小麦叶绿素酸酯a氧化酶(TaCAO)基因的克隆与表达分析

doi:10.6048/j.issn.1001-4330.2021.09.001

新疆农业科学 ›› 2021, Vol. 58 ›› Issue (9): 1569-1576.DOI: 10.6048/j.issn.1001-4330.2021.09.001

• 作物遗传育种·种质资源·分子遗传学·耕作栽培 • 上一篇下一篇

小麦叶绿素酸酯a氧化酶(TaCAO)基因的克隆与表达分析

王重^1,2, 樊哲儒^1,2, 张跃强^1,2, 李剑峰^1,2, 高新^1,3, 时佳^1,3

1.新疆农业科学院核技术生物技术研究所,乌鲁木齐 830091;
2.农业部荒漠绿洲作物生理生态与耕作重点实验室,乌鲁木齐 830091;
3.新疆农作物生物技术重点实验室,乌鲁木齐 830091

收稿日期:2020-06-23 出版日期:2021-09-20 发布日期:2021-09-29
通信作者: 樊哲儒(1964-),男,甘肃民勤人,研究员,研究方向为小麦遗传育种,(E-mail)fzr640814@qq.com
作者简介:王重(1982-),男,陕西长安人,副研究员,研究方向为小麦遗传育种,(E-mail)wangzhongac@126.com
基金资助:
自治区公益性科研院所基本科研业务经费(KY2018037/KY2019006)

Cloning and Expression Analysis of Chlorophyll a Oxygenase Gene in Wheat

WANG Zhong^1,2, FAN Zheru^1,2, ZHANG Yueqiang^1,2, LI Jianfeng^1,2, GAO Xin^1,3, SHI Jia^1,3

1. Research Institute of Nuclear and Biological Technology, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, China;
2. Key Laboratory of Desert-Oasis Crop Physiology, Ecology and Cultivation, MOARA, Urumqi 830091, China;
3. Xinjiang Key Laboratory of Crop Biotechnology, Urumqi 830091, China

Received:2020-06-23 Online:2021-09-20 Published:2021-09-29
Correspondence author: FAN Zheru(1964-), male, Minqin of Gansu Province, Researcher, Research focuses on wheat genetics and breeding, (E-mail)fzr640814@qq.com
Supported by:
Supported by Basic Scientific R &D Program of Public Welfare Research Institutions of Xinjiang Uygur Autonomous Region of China(KY2018037/KY2019006)

摘要/Abstract

摘要： 【目的】研究弱光环境下叶绿素酸酯a氧化酶(TaCAO)的表达水平变化,分析叶绿素合成途径对于弱光环境的响应。【方法】利用同源克隆,由小麦品种新冬20号克隆TaCAO并分析其序列。使用半定量PCR方法分析900、1 800、3 600和7 200 lx光照强度下TaCAO表达量。【结果】该基因ORF长度为1 653 bp,编码蛋白大小为62.12 kD,包含550个氨基酸残基,含有叶绿素转运肽和Rieske_RO_Alpha_CAO结构域,还存在一个Rieske铁硫配位中心和铁结合位点,随着光照强度的减弱,TaCAO表达量呈现上升的趋势。【结论】克隆获得的TaCAO长度为1 653 bp,具有完整的CAO活性,TaCAO的相对表达量与光照强度呈反比关系。

关键词: 小麦; 叶绿素酸酯a氧化酶; 基因克隆; 序列分析; 表达

Abstract: 【Objective】 Chlorophyll a oxidase (CAO) is the key enzyme for the synthesis of chlorophyll b. This project aims to study the changes of TaCAO expression level in low light environment and explore the response of chlorophyll synthesis pathway to low light environment. 【Methods】 TaCAO was cloned from Xindong 20 by homologous cloning, and the sequence was analyzed. The semi-quantitative PCR method was used to analyze the expression of TaCAO under 900 lx, 1,800 lx, 3,600 lx and 7,200 lx light intensity. 【Results】The ORF length of this gene was 1,653 bp, the size of the encoded protein was 62.12 kD, containing 550 amino acid residues with the chlorophyll transit peptide and Rieske_RO_Alpha_CAO domain, and there was also a Rieske iron-sulfur coordination center and iron binding site. With the decrease of light intensity, the expression of TaCAO showed an upward trend. 【Conclusion】 The cloned TaCAO is 1,653 bp in length and has complete CAO activity. The relative expression of TaCAO has an inverse relationship with the light intensity.

Key words: wheat; chlorophyll a oxidase; gene clone; sequence analysis; expression analysis

中图分类号:

S512

王重, 樊哲儒, 张跃强, 李剑峰, 高新, 时佳. 小麦叶绿素酸酯a氧化酶(TaCAO)基因的克隆与表达分析[J]. 新疆农业科学, 2021, 58(9): 1569-1576.

WANG Zhong, FAN Zheru, ZHANG Yueqiang, LI Jianfeng, GAO Xin, SHI Jia. Cloning and Expression Analysis of Chlorophyll a Oxygenase Gene in Wheat[J]. Xinjiang Agricultural Sciences, 2021, 58(9): 1569-1576.

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小麦叶绿素酸酯a氧化酶(TaCAO)基因的克隆与表达分析

Cloning and Expression Analysis of Chlorophyll a Oxygenase Gene in Wheat

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