USE OF INTER SIMPLE SEQUENCE REPEAT-POLYMERASE CHAIN REACTION (ISSR-PCR) TECHNIQUE FOR DETERMINATION OF THE EFFECT OF FAST NEUTRON IRRADIATION ON VICIA FABA SEEDS

Abstract

The effect of low doses of fast neutrons on the DNA genome of broad bean (Vicia
faba) mature seeds was determined. In this study, inter simple sequence repeat
(ISSR)-polymerase chain reaction (PCR) tool was used to analyze the genetic
alteration of treated seeds of V. faba using six ISSR primers. Results showed that
each primer gave rich and clear patterns with bands ranged from 179 to 1256 bp.
Among the 61 ISSR fragments, 21 were representing the effect of the treatment of V.
faba seed samples. ISSR markers clearly and easily distinguish all the tested samples
and also discriminate between fast neutron-treated and untreated seeds. The
dissimilarity values ranged 42 to 47% between the fast neutron-treated seeds and the
control seed (untreated) and from 27 to 38% between the fast neutron-treated seeds.
The dendrogram resulting from a UPGMA cluster analysis comprised two main
distinct clusters, the first one including the control V. faba seed (untreated) and the
second, includes all samples from the same kind of treatment divided into three
groups: the S4 (group A), S2 and S3 (group B) and S1 (group C), these groups being
separated at a similarity level.