Abstract
The basidiomycete fungus Coprinus cinereus has naturally synchronous meiosis and is amenable to analysis using an array of well-developed genetic and molecular tools. In this chapter, we explain in detail the two methods most commonly employed for C. cinereus, staining of intact gill segments and chromosome spreads, with an example of the application of each. We describe iron-hematoxylin staining of intact gill segments for the brightfield examination of meiotic progression, and the use of surface spreads and fluorescence in situ hybridization (FISH) to investigate meiotic chromosome pairing. Gill segments can alternatively be stained with DAPI for the determination of meiotic stage, or propidium iodide for the quantitation of nuclear DNA content, and the chromosome fixation and spreading techniques used for FISH are also suitable for immunolocalization studies of chromosomal proteins.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Pukkila, P. J., Yashar, B. M., and Binninger, D. M. (1984) Analysis of meiotic development in Coprinus cinereus, in Controlling Events in Meiosis (Evans, C. W., and Dickinson, H. G., eds.), Society for Experimental Biology, Cambridge, pp. 177–194.
Holm, P. B., Rasmussen, S. W., Zickler, D., Lu, B. C., and Sage, J. (1981) Chromosome pairing, recombination nodules and chiasma formation in the basidiomycete Coprinus cinereus. Carlsberg Res. Commun. 46, 305–346.
Pukkila, P. J., Skrzynia, C., and Lu, B. C. (1992) The rad3-1 mutant is defective in axial core assembly and homologous chromosome pairing during meiosis in the basidiomycete Coprinus cinereus. Dev. Genet. 13, 403–410.
Pukkila, P. J., and Lu, B. C. (1985) Silver staining of meiotic chromosomes in the fungus, Coprinus cinereus. Chromosoma 91, 108–112.
Li, L., Gerecke, E. E., and Zolan, M. E. (1999) Homolog pairing and meiotic progression in Coprinus cinereus. Chromosoma 108, 384–392.
Kanda, T., Arakawa, H., Yasuda, Y., and Takemaru, T. (1990) Basidiospore formation in a mutant of incompatibility factors and in mutants that arrest at meta-anaphase I in Coprinus cinereus. Experimental Mycology 14, 218–226.
Many, A. (2006) Meiotic roles of Mre11 and Rad50 in Coprinus cinereus. PhD thesis, Indiana University.
Lu, B. C., and Raju, N. B. (1970) Meiosis in Coprinus. II. Chromosome pairing and the lampbrush diplotene stage of meiotic prophase. Chromosoma 29, 305–316.
O'Shea, S. F., Chaure, P. T., Halsall, J. R., Olesnicky, N. S., Leibbrandt, A., Connerton, I. F., and Casselton, L. A. (1998) A large pheromone and receptor gene complex determines multiple B mating type specificities in Coprinus cinereus. Genetics 148, 1081–1090.
Acknowledgments
The authors would like to dedicate this chapter to Dr. Ben Lu, who introduced each of them to C. cinereus cytology. He first provided P.J.P. with some aged ferric acetate so she could use the optimal iron-hematoxylin method right away. (In turn, she would be happy to supply aged ferric acetate to anyone who would like to try this method.) He also taught the techniques of chromosome spreading and silver staining to two generations of Zolan lab students and post-docs, who bestowed on him the title of Zolan-lab “fairy godfather.” M.E.Z. also thanks Sasha Savytskyy for help on this manuscript and for the data presented in Fig. 8.2, and Heather Palmerini, Felicia Kennedy, Claire Burns, and Elizabeth Sierra Potchanant for helpful comments on the manuscript. The work in our laboratories is supported by NSF grant EF-0412016 (to P.J.P.) and NIH grant GM43930 (to M.E.Z.).
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Zolan, M.E., Pukkila, P.J. (2009). Meiotic Cytogenetics in Coprinus cinereus . In: Keeney, S. (eds) Meiosis. Methods in Molecular Biology, vol 558. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-103-5_8
Download citation
DOI: https://doi.org/10.1007/978-1-60761-103-5_8
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60761-102-8
Online ISBN: 978-1-60761-103-5
eBook Packages: Springer Protocols