| 紫花苜蓿MsHSP17.7基因的克隆及功能分析 |
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| 引用本文： | 栗振义,龙瑞才,张铁军,杨青川,康俊梅.紫花苜蓿MsHSP17.7基因的克隆及功能分析[J].草地学报,2016,24(1):121-128. |
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| 作者姓名： | 栗振义 龙瑞才 张铁军 杨青川 康俊梅 |
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| 作者单位： | 中国农业科学院北京畜牧兽医研究所, 北京 100193 |
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| 基金项目： | 国家重点基础研究发展计划(973)—紫花苜蓿品质性状相关基因的挖掘与功能分析(2014CB138703-2)，中国农业科学院北京畜牧兽医研究所基本科研业务费项目(2014ywf-zd-2) |
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| 摘 要： | 热激蛋白是一类普遍参与抗逆的保护性蛋白,在植物生长发育及逆境调控中起重要作用。为了解紫花苜蓿(Medicago sativaL.)热激蛋白基因特性及功能,本研究通过同源克隆法获得热激蛋白基因MsHSP17.7,该基因包含477bp开放阅读框,编码158个氨基酸,蛋白分子量为17.67kDa。序列比对发现MsHSP17.7与截形苜蓿(Medicago truncatula)和豌豆(Pisum sativum)中的2个小分子热激蛋白同源性较高,相似度分别是93.38%,83.13%。组织差异性表达显示,MsHSP17.7在茎中表达最多,叶和花次之,在根中最少;实时定量PCR分析显示,MsHSP17.7受高温、高盐诱导表达,在过氧化诱导下表达量变化不明显。对T3代转基因拟南芥(Arabidopsis thaliana)实时定量PCR分析显示,该基因在高盐和过氧化胁迫下均能大量表达,结果表明MsHSP17.7基因可能参与了高盐与过氧化逆境下的胁迫应激调控。
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| 关 键 词： | 紫花苜蓿 热激蛋白 实时定量PCR 非生物胁迫 遗传转化 |
| 收稿时间： | 2014-12-19 |
Cloning and Functional Analysis of MsHSP17.7 Gene from Alfalfa |
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| LI Zhen-yi,LONG Rui-cai,ZHANG Tie-jun,YANG Qing-chuan,KANG Jun-mei.Cloning and Functional Analysis of MsHSP17.7 Gene from Alfalfa[J].Acta Agrestia Sinica,2016,24(1):121-128. |
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| Authors: | LI Zhen-yi LONG Rui-cai ZHANG Tie-jun YANG Qing-chuan KANG Jun-mei |
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| Institution: | Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China |
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| Abstract: | Heat shock protein(HSP) is a kind of ubiquitous protective protein that plays an important role in plant development and stress adaption. In order to learn more about expression characteristics and function of small HSP gene, MsHSP17.7 gene was cloned from alfalfa (Medicago sativa L.) through homology-based cloning. MsHSP17.7 contains an open reading frame of 477-bp encoding a protein of 17.67-kDa. The amino acid sequence shares 93.38% and 83.13% identity with sHSPs in Medicago truncatula and in Pisum sativum. Quantitative RT-PCR results showed that the maximum mRNA expression of MsHSP17.7 was in stems, followed by leaves and flowers, and the least was in root. MsHSP17.7 was induced by high temperature and high salinity stress, but the expression did not chang obviously under peroxide stress. Then the quantitative RT-PCR indicated that MsHSP17.7was abundantly expressed under high salinity stress and peroxide stress in T3 transgenic Arabidopsis. In summary, results implied that MsHSP17.7 might involve in the regulation of high salinity stress and peroxide stress response. |
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| Keywords: | Medicago sativa Heat shock protein Quantitative RT-PCR Abiotic stress Genetic transformation |
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