title: Isolation and Identification of Pathogenicity Mutant of Curvularia lunata via Restriction Enzyme-Mediated Integration.
abstract: In this report, 156 hygromycin-resistant mutants were generated via restriction enzyme-mediated insertional (REMI) mutagenesis. All mutants were subjected to a bioassay on detached leaves. Five mutants (T4, T39, T71, T91, and T135) showed reduced symptom development, whereas one mutant (T120) did not exhibit any symptoms on the leaves compared with the wild type. The pathogenicity of these mutants was further assayed through the spray inoculation of whole seedlings. The results demonstrated that the pathogenicity of the T4, T39, T71, T91, and T135 mutants was reduced, whereas the T120 mutant lost its pathogenicity. Southern blot analysis revealed that the plasmids were inserted at different sites in the genome with different copy numbers. Flanking sequences approximately 550, 860, and 150 bp were obtained from T7, T91, and T120, respectively through plasmids rescue. Sequence analysis of the flanking sequences from T7 and T91 showed no homology to any known sequences in GenBank. The flanking sequence from the T120 mutant was highly homologous to MAPKK kinases, which regulates sexual/asexual development, melanization, pathogenicity from Cochliobolus heterostrophus. These results indicate that REMI and plasmids rescue have great potential for finding pathogenicity genes.
authors.affiliation: College of Life Science and Technology, Heilongjiang Bayi Agricultural University, Daqing, 163319 Heilongjiang People's Republic of China.
authors.forename: Y J
authors.initials: YJ
authors.lastname: Wang
authors.affiliation: Institute of Plant Pathology and Applied Microbiology, Heilongjiang Bayi Agricultural University, Daqing, 163319 Heilongjiang People's Republic of China ; State Key Laboratory of Crop Stress Biology in Arid Regions, Northwest A&F University, Yangling, 712100 Shaanxi People's Republic of China ; Key Laboratory of Plant Protection Resources and Pest Management (Northwest A&F University), Ministry of Education, Northwest A&F University, Yangling, 712100 Shaanxi People's Republic of China.
authors.forename: T
authors.initials: T
authors.lastname: Liu
authors.affiliation: Institute of Plant Pathology and Applied Microbiology, Heilongjiang Bayi Agricultural University, Daqing, 163319 Heilongjiang People's Republic of China.
authors.forename: J M
authors.initials: JM
authors.lastname: Hou
authors.affiliation: Institute of Plant Pathology and Applied Microbiology, Heilongjiang Bayi Agricultural University, Daqing, 163319 Heilongjiang People's Republic of China ; State Key Laboratory of Crop Stress Biology in Arid Regions, Northwest A&F University, Yangling, 712100 Shaanxi People's Republic of China ; Key Laboratory of Plant Protection Resources and Pest Management (Northwest A&F University), Ministry of Education, Northwest A&F University, Yangling, 712100 Shaanxi People's Republic of China.
authors.forename: Y H
authors.initials: YH
authors.lastname: Zuo
keywords: Curvularia lunata; Pathogenicity; Plasmids rescue; REMI