Genetic diversity among varieties and wild species accessions of pea (Pisum sativum L.) based on SSR markers

To assess the genetic relations inPisum genus and to examine putative duplicate accessions, 20 pea varieties (Pisum sativum L.) with 57 accessions from wild Pisum species fulvum, subspecies (subsp.) asiaticum, elatius, thebaicum, abyssinicum, transcaucasicum and arvense were analyzed using 10 out of 20 microsatellite primer pairs. We genotyped all accessions. In total, 59 alleles were identified in whole collection. The maximum number of alleles (8 alleles) was obtained from the PEACPLHPP, AF004843, and AA43090 loci. The maximum number of private alleles (4) in the wild collection was detected in AF004843 locus but in the cultivar collection, it was detected in AA430902 and PSBLOX13.2 loci. Cluster analysis and principal coordinate analysis located accessions in 3 groups and cultivated varieties were obviously separated from the wild accessions. Analysis of molecular variance (AMOVA) revealed that the intergroups component of variance (29%) is lower than the intragroups component of variance (71%). The lowest value of genetic differentiation ( Pisum genus and to examine putative duplicate accessions, 20 pea varieties (Pisum sativum L.) with 57 accessions from wild Pisum species fulvum, subspecies (subsp.) asiaticum, elatius, thebaicum, abyssinicum, transcaucasicum and arvense were analyzed using 10 out of 20 microsatellite primer pairs. We genotyped all accessions. In total, 59 alleles were identified in whole collection. The maximum number of alleles (8 alleles) was obtained from the PEACPLHPP, AF004843, and AA43090 loci. The maximum number of private alleles (4) in the wild collection was detected in AF004843 locus but in the cultivar collection, it was detected in AA430902 and PSBLOX13.2 loci. Cluster analysis and principal coordinate analysis located accessions in 3 groups and cultivated varieties were obviously separated from the wild accessions. Analysis of molecular variance (AMOVA) revealed that the intergroups component of variance (29%) is lower than the intragroups component of variance (71%). The lowest value of genetic differentiation (Pisum genus and to examine putative duplicate accessions, 20 pea varieties (Pisum sativum L.) with 57 accessions from wild Pisum species fulvum, subspecies (subsp.) asiaticum, elatius, thebaicum, abyssinicum, transcaucasicum and arvense were analyzed using 10 out of 20 microsatellite primer pairs. We genotyped all accessions. In total, 59 alleles were identified in whole collection. The maximum number of alleles (8 alleles) was obtained from the PEACPLHPP, AF004843, and AA43090 loci. The maximum number of private alleles (4) in the wild collection was detected in AF004843 locus but in the cultivar collection, it was detected in AA430902 and PSBLOX13.2 loci. Cluster analysis and principal coordinate analysis located accessions in 3 groups and cultivated varieties were obviously separated from the wild accessions. Analysis of molecular variance (AMOVA) revealed that the intergroups component of variance (29%) is lower than the intragroups component of variance (71%). The lowest value of genetic differentiation (PT = 0.27) of pair wise collections between wild and variety collections, was detected in ssp. elatius. Assignment test on the basis of log-likelihood to estimate the likelihood that an individual belongs to a given group, showed that 96% of accessions being assigned correctly to their groups. This study showed that genetic probability profiles of accessions can corroborate clustering analyses while providing additional information as a powerful tool for assigning accessions into their related groups

Validation of a genus-specific gene; TPS, used as internal control in quantitative Real Time PCR of transgenic cotton

Abstract Identification of genes with invariant levels of gene expression is a prerequisite for validating transcriptomic changes accompanying development. Ideally expression of these genes should be independent of the morphogenetic process or environmental condition.We report here the validation of internal control gene i.e.TPS (trehalose 6-phosphate-synthase) in cotton (Gossypium spp), using TaqMan system in quantitative Real Time PCR (qRT-PCR). The Gene expression was tested in five different G. hirsutum cultivars including Coker 312, Acala SJ, ZETA 2, Taghva, Neishabour and a diploid wild type; G. barbadense. Identical amplicons were obtained within these cultivars. No amplifications was achieved when DNA samples from barley (Hordeum vulgare), maize (Zea mays), rice (Oryza sativa)

