Validate The Function Of The Mating Type Of Relative Mat1-1 Gene Of Sclerotini Sclerotiorum

Sclerotinia sclerotiorum is belong to Ascomycotina, Helotiales, Sclerotinia, which could parasitize in 75 families 450 species plants.The main host incloud that oil grain(such as rape, sunflower and soybean) and most of cruciferae. It could do a great harm to the agriculture when it was invaded into the host. A great deal of researches were carry out in order to find out a better way to prevent and cure soybean sclerotinia rot. There were a great of researches on the plant for resistant sclerotinia rot,on the reform of species and on developing of the pesticide inhibiter. But there was only few of researches on the infextation after sexual reproduction. Sclerotinia sclerotiorum could not produce conidium in asexual stage,it could produce apothecium by homothallism and then release the thecaspore,which could disperse with wind and increase the disseminiation of the plant disease. Therefore, it is much more important to study on the sexual problem of Sclerotinia sclerotiorum. It has been reported that there were a pair of allele mat1-1 and mat1-2, which could control mating of paroecious in sexual process of Fungi. It is sure that mating type can demonstration the cortroling contidion of inherit and the derecopmed of sexual in both filamentary fungus and non-filamentary fungus.That is to say, mating type gene play a decisive role of the Fungi′s sex in the devecopment of Fungi′s sexual stage. According to the importance of the mating gene in Fungi, the present study was carried out to focas on the mating gene in Sclerotinia sclerotiorum, which will promote farther researches on the work of genetics analysis of Sclerotinia sclerotiorum.In this study,we establish the heredity transformation system of Sclerotinia sclerotiorum which mediating by Agrobacterium successfully, and improve the experimental condition to establish the gene knockout method by using Agrobacterium-mediated transformation, and carried out the knocking out contrast experiment of mat1-1 of Sclerotinia sclerotiorum, and then obtain the transformed strain of mat1-1 gene that deletioned.The right arm and left arm were cloned by PCR, which were selected the 1000 bp from twosides of mat1-1 gene that can give a great help in the construction of knock out vector. And then construet the targeting vector△PBI-G3CN-MAT1-1,which was comprise the twoside arm that correctly sequence and resitance gene of Fradiomycin. The positive recombinants was transformed into EHA105 by using hypha of Sclerotinia sclerotiorum. We carried out the study tu confirm the positive knockout strain by using PCR and also detect the physiological phenotype. We discoved that the growth speed of mat1-1 gene deleted knocking out strain is far greater than Sclerotinia sclerotiorum, approximately 2 times, but other physiological phenotype has not shown obvious different.from that .In a word, this research is the first time to use the Agrobacterium-mediated transformation to realize the homologous recombination for target gene of Sclerotinia sclerotiorum in the word. And constructed target vector△PBI-G3CN-MAT1-1 by knock out. Meanwhile, we knocked out the mating type gene mat1-1 of Sclerotinia sclerotiorum. This research was confirmed physiological phenotype of the knockout strain. This research was mainly focus on mat1-1 gene of Sclerotinia sclerotiorum. It was promoted development of new pesticide and soybean sclerotinia rot.