生物技术通报 ›› 2017, Vol. 33 ›› Issue (6): 104-111.doi: 10.13560/j.cnki.biotech.bull.1985.2016-1142

• 研究报告 • 上一篇    下一篇

稻瘟病菌不同生长期蛋白精氨酸甲基转移酶基因的表达分析

吴立叶, 王国梁, 刘文德   

  1. 中国农业科学院植物保护研究所,北京 100193
  • 收稿日期:2016-12-19 出版日期:2017-06-26 发布日期:2017-06-19
  • 作者简介:吴立叶,硕士,研究方向:分子植物病理学;E-mail:wuliye001@126.com
  • 基金资助:
    国家自然科学基金项目(31422045)

Expression Analysis of Protein Arginine Methyltransferase Genes in Magnaporthe oryzae

WU Li-ye, WANG Guo-liang, LIU Wen-de   

  1. Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193
  • Received:2016-12-19 Published:2017-06-26 Online:2017-06-19

摘要: 精氨酸甲基化是一种广泛存在于真核生物中的蛋白质翻译后修饰,其主要过程为蛋白质精氨酸甲基转移酶(Protein arginine methyltransferases,PRMTs)催化S-腺苷甲硫胺酸(S-adenosylmethionine,SAM)提供的甲基基团转移到蛋白质精氨酸侧链胍基基团上。由稻瘟病菌(Magnaporthe oryzae)侵染水稻引起的稻瘟病,每年均会为水稻生产造成严重损失。近年来稻瘟病菌和水稻已成为研究植物病原真菌与寄主植物互作的理想模式生物。生物信息学分析表明,稻瘟病菌中含有4个PRMT基因(MoPRMT1-4),经蛋白序列比对分析发现,它们均含有保守的甲基转移酶结构域;构建系统发育树,表明稻瘟病菌PRMTs在丝状真菌中高度保守;通过提取稻瘟病菌在不同生长和侵染时期的mRNA,利用实时荧光定量PCR技术,分析稻瘟病菌PRMTs基因的表达谱,发现MoPRMT1在侵染后24 h表达量达到最高峰,而其他阶段表达水平基本一致;MoPRMT2与其他3个基因相比,各阶段表达量均较低,且各时期表达水平也没有明显变化;MoPRMT3在芽管和附着胞发育阶段表达量较高,MoPRMT3和MoPRMT4在成熟附着胞时期表达量均达到最高,MoPRMT4在侵染后42 h也出现表达峰值。这些数据表明PRMTs基因对于稻瘟病菌的侵染致病可能起重要调控作用。

关键词: 稻瘟病菌, PRMTs, 表达分析

Abstract: Arginine methylation is a kind of post-translational modifications existing widely in eukaryotes,of which the main process is to transfer methyl groups provided by S-adenosylmethionine(SAM)to the guanidino group of protein arginine side chain while catalyzed by protein arginine methyltransferases(PRMTs). Rice blast fungus(Magnaporthe oryzae)infects rice under the suitable conditions,causing severe rice blast and resulting in serious loss of rice yield per year. In recent years,rice blast fungus and rice have already become the ideal model for the study of interaction between plant pathogenic fungi and their host plants. Bioinformatics analysis showed that the rice blast fungus contained four PRMTgenes(MoPRMT1-4),and PRMTs protein sequence alignment revealed that they contained conservative methyltransferase domains. The PRMTs were demonstrated to be very conservative in filamentous fungi through phylogenetic tree analysis. Extracting the mRNAs of MoPRMT1-4 at various growth and infection stages,we used real time fluorescence quantitative PCR to analyze the expression profiles of the genes,and discovered that the expression of MoPRMT1 reached peak at 24 h post infection,while almost the same at other stages. Compared with other three genes,the expression of MoPRMT2 was low and showed no obvious variation at each stage. MoPRMT3 expressed in high level at germ tube and appressorium developmental stages,while both MoPRMT3 and MoPRMT4 reached peak expressions at mature appressorium stage,and MoPRMT4 also presented peak at 42 h post infection. These data implied that PRMTs genes may play an important regulation role in the pathogenicity of M. oryzae.

Key words: Magnaporthe oryzae, PRMTs, expression analysis