffea arabica L. faces some problems, mainly in the planlet formation regenerated from explants. The objective of this experiment was to examine the effect 2,4-D and 2-ip combination on the formation of direct somatic embryogenesis of Coffea arabica L. in leaves explant. Auxin (2,4-D and cytokinin (2-ip concentrations of, respectively, 1; 5 µM and 5; 10; 15; 20 were used as treatments. This research was conducted using completely randomized design with 10 replications. Observation to induce somatic embryos was done by quantitatively on number of callus from explant and number of embryogenic callus. Beside that, observation by qualitative descriptive was also done on deve lopment of embryogenesis. The results showed that Arabica coffee leaves explant of AS 2K clones could be induced in all medium combination except 5µM 2,4-D and 20µM 2-ip combination. Arabica coffee leaves explant of S 795, Sigararutang and AS 1 varieties could be induced in all medium combination. The highest frequency of callus formation was found in AS 2K, Sigararutang and AS 1 varieties on medium containing 1µM 2,4-D in combination with 10µM 2-ip, whereas for the S 795 variety on medium containing 5µM 2,4-D in combination with 10µM 2-ip. The highest frequency of embriogenic callus in all Arabica coffee variety could be reached on medium containing 5µM 2,4-D in combination with 15µM 2-ip. Key words : Coffea arabica L., somatic embryogenesis, 2,4-D, 2-ip, tissue culture, leaves, callus embryogenic.

  4. Zebrafish yolk-specific not really started (nrs) gene is a vertebrate homolog of the Drosophila spinster gene and is essential for embryogenesis.

    Science.gov (United States)

    Young, Rodrigo M; Marty, Scott; Nakano, Yoshiro; Wang, Han; Yamamoto, Daisuke; Lin, Shuo; Allende, Miguel L

    2002-03-01

    By using retroviral insertional mutagenesis in zebrafish, we have identified a recessive lethal mutation in the not really started (nrs) gene. The nrs mutation disrupts a gene located in linkage group 3 that is highly homologous to the spinster gene identified in Drosophila and to spinster orthologs identified in mammals. In flies, spinster encodes a membrane protein involved in lysosomal metabolism and programmed cell death in the central nervous system and in the ovary. In nrs mutant fish embryos, we detect an opaque substance in the posterior yolk cell extension at approximately 1 day after fertilization. This material progressively accumulates and by 48 hr after fertilization fills the entire yolk. By day 3 of embryogenesis, mutant embryos are severely reduced in size compared with their wild-type siblings and they die a few hours later. By in situ hybridization, we show that the nrs mRNA is expressed in the yolk cell at the time the mutant phenotype becomes apparent. In wild-type embryos, nrs message is present maternally and zygotically throughout embryogenesis and is also detected in adult animals. In nrs homozygous mutant embryos, nrs transcripts are undetectable at the time the phenotype becomes apparent, indicating that the retroviral insertion has most likely abolished expression of the nrs gene. Finally, the nrs phenotype can be partially rescued by microinjection of nrs encoding DNA. These results suggest that the nrs mutation affects an essential gene encoding a putative membrane-bound protein expressed specifically in the yolk cell during zebrafish embryogenesis. Copyright 2002 Wiley-Liss, Inc.

  5. Two cDNA clones (FsDhn1 and FsClo1) up-regulated by ABA are involved in drought responses in Fagus sylvatica L. seeds.

    Science.gov (United States)

    Jiménez, Jesús Angel; Alonso-Ramírez, Ana; Nicolás, Carlos

    2008-11-28

    Two abscisic acid (ABA)-responsive clones (FsDhn1 and FsClo1) were isolated from a cDNA library of ABA-treated seeds of Fagus sylvatica L. FsDhn1 codes for type-II late embryogenesis abundant (LEA) proteins, also known as dehydrins. The corresponding transcripts were ABA-induced and expressed when seeds were artificially dried. FsClo1 codes for a caleosin, a calcium-binding protein. By expressing FsCLO1 as a fusion protein in Escherichia coli, we obtained biochemical evidence of its ability to bind calcium, a second messenger involved in ABA signaling. This cDNA clone is seed specific, shows a LEA expression pattern and is also induced by ABA and dehydration treatments. The relationship of both genes with seed responses to ABA is discussed.

