Chromosome Research

, 17:1041

Higher axial-resolution and sensitivity pachytene fluorescence in situ hybridization protocol in tetraploid cotton

  • Kai Wang
  • Zaijie Yang
  • Changshen Shu
  • Jing Hu
  • Qiuyun Lin
  • Wenpan Zhang
  • Wangzhen Guo
  • Tianzhen Zhang
Article

Abstract

Fluorescence in situ hybridization (FISH) based on pachytene chromosomes has become an important cytogenetic tool to construct high axial-resolution and sensitivity cytogenetic maps. However, the application of this technique in cotton has lagged behind due to difficulties in chromosome preparation. To date, successful FISH based on cotton pachytene chromosomes has not been reported. In this study, the first protocol developed for pachytene chromosome preparation in tetraploid cotton is presented. This protocol yielded chromosome spreads suitable for large and small DNA probe FISH labeling. Two important parameters, axial-resolution and sensitivity, of FISH on mitotic metaphase and pachytene chromosomes were systematically analyzed. The results demonstrated that DNA targets separated by 0.6 cM and low-copy targets as small as 3-kb were resolved and detected, respectively, in pachytene FISH. The application of our FISH protocol will continue to improve and provide a point of departure for constructing an integrated high axial-resolution cytogenetic map in cotton.