Mechanisms of carbon dioxide detection in the earthworm Dendrobaena veneta

IntroductionCarbon dioxide (CO2) is a critical biological signal that is noxious to many animals at high concentrations. The earthworm Dendrobaena veneta lives in subterranean burrows containing high levels of CO2 and respires through its skin. Despite the ecological and agricultural importance of earthworms, relatively little is known about how they make decisions in their environment, including their response to elevated levels of CO2.MethodsTo examine CO2 detection in this species, we designed the exudate assay, in which we placed an earthworm in a sealed container, exposed it to varying concentrations of CO2 for one minute, and recorded the amount of exudate secreted. Because earthworms excrete exudate in response to noxious stimuli, we hypothesized that the amount of exudate produced was proportional to the amount of irritation. We repeated these experiments after treatment with several blockers for molecules with potential involvement in CO2 detection, including carbonic anhydrases, guanylate cyclase, TRPA1, ASICs, and OTOP channels. We also confirmed the presence of homologous transcripts for each of these gene families in an epithelial transcriptome for D. veneta. Additionally, since organisms often detect CO2 levels indirectly by monitoring the conversion to carbonic acid (a weak acid), we used the exudate assay to evaluate aversion to additional weak acids (formic acid, acetic acid, and propionic acid).ResultsEarthworms excreted significantly more exudate in response to CO2 in a dosage-dependent manner, and this response was muted by the general carbonic anhydrase inhibitor acetazolamide, the carbonic anhydrase IX/XII inhibitor indisulam, the calcium channel blocker ruthenium red, the sodium channel blocker amiloride, and the acid-sensing ion channel blocker diminazene aceturate.DiscussionThese data provide evidence of the role of carbonic anhydrase and epithelial sodium channels in earthworm CO2 detection, establish that, similar to other subterranean-dwelling animals, earthworms are extremely tolerant of CO2, and contribute to our understanding of the mechanisms used by earthworms to detect and react to weak acids in their environment

Tagging of resistance gene(s) to rhizomania disease in sugar beet (Beta vulgaris L.)

The rhizomania disease is one of the most important diseases in Iran and some other parts of the world which potentially could play a role in decreasing sugar yield in fields. One approach to combat with thisdisease is the use of resistance varieties. This varieties have been identified which are having resistance genes to rhizomania disease (i.e. Rz1, Rz2). In order to use these genes in breeding programs(MAS) tagging these genes with molecular markers is necessary. In our study, we used infected soil which was provided from infected fields then greenhouse test was done to identify resistance and susceptible plants. Extracted DNA from leaves of resistant and susceptible plants was bulked to provide two bulks for resistance and susceptible plants. Three-hundred RAPD primers were used in analysis of the two bulks and two F2 populations. One population was obtained from a cross betweenHolly1-4 as resistance parent and an annual variety as susceptible plant. The second population was constructed by crossing between WB42 as resistance parent and L 261 as susceptible one. Finallygenes were tagged using two RAPD primers and one of the markers is OP-091150 which is 27 cM apart from Rz1 gene in coupling phase. The second marker is OP-AN9600 which is 13.7 cM apart from Rz1 geneand in repulsion phase

Phytobiosynthesis of gold nano-particles and comparison of two plant species (Canola and Alfalfa)

245-247Different varieties of two plant species, Canola and Alfalfa were grown at exactly same laboratory conditions and bio-reduction of Au (III) to Au (0) was studied. Subsequently, production of gold nano-particles of various morphologies and sizes were characterized. Plant seeds were grown in a culture medium that contained gold ions from KAuCl4. Gold nano-particle formation was analyzed by atomic absorption spectrometry and transmission electron microscopy. Results showed that the plants pull up gold ions from KAuCl4 and form gold particles in nano sizes due to chemical behaviour of the gold. Significant differences in the nature of nano-particles were observed when particles synthesized by these two plant species were compared. The size range of gold nano-particles synthesized by Canola was 20-128 nm, while it was 8-48 nm by Alfalfa

Inhibition of overactivated p38 MAPK can restore hematopoiesis in myelodysplastic syndrome progenitors

The myelodysplastic syndromes (MDSs) are collections of heterogeneous hematologic diseases characterized by refractory cytopenias as a result of ineffective hematopoiesis. Development of effective treatments has been impeded by limited insights into any unifying pathogenic pathways. We provide evidence that the p38 MAP kinase is constitutively activated or phosphorylated in MDS bone marrows. Such activation is uniformly observed in varied morphologic subtypes of low-risk MDS and correlates with enhanced apoptosis observed in MDS hematopoietic progenitors. Most importantly, pharmacologic inhibition of p38α by a novel small molecule inhibitor, SCIO-469, decreases apoptosis in MDS CD34+ progenitors and leads to dose-dependant increases in erythroid and myeloid colony formation. Down-regulation of the dominant p38α isoform by siRNA also leads to enhancement of hematopoiesis in MDS bone marrow progenitors in vitro. These data implicate p38 MAPK in the pathobiology of ineffective hematopoiesis in lowrisk MDS and provide a strong rationale for clinical investigation of SCIO-469 in MDS