《植物生理学报》 2017, 53(8): 1392-1398

巴西橡胶树HbTFIIB的克隆与表达分析

晁金泉, 陈月异, 杨署光, 田维敏^*

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/省部共建国家重点实验室培育基地-海南省热带作物栽培生理学重点实验室, 海南儋州571737

通信作者：田维敏；E-mail: wmtian@163.com

摘　要：

本研究采用RACE和RT-PCR方法, 从巴西橡胶树无性系热研7-33-97的乳管细胞中克隆到转录复合体关键成员TFIIB的同源基因, 命名为HbTFIIB。该基因全长1 472 bp, 其中开放阅读框810 bp, 编码一个由270个氨基酸残基组成的蛋白质。该蛋白质分子量为29.48 kDa, 理论等电点为8.3。实时荧光定量PCR结果表明, HbTFIIB基因受割胶上调表达, 而且在后期排出的胶乳中表达量显著高于前期排出的胶乳。Western blot分析表明, 开割树胶乳中的HbTFIIB蛋白含量高于未开割树, 而且在早期排出的胶乳中的含量明显高于后期排出的胶乳中的含量。本研究表明, 割胶促进转录复合体蛋白基因表达和转录蛋白质合成, 而且排胶会流失转录复合体蛋白。

关键词：橡胶树; 乳管细胞; 胶乳; 转录复合体; HbTFIIB

收稿：2016-12-14   修定：2017-07-03

资助：现代农业产业技术体系建设专项资金(CARS-34-GW1)和中国热带农业科学院橡胶研究所基本科研业务费(1630022014004)。

Cloning and expression analysis of HbTFIIB in rubber tree (Hevea brasiliensis)

CHAO Jin-Quan, CHEN Yue-Yi, Yang Shu-Guang, TIAN Wei-Min^*

Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Minstry of Agriculture Key Laboratory of Rubber Biology/State Key Laboratory Incubation Base for Cultivation & Physiology of Tropical Crops, Danzhou, Hainan 571737, China

Corresponding author: TIAN Wei-Min; E-mail: wmtian@163.com

Abstract:

In the present study, a homologue of TFIIB was cloned from the latex of rubber tree clone CATAS7-33-97 by the rapid amplification of cDNA ends (RACE) and RT-PCR techniques. The cDNA was named as HbTFIIB. HbTFIIB was 1 472 bp in length, and contained a 810 bp open reading frame (ORF) encoding a putative protein of 270 amino acids. The predicted molecular mass of HbTFIIB was 29.48 kDa with a pI of 8.3. Real-time RT-PCR analysis showed that HbTFIIB expression was up-regulated by tapping. Its expression level in the latex collected at the early stage of latex flow was significantly higher than that collected at the late stage of latex flow. Although the content of HbTFIIB protein increased upon tapping, it significantly decreased in the latex collected at the late stage of latex flow. The results suggested that the transcription and translation of transcriptional complex may be enhanced by tapping and the transcriptional complex could be lost during latex flow.

Key words: Hevea brasiliensis; laticifer cells; latex; transcriptional complex; HbTFIIB