Analysis of Pollen Grains by Immunostaining and FISH in Triticeae Species

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

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

Analysis of Pollen Grains by Immunostaining and FISH in Triticeae Species

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

Authors

Dan D Wu^{1

2}, Alevtina Ruban¹, Twan Rutten¹, Yong H Zhou², Andreas Houben³

Affiliations

¹ Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Stadt Seeland, Germany.
² Triticeae Research Institute, Sichuan Agricultural University, Wenjiang, China.
³ Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Stadt Seeland, Germany. houben@ipk-gatersleben.de.

PMID: 31583671
DOI: 10.1007/978-1-4939-9818-0_24

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.

Keywords: Aegilops speltoides; FISH; Hordeum vulgare; Immunostaining; Meiosis; Pollen mitosis; Secale cereale; Tissue section.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Chromosomes, Plant
Flowers / genetics
Fluorescent Antibody Technique*
In Situ Hybridization, Fluorescence*
Meiosis*
Mitosis*
Pollen / genetics*
Secale / genetics*