《植物生理学报》 2018, 54(5): 763-772
通信作者：李维国；E-mail: rekeyuanlywmm@126.com
摘 要：
CPP基因家族是广泛存在于动植物中, 成员数目较少的一类转录因子家族, 在植物生殖发育和细胞分裂过程中起着重要的调控作用。本研究以橡胶树优良品种‘热研7-33-97’为供试材料, 通过RT-PCR方法, 在橡胶树花器官中克隆到一个CPP基因, 命名为HbCPP1 (登录号为MF360959)。序列分析表明, HbCPP1基因开放阅读框为1 755 bp, 编码584个氨基酸, 蛋白质分子量为63.676 kDa,等电点(PI)为8.4。序列多重比对结果显示, Hb-CPP1蛋白具有CPP蛋白的特征基序, 含有2个保守的CXC结构域和1个比较保守R结构域。进化树分析表明, HbCPP1属于A亚族基因, 与其近缘物种木薯的MeCPP基因以及麻风树的JcCPP7基因同源性较高, 进化关系较近。氨基酸组分、理化性质以及蛋白结构预测分析显示, HbCPP1蛋白是一个不具有跨膜结构和信号肽的不稳定亲水蛋白, 可能定位于细胞核, 其二级结构主要由两个蛋白结合区域和多个无规则的卷曲螺旋组成, 其三级结构中CRC结构域构成了该蛋白的核心结构。利用QRT-PCR对HbCPP1基因在橡胶树不同组织中的表达量进行检测, 分析结果表明HbCPP1基因在橡胶树茎尖、花序以及雌花中特异性高调表达。在干旱、盐、高温胁迫以及ABA处理下, HbCPP1基因的表达量呈显著下调表达; 而在低温处理下, HbCPP1基因的表达量则呈显著上调表达, 推测HbCPP1基因可能参与了橡胶树花器官发育以及对多种非生物学胁迫的响应过程。关键词：巴西橡胶树; CPP基因; 基因克隆; 表达分析; 胁迫处理
收稿：2017-08-10 修定：2018-04-04
资助：海南省重要科技计划项目(ZDKJ2016020)和国家天然橡胶产业技术体系项目(CARS-34)。
Corresponding author: LI Wei-Guo; E-mail: rekeyuanlywmm@126.com
Abstract:
The CPP gene family is a family of transcription factors widely found in plants and animals, and plays an important role in the regulation of plant reproductive development and cell division. In this study, a CPP gene was cloned from the floral organs of by RT-PCR method using a cDNA template from rubber tree (Hevea brasiliensis) cultivar ‘Reyan7-33-97’. This gene was named as HbCPP1 (accession number MF360959). Sequence analysis showed that the open reading frame of HbCPP1 gene was 1 755 bp length, which encoding 584 amino acids, the molecular weight of protein was 63.676 kDa, and the isoelectric point (pI) was 8.4. Multiple alignments showed that HbCPP1 protein contained two conserved CXC domains and a relatively conserved R domain, which are the characterized motifs of CPP protein. Analysis of phylogenetic tree showed that HbCPP1 belonged to the A subfamily gene, and has higher similarity with the MeCPP gene in cassava and the JcCPP7 gene in Jatropha, and the phylogenetic relationship was close. The amino acid composition, physical and chemical properties, and structure prediction analysis showed that the HbCPP1 protein was an unstable hydrophilic protein with no trans-membrane and signal peptide, and which may be localized in the nucleus. The secondary structure of HbCPP1 proteins was mainly composed of two protein binding regions and a number of irregular coiled coils. In the 3D model structure, the CRC domain constituted the core structure of HbCPP1 proteins protein. The expression of HbCPP1 gene in different tissues of rubber tree was detected by QRT-PCR. The results showed that HbCPP1 gene expressed specifically in the stem tip, inflorescence and female flower. Under drought, salt, high temperature stress, and ABA treatment, the expression level of HbCPP1 gene was significantly down-regulated; while in low temperature treatment, the expression level of HbCPP1 gene was significantly up-regulated. It was speculated that HbCPP1 gene may be involved in the development of floral organs and responses to many abiotic stresses in H. brasiliensis.Key words: Hevea brasiliensis; CPP gene; gene cloning; expression analysis; stress treatment
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