苹果6-磷酸山梨醇脱氢酶基因启动子逆境诱导表达特性研究

doi:10.13523/j.cb.20170608

中国生物工程杂志

2017, Vol. 37

Issue (6): 50-55 DOI: 10.13523/j.cb.20170608

研究报告

苹果6-磷酸山梨醇脱氢酶基因启动子逆境诱导表达特性研究

夏惠^1,2, 刘磊³, 王秀¹, 沈妍秋¹, 郭雨伦¹, 梁东^1,2

1. 四川农业大学园艺学院成都 611130;
2. 四川农业大学果蔬研究所成都 611130;
3. 成都市农林科学院成都 611130

Research on Stress-inducible Expression Characteristics of Sorbitol-6- phosphate Dehydrogenase Promoter from Apple

XIA Hui^1,2, LIU Lei³, WANG Xiu¹, SHEN Yan-qiu¹, GUO Yu-lun¹, LIANG Dong^1,2

1. College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China;
2. Institute of Pomology and Olericulture, Sichuan Agricultural University, Chengdu 611130, China;
3. Chengdu Academy of Agriculture and Forestry Sciences, Chengdu 611130, China

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摘要： 为研究6-磷酸山梨醇脱氢酶（sorbitol-6-phosphate dehydrogenase，S6PDH）基因启动子（S6PDHp）的逆境诱导表达特性，利用Gateway技术构建了S6PDH基因启动子区 5'端系列缺失体与GUS基因的融合表达载体，并通过农杆菌介导法转化拟南芥。对转基因拟南芥进行低温和外源ABA处理，通过GUS蛋白活性变化分析S6PDHp的逆境诱导表达特性。研究结果发现，通过Gateway技术构建了4个S6PDHp 5'端系列缺失体与β-葡萄糖苷酸酶（GUS）基因的融合表达载体（pGWB433-S6PDHp1、pGWB433-S6PDHp2、pGWB433-S6PDHp3和pGWB433-S6PDHp4）并获得了相应的转基因拟南芥。对转基因植株进行低温处理后发现，pGWB433-S6PDHp3转基因植株中的GUS活性增幅最大，达到显著水平，而其他转基因植株中的GUS活性基本保持不变。外源ABA处理后发现，除pGWB433-S6PDHp4外，其余启动子缺失体转基因拟南芥中GUS活性显著升高。以上结果表明，低温和外源ABA能够诱导S6PDHp的表达，但不同的缺失体响应程度不同，意味着在S6PDHp序列（-2 396bp至-236bp）中可能存在着响应逆境胁迫的正负调控顺式作用元件。

关键词： 6-磷酸山梨醇脱氢酶; 启动子; 苹果; 报告基因; 遗传转化

Abstract: In order to understand stress-inducible expression characteristics of sorbitol-6-phosphate dehydrogenase (S6PDH) promoter (S6PDHp) from apple, a series of 5' deletions of S6PDH promoter were fused with GUS reporter gene by Gateway and introduced into arabidopsis byagrobacterium-mediated transformation. The expression characteristics of S6PDHp were validated by GUS activity changed in transgenetic Arabidopsis against low temperature and exogenous ABA. Molecular detection analysis revealed that four 5' deletions of S6PDHp were fused with GUS reporter gene successfully (i.e. pGWB433-S6PDHp1, pGWB433-S6PDHp2, pGWB433-S6PDHp3 and pGWB433-S6PDHp4) and relevant transgenetic Arabidopsis were obtained. In low temperature treatment, GUS activity in pGWB433-S6PDHp3 transgenetic line increased markedly, while there were obvious change in other deletion transgenetic lines. In exogenous ABA treatment, GUS activity in others deletions transgenetic lines increased dramatically except pGWB433-S6PDHp4. These results suggest that S6PDHp can be induced by low temperature or exogenous ABA, however, there were different response in every deletions. These indicated that positive and negative cis-regulatory elements in response to abiotic stress may co-existed in the promoter region (-2 396bp to -236bp) of S6PDH.

Key words: Sorbitol-6-phosphate dehydrogenase Reporter gene Promoter Transformation apple

收稿日期: 2017-03-21 出版日期: 2017-06-25

ZTFLH:

S603.2

基金资助: 国家自然科学基金资助项目（31201600）

通讯作者: 梁东 E-mail: liangeast@sina.com

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引用本文:

夏惠, 刘磊, 王秀, 沈妍秋, 郭雨伦, 梁东. 苹果6-磷酸山梨醇脱氢酶基因启动子逆境诱导表达特性研究[J]. 中国生物工程杂志, 2017, 37(6): 50-55.

XIA Hui, LIU Lei, WANG Xiu, SHEN Yan-qiu, GUO Yu-lun, LIANG Dong. Research on Stress-inducible Expression Characteristics of Sorbitol-6- phosphate Dehydrogenase Promoter from Apple. China Biotechnology, 2017, 37(6): 50-55.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20170608 或 https://manu60.magtech.com.cn/biotech/CN/Y2017/V37/I6/50

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