茶树‘黄金芽’ CsHIPP26.1基因克隆与光响应表达分析

范延艮,刘富浩,赵秀秀,王域,张丽霞*
山东农业大学园艺科学与工程学院, 作物生物学国家重点实验室, 山东泰安271018

通信作者:张丽霞;E-mail: lxzhang@sdau.edu.cn

摘 要:

本研究以茶树(Camellia sinensis) ‘黄金芽’芽下第二叶为材料, 克隆了前期蛋白组学分析获得的一个响应光强变化且在‘黄金芽’中高表达的重金属相关异戊二烯化植物蛋白(HIPP)基因, 将其命名为CsHIPP26.1, GenBank登记号: MK654903。该基因DNA全长为465 bp, 编码154个氨基酸, 蛋白分子质量为17.01 kDa; 理论等电点为9.13, 无跨膜结构域; 共有16个磷酸化位点, 位于丝氨酸、苏氨酸和酪氨酸三个氨基酸残基上; 二级结构中含α螺旋28.6%、 310螺旋3.9%、 β折叠2.5%、 β转角6.5%, 此外还含无序化区域28.6%, 且主要分布于N端和C端; 亚细胞定位于核中。以不同遮荫度‘黄金芽’新梢为材料进行实时荧光定量PCR (qRT-PCR)实验, 结果显示CsHIPP26.1表达响应光照强度变化, 且随光照强度增大表达量增加; 将CsHIPP26.1转化苹果(Malus pumila) ‘王林’愈伤组织, 能助其在强光下正常生长。

关键词: ‘黄金芽’; 重金属相关异戊二烯化植物蛋白; 基因克隆; 生物信息学分析; 表达分析

收稿:2021-01-13   修定:2021-04-20

资助:山东省“双一流”奖补资金(SYL2017YY03)和山东省现代农业产业技术体系创新团队项目(SDAIT-19-05)

Cloning of CsHIPP26.1 in Camellia sinensis ‘Huangjinya’ and analysis of its expression level in response to light intensity

FAN Yangen, LIU Fuhao, ZHAO Xiuxiu, WANG Yu, ZHANG Lixia*
College of Horticulture Science and Engineering, State Key Laboratory of Crop Biology, Shandong Agricultural University, Taian, Shandong 271018, China

Corresponding author: ZHANG Lixia; E-mail: lxzhang@sdau.edu.cn

Abstract:

In this study, a gene of heavy metal–associated isopentenylated plant protein (HIPP), named
CsHIPP26.1, was cloned from the second leaf of Camellia sinensis ‘Huangjinya’. Its GenBank accession number is MK654903. CsHIPP26.1 protein was highly expressed in ‘Huangjinya’ in response to the changes of light intensity by proteomic analysis. The full-length cDNA of CsHIPP26.1 is 465 bp, encoding 154 amino acids with a protein molecular weight of 17.01 kDa. The theoretical isoelectric point of CsHIPP26.1 is 9.13 and this protein had no transmembrane domain. Besides, there are 16 phosphorylation sites on the three amino acid residues of l-serine, l-threonine and l-tyrosine. The protein secondary structure of CsHIPP26.1 contains 28.6% α-helix, 3.9% 310-helix, 32.5% β-fold, 6.5% β-turn, and 28.6% disordered region, mainly distributed in N-terminal and C-terminal. Subcellular localization showed that the gene is located in the nucleus. The quantitative real-time PCR (qRT-PCR) experiment with different shading degrees of ‘Huangjinya’ showed that the expression of CsHIPP26.1 responded to the changes of light intensity, and the expression increased with consistent to light intensity. Besides, the  transformation of CsHIPP26.1 into apple (Malus pumila) ‘Orin’ callus could help the callus grow normally under strong light.

Key words: ‘Huangjinya’; heavy metal–associated isoprenylated plant protein; gene cloning; bioinformatics analysis; expression analysis

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