莲腐败病菌多聚半乳糖醛酸酶Fcpg1基因功能研究  

Functional studies on polygalacturonase gene Fcpg1 in Fusarium commune causing lotus rhizome rot

作　　者：叶丽芳 宋水林[1] 张连虎 林桠春 崔汝强[1] 况卫刚 YE Lifang;SONG Shuilin;ZHANG Lianhu;LIN Yachun;CUI Ruqiang;KUANG Weigang(College of Agronomy,Jiangxi Agricultural University,Nanchang 330045,China)

机构地区：[1]江西农业大学农学院,江西南昌330045

出　　处：《江西农业大学学报》2024年第1期77-85,共9页Acta Agriculturae Universitatis Jiangxiensis

基　　金：国家自然科学基金项目(32260663)。

摘　　要：【目的】共享镰刀菌(Fusarium commune)侵染莲引起的莲腐败病严重影响莲的生产,鉴定莲腐败病菌中多聚半乳糖醛酸酶基因Fcpg1,分析其在生长发育及致病过程中的作用,为进一步研究共享镰刀菌的致病机理及莲腐败病菌的防治提供依据。【方法】利用BLASTP在莲腐败病菌全基因组数据中查找PG同源蛋白,并使用ExPASy等在线软件对Fcpg1氨基酸序列进行生物信息学分析。通过基因敲除的方法,获得Fcpg1基因敲除突变体ΔFcpg1和回补菌株ΔFcpg1-Com。测定野生型菌株FCN23、突变体ΔFcpg1和回补菌株ΔFcpg1-Com的菌落形态、生长速率、产孢量及致病力。【结果】Fcpg1基因编码388个氨基酸,分子量为40.354 ku,分子式为C_(1756)H_(2775)N_(489)O_(577)S_(12),理论等电点为6.57,脂溶系数为79.9。Fcpg1为稳定的、易溶于水的外泌性蛋白,其信号肽剪切位点在第16~17个氨基酸,主要定位于胞外基质。Fcpg1蛋白与尖孢镰刀菌的多聚半乳糖醛酸酶蛋白序列具有较高的同源性。与野生型菌株FCN23和回补菌株ΔFcpg1-Com相比,ΔFcpg1突变体的菌落形态、营养生长和产孢量等方面均无明显差异,但突变体ΔFcpg1的致病力显著减弱。【结论】Fcpg1是典型的多聚半乳糖醛酸酶蛋白,参与对莲腐败病致病力的调控。[Objective]Lotus rhizome rot caused by Fusarium commune seriously affects the production of lotus in China.The polygalacturonase gene Fcpg1 in the pathogen of lotus spoilage was identified,and its role in growth,development and pathogenicity was analyzed,which provided a basis for further research on the pathogenic mechanism of F.commune and the prevention and control of the disease.[Method]BLASTP was used to find Fcpg1 homologous proteins in the whole genome data of F.commune and the amino acid sequence of Fcpg1 was analyzed using online software ExPASy.The Fcpg1 knockout mutantΔFcpg1 and the complementation strainΔFcpg1-Com were obtained.Colony morphology,growth rate,spore production,and pathogenicity were measured for the wild-type strain FCN23,the mutantΔFcpg1,and the complementation strainΔFcpg1-Com.[Result]The Fcpg1 gene encoded 388 amino acids with a molecular weight of 40.354 ku.The molecular formula was C_(1756)H_(2775)N_(489)O_(577)S_(12),the isoelectric point was 6.57 and an aliphatic index was 79.9.Fcpg1 was a stable,water-soluble exocytotic protein,mainly localized in the extracellular.Its signal peptide shear site was between amino acids 16-17.The Fcpg1 protein had high homology to the polygalacturonase protein sequence of F.oxysporum.There were no significant differences in colony morphology,vegetative growth,and spore production in theΔFcpg1 mutant compared with the wild-type strain FCN23 and the complementation strainΔFcpg1-Com,but the pathogenicity of the mutantΔFcpg1 was significantly reduced.[Conclusion]Fcpg1 is a typical polygalacturonase protein involved in the regulation of the pathogenicity of lotus rhizome rot.

关 键 词：莲 Fusarium commune 基因敲除 多聚半乳糖醛酸酶 表型 致病力 

分 类 号：S436[农业科学—农业昆虫与害虫防治]