  6. The Arabidopsis NF-YA3 and NF-YA8 genes are functionally redundant and are required in early embryogenesis.

    Directory of Open Access Journals (Sweden)

    Monica Fornari

    Full Text Available Nuclear factor Y (NF-Y is a trimeric transcription factor composed of three distinct subunits called NF-YA, NF-YB and NF-YC. In Arabidopsis thaliana, NF-Y subunits are known to play roles in many processes, such as gametogenesis, embryogenesis, seed development, drought resistance, ABA signaling, flowering time, primary root elongation, Endoplasmic Reticulum (ER stress response and blue light responses. Here, we report that the closely related NF-YA3 and NF-YA8 genes control early embryogenesis. Detailed GUS and in situ analyses showed that NF-YA3 and NF-YA8 are expressed in vegetative and reproductive tissues with the highest expression being during embryo development from the globular to the torpedo embryo stage. Plants from the nf-ya3 and nf-ya8 single mutants do not display any obvious phenotypic alteration, whereas nf-ya3 nf-ya8 double mutants are embryo lethal. Morphological analyses showed that the nf-ya3 nf-ya8 embryos fail to undergo to the heart stage and develop into abnormal globular embryos with both proembryo and suspensor characterized by a disordered cell cluster with an irregular shape, suggesting defects in embryo development. The suppression of both NF-YA3 and NF-YA8 gene expression by RNAi experiments resulted in defective embryos that phenocopied the nf-ya3 nf-ya8 double mutants, whereas complementation experiments partially rescued the abnormal globular nf-ya3 nf-ya8 embryos, confirming that NF-YA3 and NF-YA8 are required in early embryogenesis. Finally, the lack of GFP expression of the auxin responsive DR5rev::GFP marker line in double mutant embryos suggested that mutations in both NF-YA3 and NF-YA8 affect auxin response in early developing embryos. Our findings indicate that NF-YA3 and NF-YA8 are functionally redundant genes required in early embryogenesis of Arabidopsis thaliana.

  7. Integral Field Spectroscopy Surveys: Oxygen Abundance Gradients

    Science.gov (United States)

    Sánchez, S. F.; Sánchez-Menguiano, L.

    2017-07-01

    We present here the recent results on our understanding of oxygen abundance gradients derived using Integral Field Spectroscopic surveys. In particular we analyzed more than 2124 datacubes corresponding to individual objects observed by the CALIFA (˜ 734 objects) and the public data by MaNGA (˜ 1390 objects), deriving the oxygen abundance gradient for each galaxy. We confirm previous results that indicate that the shape of this gradient is very similar for all galaxies with masses above 109.5M⊙, presenting in average a very similar slope of ˜ -0.04 dex within 0.5-2.0 re, with a possible drop in the inner regions (r109.5M⊙) the gradient seems to be flatter than for more massive ones. All these results agree with an inside-out growth of massive galaxies and indicate that low mass ones may still be growing in an outside in phase.

  8. Angel lichen moth abundance and morphology data

    Science.gov (United States)

    Metcalfe, Anya; Kennedy, Theodore A.; Muehlbauer, Jeffrey D.

    2016-01-01

    Two unique datasets on the abundance and morphology of the angel lichen moth ( Cisthene angelus) in Grand Canyon, Arizona, USA were compiled to describe the phenology and life history of this common, but poorly known, species. The abundance data were collected from 2012 to 2013 through a collaboration with river runners in Grand Canyon National Park. These citizen scientists deployed light traps from their campsites for one hour each night of their expedition. Insects were preserved in ethanol on site, and returned to the Southwest Biological Science Center in Flagstaff, Arizona for analysis in the laboratory. A total of 2,437 light trap samples were sorted through, 903 of which contained C. angelus. In total, 73,841 C. angelus were identified and enumerated to create the abundance data set. The morphology dataset is based on a subset of 28 light trap samples from sampling year 2012 (14 from spring and 14 from fall.) It includes gender and forewing lengths for 2,674 individual moths and dry weights for 1,102 of those individuals.

