Abstract
We present an improved FISH strategy for differentiating the sex chromosomes of the dioecious model plant, Silene latifolia. Fixed mitotic protoplasts were dropped on a polyethylene naphthalate membrane, the X or Y chromosomes were isolated using nitrogen laser beam microdissection, catapulted by laser pressure, and amplified by DOP-PCR. A modified FAST-FISH protocol based on a short hybridization time combined with a low concentration of probe was used. The success of this approach is demonstrated by the differential labeling of the X and Y chromosomes and it could represent a quick method for comparing organization of plant genomes.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Charlesworth B (1996) The evolution of chromosomal sex determination and dosage compensation. Curr Biol 2: 149-162.
Durm M, Schussler L, Munch H et al. (1998) Fast-painting of human metaphase spreads using a chromosome-specific, repeat-depleted DNA library probe. Biotechniques 24: 820-825.
Hladilova R, Siroky J, Vyskot B (1998) A cytospine technique for spreading plant metaphases suitable for immunofluorescence studies. Biotechnic Histochem 73: 150-156.
Kubickova S, Cernohorska H, Musilova P, Rubes J (2001) The use of laser microdissection for the preparation of chromosome-specific painting probes in farm animals. Chromosome Res 10: 571-577.
Matsunaga S, Kawano S, Michimoto T et al. (1999) Semiautomatic laser beam microdissection of the Y chromosome and analysis of Y chromosome DNA in a dioecious plant, Silene latifolia. Plant Cell Physiol 40: 60-68.
Negrutiu I, Vyskot B, Barbacar N, Georgiev S, Moneger F (2001) Dioecious plants. A key to the early events of sex chromosome evolution. Plant Physiol 127: 1418-1424.
Ried T, Schrock E, Ning Y, Wienberg J (1998) Chromosome painting: a useful art. Hum Mol Genet 7: 1619-1626.
Schermelleh L, Thalhammer S, Heckl W et al. (1999) Laser microdissection and laser pressure catapulting for the generation of chromosome-specific paint probes. Biotechniques 27: 362-367.
Schmidt T, Heslop-Harrison JS (1998) Genomes, genes and junk: the large-scale organization of plant chromosomes. Trends Plant Sci 3: 195-199.
Scutt CP, Kamisugi Y, Sakai F, Gilmartin PM (1997) Laser isolation of plant sex chromosomes: studies on the DNA composition of the X and Y sex chromosomes of Silene latifolia. Genome 40: 705-715.
Siroky J, Hodurkova J, Negrutiu I, Vyskot B (1999) Functional and structural chromosome analyses in autotetraploid Silene latifolia. Ann Bot 84: 633-638.
Telenius H, Carter NP, Bebb CE, Nordenskjodl M, Ponder BAJ, Tunnacliffe A (1992) Degenerate Oligonucleotide-Primed PCR: General amplification of target DNA by a single degenerate primer. Genomics 13: 718-725.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Hobza, R., Lengerova, M., Cernohorska, H. et al. FAST-FISH with laser beam microdissected DOP-PCR probe distinguishes the sex chromosomes of Silene latifolia . Chromosome Res 12, 245–250 (2004). https://doi.org/10.1023/B:CHRO.0000021929.97208.1c
Issue Date:
DOI: https://doi.org/10.1023/B:CHRO.0000021929.97208.1c