巴西橡胶树HbbHLH1启动子克隆与序列分析.pdfVIP

分子植物育种，2009 年，第7 卷，第3 期，第531-536 页 Molecular Plant Breeding, 2009, Vol.7, No.3, 531-536 研究报告 Research Report 巴西橡胶树HbbHLH1 启动子的克隆与序列分析 1,2 2 1,2 2,3* 张全琪 朱家红 蔡元保 张治礼 1 海南大学园艺园林学院, 儋州, 571737; 2 中国热带农业科学院热带生物技术研究所, 海口, 571101; 3 海南省农业科学院, 海口, 571000 * 通讯作者, zzl_catas@ 摘 要 bHLH 转录因子在高等植物生长发育、激素应答和次生代谢调节中具有重要的功能。根据巴西橡胶 树(Hevea brasiliensis) HbbHLH1 基因设计了2 个特异性反向引物，利用GenomeWalking 方法从巴西橡胶树 叶片基因组DNA 中克隆获得了HbbHLH1 基因起始密码子上游819 bp 的调控片段。序列分析表明，该片段 除了含有一个典型的真核生物核心启动子区域(-60~-10 bp)外，还含有多个TATA-box、CAAT-box 等启动子 元件以及多种与激素和胁迫诱导相关的顺式调控元件，其中在-290 bp 和-309 bp 位点分别有一个应答Me- JA 信号反应的调控元件，但没有发现典型的乙烯响应元件。这暗示了HbbHLH1 转录因子可能在橡胶树乳 管分化和胶乳生物合成中具有调控作用。 关键词 巴西橡胶树, HbbHLH1 启动子, 顺式调控元件, 茉莉酸, 乙烯 Cloning and Sequence Analysis of HbbHLH1 Promoter from Hevea brasiliensis Zhang Quanqi 1,2 Zhu Jiahong 2 Cai Yuanbao 1,2 Zhang Zhili 2,3* 1 College of Horticulture and Landscape, Hainan University, Danzhou, 571737; 2 Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou, 571101; 3 Hainan Academy of Agricultural Sciences, Haikou, 571000 * Corresponding author, zzl_catas@ DOI: 10.3969/mpb.007.000531 Abstract The basic/helix-loop-helix (bHLH) transcription factors play an important role in the regulation of plant growth and development, hormone responses and secondary metabolism. Two gene specific reverse primers were designed from the 5 region of HbbHLH1 cDNA from Hevea brasiliensis and the 819 bp 5 regulatory sequence of HbbHLH1 was cloned from leaf genomic DNA of H. brasiliensis by GenomeWalking method. Sequence analysis showed that it consisted of a typical core promoter region of eukaryotic (-60~-10 bp), several promoter element such as TATA-box and CAAT-box, as well as many cis-regulatory elements related to hormone and stress respons- es, in which two cis-acting elements involved in the MeJA-responses were identified at -290 bp and