Abiotic stress limits crop productivity, and plays a major role in determining the distribution of plant species across different types of environments. Chickpea (Cicer arietinum L.) is an important legume crop in Iran. Genetic diversity information is crucial for the choice of proper parents to establish new breeding programs. A number of molecular techniques have been developed to unveil the genetic potentials of plant materials. One of the most important methods for studying genetic diversity is using of Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) analysis of seed protein.SDS-PAGE seems to be a reliable molecular marker to investigate the genetic diversity of chickpea genotypes in IRAN. SDS-PAGE is practically a reliable method because seed storage proteins are largely independent of environmental fluctuation. The main objectives of present research was: (1) to investigate the power of SDS-PAGE markers for estimation of genetic diversity among some chickpea genotypes in west of Iran, (2) to investigate the genetic relationships between chickpea genotypes and (3) to determine and studying main drought indexes in chickpea genotypes could be selected main cultivars in drought and non-drought condition. This information will be useful to improve techniques for sampling chickpea genetic variation which might increase efficiency of conservation of germplasm.
Field experiment was carried out with four chickpea cultivars, based on randomized complete blocks design (RCBD) with three replications at the research farm of the Payame Noor University center of Asadabad in two different conditions (Drought conditions and supplementary irrigation) during 2010 cropping season. The cultivars were taken from Dryland Agricultural Research Institute, Sararood. In this study, seed protein profile of four chickpea cultivars include Arman, Hashem, Azad and ILC-482, were analyzed by Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE using 12/5% polyacrylamide). Protein extraction and gel electrophoresis was done By Laemmle (1970) Method. The seeds were powdered separately by liquid nitrogen. Cultivars seed protein were extracted by extraction buffer. At end of electrophoresis, protein bands were revealed by Coomassie Brilliant Blue R-250 staining. Biplot, 3D Plot, coefficient jaccard, cluster analysis and other analyses were done using the statistical software MSTAT-C, NTSYS and SPSS. The data for each cultivar was collected on a plot basis.
SDS-PAGE analysis showed significant differences between four cultivars chickpea at 17 and 25 kilo Dalton (kDa) bands. There are strong and weak proteins bands differences between cultivars. On the basis of SDS-PAGE results, 28.75 protein bands were observed with the Molecular Weight (MW) of 14 to 100 kDa. The presence of protein bands about 17 (kDa) in protein pattern of ILC-482, Hashem and Azad Cultivars and the absence of protein bands with approximate molecular weight 14 (kDa) in protein pattern of Arman cultivar can be used as protein markers for these chickpea cultivars. The specific proteins bands of seed may be used as markers for identification of the genotypes. Correlation between agronomic and molecular traits was assessed using Mantel test and significant positive correlation was observed between them. The results of drought stress indices and Biplot analysis suggested Arman genotype was the most tolerant. Biplot analysis also showed HAM and YI selection indices were the best to identify drought tolerant genotypes.
In most cases, an intermediate level of polymorphism have been reported with SDS-PAGE markers, and this study showed a considerable amount of polymorphism. We recommend further studies to be conducted by using number of chickpea genotypes as well as bigger and DNA molecular markers. Results of this study can be used in germplasm management practices, developing plant geneticist and breeders for planning future explorations, and crop improvement purposes.
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