Screening and identification of mutants of Magnaporthe grisea by REMI

Autor:	Zheng Xiaobo, Liu Juan, Fan YongJian, Zhou YiJun, Xiong RuYi
Rok vydání:	2007
Předmět:	Genetics Mutant Biology biology.organism_classification Molecular biology Transformation (genetics) chemistry.chemical_compound Plasmid chemistry Remi Magnaporthe grisea Restriction fragment length polymorphism Agronomy and Crop Science Hygromycin B Gene
Zdroj:	Frontiers of Agriculture in China. 1:179-182
ISSN:	1673-744X 1673-7334
DOI:	10.1007/s11703-007-0030-y
Popis:	The plasmid pUCATPH was used to establish a transformation system in wild-type isolate M131 of Magnaporthe grisea. Six hundred and thirty-nine transfor- mants were obtained by restriction enzyme-mediated integra- tion (REMI) with hygromycin B (hyg B) resistance as a tag. Morphological analysis of two of the REMI mutants con- firmed that they produced little melanin under black light and continued for three generations. Pathogenicity identification of six mutants screened proved that they made pathogenicity changes on three sets of differential varieties with different resistance genes. Rep-PCR analyses showed that two mor- phological mutants and two pathogenicity mutants differed from wild-type isolate M131 at the molecular level. RFLP analyses were performed to study the four mutants at the molecular level and the integration sites of the plasmid DNA. The results showed that the plasmid was inserted into all four mutants and that the insertion sites were random.
Databáze:	OpenAIRE
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