Comparison of Post-transcriptional Gene Silencing (PTGS) Strategies for Developing Transgenic Plants Resistant to Tomato Leaf Curl Virus (ToLCV)

Tomato (Solanum lycopersicum L.), an economically important crop is host to many whitefly transmitted geminiviruses including tomato leaf curl virus (ToLCV). Genetically engineering resistance of pathogen through Post-transcriptional gene silencing (PTGS/RNAi) a powerful strategy that can provide alternative existing methods producing resistant plants. We cloned and characterized ToLCV-replicase (TRP) from local Dharwad, Karnataka, India ToLCV isolate for development transgenic Plant expression vectors carrying viral replicase (rep) in sense, antisense, ihp (intron spaced hairpin) HUTR (inverted repeats heterologous 3΄-untranslated region) were constructed. Transgenic plants rep different strategies when challenged with whiteflies showed varied degrees resistance. Such confirmed PCR, GUS, Dot blot, Southern blot semiquantitative PCR analysis. High degree was observed the construct both sense antisense strand interrupted by intron (ihp). Our results demonstrate that, simultaneous strands are more efficient than those expressing either or alone.