Date Received: 28-09-2016 / Date Accepted: 13-02-2017
Wild bitter melon (Momordica charantia L. var. abbreviata Ser.) is known as a valuable vegetable and herb. It contains several important bioactive substances, including glycoside, saponin, alkaloide, triterpen and steroide. This study was carried out to establish a simple and efficient transformation protocol for inducting hairy roots of wild bitter melon by using strain TR7 of Agrobacterium rhizogenes which carries pPE113 binary vector harbouring e-gfp gene. Major parameters were optimized including, the age of plant and position for injection. The results indicated that highest hairy root induction frequencies was about 69.66-73.33% when seven-day-old seedlings were used to infect with the bacterium using hypocotyl. After regeneration and selection, thirty putative root clones of 4 specific phenotypes, i.e. white hairs, fast growth and branching on hormone-free MS medium, were obtained. The average GFP expression rate in 4 specific phenotypic groups were about 46.7%. PCR analysis using specific primers for rolA gene and virD gene confirmed the presence of the rolA gene and absence of virD gene in the genome of four hairy root lines. It was suggested that this simple transformation protocol can be applied for hairy root induction of other varieties as well.
