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Multicolored Fluorescent In Situ Hybridization to Assess Pairing Configurations at Metaphase I in Brassica Hybrids

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Plant Meiosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2061))

Abstract

Genetic diversity can be introduced into polyploid crop species through meiotic recombination by exchanges between homologous or homoeologous chromosomes. Fluorescent in situ hybridization (FISH) enables the characterization of these homologous and homoeologous chromosome pairs during meiosis and identification of structural rearrangements during mitosis in metaphase I. In this chapter, we describe a protocol for the multicolored fluorescent labeling of chromosome spreads. This protocol allows the characterization of each A and C homoeologous subgenomes in a polyploid species using a genome-specific BAC combined with specific chromosome labeling BAC sequences.

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References

  1. Soltis DE, Visger CJ, Soltis PS (2014) The polyploidy revolution then … and now: Stebbins revisited. Am J Bot 101(7):1057–1078

    Article  Google Scholar 

  2. Nagaharu U (1935) Genome analysis in Brassica with special reference to the experimental formation of B. napus and its peculiar mode of fertilization. Jap J Bot 7:389–452

    Google Scholar 

  3. Schranz ME, Lysak MA, Mitchell-Olds T (2006) The ABC’s of comparative genomics in the Brassicaceae: building blocks of crucifer genomes. Trends Plant Sci 11(11):535–542

    Article  CAS  Google Scholar 

  4. Lysak MA, Cheung K, Kitschke M, Bures P (2007) Ancestral chromosomal blocks are triplicated in Brassiceae species with varying chromosome number and genome size. Plant Physiol 145(2):402–410

    Article  CAS  Google Scholar 

  5. Leflon M, Brun H, Eber F et al (2007) Detection, introgression and localization of genes conferring specific resistance to Leptosphaeria maculans from Brassica rapa into B. napus. Theor Appl Genet 115:897–906

    Article  CAS  Google Scholar 

  6. Fredua-Agyeman R, Coriton O, Huteau V, Parkin IA, Chèvre AM, Rahman H (2014) Molecular cytogenetic identification of B genome chromosomes linked to blackleg disease resistance in Brassica napus × B. carinata interspecific hybrids. Theor Appl Genet 127(6):1305–1318

    Article  Google Scholar 

  7. Stein A, Coriton O, Rousseau-Gueutin M, Samans B, Obermeier C, Parkin I, Chevre A-M, Snowdon R (2017) Mapping of homoeologous chromosome exchanges influencing quantitative trait variation in Brassica napus. Plant Biotechnol J 15:1478–1849

    Article  CAS  Google Scholar 

  8. Mason AS, Chèvre AM (2016) Optimization of recombination in interspecific hybrids to introduce new genetic diversity into oilseed rape (Brassica napus L.). In: Mason AS (ed) Polyploidy and hybridization for crop improvement. Science Publishers, pp 431–444

    Google Scholar 

  9. Pelé A, Falque M, Trotoux G, Eber F, Nègre S, Gilet M, Huteau V, Lodé M, Jousseaume T, Dechaumet S, Morice J, Poncet C, Coriton O, Martin OC, Rousseau-Gueutin M, Chèvre AM (2017) Amplifying recombination genome-wide and reshaping crossover landscapes in Brassicas. PLoS Genet 13(5):e1006794

    Article  Google Scholar 

  10. Nicolas SD, Leflon M, Monod H, Eber F, Coriton O, Huteau V, Chèvre AM, Jenczewski E (2009) Genetic regulation of meiotic cross-overs between related genomes in Brassica napus haploids and hybrids. Plant Cell 21(2):373–385

    Article  CAS  Google Scholar 

  11. Gaeta RT, Pires JC, Iniguez-Luy F, Leon E, Osborn TC (2007) Genomic changes in resynthesized Brassica napus and their effect on gene expression and phenotype. Plant Cell 19(11):3403–3417

    Article  CAS  Google Scholar 

  12. Rousseau-Gueutin M, Morice J, Coriton O, Huteau V, Trotoux G, Nègre S, Falentin C, Deniot G, Gilet M, Eber F, Pelé A, Vautrin S, Fourment J, Lodé M, Bergès H, Chèvre AM (2017) The impact of open pollination on the structural evolutionary dynamics, meiotic behavior and fertility of resynthesized allotetraploid Brassica napus L. G3 (Bethesda) 7(2):705–717

