草莓果生刺盘孢菌候选效应子基因CfE15对孢子产量的影响  

The Effect of Effector Gene CfE15 on Conidia Production of Colletotrichum fructicola in Strawberry

作　　者：张丽勍[1,2] 余永婷 方献平 李水根[1,2] 张学英[1,2] Zhang Liqing;Yu Yongting;Fang Xianping;Li Shuigen;Zhang Xueying(Forest&Fruit Research Institute,Shanghai Academy of Agriculture Science,Shanghai,201403;Shanghai Key Laboratory of Protected Horticultural Technology,Shanghai Academy of Agriculture Science,Shanghai,201403)

机构地区：[1]上海市农业科学院林木果树研究所,上海201403 [2]上海市农业科学院,上海市设施园艺技术重点实验室,上海201403

出　　处：《分子植物育种》2023年第12期3919-3927,共9页Molecular Plant Breeding

基　　金：上海市科委启明星人才计划项目(18QB1402800)资助。

摘　　要：草莓炭疽病是由炭疽菌引起的草莓生产中重要的真菌病害之一。果生炭疽菌为华东地区草莓炭疽病菌的优势种。效应子在真菌与植物互作中发挥重要作用,其致病机制是目前研究的热点。实验室前期结合转录组学结果对草莓果生炭疽菌的效应蛋白进行了预测,其中一个在侵染过程中上调表达的编码候选效应蛋白的基因被命名为CfE15。该基因编码一个小分子量富含半胱氨酸的分泌蛋白。生物信息学分析结果表明,该基因编码一条含有109个氨基酸,分子量约为11.6 kD,等电点pI为4.45的多肽。N端含有一段24个氨基酸的信号肽序列。CfE15的二级结构主要由α-螺旋(3%)、β-折叠(33%)、延伸链(17.43%)、β-转角(7.34%)和无规则卷曲(66.97%)组成。构建了CfE15的敲除载体pKH-CfE15和互补载体pKN-CfE15,并通过PEG介导的原生质体转化法获得CfE15基因的突变菌株ΔCfE15和互补菌株ΔCfE15C。野生型菌株、突变菌株ΔCfE15和互补菌株ΔCfE15C在菌落直径及孢子形态无显著差异,但CfE15的缺失导致产孢量降低,互补菌株ΔCfE15C的孢子产量与野生型菌株无显著差异。本研究结果为阐明果生炭疽菌的致病机理,深入解析果生炭疽菌与草莓互作提供了一定理论依据。Strawberry anthracnose,caused by Colletotrichum species,is one of the most serious diseases of strawberry production worldwide.C.fructicola was the dominant species of strawberry anthracnose in East China.Effectors play important roles in the interaction between fungus and plants,and the pathogenicity mechanism of effectors is a research hotspot.In our previous studies,the effectors of C.fructicola were predicted based on the results of transcriptome.One candidate effector named as CfE15,which was significantly up-regulated in strawberry infected by C.fructicola,was chosen for further study.CfE15 encodes a small secreted cysteine-rich protein.Bioinformatics analysis showed that CfE15 encodes a 109 amino acid(aa)polypeptide with the molecular mass of 11.6 kD and isoelectric point of 4.45.The N-terminal of CfE15 protein contains a 24 aa signal peptide sequence.The secondary structure of CfE15 was mainly composed ofα-helix(3%),β-strands(33%),extended strands(17.43%),β-turns(7.34%)and random coils(66.97%).CfE15 gene knockout vector pKH-CfE15 and complementary vector pKN-CfE15 were constructed.The gene deletion and complementary strains,CfE15 and△CfE15C,respectively,were obtained by PEG mediated protoplast transformation and homologous recombination.No significant differences were observed in mycelial growth,morphology and conidia morphology among wild type(WT),△CfE15 and△CfE15C strains.The amount of conidia production of△CfE15 strain was significantly lower than that of WT and△CfE15C strain,and there was no difference between the wild type and△CfE15C.Our results provide a theoretical basis for elucidating the pathogenicity mechanism of C.fructicola and the interaction between C.fructicola and strawberry.

关 键 词：果生炭疽菌(Colletotrichum fructicola) 效应子 CfE15基因 产孢量 

分 类 号：S436.68[农业科学—农业昆虫与害虫防治]