大麦条纹病菌基因Pgr 03902致病功能研究  

Study on the function of the gene Pgr03902 of Pyrenophora graminea

作　　者：祁天涛 司二静[1,2] 郭铭 孟亚雄[1,2] 姚立蓉 汪军成[1,2] 马小乐[1,2] 李葆春[1,3] 杨轲[1,2] 尚勋武[2] 王化俊[1,2] QI Tiantao;SI Erjing;GUO Ming;MENG Yaxiong;YAO Lirong;WANG Juncheng;MA Xiaole;LI Baochun;YANG Ke;SHANG Xunwu;WANG Huajun(State Key Laboratory of Aridland Crop Science,Gansu Provincial Key Laboratory of Crop Improvement and Germplasm Enhancement,Lanzhou 730070,China;College of Agronomy,Gansu Agricultural University,Lanzhou 730070,China;College of Life Science and Technology,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]省部共建干旱生境作物学国家重点实验室,甘肃省作物遗传改良与种质创新重点实验室,兰州730070 [2]甘肃农业大学农学院,兰州730070 [3]甘肃农业大学生命科学技术学院,兰州730070

出　　处：《植物保护》2023年第6期16-22,54,共8页Plant Protection

基　　金：国家自然科学基金(32160496,31960426,32160647);国家大麦青稞产业技术体系(CARS-05-03B-03);甘肃省青年科技基金计划(20JR5RA010);甘肃省教育厅创新基金(2021A-055);甘肃省教育厅产业支撑计划(2021CYZC-12);甘肃省省级大学生创新创业训练计划(S202210733005)。

摘　　要：为明确基因Pgr 03902(序列号:OP999070)是否参与调控大麦条纹病菌的致病性,为大麦条纹病菌致病机理的研究提供理论依据,本研究运用生物信息学、亚细胞定位和基因干扰技术初步研究了该基因功能。结果表明,Pgr 03902基因开放阅读框大小为348 bp,编码116个氨基酸,编码蛋白二级结构中无规则卷曲较多,编码蛋白具有酸性、不稳定性、亲水性、无信号肽结构等特性;亚细胞定位结果显示,Pgr 03902基因在细胞核和细胞膜上均有表达;采用PEG介导的原生质体转化法获得了1个干扰菌株ΔPgr 03902,RT-qPCR结果表明,ΔPgr 03902中Pgr 03902基因表达量较野生型菌株QWC下降了60.79%(P<0.05),对干扰菌株的营养生长、菌丝形态观察以及致病性研究结果显示,ΔPgr 03902生长速率和致病力均显著低于野生型菌株QWC(P<0.05)。以上结果表明,Pgr 03902参与该菌的生长发育和致病过程。本研究初步明确了Pgr 03902基因在大麦条纹病菌侵染过程中的功能,为进一步研究大麦条纹病菌与寄主之间互作奠定了基础。To study the function of Pgr 03902 gene in pathogenicity of Pyrenophora graminea and provide a theoretical basis for analysis of pathogenic mechanism of P.graminea,the secondary structure,physical and chemical properties of its expressed product were preliminarily predicted using bioinformatics,and its function was investigated through subcellular localization and RNA interference technology.The results showed that the open reading frame of Pgr 03902 gene was 348 bp,encoding 116 amino acids,with its secondary structure composed mainly of random coils.The encoded protein was acidic,unstable,hydrophilic and had no signal peptide structure.The results of subcellular localization showed that Pgr 03902 gene was expressed in both cell nucleus and cell membrane.After PEG-mediated protoplast transformation,one interference transformation strainΔPgr 03902 was obtained.The results of RT-qPCR showed that the expression of this gene inΔPgr 03902 decreased by 60.79%compared with the wild type strain QWC(P<0.05),and the results of vegetative growth,mycelium morphology observation and pathogenicity research showed that the growth rate and pathogenicity ofΔPgr 03902 was significantly lower than that of the wild type strain QWC(P<0.05),indicating that this gene was involved in the growth,development and pathogenicity of P.graminea.Our study preliminarily identified the function of Pgr 03902 gene in the process of P.graminea infection,and laid a foundation for further studies of the interaction between P.graminea and its host.

关 键 词：大麦条纹病菌 RNA干扰 遗传转化 致病性 

分 类 号：S435.123[农业科学—农业昆虫与害虫防治]