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侵染早期小麦叶锈菌相关基因的筛选及表达分析
张艺博,路亚南,刘娜,肖继斌,李闯,程琨,邢国珍,马振玲,郑文明
0
(河南农业大学 生命科学学院/小麦玉米作物学国家重点实验室, 郑州 450002)
摘要:
为挖掘与叶锈菌萌发及早期侵染相关的关键基因,利用PacBio SMRT和Illumina HiSeq测序技术对叶锈菌夏孢子萌发0、2、4、8、12和24 h的孢子和芽管进行转录组测序,通过全长转录本基因表达差异分析和生物信息学分析进行候选基因筛选,利用qRT-PCR技术对候选基因在不同毒力菌系与小麦的亲和/非亲和互作中的表达模式进行系统分析。结果表明:1)综合分析筛选出差异显著且功能相关的4个候选基因,其中,PTTG_1050为SNARE蛋白的编码基因,可能参与SNARE介导的囊泡运输;PTTG_4765属于ABC转运蛋白基因家族PDR亚族;PTTG_1823属于PKc_Like超基因家族;PTTG_1279为候选分泌蛋白编码基因。2)4个候选基因在侵染早期受到强烈诱导或抑制,且在亲和互作中表达量高于非亲和互作,说明其在叶锈菌的早期侵染和扩展中发挥了重要作用,可能是早期成功侵染的关键基因。
关键词:  小麦叶锈菌  转录组测序  夏孢子萌发  早期互作  qRT-PCR  致病基因
DOI:10.11841/j.issn.1007-4333.2022.10.02
投稿时间:2021-11-10
基金项目:河南省自然科学基金(202300410202,182300410023);河南省科技攻关项目(182102110278);河南省教育厅重点项目(17B210003)
Screening and expression analysis of candidate genes related to early infection of Puccinia triticina
ZHANG Yibo,LU Yanan,LIU Na,XIAO Jibin,LI Chuang,CHENG Kun,XING Guozhen,MA Zhenling,ZHENG Wenming
(College of Life Science/State Key Laboratory of Wheat and Maize Crop, Henan Agricultural University, Zhengzhou 450002, China)
Abstract:
To discover the key genes involved in the early infection and urediospores germination of Puccinia triticina, PacBio single molecule real-time and Illumina sequencing technology were used to sequence the transcriptome of spores and germ tubes from urediniospores germinated at 0, 2, 4, 8, 12 and 24 h. Candidate genes were screened by expression difference analysis and bioinformatics analysis of full-length transcripts, the expression patterns of candidate genes in the compatible and incompatible interaction between different virulent isolates and wheat were analyzed by qRT-PCR. The results showed that: 1)Four candidate genes with significant differences and functional correlation were identified by comprehensive analysis. Among them, PTTG_1050 is the encoding gene of SNARE protein, which may be involved in SNARE mediated vesicle transport; PTTG_4765 belongs to the PDR subfamily of ABC transporter gene family; PTTG_1823 belongs to PKc_Like supergene family; PTTG_1279 is a candidate secretory protein encoding gene. 2)The four candidate genes were strongly induced or inhibited in the early stage of infection, and the expression in the compatible interaction was higher than that in the incompatible interaction, indicating that they might play key roles in the early infection and expansion of Puccinia triticina, and be the important genes involved in the early infection.
Key words:  Puccinia triticina  transcriptome sequencing  urediniospore germination  early interaction  qRT-PCR  pathogenic gene