Keywords

pachytene FISH axial-resolution sensitivity cotton 

References

  1. Albini SM, Schwarzacher T (1992) In situ localization of two repetitive DNA sequences to surface-spread pachytene chromosomes of rye. Genome 35:551–559Google Scholar
  2. Cheng Z, Buell CR, Wing RA, Gu M, Jiang J (2001) Toward a cytological characterization of the rice genome. Genome Res 11:2133–2141CrossRefPubMedGoogle Scholar
  3. Cheng Z, Buell CR, Wing RA, Jiang J (2002) Resolution of fluorescence in-situ hybridization mapping on rice mitotic prometaphase chromosomes, meiotic pachytene chromosomes and extended DNA fibers. Chromosome Res 10:379–387CrossRefPubMedGoogle Scholar
  4. Danilova TV, Birchler JA (2008) Integrated cytogenetic map of mitotic metaphase chromosome 9 of maize: resolution, sensitivity, and banding paint development. Chromosoma 117:345–356CrossRefPubMedGoogle Scholar
  5. De Jong JH, Fransz P, Zabel P (1999) High resolution FISH in plants—techniques and applications. Trends Plant Sci 4:258–263CrossRefGoogle Scholar
  6. Fransz P, Stam M, Montijn B et al (1996) Detection of single-copy genes and chromosome rearrangements in Petunia hybrida by fluorescence in situ hybridization. Plant J 9:767–774CrossRefGoogle Scholar
  7. Fransz P, Armstrong S, Alonso-Blanco C et al (1998) Cytogenetics for the model system Arabidopsis thaliana. Plant J 13:867–876CrossRefPubMedGoogle Scholar
  8. Guo W, Cai C, Wang C et al (2008) A preliminary analysis of genome structure and composition in Gossypium hirsutum. BMC Genomics 9:314CrossRefPubMedGoogle Scholar
  9. Hanson RE, Zwick MS, Choi S et al (1995) Fluorescent in situ hybridization of a bacterial artificial chromosome. Genome 38:646–651CrossRefPubMedGoogle Scholar
  10. Hanson RE, Islam-Faridi MN, Percival EA et al (1996) Distribution of 5 S and 18 S–28 S rDNA loci in a tetraploid cotton (Gossypium hirsutum L.) and its putative diploid ancestors. Chromosoma 105:55–61CrossRefPubMedGoogle Scholar
  11. Hu Y, Guo WZ, Zhang TZ (2009) Construction of a bacterial artificial chromosome library of TM-1, a standard line for genetics and genomics in Upland cotton. J Integr Plant Biol 51:107–112CrossRefPubMedGoogle Scholar
  12. Jack Mursal I, Endrizzi JE (1976) A reexamination of the diploidlike meiotic behavior of polyploid cotton. Theor Appl Genet 47:171–178CrossRefGoogle Scholar
  13. Ji Y, De Donato M, Crane CF et al (1999) New ribosomal RNA gene locations in Gossypium hirsutum mapped by meiotic FISH. Chromosoma 108:200–207CrossRefPubMedGoogle Scholar
  14. Ji Y, Zhao X, Paterson AH, Price HJ, Stelly DM (2007) Integrative mapping of Gossypium hirsutum L. by meiotic fluorescent in situ hybridization of a tandemly repetitive sequence (B77). Genetics 176:115–123CrossRefPubMedGoogle Scholar
  15. Jiang J, Gill BS (2006) Current status and the future of fluorescence in situ hybridization (FISH) in plant genome research. Genome 49:1057–1068CrossRefPubMedGoogle Scholar
  16. Kato A, Vega JM, Han F, Lamb JC, Birchler JA (2005) Advances in plant chromosome identification and cytogenetic techniques. Curr Opin Plant Biol 8:148–154CrossRefPubMedGoogle Scholar
  17. Khrustaleva LI, Kik C (2001) Localization of single-copy T-DNA insertion in transgenic shallots (Allium cepa) by using ultra-sensitive FISH with tyramide signal amplification. Plant J 25:699–707CrossRefPubMedGoogle Scholar
  18. Koo D, Jiang J (2009) Super-stretched pachytene chromosomes for fluorescence in situ hybridization mapping and immunodetection of DNA methylation. Plant J 59:509–516CrossRefPubMedGoogle Scholar
  19. Kulikova O, Gualtieri G, Geurts R et al (2001) Integration of the FISH pachytene and genetic maps of Medicago truncatula. Plant J 27:49–58CrossRefPubMedGoogle Scholar
  20. Lawrence JB, Singer RH, McNeil JA (1990) Interphase and metaphase resolution of different distances within the human dystrophin gene. Science 249:928–932CrossRefPubMedGoogle Scholar
  21. Lichter P, Tang CJ, Call K et al (1990) High-resolution mapping of human chromosome 11 by in situ hybridization with cosmid clones. Science 247:64–69CrossRefPubMedGoogle Scholar
  22. Ohmido N, Akiyama Y, Fukui K (1998) Physical mapping of unique nucleotide sequences on identified rice chromosomes. Plant Mol Biol 38:1043–1052CrossRefPubMedGoogle Scholar
  23. Sadder MT, Ponelies N, Born U, Weber G (2000) Physical localization of single-copy sequences on pachytene chromosomes in maize (Zea mays L.) by chromosome in situ suppression hybridization. Genome 43:1081CrossRefPubMedGoogle Scholar
  24. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring HarborGoogle Scholar
  25. Schwarzacher T (2003) DNA, chromosomes, and in situ hybridization. Genome 46:953–962CrossRefPubMedGoogle Scholar
  26. Shen DL, Wang ZF, Wu M (1987) Gene mapping on maize pachytene chromosomes by in situ hybridization. Chromosoma 95:311–314CrossRefGoogle Scholar
  27. Stephens JL, Brown SE, Lapitan NL, Knudson DL (2004) Physical mapping of barley genes using an ultrasensitive fluorescence in situ hybridization technique. Genome 47:179–189CrossRefPubMedGoogle Scholar
  28. Wang CJ, Chen CC (2005) Cytogenetic mapping in maize. Cytogenet Genome Res 109:63–69CrossRefPubMedGoogle Scholar
  29. Wang CJ, Harper L, Cande WZ (2006a) High-resolution single-copy gene fluorescence in situ hybridization and its use in the construction of a cytogenetic map of maize chromosome 9. Plant Cell 18:529–544CrossRefPubMedGoogle Scholar
  30. Wang K, Song X, Han Z et al (2006b) Complete assignment of the chromosomes of Gossypium hirsutum L. by translocation and fluorescence in situ hybridization mapping. Theor Appl Genet 113:73–80CrossRefPubMedGoogle Scholar
  31. Wang K, Guo W, Zhang T (2007a) Detection and mapping of homologous and homoeologous segments in homoeologous groups of allotetraploid cotton by BAC-FISH. BMC Genomics 8:178CrossRefPubMedGoogle Scholar
  32. Wang K, Guo W, Zhang T (2007b) Development of one set of chromosome-specific microsatellite-containing BACs and their physical mapping in Gossypium hirsutum L. Theor Appl Genet 115:675–682CrossRefPubMedGoogle Scholar
  33. Wang K, Zhang YJ, Guan B, Guo W, Zhang T (2007c) Fluorescence in situ hybridization of bacterial artificial chromosome in cotton. Prog Biochem Biophys 34:1216–1222Google Scholar
  34. Wang K, Guan B, Guo W et al (2008) Completely distinguishing individual A-genome chromosomes and their karyotyping analysis by multiple bacterial artificial chromosome-fluorescence in situ hybridization. Genetics 178:1117–1122CrossRefPubMedGoogle Scholar
  35. Yin J, Guo W, Yang L, Liu L, Zhang T (2006) Physical mapping of the Rf 1 fertility-restoring gene to a 100 kb region in cotton. Theor Appl Genet 112:1318–1325CrossRefPubMedGoogle Scholar
  36. Zhong XB, Fransz P, Wennekes-van Eden J et al (1996a) High-resolution mapping on pachytene chromosomes and extended DNA fibers by fluorescence in-situ hybridisation. Plant Mol Biol Report 14:232–242CrossRefGoogle Scholar
  37. Zhong XB, Hans de Jong J, Zabel P (1996b) Preparation of tomato meiotic pachytene and mitotic metaphase chromosomes suitable for fluorescence in situ hybridization (FISH). Chromosome Res 4:24–28CrossRefPubMedGoogle Scholar
  38. Zwick MS, Hanson RE, McKnight TD et al (1997) A rapid procedure for the isolation of C0t–1 DNA from plants. Genome 40:138–142CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Kai Wang
    • 1
  • Zaijie Yang
    • 1
  • Changshen Shu
    • 1
  • Jing Hu
    • 1
  • Qiuyun Lin
    • 1
  • Wenpan Zhang
    • 1
  • Wangzhen Guo
    • 1
  • Tianzhen Zhang
    • 1
  1. 1.National Key Laboratory of Crop Genetics and Germplasm Enhancement, Cotton Research InstituteNanjing Agricultural UniversityNanjingChina

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