分子植物育种2024,Vol.22Issue(20) :6653-6660.DOI:10.13271/j.mpb.022.006653

高温胁迫下茄子小热激蛋白基因SmsHSP20的克隆与表达分析

Cloning and Expression Analysis of Small Heat Shock Protein Gene Sms-HSP20 from Eggplant under High Temperature Stress

周亚东 赵恩鹏 申磊 杜丽慧 姜政 胡雪 夏昕 杨旭
分子植物育种2024,Vol.22Issue(20) :6653-6660.DOI:10.13271/j.mpb.022.006653

高温胁迫下茄子小热激蛋白基因SmsHSP20的克隆与表达分析

Cloning and Expression Analysis of Small Heat Shock Protein Gene Sms-HSP20 from Eggplant under High Temperature Stress

周亚东 1赵恩鹏 1申磊 1杜丽慧 1姜政 1胡雪 1夏昕 1杨旭1
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作者信息

  • 1. 扬州大学园艺园林学院,扬州,225009
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摘要

利用茄子SmsHSP20(基因编号>Smechr0302276.1)在杭茄数据库中搜索得到其ORF(open reading frame)序列,然后参考此序列设计特异引物及荧光定量PCR引物.以茄子品种'AC013'叶片cDNA为模板克隆获得了该基因,该基因大小为714bp,编码237个氨基酸,C端含有一个Hsp20功能域.对SmsHSP20进行生物信息学分析,发现SmsHSP20蛋白与马铃薯、辣椒、番茄、枸杞等茄科植物有较高的同源性.对SmsHSP20的启动子序列进行分析,发现其包含2个光反应顺式作用元件、多个植物激素响应顺式作用元件以及多个转录因子结合位点.采用RT-qPCR研究了SmsHSP20在高温胁迫下茄子中的表达情况,结果显示该基因在茄子受到高温胁迫时其表达量急剧升高,在0.5 h达到最高值,说明该基因可能在茄子响应高温胁迫的过程中起正调节作用.

Abstract

The ORF(open reading frame)sequence of eggplant SmsHSP20(gene number>Smechr0302276.1)was searched in Hangzhou-eggplant database,and the specific primers and fluorescence quantitative real-time PCR primers were designed according to this sequence.This gene,encoded 237 amino acids with 714 bp,was cloned from the cDNA of eggplant variety'AC013'leaves,and harbored a HSP20 domain.Bioinformatics analysis of SmsHSP20 revealed that SmsHSP20 protein had high homology with Solanaceae species including potato,pepper,tomato and Chinese wolfberry.Sequence analysis of the SmsHSP20 promoter revealed that it contained two light-responsive cis-elements,multiple plant hormones responsive cis-elements and multiple transcription factor binding sites.The RT-qPCR assay was performed to detect the expression pattern analysis of SmsHSP20 in egg-plant variety'AC013'leaves in the condition of high temperature stress and the result showed that the transcript ex-pression level of SmsHSP20 was sharply up-regulated to peak at 0.5 hours post treatment,indicating that SmsHSP20 may act a positive role in eggplant response to high temperature stress.

关键词

茄子/HSP20/高温胁迫/生物信息学分析/荧光定量PCR

Key words

Eggplant/HSP20/High temperature stress/Bioinformatics analysis/RT-qPCR

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基金项目

国家重点研发计划项目(2019YFD1000300)

出版年

2024
分子植物育种
海南省生物工程协会

分子植物育种

CSTPCD北大核心
影响因子:0.765
ISSN:1672-416X
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