摘要
热激蛋白(HSP)以分子伴侣形式参与蛋白质组装、折叠、跨膜传导等生物过程,可以提高植物对多种逆境胁迫的耐受性。我们前期通过转录组测序技术(RNA-seq)研究发现甘蔗(Saccharum officinarum)HSP20基因在宿根矮化病菌(Leifsonia xyli subsp.xyli,Lxx)诱导下差异表达。为了探讨HSP20基因与甘蔗抗逆性的关系,本研究从甘蔗品种‘Badila’中克隆了HSP20基因,分析了甘蔗HSP20蛋白原核表达诱导条件以及HSP20基因在不同非生物胁迫[4℃(低温)、聚乙二醇(PEG;干旱)、NaCl(高盐)]及脱落酸(ABA)处理下的诱导表达情况。结果表明,甘蔗HSP20基因(GenBank登记号:MZ220548)全长735 bp,编码244个氨基酸,与高粱(Sorghum bicolor)亲缘关系最近;该基因编码蛋白分子质量为26.79 kDa,有较好的脂溶性,为不稳定亲水蛋白,且亚细胞定位预测HSP20蛋白位于叶绿体。HSP20蛋白在大肠杆菌(Escherichia coli)感受态Transetta(DE3)中诱导表达效果好于大肠杆菌感受态BL21(DE3),使用0.05 mmol·L^(-1)异丙基硫代半乳糖苷(IPTG)在28℃下诱导4 h,即可获得较好的诱导表达效果。实时荧光定量PCR(RT-qPCR)分析表明,低温、干旱、高盐胁迫和脱落酸处理均能诱导甘蔗茎、叶中HSP20基因表达,茎、叶中HSP20基因通过不同表达模式响应这些非生物胁迫以及ABA处理,说明甘蔗HSP20基因在响应逆境胁迫中有重要作用。研究结果为甘蔗HSP20基因生物学功能研究以及甘蔗多重抗逆性研究提供参考。
Heat shock protein(HSP)participates in biological process such as protein assembly,folding and transmembrane conduction in the form of molecular chaperones,which is very important to improve the tolerance of plants to various adversity stresses.Our previous study by RNA sequencing(RNA-seq)found that sugarcane(Saccharum officinarum)HSP20 gene was differentially expressed under Leifsonia xyli subsp.xyli(Lxx)induction.In order to explore the relation between HSP20 gene and stress resistance of sugarcane,a HSP20 gene was cloned from sugarcane variety‘Badila’.The induction conditions of HSP20 in prokaryotic expression was analyzed,as well as the expression profile of HSP20 gene under abiotic stresses[4℃(low temperature),polyethylene glycol(PEG;drought),and NaCl(high salt)]and abscisic acid(ABA)treatment were analyzed.The results show that HSP20 gene(GenBank accession number:MZ220548)is 735 bp in length,encoding 536 amino acids,which is closest to sorghum(Sorghum bicolor).The molecular mass of the protein encoded by this gene is 26.79 kDa,which is an unstable hydrophilic protein with good lipid solubility.It was predicted to be located in chloroplast.The induced expression of HSP20 protein in Escherichia coli Transetta(DE3)was better than E.coli BL21(DE3).A better induced expression effect can be obtained by using isopropylβ-D-thiogalactoside(IPTG)at 28℃ for 4 hours.Real time fluorescence quantitative PCR(RT-qPCR)analysis showed that low temperature,drought,high salt stresses and ABA treatment could induce the expression of HSP20 gene in sugarcane stems and leaves.HSP20 gene in stems and leaves had different expression patterns in response to low temperature,drought,high salt stresses and ABA treatment,indicating that sugarcane HSP20 gene plays an important role in response to stress.This study is intended to provide a reference for the study of the biological function of sugarcane HSP20 gene and the multiple stress resistance of sugarcane.
作者
宋奇琦
张小秋
宋修鹏
颜梅新
王泽平
雷敬超
黄冬梅
李秋芳
梁永检
SONG Qiqi;ZHANG Xiaoqiu;SONG Xiupeng;YAN Meixin;WANG Zeping;LEI Jingchao;HUANG Dongmei;LI Qiufang;LIANG Yongjian(Guangxi South Subtropical Agricultural Science Research Institute,Longzhou,Guangxi 532415,China;Guangxi Subtropical Crops Research Institute,Nanning 530001,China;Sugarcane Research Institute,Guangxi Academy of Agricultural Sciences,Sugarcane Research Center,Chinese Academy of Agricultural Sciences,Key Laboratory of Sugarcane Biotechnology and Genetic Improvement(Guangxi),Ministry of Agriculture and Rural Affairs,Nanning 530007,China)
出处
《植物生理学报》
CAS
CSCD
北大核心
2022年第2期371-380,共10页
Plant Physiology Journal
基金
国家自然科学基金(32001606)
广西自然科学基金(2018GXNSFAA294041和2020GXNSFBA297040)
广西农业科学院基本科研业务专项(桂农科2020YM01和桂农科2021YT007)
中国科学院科技服务网络计划(STS计划)(KFJ-STS-QYZD-199-2)。
关键词
甘蔗
HSP20
基因克隆
原核表达
逆境胁迫
sugarcane
HSP20
gene cloning
prokaryotic expression
adversity stress
