滇水金凤花距发育相关基因<i>GRF</i>的克隆及表达分析

魏春梅; 蔡斌; 陶宇蝶; 李林菊; 赵秋燕; 周敏; 黄美娟; 黄海泉

doi:10.11913/PSJ.2095-0837.2022.50669

滇水金凤花距发育相关基因GRF的克隆及表达分析

西南林业大学园林园艺学院, 国家林业和草原局西南风景园林工程技术工程研究中心, 云南省功能性花卉资源及产业化技术工程研究中心, 西南林业大学园林园艺花卉研发中心, 昆明 650224

基金项目:

云南省重大科技专项(202102AE090052)；国家自然科学基金项目(32060364，32060366，31860230，31560228)；云南省高校园林植物与观赏园艺科技创新团队(51700204)；云南省园林植物遗传改良与高效繁育博士生导师团队和云南省中青年学术和技术带头人培养项目(2018HB024)。

详细信息

作者简介:
魏春梅(1998-),女,硕士研究生,研究方向为园林植物资源与应用(E-mail:weichunmei@swfu.edu.cn)。

通讯作者:
黄美娟,E-mail:xmhhq2001@163.com

黄海泉,E-mail:haiquanl@163.com

中图分类号: Q943.2
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- 文章访问数: 190
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出版历程
- 收稿日期: 2022-03-16
- 修回日期: 2022-05-14
- 网络出版日期: 2022-12-08

Cloning and expression analysis of GRF genes related to spur development in Impatiens uliginosa Franch.

College of Landscape Architecture and Horticulture Sciences, Southwest Research Center for Engineering Technology of Landscape Architecture (State Forestry and Grassland Administration), Yunnan Engineering Research Center for Functional Flower Resources and Industrialization, Research and Development Center of Landscape Plants and Horticulture Flowers, Southwest Forestry University, Kunming 650224, China

Funds:

This work was supported by grants from the Key Research and Development Plan Program of Yunnan Province (202102AE090052),National Natural Science Foundation of China (32060364,32060366,31860230,31560228),Program for Innovative Research Team (in Science and Technology) in University of Yunnan Province (51700204),Team of Doctoral Supervisors for Genetic Improvement and Efficient Breeding of Garden Plants in Yunnan Province and Young and Middle-aged Academic and Technical Leadership Training Project of Yunnan (2018HB024).

摘要

摘要: 采用RT-PCR技术对滇水金凤(Impatiens uliginosa Franch.)GRF基因进行克隆，对其编码蛋白序列进行同源性分析和系统进化分析，并利用qRT-PCR方法分析GRF基因的时空表达模式。结果显示，从滇水金凤中成功获得两个GRF基因，其 cDNA全长分别为1137和903 bp，分别编码378和300 aa，分别命名为IuGRF1和IuGRF2，二者均具有QLQ和WRC结构域。除了保守区域外，滇水金凤GRF基因编码产物序列与其他物种的同源性较低。系统进化分析结果表明，IuGRF1和IuGRF2分别位于两个不同的进化分支。基因表达分析发现，IuGRF1在花苞期和盛花期距部表达量最高，而在檐部的表达随花距的发育逐渐降低。IuGRF2在花苞期檐部表达量最高，随着花距的发育逐渐降低。此外，IuGRF1和IuGRF2基因均在始花期花距弯部表达量最高，且随着花距的发育，在尖部和弯部的表达量逐渐降低，而在基部的表达量逐渐升高。推测滇水金凤的两个基因为同源基因，IuGRF1主要参与花距距部的生长和发育，而IuGRF2主要参与檐部的生长和发育。
- 滇水金凤 /
- 花距发育 /
- GRF基因 /
- 基因克隆 /
- 表达分析
Abstract: GRF genes related to spur development in Impatiens uliginosa Franch. were cloned using reverse transcription polymerase chain reaction (RT-PCR), with homology and phylogenetic analysis of protein sequences then investigated using DNAMAN and MEGA. In addition, the spatiotemporal expression patterns of GRF genes were investigated by quantitative RT-PCR(qRT-PCR). Results showed that two GRF genes of I. uliginosa were successfully cloned, whose full-length cDNA sequences were 1137 bp and 903 bp, encoding 378 amino acids (aa) and 300 aa and named IuGRF1 and IuGRF2, respectively. Both contained the QLQ and WRC structural domains. The GRF genes of I. uliginosa showed low homology with GRF genes from other species, except for the conserved domains, which may be correlated with the diversity of the C-terminal domain in GRF genes. Based on phylogenetic analysis, IuGRF1 and IuGRF2 were separated into different branches. The qRT-PCR results showed that the expression of IuGRF1 was the highest in the spur cup at both the bud and blooming stages but decreased in the blade with the development of the spur. The expression of IuGRF2 was the highest in the blade at the bud stage but declined gradually with spur growth and was low in the spur cup. The expression levels of IuGRF1 and IuGRF2 were the highest in the cup curve at the beginning of the flowering stage but decreased gradually in the cup mucro and curve with spur development and increased gradually in the cup base. Based on the above results, we speculated that the two GRF genes in I. uliginosa were paralogous, with IuGRF1 playing a major role in spur cup development and IuGRF2 playing a major role in blade development.
- Impatiens uliginosa /
- Spur development /
- GRF genes /
- Gene cloning /
- Expression analysis

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