| 小麦热激蛋白基因TaHSP90-1的克隆与表达分析 |
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| 引用本文： | 陆青,梁婷,王伟伟,汪德州,吴娴,王小燕,唐益苗.小麦热激蛋白基因TaHSP90-1的克隆与表达分析[J].中国农业科技导报,2022,24(8):44-54. |
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| 作者姓名： | 陆青 梁婷 王伟伟 汪德州 吴娴 王小燕 唐益苗 |
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| 作者单位： | 1.长江大学农学院，湖北 荆州 434025;2.北京市农林科学院杂交小麦研究所，北京 100097 |
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| 基金项目： | 北京市农林科学院科技创新能力建设专项(KJCX20220421);北京市农林科学院协同创新中心资助项目(KJCX201907-2) |
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| 摘 要： | 热激蛋白（heat shock protein，HSPs）是一类广泛存在于植物体内的应激蛋白，在植物响应逆境过程中发挥重要作用。为进一步了解小麦HSP90-1基因并探究其相关生物学功能，搜索小麦基因组序列数据库，获得了小麦A、B、D 3个基因组上的同源序列，根据其染色体位置分别命名为TaHSP90-1-A、TaHSP90-1-B和TaHSP90-1-D，通过同源克隆从12个小麦品种中得到TaHSP90-1 DNA序列。序列分析表明，TaHSP90-1-A、TaHSP90-1-B和TaHSP90-1-D分别含有2 121、2 136和2 130 bp的完整开放阅读框，分别编码707、712和710个氨基酸；其蛋白序列C端均含有1个HSP90结构域，为HSP家族HSP90亚家族成员。TaHSP90-1在不同小麦品种中存在SNP位点，分别位于TaHSP90-1-B第2个内含子区域和TaHSP90-1-D启动子区域。系统进化分析表明，TaHSP90-1-A与二粒小麦处于同一分支；TaHSP90-1-B和TaHSP90-1-D处于同一分支。顺式作用元件分析发现，TaHSP90-1-A、TaHSP90-1-B和TaHSP90-1-D启动子区域均含有干旱响应元件（MBS element）和热响应元件（HSE element）。RT-PCR结果显示，TaHSP90-1-A、TaHSP90-1-B和TaHSP90-1-D在苗期和成熟期的叶片中表达量较高。在干旱胁迫条件下，TaHSP90-1表达量在耐旱小麦品种中较高，为旱敏感小麦品种表达量的150倍；在热胁迫条件下，TaHSP90-1在热敏感与耐热小麦品种中均上调表达，其中，TaHSP90-1-A和TaHSP90-1-D在耐热小麦品种中上调表达倍数更高。
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| 关 键 词： | 小麦 HSP90 干旱胁迫 热胁迫 SNP |
| 收稿时间： | 2022-05-11 |
Cloning and Expression Analysis of Wheat Heat Shock Protein Gene TaHSP90-1 |
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| Qing LU,Ting LIANG,Weiwei WANG,Dezhou WANG,Xian WU,Xiaoyan WANG,Yimiao TANG.Cloning and Expression Analysis of Wheat Heat Shock Protein Gene TaHSP90-1[J].Journal of Agricultural Science and Technology,2022,24(8):44-54. |
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| Authors: | Qing LU Ting LIANG Weiwei WANG Dezhou WANG Xian WU Xiaoyan WANG Yimiao TANG |
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| Institution: | 1.College of Agriculture，Yangtze University，Hubei Jingzhou 434025，China;2.Institute of Hybrid Wheat，Beijing Academy of Agriculture and Forestry Sciences，Beijing 100097，China |
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| Abstract: | Heat shock proteins （HSPs） are a kind of stress proteins widely exist in plants and play important role in the response of plants to stress. In order to further understand the wheat HSP90-1 gene and explore its related biological functions，this paper searched the wheat genome sequence database， and obtained the homologous gene sequences on wheat A， B and D subgenomes， which were named as TaHSP90-1-A， TaHSP90-1-B and TaHSP90-1-D， respectively. TaHSP90-1 DNA sequences were obtained from 12 wheat varieties through homologous cloning and sequence analysis showed that the complete open reading frames of TaHSP90-1-A， TaHSP90-1-B and TaHSP90-1-D were 2 121， 2 136 and 2 130 bp， encoding 707， 712 and 710 amino acids， respectively. The C-terminal sequence of TaHSP90-1-A， TaHSP90-1-B and TaHSP90-1-D contained an HSP90 domain， which were members of HSP90 subfamily in HSP family. SNP loci were found among different wheat varieties， which were located in the second intron region of TaHSP90-1-B and the promoter region of TaHSP90-1-D. Phylogenetic analysis showed that TaHSP90-1-A and two grain wheat were in the same branch， and TaHSP90-1-B and TaHSP90-1-D were in the same branch. Cis acting element analysis showed that the promoter regions of TaHSP90-1-A， TaHSP90-1-B and TaHSP90-1-D contained drought response element （MBS element） and thermal response element （HSE element）. RT-PCR showed that TaHSP90-1-A， TaHSP90-1-B and TaHSP90-1-D had high expression in leaves at seedling and mature stages of wheat. Under drought stress， the expression level of TaHSP90-1 was higher in drought tolerant wheat varieties， which was 150 times compared with varieties of drought sensitive. Under heat stress， TaHSP90-1 was up-regulated in both heat sensitive and heat resistant varieties， while TaHSP90-1-A and TaHSP90-1-D were significantly up-regulated in heat-resistant varieties of wheat. |
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| Keywords: | wheat HSP90 drought stress heat stress SNP |
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