ASSESSMENT OF GENETIC DIVERSITY IN COWPEA (Vigna unguicalata) USING SDS-PAGE, RANDOM AMPLIFIED POLYMORPHIC DNA (RAPD) AND INTER-SIMPLE SEQUENCE REPEAT (ISSR) MARKERS
Abstract
Genetic diversity of four cultivars of Egyptian cowpea (Vigna unguicalata) was studied by SDS-PAGE, RAPD and ISSR-PCR. SDS-PAGE recorded a low degree of polymorphism among the four cultivars of cowpea. Only four polymorphic bands were recorded with a percentage of 22.22%. On the other hand, RAPD-PCR generated polymorphism among the DNA samples of the studied cultivars. The ten primers generated a sum of 23 polymorphic bands in the cultivars under study. Nine unique bands were identified out of the polymorphic ones. The percentage of polymorphism ranged from 16.67% to 55.56% with an average of 26.37%. ISSR bands generated higher level of polymorphism than RAPD among the DNA samples of the studied cowpea cultivars. The ten ISSR primers generated a sum of 32 polymorphic bands in the cultivars under study. Nineteen unique bands were identified out of the polymorphic ones. The percentage of polymorphism ranged between 16.67% and 75% with an average of 28.27%.
A dendrogram obtained from UPGMA cluster analysis based on protein analysis classified the four cultivars into two groups. Only cultivar Kareem 7 was isolated in a single group, while the second group contained the other 3 cultivars. Dendrogram based on the ten RAPD primers revealed two main genetic clusters. The first cluster comprises the cultivar Kareem 7 and Dokki 331, while the second cluster includes Kaha 1 and Kafer El-Sheikh 1. On the other hand, a dendrogram based on the ten ISSR data revealed two main genetic clusters. Cultivar Kafer El-Sheikh 1 separated in a single group, while the second cluster includes the other three cultivars. Dendrogram obtained from combined data are inagreement with that of ISSR dendrogram in which cultivar Kafer El-Sheikh-1 was separated in a single group, while the SDS-PAGE and RAPD dendrograms showed some variations in the clustering of cowpea cultivars.
Considering all the gained data, it is evident that molecular detection of RAPD and ISSR are suitable tools than SDS-PAGE in assessing genetic variation among the four cultivars of cowpea. These markers provide interesting tools for breeding new varieties of Egyptian cowpea.
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