杜鹃兰侧芽萌发及其相关基因表达分析

吕享, 李小兰, 阙云飞, 田海露, 王倩, 叶睿华, 张明生*
贵州大学生命科学学院/国家喀斯特石漠化防治工程技术研究中心, 山地植物资源保护与种质创新教育部重点实验室, 贵阳550025

通信作者:张明生;E-mail: mszhang@gzu.edu.cn

摘 要:

以打顶诱导杜鹃兰侧芽萌发过程中其水分组成、细胞形态结构等的变化为基础, 探讨杜鹃兰侧芽萌发与其相关基因表达量的关系。本文采用光镜和扫描电镜观察侧芽基部细胞的变化过程, 检测从休眠到萌发期间侧芽自由水和束缚水的含量, 并通过qRT-PCR分析细胞壁扩张蛋白和水通道蛋白相关基因的表达量。结果发现, 打顶处理诱导了杜鹃兰侧芽的萌发; 与对照相比, 侧芽基部的薄壁组织细胞的细胞壁由皱缩状逐渐伸展, 细胞变大。在此过程中, 细胞壁扩张蛋白基因αEXPA1EXPA2EXPA6以及水通道蛋白基因PIP1-2PIP2表达上调, 而液泡膜上的水通道蛋白基因δTIP表达下调; 总含水量和自由水含量逐渐升高, 束缚水含量降低。结果表明, 杜鹃兰侧芽萌发与自由水含量升高有关, 而自由水含量升高可能是因细胞壁扩张蛋白和水通道蛋白基因上调所致。

关键词:杜鹃兰; 侧芽萌发; 细胞壁扩张蛋白基因; 水通道蛋白基因; 自由水

收稿:2018-05-12   修定:2018-07-26

资助:国家自然科学基金(81660627和81360613)、国家重点研发计划课题(2016YFC0502604)、贵州省科技计划重大专项课题(黔科合平台人才[2017]5411-06)、国家喀斯特石漠化防治工程技术研究中心建设(2012FU125X13)、贵州省科技创新人才团队建设专项资金(黔科合平台人才[2016]5624)、贵州省教育厅创新群体重大研究项目(黔教合KY字[2016]023)、贵州省研究生科研基金(黔教研合KYJJ字[2016]011)和贵州大学国家级重点培育项目(黔科合平台人才[2017]5788)。

Lateral buds germination of Cremastra appendiculata and its related gene expression analysis

LÜ Xiang, LI Xiao-Lan, QUE Yun-Fei, TIAN Hai-Lu, WANG Qian, YE Rui-Hua, ZHANG Ming-Sheng*
College of Life Sciences/State Engineering Technology Institute for Karst Desertification Control, Key Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountainous Region (Ministry of Education), Guizhou University, Guiyang 550025, China

Corresponding author: ZHANG Ming-Sheng; E-mail: mszhang@gzu.edu.cn

Abstract:

In order to investigate the relationship between the lateral bud germination of Cremastra appendiculata and the related genes of expression levels, this research was based on the changes of water composition and cell morphological structure in the lateral buds after decapitation. Basal cells of lateral buds were observed using light microscope and scanning electron microscope; free water and bound water contents were detected and the related expansion genes and water channels genes of expression levels were analyzed using qRT-PCR from dormancy to sprouting. The results showed that decapitation induced the C. appendiculata lateral buds of growth. Compared with the control, cell wall of parenchyma cell in the buds basal region recovered extension from shrinkage shape and these cells got bigger; αEXPA1, EXPA2, EXPA6, PIP1-2 and PIP2 expression levels were up-regulated, but δTIP was down-regulated. Water and free water contents were gradually increased, and bound water content was decreased in this process. In other words, the germination of C. appendiculata was related to the increase of free water content, and its increasing might be due to up-regulation of the related expansion genes and water channels genes.

Key words: Cremastra appendiculata; lateral bud germination; cell wall expansion genes; aquaporin genes; free water

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