Marker assisted characterization of chickpea genotypes for wilt resistance

Abstract

Marker assisted characterization of six chickpea genotypes differing for Fusarium wilt reaction was carried out using seven molecular markers reported by earlier workers linked to disease resistant/susceptibility. In the present study, four different markers (namely, CS-27, UBC-170, CS-27A and UBC-825) linked to susceptibility and three microsatellite based markers (TA-59, TA-96 and TR-19) linked to resistance allele were validated. It was observed that two Random Amplified Polymorphic DNA (RAPD) markers, CS-27 and UBC-170 and one sequenced characterized amplified region (SCAR) CS-27A₇₀₀ gave amplification of 700, 550 and 700 bp, respectively in susceptible genotype only as reported by earlier worker. The inter simple sequence repeat (ISSR) marker UBC-825 produced amplification of 1200 bp in susceptible genotypes (JG-62 and GG 4) and intermediate genotype (Chaffa). Three sequence tagged microsatellites site (STMS) primers (TA-59, TA-96 and TR-19) gave specific allele in wilt resistant genotypes. The PCR amplification of TA-59 primer generated two alleles, out of which the allele of 258 bp was present only in resistance genotypes. The alleles of 265 bp amplified by primer TA-96 was present only in resistance genotypes and absent in other genotypes. The marker TR-19 amplified allele of 227 bp in resistant genotypes. Further, the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) analysis of seed storage protein showed a difference in protein profile among studied genotypes but none of polypeptide fragment was specific to wilt resistance or susceptibility. In present study, the reported markers linked to susceptibility and resistance proved their effectiveness and further can be exploited for maker assisted selection (MAS) of wilt resistance breeding in chickpea.

Keywords: Chickpea, Fusarium wilt, molecular markers, resistance, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE).

Afr. J. Biotechnol. Vol. 12 No. 50