荔枝霜疫霉自噬相关基因的鉴定与表达分析  

Identification and expression analysis of autophagy-related genes in Peronophythora litchii

作　　者：王荣波[1] 陈姝樽 郭朦朦 李文强 刘裴清[1] 李本金[1] 翁启勇[1] 陈庆河[1,2] WANG Rongbo;CHEN Shuzun;GUO Mengmeng;LI Wenqiang;LIU Peiqing;LI Benjin;WENG Qiyong;CHEN Qinghe(Fujian Key Laboratory for Monitoring and Integrated Management of Crop Pests,Institute of Plant Protection,Fujian Academy of Agricultural Sciences,Fuzhou 350013,China;Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests,Ministry of Education,College of Plant Protection,Hainan University,Haikou 570228,China)

机构地区：[1]福建省农业科学院植物保护研究所,福建省作物有害生物监测与治理重点实验室,福州350013 [2]海南大学植物保护学院,热带农林生物灾害绿色防控教育部重点实验室,海口570228

出　　处：《植物病理学报》2022年第3期310-320,共11页Acta Phytopathologica Sinica

基　　金：国家自然科学基金(31801683、31772141);福建省自然科学基金(2018J05054);福建省属公益类科研院所基本科研专项(2018R1025-8);福建省农业科学院国家基金延伸项目(AGY2018-2)。

摘　　要：细胞自噬(Autophagy)是一种真核生物中高度保守的胞内物质降解和循环再利用的生理过程,其调控细胞动态平衡、压力适应和细胞程序性死亡,与植物病原真菌生长发育、孢子形成、侵染等一系列过程密切相关。荔枝霜疫霉(Peronophythora litchii)侵染引起的荔枝霜疫病严重危害荔枝生产及采后贮藏。本研究利用模式物种中已知的自噬相关蛋白,采用同源比对的方法在荔枝霜疫霉基因组数据库中鉴定到19个同源蛋白,分属16种蛋白类型。分析基因结构和蛋白保守功能域,发现荔枝霜疫霉自噬相关基因均具有内含子,且数目差异明显;所鉴定候选蛋白都具有相应分组的保守功能域,而PlATG1a在C端具有2个额外的ATG11功能域,PlATG11则在其N端包含1个APG17功能域。进一步对具有核心功能且多成员的自噬相关蛋白ATG1、ATG6、ATG8和ATG18进行系统进化和序列模块分析发现,这些蛋白在不同物种之间十分保守,但是其同源蛋白间存在分化。通过转录组数据分析和qRT-PCR验证结果显示,荔枝霜疫霉中自噬相关基因在生长发育和侵染阶段均有表达,与菌丝阶段相比,PlATG1a、PlATG2、PlATG3、PlATG6a、PlATG8、PlATG18a基因在游动孢子阶段特异上调表达,其中PlATG8最为显著;PlVPS34在孢子囊阶段特异诱导表达,PlATG1b、PlATG12、PlATG17在孢子囊与游动孢子阶段上调表达但是在侵染阶段下调表达;PlATG6b、PlATG9、PlATG10、PlATG13、PlATG14及PlVPS15基因在生长发育和侵染阶段均表现出不同程度的上调表达,PlATG7、PlATG11、PlATG18b则在侵染阶段显著下调表达。结果表明,细胞自噬可能参与调控荔枝霜疫霉的生长发育及致病过程。Autophagy is a highly conserved degradation and recycling process that controls cellular homeostasis, stress adaptation, and programmed cell death in eukaryotes. Emerging evidence indicates that autophagy is important for fungal growth, development, sporulation and pathogenicity. Litchi downy blight caused by Peronophythora litchii greatly threatens the healthy development of the Chinese litchi industry, which is the most serious disease affecting litchi production and storage. In this study, a total of 19 autophagy-related genes belonging to 16 different groups were identified in the plant pathogenic oomycete Pe. litchii using a genome-wide survey. Gene structures analysis revealed that the distribution, number, length and splicing phase of intron/exon organization varies significantly between orthologous ATGs of Pe. litchii. Moreover, all candidate proteins contained the conserved functional domains, while two additional ATG11 domains have been identified at the C-terminal of PlATG1 a. In addition, an extra APG17 domain existed in PlATG11. Phylogenetic analysis of the multi-member subfamilies ATG1, ATG6, ATG8 and ATG18 showed that the ATG proteins are very conserved among different species with some differences in individual homologous proteins. Transcriptome data and real-time PCR analysis revealed that the ATGs are broadly expressed during the development and virulence of Pe. litchii. PlATG1 a, 2, 3, 6 a, 8 and 18 a were specifically up-regulated in zoospores, while PlVPS34 showed a dramatically higher expression during sporangia stage. Interestingly, PlATG1 b, PlATG12 and PlATG17 were up-regulated at sporangia and zoospore stages but down-regulated during infection stages. Moreover, the relative expression levels of 6 ATGs(PlATG6 b, PlATG9, PlATG10, PlATG13, PlATG14 and PlVPS15) were up-regulated in different degrees during development and infection stages, whereas PlATG7, PlATG11 and PlATG18 b exhibited a low expression level during infection stages. These results suggested that autophagy might play essenti

关 键 词：荔枝霜疫霉 细胞自噬 基因鉴定 表达分析 

分 类 号：S436.67[农业科学—农业昆虫与害虫防治]