  9. Abundance effects on the Cepheid distance scale

    Science.gov (United States)

    Stothers, Richard B.

    1988-06-01

    A new investigation is made of the dependence of the period-luminosity (PL) and period-luminosity-color (PLC) relations on the helium (Y) and metal (Z) abundances in classical Cepheids. For consistency in overall approach and in error estimation, theoretical data alone have been used. These data come from stellar evolution, atmosphere, and pulsation theory. Composition dependences have been assigned to all quantities. A major quantitative result is that the PLC relation in B, V magnitudes is at least 5 times more sensitive to chemical composition than is the PL relation. If the PL relation is adopted and the helium abundance is held fixed, a Cepheid with Z = 0.005 should be fainter than one with Z = 0.02 by ≡0.04 mag. Since the slope of the predicted PL relation is independent of chemical composition, the PL relation can be considered to be effectively universal. Theoretical results are also presented for the bolometric PL relation, whose slope and dependence on metal abundance should be roughly applicable to infrared observations of Cepheids.

  10. A global database of ant species abundances

    Science.gov (United States)

    Gibb, Heloise; Dunn, Rob R.; Sanders, Nathan J.; Grossman, Blair F.; Photakis, Manoli; Abril, Silvia; Agosti, Donat; Andersen, Alan N.; Angulo, Elena; Armbrecht, Ingre; Arnan, Xavier; Baccaro, Fabricio B.; Bishop, Tom R.; Boulay, Raphael; Bruhl, Carsten; Castracani, Cristina; Cerda, Xim; Del Toro, Israel; Delsinne, Thibaut; Diaz, Mireia; Donoso, David A.; Ellison, Aaron M.; Enriquez, Martha L.; Fayle, Tom M.; Feener Jr., Donald H.; Fisher, Brian L.; Fisher, Robert N.; Fitpatrick, Matthew C.; Gomez, Cristanto; Gotelli, Nicholas J.; Gove, Aaron; Grasso, Donato A.; Groc, Sarah; Guenard, Benoit; Gunawardene, Nihara; Heterick, Brian; Hoffmann, Benjamin; Janda, Milan; Jenkins, Clinton; Kaspari, Michael; Klimes, Petr; Lach, Lori; Laeger, Thomas; Lattke, John; Leponce, Maurice; Lessard, Jean-Philippe; Longino, John; Lucky, Andrea; Luke, Sarah H.; Majer, Jonathan; McGlynn, Terrence P.; Menke, Sean; Mezger, Dirk; Mori, Alessandra; Moses, Jimmy; Munyai, Thinandavha Caswell; Pacheco, Renata; Paknia, Omid; Pearce-Duvet, Jessica; Pfeiffer, Martin; Philpott, Stacy M.; Resasco, Julian; Retana, Javier; Silva, Rogerio R.; Sorger, Magdalena D.; Souza, Jorge; Suarez, Andrew V.; Tista, Melanie; Vasconcelos, Heraldo L.; Vonshak, Merav; Weiser, Michael D.; Yates, Michelle; Parr, Catherine L.

    2017-01-01

    What forces structure ecological assemblages? A key limitation to general insights about assemblage structure is the availability of data that are collected at a small spatial grain (local assemblages) and a large spatial extent (global coverage). Here, we present published and unpublished data from 51,388 ant abundance and occurrence records of more than 2693 species and 7953 morphospecies from local assemblages collected at 4212 locations around the world. Ants were selected because they are diverse and abundant globally, comprise a large fraction of animal biomass in most terrestrial communities, and are key contributors to a range of ecosystem functions. Data were collected between 1949 and 2014, and include, for each geo-referenced sampling site, both the identity of the ants collected and details of sampling design, habitat type and degree of disturbance. The aim of compiling this dataset was to provide comprehensive species abundance data in order to test relationships between assemblage structure and environmental and biogeographic factors. Data were collected using a variety of standardised methods, such as pitfall and Winkler traps, and will be valuable for studies investigating large-scale forces structuring local assemblages. Understanding such relationships is particularly critical under current rates of global change. We encourage authors holding additional data on systematically collected ant assemblages, especially those in dry and cold, and remote areas, to contact us and contribute their data to this growing dataset.

