Plant & Animal Genomes XIV Conference Plant & Animal Genomes XIV ConferenceJanuary 14-18, 2006
Town & Country Convention Center
San Diego, CA
RNA silencing provides potentially versatile reverse genetic tools for exploring gene function in the post-genomics era. We have developed two types of silencing vectors, pSilent-1 and pSilent-dual for the rice blast fungus, Magnaporthe oryzae. pSilent-1 is a pHANNIBAL-like vector, which carries a transcriptional unit for hairpin RNA expression with a spacer of a cutinase gene intron from M. oryzae, and pSilent-Dual is a high throughput silencing vector, which carries two opposing Pol II promoters. The latter vector allows non-orientated one step cloning for generation of silencing constructs; thus offers high throughput system for gene function analysis. Application of pSilent-1 and pSilent-dual to M. oryzae endogenous genes such as MPG1, polyketide synthase-like gene and xylanase in addition to the model gene GFP, resulted in efficient silencing of the genes in a dicer-dependent manner, therefore through the RNA silencing pathway. The efficiency of silencing induced by pSilent-1-based vectors was generally higher than that by pSilent-dual-based vectors. Therefore, pSilent-1 should be useful for inducing high efficiency gene silencing on a small or moderate scale, and pSilent-dual provides a potential reverse genetic tool for functional genomics in M.oryzae as a feasible alternative to other systems.