    Article  CAS  Google Scholar 

  13. Jenczewski E, Chèvre AM, Alix K (2013) Chromosomal and gene expression changes in Brassica allopolyploids. In: Chen ZJ, Birchler JA (eds) Polyploid and hybrid genomics. John Wiley & Sons, Inc., Oxford, pp 171–186

    Chapter  Google Scholar 

  14. Chalhoub B, Denoeud F, Liu S, Parkin IA, Tang H, Wang X, Chiquet J, Belcram H, Tong C, Samans B, Corréa M, Da Silva C, Just J, Falentin C, Koh CS, Le Clainche I, Bernard M, Bento P, Noel B, Labadie K, Alberti A, Charles M, Arnaud D, Guo H, Daviaud C, Alamery S, Jabbari K, Zhao M, Edger PP, Chelaifa H, Tack D, Lassalle G, Mestiri I, Schnel N, Le Paslier MC, Fan G, Renault V, Bayer PE, Golicz AA, Manoli S, Lee TH, Thi VH, Chalabi S, Hu Q, Fan C, Tollenaere R, Lu Y, Battail C, Shen J, Sidebottom CH, Wang X, Canaguier A, Chauveau A, Bérard A, Deniot G, Guan M, Liu Z, Sun F, Lim YP, Lyons E, Town CD, Bancroft I, Wang X, Meng J, Ma J, Pires JC, King GJ, Brunel D, Delourme R, Renard M, Aury JM, Adams KL, Batley J, Snowdon RJ, Tost J, Edwards D, Zhou Y, Hua W, Sharpe AG, Paterson AH, Guan C, Wincker P (2014) Plant genetics. Early allopolyploid evolution in the post-Neolithic Brassica napus oilseed genome. Science 345(6199):950–953

    Article  CAS  Google Scholar 

  15. Suay L, Zhang D, Eber F, Jouy H, Lodé M, Huteau V, Coriton O, Szadkowski E, Leflon M, Martin OC, Falque M, Jenczewski E, Paillard S, Chèvre AM (2014) Crossover rate between homologous chromosomes and interference are regulated by the addition of specific unpaired chromosomes in Brassica. New Phytol 201(2):645–656

    Article  CAS  Google Scholar 

  16. Howell EC, Barker GC, Jones GH, Kearsey MJ, King GJ, Kop EP et al (2002) Integration of the cytogenetic and genetic linkage maps of Brassica oleracea. Genetics 161(3):1225–1234

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Mun JH, Kwon SJ, Yang TJ, Kim HS, Choi BS et al (2008) The first generation of a BAC-based physical map of Brassica rapa. BMC Genomics 9:280

    Article  Google Scholar 

  18. Xiong Z, Gaeta RT, Pires JC (2011) Homoeologous shuffling and chromosome compensation maintain genome balance in resynthesized allopolyploid Brassica napus. Proc Natl Acad Sci U S A 108(19):7908–7913

    Article  CAS  Google Scholar 

  19. Szadkowski E, Eber F, Huteau V, Lodé M, Huneau C, Belcram H, Coriton O, Manzanares-Dauleux MJ, Delourme R, King GJ, Chalhoub B, Jenczewski E, Chèvre AM (2010) The first meiosis of resynthesized Brassica napus, a genome blender. New Phytol 186(1):102–12

    Google Scholar 

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Acknowledgments

We thank Anne-Marie Chèvre and Julie Ferreira-De-Carvalho for constructive reading of the manuscript.

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Authors and Affiliations

  1. Institut de Génétique, Environnement et Protection des Plantes, INRA, UMR 1349 IGEPP, Molecular cytogenetics Platform, BP35327, F-35653, Le Rheu Cedex, France

    Virginie Huteau & Olivier Coriton

Authors
  1. Virginie Huteau
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  2. Olivier Coriton
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Corresponding author

Correspondence to Olivier Coriton .

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Editors and Affiliations

  1. Departamento de Genética, Fisiología y Microbiología, Facultad de Biología, Universidad Complutense de Madrid, Madrid, Spain

    Mónica Pradillo

  2. Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) OT Gatersleben, Stadt Seeland, Germany

    Stefan Heckmann

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Huteau, V., Coriton, O. (2020). Multicolored Fluorescent In Situ Hybridization to Assess Pairing Configurations at Metaphase I in Brassica Hybrids. In: Pradillo, M., Heckmann, S. (eds) Plant Meiosis. Methods in Molecular Biology, vol 2061. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9818-0_12

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  • DOI: https://doi.org/10.1007/978-1-4939-9818-0_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9817-3

  • Online ISBN: 978-1-4939-9818-0

  • eBook Packages: Springer Protocols

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