假禾谷镰孢中三角状五肽重复蛋白的全基因组鉴定与表达分析  

Genome-wide identification and expression analysis of pentatricopeptide repeat proteins in Fusarium pseudograminearum

作　　者：王利民[1] 孟家兴 庄训宇 张媛 李海洋 陈琳琳[1] 汪敏[1] 丁胜利[1] 李洪连[1] WANG Limin;MENG Jiaxing;ZHUANG Xunyu;ZHANG Yuan;LI Haiyang;CHEN Linlin;WANG Min;DING Shengli;LI Honglian(College of Plant Protection,Henan Agricultural University/State Key Laboratory of Wheat and Corn Crop Science/Henan Food Crops Collaborative Innovation Center,Zhengzhou 450002,China)

机构地区：[1]河南农业大学植物保护学院/小麦玉米作物学国家重点实验室/河南粮食作物协同创新中心,郑州450002

出　　处：《植物病理学报》2022年第3期341-351,共11页Acta Phytopathologica Sinica

基　　金：国家自然科学基金项目(31871922);国家自然科学基金国际(地区)合作与交流项目(31961143018)。

摘　　要：假禾谷镰孢(Fusarium pseudograminearum,Fp)是引起小麦茎基腐病的优势病原菌,但关于该病原菌致病机理的研究报道很少。三角状五肽重复蛋白(Pentatricopeptide repeat protein,Ppr)为一类核编码的RNA结合蛋白,通过结合特定的RNA序列调控靶标基因RNA的成熟过程,参与线粒体基因组转录和翻译。Ppr蛋白在植物、动物(包括人)及酵母中有一些研究报道,但在丝状真菌特别是植物病原真菌中鲜见报道。为了明确假禾谷镰孢中PPR基因家族的特征,本研究通过全基因组Blastp分析,共鉴定出8个PPR候选基因序列FpPPR1~FpPPR8,主要分布在1号、3号、4号染色体上,基本不含有内含子。理化性质与功能预测显示,Ppr为亲水性蛋白,氨基酸长度510~1353 aa,分子量59.09~152.23 kDa,等电点为5.23~9.69,其中FpPpr3和FpPpr7为稳定蛋白,其他6个蛋白为不稳定蛋白。亚细胞定位预测显示,Ppr蛋白主要位于线粒体。FpPpr1、FpPpr3、FpPpr5、FpPpr6和FpPpr7的3D结构预测形成明显的超螺旋结构。在假禾谷镰孢营养阶段和侵染过程的转录组数据中分析了8个基因表达模式,通过qRT-PCR进行验证,FpPPR1、FpPPR3、FpPPR5在菌丝阶段高表达,FpPPR1、FpPPR5在侵染后期显著下调。本研究结果为进一步研究假禾谷镰孢中PPRs基因的生物学功能奠定基础。Fusarium pseudograminearum(Fp)is a predominant pathogen causing Fusarium crown rot in wheat,but there are few reports on the growth regulation and pathogenic mechanism.Pentatricopeptide repeat proteins(Ppr)are nuclear-encoded RNA binding proteins regulating the process of RNA maturation,which participate in the transcription and translation of mitochondrial genome.Many Ppr proteins have been well-studied in plant,animals(including human)and yeast systems.Nevertheless,the Ppr components and functions in filamentous fungi,especially plant pathogenic fungi remain largely unknown.To clarify the characteristics of the PPR gene family in F.pseudograminearum,a total of 8 PPR candidate genes were identified through genome-wide Blastp analysis,which are mainly distributed on chromosome 1,3,and 4 with almost no introns.The Ppr proteins predicted were all hydrophilic based on the physical and chemical properties analysis ranging from 510 to 1353 aa in length,with molecular weight of 59.09 to 152.23 kDa and isoelectric point of 5.23 to 9.69,respectively.FpPpr3 and FpPpr7 are stable proteins,while the others are unstable.The Ppr proteins were predicted to be mainly localized in mitochondria.FpPpr1,FpPpr3,FpPpr5,FpPpr6,and FpPpr7 folded obvious super-helical in 3 D model.We analyzed the expression patterns of PPRs from transcriptome of the fungal vegetative and infection stages along with validation by qRT-PCR.FpPPR1,FpPPR3,and FpPPR5 were highly expressed at the vegetative hyphae stage,while FpPPR1 and FpPPR5 were significantly down-regulated at the later stage of infection.Collectively,our findings lay the base for deep understanding the roles of PPR genes in F.pseudograminearum.

关 键 词：小麦茎基腐病 假禾谷镰孢 三角状五肽重复蛋白(Ppr) Blastp分析 表达模式 

分 类 号：S432.1[农业科学—植物病理学]