A Cytological Analysis of Wheat Meiosis Targeted by Virus-Induced Gene Silencing (VIGS)
Virus-induced gene silencing (VIGS) is a rapid and cost-effective reverse genetic technology that can be used to assess gene function in wheat. This chapter contains a detailed description of how to target wheat meiotic genes by VIGS. The timing of this technique is critical and has been optimized to silence meiotic genes at peak expression, evidenced by silencing of Triticum aestivum disrupted meiotic cDNA1 (TaDMC1). We also describe cytological techniques that have been adapted for the preparation and analysis of meiocytes in wheat, including fluorescent in situ hybridization (FISH) with directly labeled, synthetic oligonucleotide probes, and immunolocalization on spread material.
Key wordsVIGS Meiosis Cytology Wheat FISH Chromosomes Immunolocalization
This work has been supported by the UK Research and Innovation (UKRI) Biotechnology and Biological Sciences Research Council (BBSRC) grant BB/N002628/1. Technical assistance was provided by Daisy Ogle, Inna Guterman, and Wing-Sham Lee.
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