巴西橡胶树一个AP2/EREBP转录因子的克隆及表达分析

杜磊1,2, 张德春1,*, 邓治2, 刘向红2,3, 李德军2,**
1三峡大学化学与生命科学学院, 湖北宜昌443002; 2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; 3海南大学农学院, 海口570228

通信作者:张德春;E-mail: djli.rricatas@gmail.com;Tel: 0898-23301174

摘 要:

本文采用RACE技术从巴西橡胶树中鉴定出一个AP2/EREBP转录因子。该转录因子全长cDNA为1 225 bp, 最长开放阅读框699 bp, 预测编码蛋白包含232个氨基酸; 序列比对分析发现, 该转录因子具有一段保守的AP2/EREBP结构域, 与拟南芥Soloist (At4g13040)类蛋白具有较高的相似性, 将该基因命名为HbSoloist。半定量RT-PCR分析结果表明, HbSoloist在巴西橡胶树的胶乳、叶片、树皮和花中均有表达。与健康橡胶树相比, 死皮树胶乳中HbSoloist表达量明显下降。研究发现HbSoloist表达受乙烯利、茉莉酸和低温处理调控, 表明HbSoloist基因可能在巴西橡胶树乙烯、茉莉酸和低温反应中发挥作用。

关键词:巴西橡胶树; AP2/EREBP; 转录因子; 基因克隆; 表达分析

收稿:2013-07-22   修定:2013-09-25

资助:海南省自然科学基金(311087)、国家自然科学基金(31270651和31200514)和国家天然橡胶现代农业产业技术体系建设项目(CARS-34-GW1)。

Cloning and Expression Analysis of an AP2/EREBP Gene in Hevea brasiliensis Muell. Arg.

DU Lei1,2, ZHANG De-Chun1,*, DENG Zhi2, LIU Xiang-Hong2,3, LI De-Jun2,**
1College of Chemistry and Life Science, China Three Gorges University, Yichang, Hubei 443002, China; 2Key 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; 3College of Agriculture, Hainan University, Haikou 570228, China

Corresponding author: ZHANG De-Chun; E-mail: djli.rricatas@gmail.com; Tel: 0898-23301174

Abstract:

In this study, a full-length cDNA sequence of AP2/EREBP transcription factor was obtained from Hevea brasiliensis with the RACE method, which was named as HbSoloist. The full-length cDNA of HbSoloist is 1 225 bp with a 699 bp open reading frame, encoding a deduced polypeptide of 232 amino acids. The deduced HbSoloist contains an AP2 domain, and shows high identity to ethylene-responsive transcription factor-like protein At4g13040-like (Soloist). Semi-quantitative RT-PCR analysis indicated that HbSoloist was ubiquitously expressed in latex, leaves, barks and flowers. Compared with tapping panel dryness (TPD) rubber tree, HbSoloist was upregulated in healthy rubber tree. The expression of HbSoloist was regulated by ethephon, jasmonic acid and low temperature treatments, suggesting that HbSoloist might play key roles in ethylene, jasmonic acid and low temperature responses in Hevea brasiliensis.

Key words: Hevea brasiliensis; AP2/EREBP; transcription factor; gene cloning; expression analysis

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