Micropropagation and Agrobacterium: mediated transformation of GUS gene in to Gymnema sylvestre antidiabetic plants
Ram Krishna Sahu, Rajesh T.S, Pavithra K
Gymnema sylvestre is an herb plant of the tropical forest of Southern and Central India where it has been used as a naturopathic treatment for diabetes for nearly two millennia. Sanskrit Name: Meshasringi, Madhunasini or madhoolika, Hindi: Gurmar, Tamil and malayalam Name Sirukurinchaan, Amudhapushpam, Chakkarakkolli. Gymnema sylvestre is a stomachic, diuretic, refrigerant, astringent, and tonic. It has been found to increase urine output and reduce hyperglycemia in both animal and human studies. Gymnemic acids have anti-diabetic, anti-sweetener and anti-inflammatory activities. Gymnema is propagated by seed germination. One of the constraints in this conventional propagation is the very short span of seed viability. No alternative mode of multiplication is available to propagate and to conserve genetic stock of this useful plant. Tissue culture offers an effective alternative method for rapid multiplication of desirable clones. Gymnema was used for in vitro callus induction and Agrobacterium mediated transformation experiment. Which showed fast and high growth rate in the MS medium at 0.03mg/l concentration of 2, 4-D. The Agrobacterium tumefaciense strain LBA4404 has carrying Ti plasmid pK1W1105 was used for infection of callus. 1µl of 24hr culture of Agrobacterium was used for 30 sec. for the infection. Successful delivery of the GUS reporter gene was confirmed by 1mM X-gluc as histochemical substrate for staining of the callus. DNA was isolated from the infected callus by using 1.2%SDS (65°C) method and then presence of GUS gene was reconfirmed by using GUS gene specific marker in PCR amplification technique
Ram Krishna Sahu, Rajesh T.S, Pavithra K. Micropropagation and Agrobacterium: mediated transformation of GUS gene in to Gymnema sylvestre antidiabetic plants. Journal of Medicinal Plants Studies. 2016; 4(4): 18-24.