《植物生理学报》 2015, 51(10): 1735-1742

巴西橡胶树TIM23-1基因克隆、进化及表达分析

陈江淑^1,2, 邓治², 刘辉², 范玉龙¹, 姜达¹, 夏立琼¹, 夏志辉^1,*, 李德军^2,*

¹海南大学农学院, 海口570228; 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

通信作者：夏志辉；E-mail: zhxia-111@163.com; dragonldj@163.com；Tel: 0898-23301174

摘　要：

本研究采用RACE技术, 从巴西橡胶树中鉴定出一个线粒体内膜转位因子TIM23基因。该基因全长cDNA为898 bp, 最长开放阅读框567 bp, 预测编码蛋白包含188个氨基酸; 序列比对分析发现, 该基因编码蛋白具有转膜区和PRAT结构域, 与拟南芥TIM23-1蛋白具有较高的相似性, 将该基因命名为HbTIM23-1。实时定量RT-PCR分析结果表明, HbTIM23-1在巴西橡胶树胶乳、叶片、树皮、雄花、雌花、花药中均有表达。在橡胶树叶片不同发育时期, HbTIM23-1表达存在变化。与健康橡胶树相比, 死皮橡胶树胶乳中HbTIM23-1表达量明显下降。研究发现HbTIM23-1表达受干旱和低温处理调控, 表明Hb-TIM23-1可能在巴西橡胶树干旱和低温胁迫应答及死皮中发挥作用。

关键词：巴西橡胶树; 线粒体内膜转位因子23; 死皮; 基因克隆; 表达分析

收稿：2015-05-19   修定：2015-09-09

资助：国家自然科学基金(31270651和31200514)和中国热带农业科学院橡胶研究所基本科研业务费专项(163002 -2015003)。

Cloning, Phylogenetic and Expression Analyses of TIM23-1 Gene in Hevea brasiliensis

CHEN Jiang-Shu^1,2, DENG Zhi², LIU Hui², FAN Yu-Long¹, JIANG Da¹, XIA Li-Qiong¹, XIA Zhi-Hui^1,*, LI De-Jun^2,*

¹College of Agriculture, Hainan University, Haikou 570228, 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

Corresponding author: XIA Zhi-Hui; E-mail: zhxia-111@163.com; dragonldj@163.com; Tel: 0898-23301174

Abstract:

In this study, a full-length cDNA sequence of TIM23 gene was cloned from Hevea brasiliensis with the rapid-amplification of cDNA ends (RACE) method, and the gene was named as HbTIM23-1. The full length cDNA of HbTIM23-1 is 898 bp in size with a 567 bp open reading frame, encoding a deduced polypeptide of 188 amino acids. The deduced HbTIM23-1 contains a predicted transmembrane region and preprotein and amino acid transporter (PRAT) domain and indicates high identity to AtTIM23-1 protein. Real-time RT-PCR analyses indicated that HbTIM23-1 was expressed in latex, barks, leaves, barks, male flowers, female flowers and anthers. With the development of leaves, the HbTIM23-1 expression showed a significant change. Compared with healthy rubber tree, HbTIM23-1 was down-regulated in tapping panel dryness (TPD) rubber tree latex. The expression of HbTIM23-1 was regulated by drought and low temperature treatments, suggesting that HbTIM23-1 might play important roles in drought and low temperature responses as well as TPD in H. brasiliensis.

Key words: Hevea brasiliensis; TIM23; tapping panel dryness; gene cloning; expression analysis