Using Genome In Situ Hybridization (GISH) to Distinguish the Constituent Genomes of Brassica nigra and B. rapa in the Hybrid B. juncea
The genome in situ hybridization (GISH) technique has become important for deciphering the organization of the constituent genomes in the allopolyploid plants that comprise many of the crop species. This technique comprises using the nuclear DNA from the constituent genomes as probes that have been labeled separately with different nucleotides that can be identified by using secondary antibodies. The Brassica family includes a range of mustard species with diverse phytochemical and morphological profile, hence making it an important plant family in agriculture. Meiosis is a specialized cellular division which brings the homologous chromosomes together and creates recombinants via pairing and synapsis during its early phase. Transfer of the genetic material within homoelog pairs creates novelty in subsequent generations which hold promise for improving the agriculture sector. This chapter is concerned with developing a GISH technique that discriminates between the constituent genomes in the allopolyploid B. juncea, in order to study meiosis.
Key wordsDNA probe In situ hybridization Chromosomes Allopolyploid Brassica Meiosis
We would like to thank Eugenio Sanchez Moran and Elaine Howell for the help and discussion in the laboratory. For technical assistance we would thank Steve Price and Karen Staples.
- 2.Stevenson M, Armstrong SJ, Ford-Lloyd BV, Jones GH (1998) Comparative analysis of crossover exchanges and chiasmata in Allium cepa x fistulosum after genomic in situ hybridization (GISH). Chromosome Research 6: 567–574.Google Scholar
- 4.Howell EC and Armstrong SJ (2013) Using sequential fluorescence and genomic in situ hybridization (FISH and GISH) to distinguish the A and C genomes in Brassica napus Wojciech. In: Pawlowski P et al. (eds) Plant meiosis: methods and protocols, methods in molecular biology, vol. 990. Springer Science+Business Media, New York, pp 38–47. https://doi.org/10.1007/978-1-62703-333-6_3