Molecular characterization of the flanking gene of T-DNA insertional, pathogenicity-defective Colletotrichum gloeosporioides mutant strain 1869

doi:10.11686/cyxb2015094

Abstract

Abstract: Colletotrichum gloeosporioides is a fungal pathogen of Stylosanthes guianensis causing anthracnose disease. Pathogenicity defective mutant strains of C. gloeosporioides were screened in the laboratory and strain 1869 selected for study. The mutant strain was compared with wild type strain CH008 for phenotypic characteristics, including colony diameter, colony morphology, sporulation ability and spore germination rate. Colony diameter of CH008 and strain 1869 differed, but the other traits did not. DNA of the mutant strain was extracted, and using thermal asymmetric interlaced PCR (TAIL-PCR) a T-DNA insertion mutation site believed responsible for loss of pathogenicity was identified. The RB and LB flanking sequences at the insertion site were cloned, also using TAIL-PCR. The RB flanking sequence was 467 bp in length, while the LB flanking sequence was 388 bp. The BLAST result showed that the sequence had 92%-96% homology to gene Pac1. By predicting the function of a protein coded by the flanking sequence,we inferred that the gene in question regulated the sensitivity to pH, and in this way affected the pathogenic potential of C. gloeosporioides.

XU Pei-Dong, ZHENG Xiao-Lan, ZHAO Yan, LI Qiu-Jie, WU Wei-Huai, HE Chun-Ping, XI Jin-Gen, LIANG Yan-Qiong, ZHENG Jin-Long, QI Shan-Jiang, ZHANG Xiao-Bo, YI Ke-Xian. Molecular characterization of the flanking gene of T-DNA insertional, pathogenicity-defective Colletotrichum gloeosporioides mutant strain 1869[J]. Acta Prataculturae Sinica, 2015, 24(8): 142-149.

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