  11. 2015-2016 Palila abundance estimates

    Science.gov (United States)

    Camp, Richard J.; Brinck, Kevin W.; Banko, Paul C.

    2016-01-01

    The palila (Loxioides bailleui) population was surveyed annually during 1998−2016 on Mauna Kea Volcano to determine abundance, population trend, and spatial distribution. In the latest surveys, the 2015 population was estimated at 852−1,406 birds (point estimate: 1,116) and the 2016 population was estimated at 1,494−2,385 (point estimate: 1,934). Similar numbers of palila were detected during the first and subsequent counts within each year during 2012−2016; the proportion of the total annual detections in each count ranged from 46% to 56%; and there was no difference in the detection probability due to count sequence. Furthermore, conducting repeat counts improved the abundance estimates by reducing the width of the confidence intervals between 9% and 32% annually. This suggests that multiple counts do not affect bird or observer behavior and can be continued in the future to improve the precision of abundance estimates. Five palila were detected on supplemental survey stations in the Ka‘ohe restoration area, outside the core survey area but still within Palila Critical Habitat (one in 2015 and four in 2016), suggesting that palila are present in habitat that is recovering from cattle grazing on the southwest slope. The average rate of decline during 1998−2016 was 150 birds per year. Over the 18-year monitoring period, the estimated rate of change equated to a 58% decline in the population.

  12. Abundant SAR11 viruses in the ocean.

    Science.gov (United States)

    Zhao, Yanlin; Temperton, Ben; Thrash, J Cameron; Schwalbach, Michael S; Vergin, Kevin L; Landry, Zachary C; Ellisman, Mark; Deerinck, Tom; Sullivan, Matthew B; Giovannoni, Stephen J

    2013-02-21

    Several reports proposed that the extraordinary dominance of the SAR11 bacterial clade in ocean ecosystems could be a consequence of unusual mechanisms of resistance to bacteriophage infection, including 'cryptic escape' through reduced cell size and/or K-strategist defence specialism. Alternatively, the evolution of high surface-to-volume ratios coupled with minimal genomes containing high-affinity transporters enables unusually efficient metabolism for oxidizing dissolved organic matter in the world's oceans that could support vast population sizes despite phage susceptibility. These ideas are important for understanding plankton ecology because they emphasize the potentially important role of top-down mechanisms in predation, thus determining the size of SAR11 populations and their concomitant role in biogeochemical cycling. Here we report the isolation of diverse SAR11 viruses belonging to two virus families in culture, for which we propose the name 'pelagiphage', after their host. Notably, the pelagiphage genomes were highly represented in marine viral metagenomes, demonstrating their importance in nature. One of the new phages, HTVC010P, represents a new podovirus subfamily more abundant than any seen previously, in all data sets tested, and may represent one of the most abundant virus subfamilies in the biosphere. This discovery disproves the theory that SAR11 cells are immune to viral predation and is consistent with the interpretation that the success of this highly abundant microbial clade is the result of successfully evolved adaptation to resource competition.

  13. BOND: Bayesian Oxygen and Nitrogen abundance Determinations

    Science.gov (United States)

    Vale Asari, N.; Stasinska, G.; Morisset, C.; Cid Fernandes, R.

