Screening of pathogenicity defect mutant in Colletotrichum gloeosporioides of Stylosanthes guianensias and molecular cloning of T-DNA integration flanking sequence

Several inoculation methods were compared to test their effect of pathogenicity to Stylosanthes guianensias. In all 1 230 mutants, 23 had decreased weak pathogenicity, 5 even completely losted its ability to infect S. guianensias. TAIL-PCR was used to identify the T-DNA integration site in the mutant that was coded t-2430 and the sequence of 476 bp was gotten. The flanking sequence of T-DNA was compared with Colletotrichum genome by blast and 401nt of sequence was found in Contig464 of Colletotrichum genome. By predicting the function of flanking sequence, we found T-DNA insertion in the promoter region of putative gene, which had 79% homology to Magnaporthe oryzae aspartate aminotransferase gene (XP_003719674.1).