《植物生理学报》 2014, 50(11): 1707-1716

番茄低温响应转录因子SlNAC41克隆及表达分析

刘辉¹, 王涛涛², 张俊红², 欧阳波², 李汉霞^2,*

¹中国热带农业科学院橡胶研究所, 农业部橡胶树生物学与遗传资源利用重点实验室, 海南儋州571737; ²华中农业大学园艺林学学院, 园艺植物生物学教育部重点实验室, 武汉430070

通信作者：李汉霞；E-mail: hxli@mail.hzau.edu.cn；Tel: 027-87286867

摘　要：

NAC转录因子在调控植物生长发育、生物及非生物逆境应答中发挥着重要作用。前期, 我们通过对番茄幼苗在低温胁迫下的基因表达谱进行分析, 发现Unigene SGN-U212711受低温诱导表达强烈。本研究从番茄中克隆了该基因, 命名为SlNAC41, 其开放阅读框(ORF) 1 173 bp, 编码390个氨基酸, 蛋白N端具有典型的NAM结构域, 属于NAC转录因子家族成员。预测SlNAC41蛋白分子量为43.5 kDa, 等电点为5.2。实时荧光定量PCR分析表明, SlNAC41在番茄各组织均有表达, 在花中的表达量最高, 在红熟果中的表达量最低。低温、干旱、高盐、甲基紫精(MV)、脱落酸(ABA)及乙烯利(ETH)处理均能诱导该基因的表达, 其中, 以低温和干旱诱导表达最为强烈。利用PLACE和PlantCARE对启动子序列进行预测分析发现, SlNAC41启动子区含有大量响应光、病原菌侵染、激素、低温、脱水及盐胁迫的顺式作用元件。这些结果表明, SlNAC41可能在番茄生物及非生物胁迫应答中发挥重要调控作用。

关键词：番茄; NAC转录因子; 基因克隆; 表达分析; 非生物胁迫

收稿：2014-09-09   修定：2014-10-14

资助：国家自然科学基金(31301789)和国家“863”计划(2012AA-100104)。

Cloning and Expression Analysis of a Cold-Responsive Transcription Factor SlNAC41 in Tomato

LIU Hui¹, WANG Tao-Tao², ZHANG Jun-Hong², OUYANG Bo², LI Han-Xia^2,*

¹Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan 571737, China; ²Key Laboratory of Horticultural Plant Biology, Ministry of Education, College of Horticulture and Forestry, Huazhong Agricultural University, Wuhan 430070, China

Corresponding author: LI Han-Xia; E-mail: hxli@mail.hzau.edu.cn; Tel: 027-87286867

Abstract:

NAC transcription factors play vital roles in regulating plant growth and developmental processes as well as in response to biotic and abiotic stresses. In our previous study, Unigene SGN-U212711 was found to be strongly induced by low temperature through analyzing the gene expression profiles of tomato seedlings under cold stress. In this study, we cloned this gene (designated as SlNAC41) from tomato. SlNAC41 had an open reading frame (ORF) of 1 173 bp encoding 390 amino acids with a typical NAM domain in its N-terminal, suggesting that it belonged to NAC transcription factor family. The molecular weight and isoelectric point of SlNAC41 protein was 43.5 kDa and 5.2, respectively. Quantitative real-time PCR analysis showed that SlNAC41 was expressed in all tested tomato tissues, with the highest expression in flowers and the lowest expression in mature red fruits. Expression of SlNAC41 was induced by cold, drought, salt, methyl viologen (MV), abscisic acid (ABA) and ethephon (ETH) treatments, especially was strongly induced by cold and drought stresses. Promoter sequence analyzed by PLACE and PlantCARE showed that there were many cis-acting regulatory elements involved in response to light, pathogen infection, hormones, cold, dehydration and salt stresses. These results suggest that SlNAC41 may play important roles in regulating biotic and abiotic stress responses in tomato.

Key words: tomato; NAC transcription factor; gene cloning; expression analysis; abiotic stress