晚疫病抗性相关基因NbMybl的克隆与功能分析
Cloning and functional analysis of the Nicotiana benthamiana NbMybl gene related to late blight resistance
赵迪 1陈胜男 1张蝶 1王洪洋 1陈爱娥2
作者信息
- 1. 云南省马铃薯生物学重点实验室,云南师范大学,昆明 650500
- 2. 云南师范大学教务处,云南师范大学,昆明 650500
- 折叠
摘要
Myb转录因子广泛参与调控植物生长发育以及应对环境胁迫,但有关其对晚疫病抗性调控机制的研究较少.本研究从本氏烟草中克隆获得一个含Myb-like结构域的蛋白编码基因,命名为NbMybl.该基因开放阅读框全长为753 bp,编码250 aa.实时荧光定量PCR(qPCR)结果显示NbMybl受致病疫霉(Phytophthora infestans)侵染诱导表达.亚细胞定位表明NbMybl定位在植物细胞核和细胞质中.利用病毒诱导的基因沉默技术,发现沉默NbMybl能够明显降低本氏烟草对致病疫霉的抗性.分析比较NbMybl沉默和非沉默对照烟草株系应对P.infestans侵染的转录组数据,发现差异表达基因(Dif-ferentially expressed genes,DEGs)8486个,其中上调基因 4202 个,下调基因 4284个.使用 KEGG(Kyoto Encyclopedia of Genes and Genomes)数据库进行基因富集分析,发现鉴定得到的DEGs中共有373个参与植物-病原菌互作,308个参与植物激素信号转导,216个参与植物MAPK信号途径.推测这些DEGs可能与沉默NbMybl降低本氏烟草抗病性有密切关联.qPCR检测部分DEGs的表达,发现其表达趋势与转录组数据一致.本研究为阐明NbMybl参与对抗致病疫霉侵染提供了理论依据.
Abstract
Myb transcription factors play important roles in the regulation of various biological processes in plants.However,the molecular mechanism underlying their roles in regulating late blight resistance remains elu-sive.Here,we report the cloning of NbMybl,a Myb-like gene from Nicotiana benthamiana,which has an open reading frame of 753 bp and encodes a protein of 250 aa.NbMybl contains a Myb-like DNA-binding domain.Real-time quantitative PCR(qPCR)revealed that NbMybl was induced by infection with Phytophthora infestans.Subcellular localization analysis showed that NbMybl is located in both the nucleus and the cytoplasm.Silencing of NbMybl by virus-induced gene silencing(VIGS)significantly increased the susceptibility of plants to P.infes-tans.Transcriptome profiling by RNA sequencing identified 8468 differentially expressed genes(DEGs)with fold change ≥ 2 and FDR＜0.01 between NbMybl silenced and non-silenced control lines in response to P.in-festans infection,and the result of RNA-seq was further validated by qPCR with 10 randomly selected DEGs.KEGG(Kyoto Encyclopedia of Genes and Genomes)enrichment analysis revealed that a total of 373 DEGs were involved in plant-pathogen interaction,308 DEGs and 216 DEGs were involved in MAPK signaling path-way and plant hormone signal transduction,respectively.We speculated that these DEGs might be closely related to the reduced resistance of NbMybl-silenced N.benthamiana lines to P.infestans.Our study provides valuable insights into the molecular mechanisms of NbMybl in regulating resistance to P.infestans.
关键词
本氏烟草/致病疫霉/NbMybl/病毒诱导的基因沉默/转录组Key words
Nicotiana benthamiana/Phytophthora infestans/NbMybl/virus-induced gene silencing/tran-scriptome引用本文复制引用
基金项目
国家自然科学基金(32060499)
国家自然科学基金(31800134)
云南省基础研究计划(202201AT070034)
广东省基础与应用基础研究重大项目(2021B0301030004)
兴滇英才支持计划()
出版年
2024
NETL
NSTL
万方数据