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Towards Positional Cloning in Brassica napus: Generation and Analysis of Doubled Haploid B. rapa Possessing the B. napus pol CMS and Rfp Nuclear Restorer Gene

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Abstract

The Polima (pol) system of cytoplasmic male sterility (CMS) and its fertility restorer gene Rfp are used in hybrid rapeseed production in Brassica napus. To facilitate map-based cloning of the Rfp gene, we have successfully transferred the pol cytoplasm and Rfp from the amphidiploid B. napus to the diploid species B. rapa and generated a doubled haploid pol cytoplasm B. rapa population that segregates for the Rfp gene. This was achieved through interspecific crosses, in vitro rescue of hybrid embryos, backcrosses, and microspore culture. Male fertility conditioned by Rfp was shown to co-segregate in this population with Rfp-specific mitochondrial transcript modifications and with DNA markers previously shown to be linked to Rfp in B. napus. The selfed-progeny of one doubled haploid plant were confirmed to be characteristic B. rapa diploids by cytogenetic analysis. Clones recovered from a genomic library derived from this plant line using the RFLP probe cRF1 fell into several distinct physical contigs, one of which contained Rfp-linked polymorphic restriction fragments detected by this probe. This indicates that chromosomal DNA segments anchored in the Rfp region can be recovered from this library and that the library may therefore prove to be a useful resource for the eventual isolation of the Rfp gene.

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Abbreviations

CMS:

cytoplasmic male sterility

DH:

doubled haploid

h:

hour

min:

minute

kb:

kilobase

PPR:

pentatricopeptide

Rf:

restorer of fertility

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Authors and Affiliations

  1. Department of Biology, McGill University, H3A 1B1, Montreal, Quebec, Canada

    Nataša Formanová, Xiu-Qing Li, Mary DePauw & Gregory G. Brown

  2. Plant Biotechnology Institute, National Research Counci, 110 Gymnasium Place, S7N OW9, Saskatoon, Saskatchewan, Canada

    Alison M. R. Ferrie & Wilf A. Keller

  3. DNA LandMarks Inc., 84 rue Richelieu, St-Jean-sur-Richelieu, Quebec, Canada

    Benoit Landry

  4. Max Planck Institut fuer Chemische Oekologie, Abteilung fuer Genetik und Evolution, Winzerlaer Strasse 10, D07745, Jena, Germany

    Nataša Formanová

  5. Agriculture and Agri-Food Canada, 850 Lincoln Rd, P.O. Box 20280, E3B 4Z7, Fredericton, New Brunswick, Canada

    Xiu-Qing Li

  6. Department of Biological Sciences, Biological Sciences Centre, University of Alberta, T6G 2E9, Edmonton, Alberta, Canada

    Mary DePauw

Authors
  1. Nataša Formanová
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  2. Xiu-Qing Li
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  3. Alison M. R. Ferrie
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  4. Mary DePauw
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  5. Wilf A. Keller
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  6. Benoit Landry
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  7. Gregory G. Brown
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Corresponding author

Correspondence to Gregory G. Brown.

Additional information

Nataša Formanová and Xiu-Qing Li: These two authors contributed equally to this paper.

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Formanová, N., Li, XQ., Ferrie, A.M.R. et al. Towards Positional Cloning in Brassica napus: Generation and Analysis of Doubled Haploid B. rapa Possessing the B. napus pol CMS and Rfp Nuclear Restorer Gene. Plant Mol Biol 61, 269–281 (2006). https://doi.org/10.1007/s11103-006-0008-9

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  • DOI: https://doi.org/10.1007/s11103-006-0008-9

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