北京大学学报自然科学版 ›› 2022, Vol. 58 ›› Issue (5): 795-800.DOI: 10.13209/j.0479-8023.2022.020

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植物生长素整体原位定位技术

王东辉   

  1. 北京大学生命科学学院, 生物学国家级实验教学示范中心, 蛋白质与植物基因研究国家重点实验室, 北京 100871
  • 收稿日期:2021-09-17 修回日期:2021-10-22 出版日期:2022-09-20 发布日期:2022-09-20
  • 通讯作者: 王东辉，E-mail: wangdh(at)pku.edu.cn
  • 基金资助:
    蛋白质与植物基因研究国家重点实验室开放运行费(8102000374/006)资助 

Whole Mount in situ Localization Technology of Plant Hormones

WANG Donghui   

  1. School of Life Sciences, National Teaching Center for Experimental Biology, State Key Laboratory of Protein and Plant Gene Research, Peking University, Beijing 100871
  • Received:2021-09-17 Revised:2021-10-22 Online:2022-09-20 Published:2022-09-20
  • Contact: WANG Donghui，E-mail: wangdh(at)pku.edu.cn

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摘要：

针对现有植物激素测定方法比较繁琐, 缺少胞内激素浓度测量方法的问题, 建立拟南芥侧根和花发育过程中生长素IAA分布整体原位检测方法。利用抗体对抗原的特异性反应, 通过与酶和荧光分子等连接的二抗结合, 利用光学显微镜和电子显微镜对组织或器官内的生长素进行检测, 实现植物生长素的整体原位定位。该方法不仅适用于生长素IAA, 也适用于脱落酸ABA等激素快速和可靠的组织或器官定位。整个检测流程通常在3~4天内完成, 是一种高度敏感、特异且简单的激素原位定位方法, 适用于各种植物幼嫩的器官和组织。

关键词: 整体原位定位技术, 抗体, 植物激素, 生长素

Abstract:

The existing phytohormones measurement methods are relatively cumbersome and difficult to measure the concentration of intracellular hormones. The paper describes whole-mount “immunolocalization” technique for the overall distribution of auxin (IAA) during lateral root and flower development in Arabidopsis. Based on the specific reaction of the antibody to the antigen, this method binds the antibody to the secondary antibody linked to enzymes, fluorescent molecules, etc. to detect the auxin in tissues or organs by light and electron microscopy, realizing the overall in situ localization of auxin (whole mount in situ localization of Hormone, WISIH). This method is not only suitable for growth hormone IAA, but also for rapid and reliable tissue or organ localization of hormones such as abscisic acid (ABA). The protocol is usually done within three to four days. It is a highly sensitive, specific and simple hormone in situ localization method, and is suitable for various plant tender organs and tissues.

Key words: whole mount in situ localization, antibody, hormone, IAA

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