    2018-01-01

    BOND determines oxygen and nitrogen abundances in giant H II regions by comparison with a large grid of photoionization models. The grid spans a wide range in O/H, N/O and ionization parameter U, and covers different starburst ages and nebular geometries. Unlike other statistical methods, BOND relies on the [Ar III]/[Ne III] emission line ratio to break the oxygen abundance bimodality. By doing so, it can measure oxygen and nitrogen abundances without assuming any a priori relation between N/O and O/H. BOND takes into account changes in the hardness of the ionizing radiation field, which can come about due to the ageing of H II regions or the stochastically sampling of the IMF. The emission line ratio He I/Hβ, in addition to commonly used strong lines, constrains the hardness of the ionizing radiation field. BOND relies on the emission line ratios [O III]/Hβ, [O II]/Hβ and [N II]/Hβ, [Ar III]/Hβ, [Ne III]/Hβ, He I/Hβ as its input parameters, while its output values are the measurements and uncertainties for O/H and N/O.

  14. Abundant Solar Nebula Solids in Comets

    Science.gov (United States)

    Messenger, S.; Keller, L. P.; Nakamura-Messenger, K.; Nguyen, A. N.; Clemett, S.

    2016-01-01

    Comets have been proposed to consist of unprocessed interstellar materials together with a variable amount of thermally annealed interstellar grains. Recent studies of cometary solids in the laboratory have shown that comets instead consist of a wide range of materials from across the protoplanetary disk, in addition to a minor complement of interstellar materials. These advances were made possible by the return of direct samples of comet 81P/Wild 2 coma dust by the NASA Stardust mission and recent advances in microscale analytical techniques. Isotopic studies of 'cometary' chondritic porous interplanetary dust particles (CP-IDPs) and comet 81P/Wild 2 Stardust samples show that preserved interstellar materials are more abundant in comets than in any class of meteorite. Identified interstellar materials include sub-micron-sized presolar silicates, oxides, and SiC dust grains and some fraction of the organic material that binds the samples together. Presolar grain abundances reach 1 weight percentage in the most stardust-rich CP-IDPs, 50 times greater than in meteorites. Yet, order of magnitude variations in presolar grain abundances among CP-IDPs suggest cometary solids experienced significant variations in the degree of processing in the solar nebula. Comets contain a surprisingly high abundance of nebular solids formed or altered at high temperatures. Comet 81P/Wild 2 samples include 10-40 micron-sized, refractory Ca- Al-rich inclusion (CAI)-, chondrule-, and ameboid olivine aggregate (AOA)-like materials. The O isotopic compositions of these refractory materials are remarkably similar to their meteoritic counterparts, ranging from 5 percent enrichments in (sup 16) O to near-terrestrial values. Comet 81P/Wild 2 and CP-IDPs also contain abundant Mg-Fe crystalline and amorphous silicates whose O isotopic compositions are also consistent with Solar System origins. Unlike meteorites, that are dominated by locally-produced materials, comets appear to be composed of

  15. Somatic embryogenesis, tetraploidy, and variant leaf morphology in transgenic diploid strawberry (Fragaria vesca subspecies vesca 'Hawaii 4').

    Science.gov (United States)

    Zhang, Qian; Folta, Kevin M; Davis, Thomas M

    2014-01-13

    The diploid (2n = 2x = 14) strawberry model plant Fragaria vesca ssp. vesca 'Hawaii 4' was employed for functional analysis of expressed DNA sequences initially identified as being unique to Fragaria and of unknown or poorly understood function. 'Hawaii 4' is prominent in strawberry research due to its ease of Agrobacterium-mediated transformation and regenerability, and its status as the source of the first complete strawberry genomic sequence. Our studies of a set of transformants have documented intriguing, construct-associated effects on leaf morphology, and provide important and unexpected insights into the performance of the 'Hawaii 4' transformation and regeneration system. Following Agrobacterium-mediated transformation of leaf explants with gene constructs carried by Gateway® vectors, plants were regenerated using a modified version of an established 'Hawaii 4' protocol. Expanding upon the findings of prior studies, we documented that plantlet regeneration was occurring via a somatic embryogenic rather than an organogenic developmental pathway. Among transformants, several variations in leaf morphology were observed. Unexpectedly, a particular leaf variant type, occurring in ~17% of all regenerants independent of construct type, was found to be attributable to tetraploidy. The tetraploidy-associated alteration in leaf morphology could be differentiated from the leaf morphology of diploid regenerants on the basis of a quantitative ratio of leaf dimensions: B/A, where B is the width of the central leaflet and A is the overall width of the trifoliate leaf. Variant effects on leaf morphology of four different transgenic constructs were also documented, and were in all cases distinguishable from the effects of tetraploidy. These results define opportunities to optimize the existing 'Hawaii 4' protocol by focusing on treatments that specifically promote somatic embryogenesis. The reported morphological metric and descriptions will guide future transgenic

  16. 2,4,5-Trichlorophenoxyacetic acid promotes somatic embryogenesis in the rose cultivar "Livin' Easy" (Rosa sp.).

