中国水稻科学
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  2006, Vol. 20 Issue (3): 231-237     DOI:
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Agrobacterium tumefaciens-Mediated Transformation of Magnaporthe grisea and Identification of Pathogenicity Defective Mutant
LIU Peng-juan , WANG Zheng-yi , WANG Qiu-hua , LI De-bao
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Abstract Based on the construction of two binary vectors containing hygromycin B, the use of Agrobacterium tumefaciens-mediated transformation as a successful method for insertional mutagenesis in rice blast fungus Magnaporthe grisea was reported. A library including more than 4000 transformants with a high transformation efficiency of over 300 hygromycin B resistant transformants per 1×106 conidia of M. grisea was generated. All of the hygromycin B resistant transformants tested were mitotically stable after several subcultures onto complete medium without hygromycin. Genomic Southern blot analysis showed that about two thirds of the transformants were single T-DNA insertional events. Through testing pathogenicity of mutational transformants by a rapid barley leaf assay method, an interesting mutant, A1-412, which was completely nonpathogenic to both barley and rice and also lost the ability to undergo infectious growth through abraded leaves was identified. Phenotypic analysis showed that the mutant A1-412 was significantly reduced in conidiation only accounting for 7% conidia of the wild type strain, and was unable to form appressorium on hydrophobic surfaces and its germination was slightly delayed. Southern blot analysis showed that T-DNA inserted into the A1-412 genome was single copy. These data suggest that an important biological process blocked in A1-412 was likely to be due to the insertion of T-DNA and the subsequent disruption of gene function.
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LIU Peng-juan
WANG Zheng-yi
WANG Qiu-hua
LI De-bao
Key wordsMagnaporthe grisea   Agrobacterium tumefaciens-mediated transformation   insertional mutagenesis   pathogenicity defective mutant     
Received: 1900-01-01;
Cite this article:   
LIU Peng-juan ,WANG Zheng-yi ,WANG Qiu-hua et al. Agrobacterium tumefaciens-Mediated Transformation of Magnaporthe grisea and Identification of Pathogenicity Defective Mutant[J]. , 2006, 20(3): 231-237 .
 
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