摘要
植物半胱氨酸蛋白酶(cysteine proteases,CPs)广泛影响着植物的生理过程及抗病调控。为研究烟草半胱氨酸蛋白酶在调控马铃薯Y病毒(potato virus Y,PVY)中的功能机理,本研究在全基因组水平上鉴定栽培烟草CPs基因家族,使用生物信息学方法分析CPs基因家族的进化关系、Motif基序及启动子调控元件;筛选CPs对应的mi RNAs,分析CPs基因及其对应的miRNAs在PVY感染后的表达特性。结果表明,栽培烟草中共鉴定到70个CPs基因家族成员,根据其系统发育特征和结构可分为5个亚家族,其中C1A 39个、C2A 2个、C125个,C138个,C14A 16个;Motif分析表明,同一亚家族CPs具有相似的motif分布;启动子顺式作用元件分析表明,CPs家族基因含有光应答元件、激素响应元件以及低温、干旱、高温、盐胁迫等响应元件。基于转录组和small RNA测序结果发现,PVY感染后有38个CPs基因上调表达,18个CPs基因下调表达。有28个CPs基因受51个miRNAs调控,PVY感染后,38个miRNAs表达上调,13个miRNAs表达下调,其中7个在PVY感染后与miRNAs负相关。RT-qPCR结果证明PVY感染后,有15个CPs基因表达量显著上调。研究结果有助于深入理解烟草CPs基因家族功能,及其与相应miRNAs通过互作调控烟草对PVY的响应,为研究茄科作物抗PVY分子机制提供理论依据。
Plant cysteine proteases(CPs)widely affect plant physiological processes and disease resistance regulation.To study the functional mechanism of tobacco cysteine proteases in regulating potato virus Y(PVY),the CPs family genes of Nicotiana tabacum L.at the genome-wide level were identifed.The evolutionary relationships,motifs and promoter cis-acting elements of CPs family genes were analyzed by bioinformatics.Then the miRNAs corresponding to CPs were screend,the expression characteristics of CPs genes and their corresponding miRNAs after PVY infection were analyzed.The results showed that a total of 70 CPs genes were identified in Nicotiana tabacum L.genome,which were divided into five subfamilies,of which C1A contained 39 CPs,C2A contained 2 CPs,C12 contained 5 CPs,C13 contained 8 CPs and C14A contained 16 CPs.Motif analyses showed that the CPs of subfamily had similar motif distribution.Promoter cis-acting elements analysis showed that several CPs family gene promoters contained light response elements,hormone response elements and low temperature,drought,high temperature,salt stress response elements.Based on the sequencing results of transcriptome and small RNA,it was found that 38 CPs were up-regulated while 18 CPs were down-regulated after PVY infection.Total 51 miRNAs were found to be targeted to 28 members of cysteine protease gene family,38 miRNAs were up-regulated while 13 were down-regulated after PVY infection,7 of them were negatively correlated with miRNAs.The results of RT-q PCR showed that 15 CPs genes were significantly up-regulated after PVY infection.This study is conducive to better understanding the functions of CPs genes and their corresponding miRNAs in regulating tobacco PVY responses,which may provide evidence for the molecular mechanism of anti-PVY in Solanaceae crops.
作者
尹国英
刘畅
常永春
羽王洁
王兵
张盼
郭玉双
YIN Guo-ying;LIU Chang;CHANG Yong-chun;YU Wang-jie;WANG Bing;ZHANG Pan;GUO Yu-shuang(Guizhou Academy of Tobacco Science,Guiyang 550081;College of Agronomy and Biotechnology,Southwest University,Chongqing 400715;College of Agriculture,Shanxi Agricultural University,Jinzhong 030800;College of Advanced Agriculture and Ecological Environment of Heilongjiang University,Harbin 150006)
出处
《生物技术通报》
CAS
CSCD
北大核心
2023年第10期184-196,共13页
Biotechnology Bulletin
基金
中国烟草总公司贵州省公司科技项目(2022XM01)
中国烟草总公司贵州省公司科技项目(2021XM08)
贵州省烟草科学研究院科技项目(GZYKY2021-02)。
