芒果胶孢炭疽菌(Colletotrichum gloeosporioides)CgFAE基因对孢子产量及菌丝生长的影响  

Gene Knockout and Preliminary Functional Analysis of CgFAE Gene of Colletotrichum gloeosporioides in Mango

作　　者：卢梦瑶 徐祥彬[1] 孟兰环 张贝[2] 安邦[2] 史学群[1] Lu Mengyao;Xu Xiangbin;Meng Lanhuan;Zhang Bei;An Bang;Shi Xuequn(College of Food Science and Engineering,Hainan University,Haikou,570228;College of Tropical Crops,Hainan University,Haikou,570228)

机构地区：[1]海南大学食品科学与工程学院,海口570228 [2]海南大学热带作物学院,海口570228

出　　处：《分子植物育种》2020年第22期7342-7347,共6页Molecular Plant Breeding

基　　金：海南省科技厅创新团队项目(2019CXTD399)资助。

摘　　要：炭疽病是芒果的重要病害之一,由胶孢炭疽菌引起。阿魏酸酯酶为细胞壁降解酶,在致病机理中起着越来越重要的作用。本研究克隆了芒果胶孢炭疽菌的一个阿魏酸酯酶基因FAE,命名为CgFAE,该基因编码一个含有539个氨基酸的蛋白,分子质量约为58.3 k D。为了进一步研究该基因的功能,根据同源重组原理,构建了CgFAE基因的敲除载体pCB1532-CgFAE,通过PEG介导的原生质体转化法将其转入到胶孢炭疽菌原生质体细胞中进行同源交换,经过对转化子进行抗性筛选和分子鉴定,获得CgFAE基因的敲除突变体ΔCgfae,并对其孢子产量和菌落生长情况进行分析,发现胶孢炭疽菌CgFAE基因的敲除突变体ΔCgfae的孢子产量低于野生型菌株,菌落生长慢于野生型菌株。以上结果表明,该基因与胶孢炭疽菌的产孢能力与菌丝生长有关。Aanthracnose is one of the most important diseases of mango, mainly caused by Colletotrichum gloeosporioides Penz. As a part of cell wall degrading enzymes, ferulic acid esterase plays an increasingly important role in pathogenesis. In this study, we cloned a ferulic acid esterase gene FAE of anthrax mangiana, named CgFAE, which encodes a protein containing 539 amino acids with a molecular mass of approximately 58.3 kD. In order to study further on the function of this gene and based on the principle of homologous recombination, the knockout vector pCB1532-CgFAE of the CgFAE gene was constructed, and it was transformed into the hypha protoplast of C.gloeosporioides by PEG-mediated protoplast transformation. After homologous exchange, the chlorimuron ethylresistant transformants were confirmed by PCR, the C. gloeosporioides knock-out mutant of CgFAE gene, ΔCgfae,was generated. Then the fungal growth rate and conidiation ability of ΔCgfae were analyzed. The conidia production of ΔCgfae was reduced to less than a-half that of the WT and the colony growth was slower than that of the wild type. The above results indicate that this gene is related to the spore-producing ability and mycelial growth of anthracnose.

关 键 词：胶孢炭疽菌(colletotrichum gloeosporioides) 细胞壁降解酶 基因敲除 CgFAE 生长发育 

分 类 号：S436.67[农业科学—农业昆虫与害虫防治] Q943.2[农业科学—植物保护]