巴西橡胶树HbAIH基因的克隆及表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2019-0940

生物技术通报 ›› 2020, Vol. 36 ›› Issue (5): 120-129.doi: 10.13560/j.cnki.biotech.bull.1985.2019-0940

巴西橡胶树HbAIH基因的克隆及表达分析

杨洪¹, 岳镒繁^1,2, 胡燕玲^1,2, 邓治¹, 代龙军¹, 李德军¹

1.中国热带农业科学院橡胶研究所农业农村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,海口 571101;
2.南京农业大学农学院,南京 210095

收稿日期:2019-10-07 出版日期:2020-05-26 发布日期:2020-06-03
作者简介:杨洪,男,助理研究员,研究方向：植物生理及分子生物学;E-mail：yang_hong0317@126.com;岳镒繁为共同第一作者
基金资助:
国家自然科学基金项目（31570684）,国家重点研发计划（2018YFD1000502）

Cloning, Expression Analyses of HbAIH Gene from Hevea brasiliensis

YANG Hong¹, YUE Yi-fan^1,2, HU Yan-ling^1,2, DENG Zhi¹, DAI Long-jun¹, LI De-jun¹

1. Rubber Research Institute，Chinese Academy of Tropical Agricultural Sciences，Key Laboratory of Biology and Genetic Resources of Rubber Tree for Ministry of Agriculture and Rural Affairs，State Key Laboratory Incubation Base for Cultivation and Physiology for Tropical Crops，Hainan Provincial Key Laboratory of Tropical Crops Cultivation and Physiology，Haikou 571101；
2. College of Agriculture，Nanjing Agricultural University，Nanjing 210095

Received:2019-10-07 Published:2020-05-26 Online:2020-06-03

摘要/Abstract

摘要： 鲱精胺亚胺水解酶（Agmatine iminohydrolase，AIH）是植物通过精氨酸途径合成多胺过程中的重要酶之一。本研究首次从巴西橡胶树中克隆了AIH基因HbAIH。序列分析结果表明HbAIH全长1 462 bp，开放阅读框长为1 137 bp，编码378个氨基酸。预测 HbAIH 蛋白的分子量是42.28 kD，等电点为5.09，是一个亲水性蛋白。氨基酸相似性比对结果表明HbAIH与大戟科木薯MeAIH、蓖麻RcAIH和麻风树JcAIH等具有很高相似性，且含有植物AIH中高度保守的与酶活性位点形成和参与底物结合的11个氨基酸位点。qRT-PCR 分析表明，HbAIH表达无组织特异性，在雌花中表达量最高，树皮、茎尖、胶乳、叶片和雄花中表达量依次降低。HbAIH在不同发育时期叶片中的表达存在变化，稳定期表达量最低，淡绿期最高，说明HbAIH可能参与橡胶树叶片发育过程。此外，低温、干旱、高盐、过氧化氢、乙烯利、茉莉酸甲酯处理以及橡胶树死皮的发生均能引起HbAIH表达变化。这些结果说明HbAIH可能参与了橡胶树生长发育、产排胶调控及逆境应答过程。

关键词: 巴西橡胶树, 鲱精胺亚胺水解酶, 胁迫响应, 表达分析

Abstract: Agmatine iminohydrolase（AIH）in plant is one of the most important enzymes in the arginine decarboxylase pathway in which arginine is subsequently converted to poluamine. This study is the first time for an AIH gene，HbAIH，to be cloned from Hevea brasiliensis. The full-length sequence of HbAIH was 1 462 bp with a 1 137 bp open reading frame，encoding a sequence with 378 amino acid residues. HbAIH was a hydrophilic protein，with a calculated molecular weight of 42.28 kD and an isoelectric point of 5.09. Sequence alignment revealed that the deduced amino acid sequence of HbAIH presented the high similarity with those of MeAIH in Manihot esculenta，RcAIH in Ricinus communis and JcAIH in Jatropha curcas from Euphorbiaceae，and also contained 11 highly conservative amino acid residues that formed the active sites and participated in the binding with the substrate. qRT-PCR analyses demonstrated that HbAIH expressed with no tissue specificity，the highest in female flowers，followed by barks，stem apexes，latex，leaves and male flowers. HbAIH expression significantly varied in different developmental stages of rubber tree leaves，with the highest expression in light young stage and the lowest one in mature stage，suggesting that HbAIH might be associated with leaf development. In addition，the expression of HbAIH was induced by wounding，low temperature，drought，high salt，hydrogen peroxide，ethrel，MeJA treatments，and tapping panel dryness. All these results suggest that HbAIH is likely involved in growth and development of H. brasiliensis，regulating latex biosynthesis and latex flow，as well as stress responses.

Key words: Hevea brasiliensis, agmatine iminohydrolase, stress responses, expression analysis

杨洪, 岳镒繁, 胡燕玲, 邓治, 代龙军, 李德军. 巴西橡胶树HbAIH基因的克隆及表达分析[J]. 生物技术通报, 2020, 36(5): 120-129.

YANG Hong, YUE Yi-fan, HU Yan-ling, DENG Zhi, DAI Long-jun, LI De-jun. Cloning, Expression Analyses of HbAIH Gene from Hevea brasiliensis[J]. Biotechnology Bulletin, 2020, 36(5): 120-129.

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巴西橡胶树HbAIH基因的克隆及表达分析

Cloning, Expression Analyses of HbAIH Gene from Hevea brasiliensis

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