通过免疫染色和荧光原位杂交技术对小麦族物种花粉粒的分析

Analysis of Pollen Grains by Immunostaining and FISH in Triticeae Species.

作者信息

Wu Dan D, Ruban Alevtina, Rutten Twan, Zhou Yong H, Houben Andreas

机构信息

Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Stadt Seeland, Germany.

Triticeae Research Institute, Sichuan Agricultural University, Wenjiang, China.

出版信息

Methods Mol Biol. 2020;2061:347-358. doi: 10.1007/978-1-4939-9818-0_24.

DOI:10.1007/978-1-4939-9818-0_24

PMID:31583671

Abstract

In this chapter we describe protocols for immunolabeling and FISH of pollen grains undergoing postmeiotic mitosis using Aegilops speltoides, Secale cereale, and Hordeum vulgare as models. Tissue sectioning of pollen overcomes the problem of the pollen grain wall impermeability that interferes with immunolocalization and in situ hybridization. The crucial element of the protocol is the generation and immobilization of tissue sections. Our method facilitates the investigation of the microspore cell divisions and pollen grain maturation.

摘要

在本章中，我们描述了以节节麦、黑麦和大麦为模型，对经历减数分裂后有丝分裂的花粉粒进行免疫标记和荧光原位杂交的实验方案。花粉的组织切片克服了花粉粒壁不透性的问题，这种不透性会干扰免疫定位和原位杂交。该实验方案的关键要素是组织切片的制备和固定。我们的方法有助于对小孢子细胞分裂和花粉粒成熟进行研究。