Transformation of Cotton with siRNA based Construct Targeting CLCuD

Cotton is an important crop. During past few years, cotton leaf curl disease (CLCuD) has emerged as an endemic. It has provided huge hindrance in production of high cotton yields. This disease is caused by cluster of begomovirus namely Cotton Leaf Curl Disease (CLCuD). Several breeding and transgenic approaches have been tried to overcome the problem. Among these RNA interference (RNAi) proved to be more promising. Use of RNA interference is favorable as the technique is environment friendly. In current research work RNAi based gene construct was transformed in two elite cotton verities MNH-786 and VH-289 to enhance viral resistance against CLCuD. The transformation of construct in plants was done by Agrobacterium mediated shoot apex method. The successful transformation and integration of the amplicon based RNAi construct was confirmed through PCR in T1 and T2 generation by using gene specific primers. Knock down ability of C2 construct was confirmed through real time CLCuD titer PCR. Comparison of virus disease index and Virus titer assured that the plants showing positive RNAi gene integration with minimum symptoms also had low viral titer.