Microsatellite analysis of chickpea (Cicer arietinum L.) genotypes

Purpose. To investigate the polymorphism of chickpea genotypes at microsatellite loci within the QTL-hotspot region of linkage group 4, associated with drought tolerance. Methods. DNA extraction and purification from seedlings using the CTAB method; polymerase chain reaction; horizontal gel electrophoresis; determination of amplification product sizes using the “GelAnalyzer” software; cluster analysis using the “MEGA12” software. Results. Of the 26 samples analyzed from the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT) collection, one to eight alleles were identified at the following microsatellite (SSR) loci within the QTL-hotspot region of linkage group 4 in the chickpea genome: ICCM0249, NCPGR127, TAA170, NCPGR21, TA130 and STMS11. The distribution of SSR locus alleles in the samples under study was compared with that in chickpea samples from various breeding centers, including those in Ukraine. Genetic distances were calculated for the 26 ICRISAT samples and seven Ukrainian varieties. A dendrogram was constructed which grouped the samples into seven clusters; the Ukrainian chickpea varieties formed a separate cluster. Conclusions. The chickpea samples from the ICRISAT collection were found to be polymorphic at SSR loci ICCM0249, TAA170, and TAA130, with three, five, and eight alleles respectively, and monomorphic at three SSR loci: STMS11, NCPGR127, and NCPGR21. This distribution of polymorphic and monomorphic alleles corresponded to that observed in the Ukrainian chickpea varieties. Cluster analysis revealed that the Ukrainian varieties formed a distinct group, suggesting differences in genetic origin and breeding approaches compared to the ICRISAT collection.