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Chromosome Research

, Volume 20, Issue 3, pp 353–361 | Cite as

Size and location of radish chromosome regions carrying the fertility restorer Rfk1 gene in spring turnip rape

  • Tarja NiemeläEmail author
  • Mervi Seppänen
  • Farah Badakshi
  • Veli-Matti Rokka
  • J. S. (Pat) Heslop-Harrison
Article

Abstract

In spring turnip rape (Brassica rapa L. spp. oleifera), the most promising F1 hybrid system would be the Ogu-INRA CMS/Rf system. A Kosena fertility restorer gene Rfk1, homolog of the Ogura restorer gene Rfo, was successfully transferred from oilseed rape into turnip rape and that restored the fertility in female lines carrying Ogura cms. The trait was, however, unstable in subsequent generations. The physical localization of the radish chromosomal region carrying the Rfk1 gene was investigated using genomic in situ hybridization (GISH) and bacterial artificial chromosome–fluorescence in situ hybridization (BAC–FISH) methods. The metaphase chromosomes were hybridized using radish DNA as the genomic probe and BAC64 probe, which is linked with Rfo gene. Both probes showed a signal in the chromosome spreads of the restorer line 4021–2 Rfk of turnip rape but not in the negative control line 4021B. The GISH analyses clearly showed that the turnip rape restorer plants were either monosomic (2n = 2x = 20 + 1R) or disomic (2n = 2x = 20 + 2R) addition lines with one or two copies of a single alien chromosome region originating from radish. In the BAC–FISH analysis, double dot signals were detected in subterminal parts of the radish chromosome arms showing that the fertility restorer gene Rfk1 was located in this additional radish chromosome. Detected disomic addition lines were found to be unstable for turnip rape hybrid production. Using the BAC–FISH analysis, weak signals were sometimes visible in two chromosomes of turnip rape and a homologous region of Rfk1 in chromosome 9 of the B. rapa A genome was verified with BLAST analysis. In the future, this homologous area in A genome could be substituted with radish chromosome area carrying the Rfk1 gene.

Keywords

turnip rape (Brassica raparadish (Raphanus sativusfertility restoration gene Rfk1 Rfo GISH BAC–FISH 

Abbreviations

BAC–FISH

Bacterial artificial chromosome–fluorescence in situ hybridization

CMS

Cytoplasmic male sterility

GISH

Genomic in situ hybridization

Rf

Restor fertility

Notes

Acknowledgments

We thank Dr. Regine Delourme for guiding us to get the BAC64 clone for this study. We also thank Xian-Hong Ge for discussions and assistance. The study was supported by the National Emergency Supply Agency, the Finnish Ministry of Agriculture and Forestry, and the Foundation of August Johannes and Aino Tiura.

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Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Tarja Niemelä
    • 1
    Email author
  • Mervi Seppänen
    • 1
  • Farah Badakshi
    • 2
  • Veli-Matti Rokka
    • 3
  • J. S. (Pat) Heslop-Harrison
    • 2
  1. 1.Department of AgricultureUniversity of HelsinkiHelsinkiFinland
  2. 2.Department of BiologyUniversity of LeicesterLeicesterUK
  3. 3.MTT Agrifood Research Finland, Biotechnology and Food ResearchJokioinenFinland

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