Induction of Ced9 mediated anti-apoptosis in commercial banana cultivar Rasthali for stable resistance against Fusarium wilt.

Anti-apoptotic gene Ced-9 enhanced resistance against Fusarium oxysporum f. sp. cubense (Foc) in the susceptible banana cultivar Rasthali by arresting tissue necrosis. The embryogenic cell suspension of banana cultivar Rasthali was stably transformed with Ced-9 gene and transformed lines were regenerated independently. The putative transgenic lines were analyzed with PCR using gene primers and further subjected to Southern blot to estimate copy number. The root-challenge bioassay with Foc showed 17-51% Vascular Discoloration Index in independent transformants compared to untransformed banana cv Rasthali (98% VDI). Four transgenic events showed a higher level of resistance over a period of 6 months. Overcoming tissue necrosis is the most ideal method to avoid Fusarium multiplication and spread in banana. Oxidative stress-induced cell necrosis is prevented by the activation of antiapoptotic pathways by Ced-9 and is proving to be an effective method to control this dreaded disease. This is the first report from India on the generation of transgenic banana cultivar Rasthali expressing antiapoptotic Ced-9 gene for resistance to Fusarium wilt.

Deployment of Stacked Antimicrobial Genes in Banana for Stable Tolerance Against Fusarium oxysporum f.sp. cubense Through Genetic Transformation.

Enhanced tolerance to wilt disease (Fusarium oxysporum f.sp. cubense) was achieved in banana variety Rasthali (AAB) by the transformation of embryogenic cells with two antimicrobial genes viz., Ace-AMP1 and pflp using Agrobacterium mediated transformation. The transgene copy numbers in stable transformants were confirmed by Southern analysis. The expression of stacked genes in the transgenic lines was validated by RT-PCR as well as Northern analysis. Bioassay using Foc race 1 in pot culture experiments demonstrated enhanced tolerance after 180 days of planting. Two independent transformants showed 10-20% Vascular Discoloration Index compared to untransformed banana cv. Rasthali (96%). The stacked lines revealed higher activity of Super Oxide Dismutase and Peroxidase compared to untransformed control which depicted higher tolerance to oxidative stress caused by Foc infection.