Detection of DNA polymorphisms induced by ion beam irradiation on three orchid species

Affrida, A. H.*; Ahmad, Z.*; Idris, N. A.*; Mohd Randzan, N.*; Idris, N.*; Basiran, M. N.*; Tanaka, Atsushi; Shikazono, Naoya; Ono, Yutaka; Hase, Yoshihiro

Mutation induction by ion beam provides an alternative for improvement of orchids. The protocorm-like bodies (PLBs) of three orchid species; , and were irradiated with C ions, accelerated by AVF Cyclotron at Japanese Atomic Energy Agency (JAEA), Japan, at various doses ranging from 0 to 12.0 Gray (Gy). Since no morphological change was detected, Amplified Fragment Length Polymorphism (AFLP) technique was used to analyse the irradiated orchids. This PCR based DNA fingerprinting technique involves digestion of DNA with restriction enzymes, ligation of adapter and selective amplification using primer with one (pre-amplification) and three (selective amplification) arbitrary nucleotides. The fingerprinting of the control and mutant lines were compared in order to detect their genetic difference. Polymorphic fragments ranging from 250 to 1200 bp length were detected.