    Science.gov (United States)

    Estabrooks, Tammy; Browne, Robin; Dong, Zhongmin

    2007-02-01

    Somatic embryogenesis (SE) offers vast potential for the clonal propagation of high-value roses. However, some recalcitrant cultivars unresponsive to commonly employed SE-inducing agents and low induction rates currently hinder the commercialization of SE technology in rose. Rose SE technology requires improvement before it can be implemented as a production system on a commercial scale. In the present work, we assessed 2,4,5-trichlorophenoxyacetic acid (2,4,5-T), a synthetic auxin not previously tested in rose, for its effectiveness to induce SE in the rose cultivar "Livin' Easy" (Rosa sp.). We ran a parallel comparison to the commonly used 2,4-dichlorophenoxyacetic acid (2,4-D). We tested each auxin with two different basal media: Murashige and Skoog (MS) basal medium and woody plant medium (WPM). MS medium resulted in somatic embryo production, whereas WPM did not. 2,4,5-T induced SE over a greater concentration range than 2,4-D's and resulted in significantly greater embryo yields. 2,4,5-T at a concentration of 10 or 25 microM was better for embrygenic tissue initiation than 2,4,5-T at 5 microM. Further embryo development occurred when the tissue was transferred to plant growth regulator (PGR) free medium or media with 40% the original auxin concentration. However, the PGR-free medium resulted in a high percentage of abnormal embryos (32.31%) compared to the media containing auxins. Upon transfer to germination medium, somatic embryos successfully converted into plantlets at rates ranging from 33.3 to 95.2%, depending on treatment. Survival rates 3 months ex vitro averaged 14.0 and 55.6% for 2,4-D- and 2,4,5-T-derived plantlets, respectively. Recurrent SE was observed in 60.2% of the plantlets growing on germination medium. This study is the first report of SE in the commercially valuable rose cultivar 'Livin' Easy' (Rosa sp.) and a suitable methodology was developed for SE of this rose cultivar.

  17. Involvement of peroxidase activity in developing somatic embryos of Medicago arborea L. Identification of an isozyme peroxidase as biochemical marker of somatic embryogenesis.

    Science.gov (United States)

    Gallego, Piedad; Martin, Luisa; Blazquez, Antonio; Guerra, Hilario; Villalobos, Nieves

    2014-01-15

    The legume Medicago arborea L. is very interesting as regards the regeneration of marginal arid soils. The problem is that it does not have a good germinative yield. It was therefore decided to regenerate via somatic embryogenesis and find a marker of embryogenic potential. In this study, peroxidase activity was evaluated in non-embryogenic and embryogenic calli from M. arborea L. A decrease in soluble peroxidase activity is observed in its embryonic calli at the time at which the somatic embryos begin to appear. This activity is always lower in embryonic calli than in non-embryonic ones (unlike what happens in the case of wall-bound peroxidases). These results suggest that peroxidases can be considered to be enzymes involved in somatic embryogenesis in M. arborea. In addition, isozyme analyses were carried out on protein extracts using polyacrylamide gel electrophoresis. The band called P5 was detected only in embryogenic cultures at very early stages of development. This band was digested with trypsin and analyzed using linear ion trap (LTQ) mass spectrometer. In P5 isoform a peroxidase-L-ascorbate peroxidase was identified. It can be used as a marker that allows the identification of embryological potential. Copyright © 2013 Elsevier GmbH. All rights reserved.

