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丝颖针茅ScTIP1;1基因的克隆及对非生物胁迫的应答
引用本文:董超,杨云强,孙旭东,李雄,杨时海,黄蓉,杨永平. 丝颖针茅ScTIP1;1基因的克隆及对非生物胁迫的应答[J]. 植物科学学报, 2016, 34(1): 99-108. DOI: 10.11913/PSJ.2095-0837.2016.10099
作者姓名:董超  杨云强  孙旭东  李雄  杨时海  黄蓉  杨永平
作者单位:1. 中国科学院昆明植物研究所东亚植物多样性与生物地理学重点实验室, 昆明 650201;
2. 中国科学院昆明植物研究所西南 野生生物种质资源库, 昆明 650201;
3. 中国科学院大学, 北京 100049;
4. 中国科学院青藏高原研究所, 北京 100101
基金项目:国家自然科学基金项目(41271058);国家重点基础研究发展计划(2010CB951704);中国科学院寒旱区陆面过程与气候变化重点实验室开放课题(Y5203511Y1).
摘    要:利用丝颖针茅(Stipa capillacea Keng)一条EST序列并结合cDNA末端快速扩增(RACE)技术,克隆了丝颖针茅的一个液泡膜内在蛋白(tonoplast intrinsic proteins,TIPs)基因ScTIP1;1的全长编码区序列。该基因开放阅读框长度为753 bp,编码250个氨基酸,其蛋白质分子量为25.8 kD,理论等电点为6.16;序列比对及系统进化分析表明,ScTIP1;1和拟南芥(Arabidopsis thaliana)AtTIP1;1蛋白的亲缘关系较近;亚细胞定位结果显示,该蛋白位于液泡膜上;实时荧光qRT-PCR检测表明,盐、干旱及低温胁迫可诱导ScTIP1;1基因的表达,且低温处理后基因的表达量变化最为明显。本研究结果为理解丝颖针茅的生态适应性提供了理论依据。

关 键 词:丝颖针茅  液泡膜内在蛋白  分子克隆  基因表达  非生物胁迫
收稿时间:2015-09-02

Molecular Cloning and Expression of ScTIP1;1 in Stipa capillacea under Abiotic Stress
DONG Chao,YANG Yun-Qiang,SUN Xu-Dong,LI Xiong,YANG Shi-Hai,HUANG Rong,YANG Yong-Ping. Molecular Cloning and Expression of ScTIP1;1 in Stipa capillacea under Abiotic Stress[J]. Plant Science Journal, 2016, 34(1): 99-108. DOI: 10.11913/PSJ.2095-0837.2016.10099
Authors:DONG Chao  YANG Yun-Qiang  SUN Xu-Dong  LI Xiong  YANG Shi-Hai  HUANG Rong  YANG Yong-Ping
Affiliation:1. Key Laboratory for Plant Diversity and Biogeography of East Asia, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, 650204, China;
2. Plant Germplasm and Genomics Center, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming 650201 China;
3. University of Chinese Academy of Sciences, Beijing, 100049 China;
4. Institute of Tibetan Plateau Research, Chinese Academy of Sciences, Beijing, 10010 China
Abstract:Based on EST sequence and RACE experiments, the full-length coding sequence of a tonoplast intrinsic protein (TIP) gene termed ScTIP1;1 was cloned from Stipa capillacea. Sequence analysis showed that the opening reading frame (ORF) of ScTIP1;1 was 753 bp, encoding a 250 amino acid peptide with a protein molecular weight of 25.8 kD and isoelectric point of 6.16. Phylogenetic analysis and sequence alignment results indicated that ScTIP1;1 had a close relationship with AtTIP1;1 of Arabidopsis thaliana. Subcellular localization analysis indicated that ScTIP1;1 was localized in the tonoplast membrane. The expression level of ScTIP1;1 was induced by salt, drought and especially low temperature in the leaves of S. capillacea. These results provide new insight into understanding the ecological adaptations of S.capillacea.
Keywords:Stipa capillacea Keng  Tonoplast intrinsic proteins  Molecular cloning  Gene expression  Abiotic stress
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