title: Agrobacterium tumefaciens-mediated transformation as an efficient tool for insertional mutagenesis of Cercospora zeae-maydis.
abstract: An efficient Agrobacterium tumefaciens-mediated transformation (ATMT) approach was developed for the plant pathogenic fungus, Cercospora zeae-maydis, which is the causative agent of gray leaf spot in maize. The transformation was evaluated with five parameters to test the efficiencies of transformation. Results showed that spore germination time, co-cultivation temperature and time were the significant influencing factors in all parameters. Randomly selected transformants were confirmed and the transformants were found to be mitotically stable, with single-copy T-DNA integration in the genome. T-DNA flanking sequences were cloned by thermal asymmetric interlaced PCR. Thus, the ATMT approach is an efficient tool for insertional mutagenesis of C. zeae-maydis.
authors.affiliation: College of Plant Protection, Shenyang Agricultural University, Shenyang, Liaoning Province 110866, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Lu
authors.affiliation: College of Plant Protection, Shenyang Agricultural University, Shenyang, Liaoning Province 110866, China.
authors.forename: Shuqin
authors.initials: S
authors.lastname: Xiao
authors.affiliation: College of Plant Protection, Shenyang Agricultural University, Shenyang, Liaoning Province 110866, China.
authors.forename: Fen
authors.initials: F
authors.lastname: Wang
authors.affiliation: College of Plant Protection, Shenyang Agricultural University, Shenyang, Liaoning Province 110866, China.
authors.forename: Jiaying
authors.initials: J
authors.lastname: Sun
authors.affiliation: College of Plant Protection, Shenyang Agricultural University, Shenyang, Liaoning Province 110866, China.
authors.forename: Likun
authors.initials: L
authors.lastname: Zhao
authors.affiliation: College of Plant Protection, Shenyang Agricultural University, Shenyang, Liaoning Province 110866, China.
authors.forename: Libin
authors.initials: L
authors.lastname: Yan
authors.affiliation: College of Plant Protection, Shenyang Agricultural University, Shenyang, Liaoning Province 110866, China. Electronic address: chunshengxue@hotmail.com.
authors.forename: Chunsheng
authors.initials: C
authors.lastname: Xue
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics*; D001203:Ascomycota / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D018920:Coculture Techniques; D004269:DNA, Bacterial / Q000235:genetics; D006921:Hygromycin B / Q000378:metabolism; D016254:Mutagenesis, Insertional; D010935:Plant Diseases / Q000382:microbiology*; D012680:Sensitivity and Specificity; D014170:Transformation, Genetic; D003313:Zea mays / Q000382:microbiology
chemical_list: D004269:DNA, Bacterial; C036483:T-DNA; D006921:Hygromycin B
keywords: Agrobacterium tumefaciens-mediated transformation; Cercospora zeae-maydis; TAIL-PCR