Construction Of Wheat Mlo Gene RNAi Vector And Molecular Screening Of Wheat Mlo Anti-sense Gene Transformation

Wheat mildew powdery caused by Erysiphe graminis f.sp. Tritici is a serious disease worldwide and have great impact on the wheat yield. The application of agricultural fungicides could reduce the disease to some extent with the cost of the damage of the environment and the pollution of the product. Due to the existing numerous physiological races of Erysiphe graminis f.sp.Tritici and their rapid variations, the most economical and effective method to control the disease is to select wheat with broad spectrum and durable resistance. It is proved that the mlo recessive allele, a mutant type of barley Mlo, confers a broad spectrum and durable resistance to almost known isolates of the Erysiphe graminis. Wheat,an intimate relative of barley,shares similar mechanism in the fuction of mlo genes. The wheat Mlo gene cDNA fragment(1434bp) has already been cloned in our lab, so we can use it to inhibit or down regulate the expression of wheat Mlo gene through RNAi (RNA interference) or anti-sense technique and increase the resistance of wheat to powdery mildew.RNAi is a efficient approach to inhibit gene expression with specificity, efficiency, stabilization and it will not change the genetic composition of the genome. With these advantage, RNAi becomes a new reverse genetics method to study the genes with unknown functions.At the mean time, it is becoming more and more important in the application of research. We chose the sequences with high inhibition efficiency of the 5' end and the 3' end of the wheat Mlo gene cDNA coding region respectively (encode N-end and C-end of wheat MLO protein respectively), constructed RNAi expression vector pRI-Nmlo and pRI-Cmlo and transformed wheat cultivar Yanyou 361 with these vectors.In our research the Mlo cDNA anti-sense transgenic plants were detected and...