橡胶树ADC1的克隆、表达及生物信息学分析

doi:10.13560/j.cnki.biotech.bull.1985.2018-0498

摘要/Abstract

摘要： 精氨酸脱羧酶（Arginine decarboxylase,ADC）是高等植物多胺合成的重要酶。为对橡胶树ADC基因表达模式及生物信息学进行分析,采用PCR和测序技术从橡胶树中克隆了一个ADC基因（HbADC1）,获得的HbADC1序列长2 594 bp,开放阅读框为2 175 bp,编码724个氨基酸。生物信息学预测HbADC1理论分子量和等电点分别为77.7 kD和5.14。进化分析表明,HbADC1与大戟科植物木薯、麻风树的ADC聚为一支。qRT-PCR分析表明,HbADC1表达无组织特异性,在雌花中表达量最高,茎尖、雄花、叶片、树皮和胶乳次之。HbADC1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HbADC1可能参与橡胶树叶片发育过程。此外,HbADC1表达受伤害、低温、干旱、高盐、过氧化氢及乙烯调控,表明HbADC1可能参与橡胶树逆境胁迫和乙烯应答过程。

关键词: 橡胶树, 精氨酸脱羧酶, 基因克隆, 生物信息学, 实时荧光定量PCR

Abstract: Arginine decarboxylase（ADC）is an important enzyme involved in polyamine biosynthesis in higher plants. In order to study the expression and bioinformatics of ADC gene from rubber tree,HbADC1 from Hevea brasiliensis was cloned with PCR and sequencing methods. The sequence length of HbADC1 was 2 594 bpwith a 2 175 bp open reading frame,encoding 724 amino acids. Bioinformatics results indicated that the deduced molecular weight and isoelectric point of HbADC1 were 77.7 kD and 5.14,respectively. Phylogenetic analyses showed that HbADC1 and ADCs from Manihot esculenta and Jatropha curcas was clustered into one. qRT-PCR analyses demonstrated that HbADC1 expressed without tissue specificity,with the highest in female flowers,followed by stem apexes,male flowers,leaves,barks,and latex. HbADC1 expression significantly varied in different developmental stages of rubber tree leaf,with the highest expression in light young stage and the lowest one in mature stage,suggesting that HbADC1 might be associated with leaf development. In addition,HbADC1 expression were regulated by wounding,low temperature,drought,high salt,hydrogen peroxide,and ethylene treatments,indicating that HbADC1 might be involved in stress and ethylene responses in rubber tree.

Key words: Hevea brasiliensis, arginine decarboxylase, gene cloning, bioinformatics analysis, qRT-PCR

侯岚菲, 杨洪, 邓治, 代龙军, 门中华, 李德军. 橡胶树ADC1的克隆、表达及生物信息学分析[J]. 生物技术通报, 2018, 34(11): 111-119.

Hou Lan-fei, Yang Hong, Deng Zhi, Dai Long-jun, Men Zhong-hua, Li De-jun. Cloning,Expression and Bioinformatics Analyses of an ADC1 Gene in Hevea brasiliensis[J]. Biotechnology Bulletin, 2018, 34(11): 111-119.

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