甘薯苯丙氨酸解氨酶基因<i>IbPAL</i>的克隆与表达分析

饶莉萍; 苏文瑾; 刘意; 宋天晓; Soviguidi Deka Reine Judesse; 杨新笋; 张文英

doi:10.11913/PSJ.2095-0837.2020.30360

甘薯苯丙氨酸解氨酶基因IbPAL的克隆与表达分析

1. 长江大学农学院, 湖北荆州 434025;
2. 湖北省农业科学院粮食作物研究所, 武汉 430064;
3. 海南大学园艺学院, 海口 570228

基金项目:

国家重点研发计划（2018YFD1000700，2018YFD1000705）；湖北省技术创新专项（2018AHB012）；农业部甘薯生物学与生物技术重点实验室开放课题；湖北省农业科技创新中心资助项目（2007-620-001-03）；湖北省技术创新专项（2017ABA149）；国家现代甘薯产业技术体系建设项目（CARS-11-C-15）；湖北省农业科学院甘薯特色学科项目（2019TSXK06）；湖北省农业科学院青年拔尖人才项目。

详细信息

作者简介:
饶莉萍(1996-),女,硕士研究生,研究方向为甘薯遗传育种(E-mail:1144023471@qq.com)。

通讯作者:
杨新笋,E-mail:yangxins013@163.com

张文英,E-mail:wyzhang@yangtzeu.edu.cn

中图分类号: Q943.2
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出版历程
- 收稿日期: 2019-09-22
- 修回日期: 2019-12-02
- 网络出版日期: 2022-10-31
- 发布日期: 2020-06-27

Cloning and expression analysis of Ipomoea batatas (L.) Lam. gene IbPAL

1. College of Agriculture, Yangtze University, Jingzhou, Hubei 434025, China;
2. Food Crops Institute, Hubei Academy of Agricultural Sciences, Wuhan 430064, China;
3. Horticulture College, Hainan University, Haikou 570228, China

Funds:

This work was supported by grants from the National Key R & D Program of China (2018YFD1000700, 2018YFD1000705), Hubei Technology Innovation Project (2018AHB012), Key Laboratory of Sweetpotato Biology and Biotechnology of Ministry of Agriculture and Rural Affairs, Hubei Agricultural Science and Technology Innovation Center (2007-620-001-03), Major Project of Hubei Technology Innovation (2017ABA149), China Agriculture Research System (CARS-11-C-15), Characteristic Subject of Sweetpotato of Hubei Academy of Agricultural Sciences (2019TSXK06), and Young Talent Cultivation Program of Hubei Academy of Agricultural Sciences.

摘要

摘要: 本研究基于甘薯（Ipomoea batatas（L.）Lam.）与绿原酸合成代谢相关的转录组数据克隆了苯丙氨酸解氨酶基因IbPAL，通过蛋白序列比对、系统进化分析、二级和三维蛋白结构预测、跨膜结构域以及启动子分析，对该基因的氨基酸序列和启动子序列的结构特征进行了解析；通过qRT-PCR技术分析该基因在不同甘薯品种的根、茎、茎尖和叶中的表达情况，并对其进行亚细胞定位验证。结果显示，IbPAL的CDS序列全长2130 bp，编码709个氨基酸。其氨基酸序列与已知物种的氨基酸序列同源性在80%以上。IbPAL蛋白以α螺旋和随机卷曲为主，具有1个可能的跨膜螺旋区。该基因启动子中包含多个顺式作用元件。表达模式分析结果表明，IbPAL主要在茎和茎尖中表达，其在甘薯品种‘EC16’4个组织器官中的表达量均显著高于其他品种。亚细胞定位结果显示该基因在细胞膜和细胞核上均可以表达。
- 甘薯 /
- IbPAL /
- 克隆 /
- qRT-PCR /
- 亚细胞定位
Abstract: In this research, the phenylalanine ammonia-lyase gene IbPAL was cloned based on transcriptome information related to chlorogenic biosynthesis metabolism in Ipomoea batatas (L.) Lam. Its amino acid (aa) structure was also analyzed by means of protein sequence alignment, phylogenetic analysis, secondary and three-dimensional protein structure prediction, and transmembrane domain and promotor analysis. The expression patterns of IbPAL in roots, stems, meristems, and leaves were further determined by qRT-PCR, with expression locations validated by subcellular experiments. Results showed that the CDS of IbPAL was 2130 bp in length, encoding 709 amino acids. Homologous alignment of the amino acid sequences indicated that IbPAL had high similarity (> 80%) with other reported species. Its protein mainly consisted of α-helixes and random coils and was predicated to have a transmembrane domain. Its promoter contained several cis-elements. IbPAL was mainly expressed in stems and meristems and its expression levels in the four tissues of sweet potato variety ‘EC16’ were significantly higher than that in other varieties and lines. The subcellular experiments indicated that the gene was expressed in the cell nucleus and cytomembrane.
- Ipomoea batatas /
- IbPAL /
- Clone /
- qRT-PCR /
- Subcellular location

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