胶孢炭疽菌CgFADBD基因敲除突变体的构建及其表型分析  

Construction and Phenotypic Analysis of CgFADBD Gene Knockout Mutant of Colletotrichum gloeosporioides

作　　者：张琦玮 张贝[1,2] 赵丽[1] 罗红丽[1,2] Zhang Qiwei;Zhang Bei;Zhao Li;Luo Hongli(College of Tropical Crops,Hainan University,Haikou,570228;Sanya Nanfan Research Institute of Hainan University,Sanya,572025)

机构地区：[1]海南大学热带作物学院,海口570228 [2]海南大学三亚南繁研究院,三亚572025

出　　处：《分子植物育种》2022年第22期7461-7468,共8页Molecular Plant Breeding

基　　金：海南省自然科学基金青年基金项目(319QN167);国家自然科学基金项目(32060591)共同资助。

摘　　要：胶孢炭疽菌引发的橡胶树炭疽病是橡胶树减产的重要原因之一。为了解析橡胶树胶孢炭疽菌的致病机理,在前期研究中对胶孢炭疽菌进行了基因组和转录组测序,预测了一个编码黄素腺嘌呤二核苷酸(FAD)结合蛋白的基因,命名为CgFADBD。本研究中,通过RT-PCR技术对CgFADBD基因全长编码区进行了克隆和生物信息学分析。结果表明,该基因编码一条由505个氨基酸组成的多肽,预测分子质量为54.39 kD。该蛋白含有典型的信号肽序列(第1~26位氨基酸)和一个保守的FAD结合域(第69~203位氨基酸),此外,不含任何跨膜结构域。根据融合PCR和同源重组原理,构建了CgFADBD基因敲除突变体ΔCgFADBD。菌落生长和孢子产量统计结果表明野生型菌株WT与ΔCgFADBD没有明显差异。致病力分析结果表明,ΔCgFADBD对橡胶树叶片的致病力明显小于WT,综合以上结果表明,CgFADBD与胶孢炭疽菌对橡胶树的致病力有关,与菌丝生长和孢子产量关系不大。本研究结果对解析胶孢炭疽菌对橡胶树的致病机制提供一定的参考。Rubber tree anthracnose caused by Colletotrichum gloeosporioides is one of the major factor for losses of natural rubber production.In order to elucidate the pathogenesis of Colletotrichum gloeosporioides,the genome and transcriptome sequencing of C.gloeosporioides were performed previously and a gene encoding a flavin adenine dinucleotide(FAD)binding protein named as CgFADBD was predicted.In this study,the full-length coding region of CgFADBD was amplified by RT-PCR technique and was bioinformatically analyzed.The results showed that CgFADBD encoded a polypeptide consisting of 505 amino acids with a molecular mass of about 54.39 kD.The CgFADBD protein contained a typical signal peptide sequence(amino acids 1~26)and a conserved FAD binding domain(amino acids 69~203),but does not contain any transmembrane structural domains.The CgFADBD knockout mutantΔCgFADBD was constructed based on the principles of fusion PCR and homologous recombination.Colony growth and conidia production statistics showed no significant difference between wild-type strain(WT)andΔCgFADBD.Pathogenicity analysis showed that the pathogenicity ofΔCgFADBD on rubber tree leaves was significantly lower than that of WT.The above data indicated that CgFADBD contributed to the pathogenicity of C.gloeosporioides on rubber trees,but not to colony growth and conidia production.The results of this study provide some reference for resolving the pathogenic mechanism of C.gloeosporioides on rubber trees.

关 键 词：胶孢炭疽菌(Colletotrichum gloeosporioides) FAD结合蛋白 致病能力 

分 类 号：S763.7[农业科学—森林保护学]