VIGS 介导的番茄转录因子SlDREB2 基因的耐旱特征

生物技术通报 ›› 2012, Vol. 0 ›› Issue (12): 93-100.

VIGS 介导的番茄转录因子SlDREB2 基因的耐旱特征

于国红, 马雪峰, 徐娜, 辛世超, 程宪国

农业部植物营养与肥料重点实验室中国农业科学院农业资源与农业区划研究所植物营养分子生物学实验室，北京 100081

收稿日期:2012-06-18 修回日期:2013-01-25 出版日期:2012-12-26 发布日期:2013-02-06
作者简介:于国红, 女, 硕士研究生, 研究方向:植物抗逆分子机理;E-mail:guangwen19840104@163.com
基金资助:
: 国家转基因重大专项（2011ZX002-5）

Profile of Drought Resistance of Tomato Transcription FactorSlDREB2 Gene Mediated by VIGS

Yu Guohong, Ma Xuefeng, Xu Na, Xin Shichao, Cheng Xianguo

The Key Laboratory of Plant Nutrition and Fertilizers，Ministry of Agriculture，Chinese Academy of Agricultural Sciences Institute ofAgricultural Resources and Regional Planning，Beijing 100081

Received:2012-06-18 Revised:2013-01-25 Published:2012-12-26 Online:2013-02-06

摘要/Abstract

摘要： SlDREB2 基因是利用rd29A 基因启动子的DRE 元件通过酵母单杂交技术从番茄幼苗（丽春）cDNA 文库中筛选得到的属于EREBP 家族的一个转录因子基因。利用构建病毒VIGS 载体，分别用含pBINTRB6、pTV00∷SlDREB2-1 和pTV00∷SlDREB2-2 重组载体的农杆菌GV3101 菌液灌根与叶片注射法侵染番茄植株，转化7 d 后进行病毒DNA 检测与验证。测定了干旱胁迫下侵染成功的番茄植株与野生型株系的生理指标，并通过Real-time qPCR 分析了VIGS 介导的SlDREB2 转录因子基因的表达特征。番茄植株干旱3 周胁迫处理后测定结果表明，野生番茄中脯氨酸和可溶性糖水平要高于被VIGS 病毒侵染的番茄植株，而丙二醛水平要低于被侵染植株，并且指标显示被pTV00∷SlDREB2-2 重组载体的农杆菌GV3101 菌液侵染的番茄植株抗旱性明显降低；复水1 周后，受VIGS 浸染株系丙二醛含量明显高于野生型，而可溶性糖与脯氨酸累积却低于野生型。Real-time qPCR 结果表明，经VIGS 病毒侵染过的番茄植株在胁迫处理时期SlDREB2 基因的相对表达量都明显低于野生型株系，表明番茄SlDREB2基因保守域末端459 bp 的特异片段具有较显著的沉默效果，说明VIGS 介导的SlDREB2 基因的表达受干旱诱导，该基因可以作为抗旱型作物品种改良的有价值基因。

关键词: VIGS, 番茄, DREB, 干旱胁迫, Real-time qPCR

Abstract: SlDREB2 gene was isolated by yeast one-hybrid system using DRE element of rd29A promoter through hybridization withtomato seedlings（Lichun）cDNA library, and it is a transcription factor belonging to EREBP family. In this study, the VIGS virus vectors wereconstructed, and transformed into the GV3101 Agrobacterium strains, and the resulting recombinants carrying pBINTRB6, pTV00-SlDREB2-1and pTV00-SlDREB2-2 vectors were used to infect leaves of tomato plants respectively by injection of liquid filling respectively. All the plantsinfected after 7 days were identified by viral DNA testing and the physiological index of wild-type strains of tomato plants as well as infectedplants successfully were determined under drought stress for three weeks, and Real-time qPCR was performed to analyze expression profileof the SlDREB2 transcription factor. Data showed that the contents of proline and soluble sugar in wild tomato plants were higher than that oftomato plants infected by VIGS virus, and malondialdehyde levels in wild plants were lower than that of plants infected by VIGS virus. Whenall tomato plants stressed by drought for three weeks were re-watered for one week, the contents of proline and soluble sugar in wild tomatoplants were lower than that of tomato plants by VIGS virus infection, and the contents of malondialdehyde in wild plants were higher than that ofplants infected by VIGS virus infection. Real-time qPCR results indicates that the expression of SlDREB2 in plants by VIGS virus infection waslower significantly than that of in wild type plants under drought stress, indicating that 500 bp of specific fragment at the end of SlDREB2 geneconservative domain has better silence effect, and the expression of SlDREB2 mediated by VIGS was inhibited by drought stress, and indirectlyimplying that the SlDREB2 gene can be a valuable candidate gene for improving crop varieties with drought resistance.

于国红, 马雪峰, 徐娜, 辛世超, 程宪国. VIGS 介导的番茄转录因子SlDREB2 基因的耐旱特征[J]. 生物技术通报, 2012, 0(12): 93-100.

Yu Guohong, Ma Xuefeng, Xu Na, Xin Shichao, Cheng Xianguo . Profile of Drought Resistance of Tomato Transcription FactorSlDREB2 Gene Mediated by VIGS[J]. Biotechnology Bulletin, 2012, 0(12): 93-100.

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