UePep1对菰黑粉菌致病力的调控作用  

Regulation Effect of UePep1 in the Pathogenicity of Ustilago esculenta

作　　者：黄琳钰 汤近天 夏文强 崔海峰 叶子弘 张雅芬 HUANG Lin-Yu;TANG Jin-Tian;XIAWen-Qiang;CUI Hai-Feng;YE Zi-Hong;ZHANG Ya-Fen(Zhejiang Provincial Key Laboratory of Biometrology and Inspection and Quarantine/College of Life Sciences,China Jiliang University,Hangzhou 310018,China)

机构地区：[1]中国计量大学生命科学学院/浙江省生物计量与检验检疫技术重点实验室,杭州310018

出　　处：《农业生物技术学报》2024年第7期1614-1625,共12页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(U20A2043)。

摘　　要：菰黑粉菌(Ustilago esculenta)专性侵染菰(Zizania latifolia)后可诱导形成膨大肉质茎—茭白,且菰黑粉菌的致病力与茭白的表型、产量等密切相关。穿透过程必需蛋白1(protein essential during penetration 1,Pep1)作为真菌效应因子,在黑粉菌属(Ustilago)真菌的致病力调控上起着重要作用。本研究先通过BLASTP定位及基因克隆分析,获得了菰黑粉菌中Pep1的同源蛋白编码基因UePep1(GenBank No.OR088111),其开放阅读框为528 bp,编码175个氨基酸,无内含子序列。进一步通过PEG介导的原生质体转化方法构建UePep1的敲除突变体,进行体外生长及侵染试验,发现该基因的缺失不影响菰黑粉菌单倍体形态与生长速率,也不影响体外菌丝形成与丝状生长能力,但显著减缓了菰黑粉菌入侵菌丝的增殖,无法诱导寄主形成膨大肉质茎。进一步将UePep1回补至玉米瘤黑粉菌(Ustilago maydis)SG200ΔPep1菌株中,部分恢复了SG200ΔPep1菌株的侵染力,表明UePep1是玉米瘤黑粉菌Pep1的功能性互补蛋白。上述研究明确了UePep1在菰黑粉菌致病力调控中的作用,同时进一步证实了该效应因子在黑粉菌属真菌中功能的保守性,为研究菰黑粉菌与菰互作机制进而解析膨大肉质茎形成机理提供借鉴和理论支持。Ustilago esculenta can induce the formation of the enlarged fleshy stem of Zizania latifolia after obligate infection,and its pathogenicity is closely related to the phenotype and yield of the swollen stem called Jiaobai.Pep1(protein essential during penetration 1),as a fungal effector,plays an important role in the pathogenicity of Ustilago fungi.In this study,UePep1(GenBank No.OR088111),the homologous protein coding gene of Pep1 in U.esculenta,was cloned by BLASTP mapping and sequence analysis,with an ORF of 528 bp,encoding 175 amino acids and no intronic sequences.The deletion of UePep1 gene did not affect the haploid morphology,growth rate,mycelial formation and filamentous growth of U.esculenta in vitro.However,it significantly inhibited the invaded mycelial proliferation of the U.esculenta in vivo,and could not induce the formation of the enlarged fleshy stem.Further supplementation of UePep1 into the SG200ΔPep1 strain of Ustilago maydis partially restored the infectivity of the SG200ΔPep1 strain,indicating that UePep1 is a functional complementary protein of the Pep1 in the smut fungi.The results provide a reference and theoretical support for further understanding of the interaction mechanism between U.esculenta and Z.latifolia and the formation mechanism of enlarged fleshy stem.

关 键 词：菰黑粉菌 穿透过程必需蛋白1(Pep1) 菰 致病性 

分 类 号：S961.6[农业科学—水产养殖] S182[农业科学—水产科学] S188[生物学—生物工程] Q81