  18. Molecular characterization and evolution studies of a SERK like gene transcriptionally induced during somatic embryogenesis in Phoenix Dactylifera L v Deglet Nour

    Directory of Open Access Journals (Sweden)

    Rekik Imen

    2015-01-01

    Full Text Available A somatic embryogenesis receptor kinase like (SERKL cDNA, designated PhSERKL, was isolated from date palm (Phoenix Dactylifera L using RACE PCR. PhSERKL protein shared all the characteristic domains of the SERK family, including five leucine-rich repeats, one proline-rich region motif, a transmembrane domain, and kinase domains. Phylogenetic analyses using PHYLIP and Notung 2.7 programs suggest that the SERK proteins of some plant species resulted from relatively ancient duplication events. We predict an ancestor protein of monocots and dicots SERK using FASTML program. Somatic embryogenic cultures of date palm were established following transfer of callus cultures to medium containing 2, 4-dichlorophenoxyacetic acid. The role of PhSERKL gene during establishment of somatic embryogenesis in culture was investigated using quantitative real-time PCR. PhSERKL gene was highly expressed during embryogenic competence acquisition and globular embryo formation in culture. Overall, levels of expression of PhSERKL gene were lower in nonembryogenic tissues and organs than in embryogenic callus.

  19. GSM 900 MHz cellular phone radiation can either stimulate or depress early embryogenesis in Japanese quails depending on the duration of exposure.

    Science.gov (United States)

    Tsybulin, Olexandr; Sidorik, Evgeniy; Brieieva, Olga; Buchynska, Lyubov; Kyrylenko, Sergiy; Henshel, Diane; Yakymenko, Igor

    2013-09-01

    Our study was designed to assess the effects of low intensity radiation of a GSM (Global System for Mobile communication) 900 MHz cellular phone on early embryogenesis in dependence on the duration of exposure. Embryos of Japanese Quails were exposed in ovo to GSM 900 MHz cellular phone radiation during initial 38 h of brooding or alternatively during 158 h (120 h before brooding plus initial 38 h of brooding) discontinuously with 48 sec ON (average power density 0.25 μW/cm(2), specific absorption rate 3 μW/kg) followed by 12 sec OFF intervals. A number of differentiated somites were assessed microscopically. Possible DNA damage evoked by irradiation was assessed by an alkaline comet assay. Exposure to radiation from a GSM 900 MHz cellular phone led to a significantly altered number of differentiated somites. In embryos irradiated during 38 h the number of differentiated somites increased (p GSM 900 MHz cellular phone radiation on early embryogenesis can be either stimulating or deleterious depending on the duration of exposure.

  20. Molecular characterization of cathepsin L cDNA and its expression during oogenesis and embryogenesis in the oriental river prawn Macrobrachium nipponense (Palaemonidae).

    Science.gov (United States)

    Zhao, W; Chen, L; Zhang, F; Wu, P; Li, E; Qin, J

    2013-10-30

    We identified the cDNA sequence of cathepsin L (CatL) in Macrobrachium nipponense, designated as MnCatL, for the first time. The MnCatL cDNA, isolated from the ovary, was 1710 bp in length, containing a 31-bp 5'-untranslated region, a 650-bp 3'-untranslated region, and an open reading frame of 1029 bp, encoding a 342-amino acid polypeptide with a predicted molecular mass of 37.7 kDa. The polypeptide is composed of an 18-amino acid signal peptide, a 106-amino acid propeptide and a 218-amino acid mature peptide. MnCatL mRNA was detected in all tissues that we examined, including the thoracic ganglia, heart, muscle, intestine, hemocytes, ovary, testis, gills, and hepatopancreas. MnCatL expression reached a maximum value in both hepatopancreas and ovaries at the later stages of vitellogenesis, suggesting that MnCatL is involved in ovarian maturation of the oriental river prawn. During embryogenesis, MnCatL expression decreased as the embryo developed. The expression of MnCatL in the ovary and embryo suggest that MnCatL plays an important role in the uptake of vitellogenin and yolk protein, which are deposited in the oocyte for ovary maturation and embryo development, during oogenesis and embryogenesis of M. nipponense.