Abstract:

We established the gene knockout method using agrobacteriummediated transformation, and carried out contrast experiment of mat 1-1 of Sclerotinia sclerotiorum, and then obtained the transformed strain of mat 1-1 gene that was knocked out. First, the right arm and left arm were cloned by PCR respectively, which were selected from two sides of mat 1-1 gene that gave a great help in the construction of knockout vector, and then constructed the targeting vector ΔPBI-G3CN-mat 1-1, which comprised the two side arms and correctly sequences and resistance gene of fradiomycin.
ΔPBI-G3CN-mat 1-1 was transformed into EHA105. Using hypha of Sclerotinia sclerotiorum, we confirmed the positive knockout strain via PCR. By means of detecting the physiological phenotype, no obvious difference was shown between mat 1-1 gene deleted strain and wild strain in growth speed. But sclerotium and apothecium were not formed in the mat 1-1 gene deleted strain.

Key words: Sclerotinia sclerotiorum, mat 1-1 gene, knockout vector, agrobacteriummediated transformation