Advertisement

Euphytica

, Volume 175, Issue 1, pp 1–12 | Cite as

Transfer of the Kosena Rfk1 gene, required in hybrid seed production, from oilseed rape to turnip rape

  • Tarja Niemelä
  • Mervi Seppänen
  • Lauri Jauhiainen
  • Unto Tulisalo
Review

Abstract

The Ogura cytoplasmic male sterility and fertility restoring (cms/Rf) system is one of the most promising hybrid systems in Brassica species. The Ogura cms has been introduced earlier into spring turnip rape (Brassica rapa L.) and the sterility has been stable. To enable the production of F1 hybrids in turnip rape, the Kosena fertility restoring gene (Rfk1), a homologue of the Ogura fertility restoring gene (Rfo), was transferred from spring oilseed rape (Brassica napus L.) into spring turnip rape by the traditional backcross method. The male fertility, measured here as a pollen production, of the F1 hybrids between turnip rape and oilseed rape was good (83%). The transmission rate of the Rfk1 gene during backcrossing was not essentially different between the pollen (35%) and egg (33%) cells. For the selection of homozygous (Rfk1/Rfk1) turnip rape plants, both testcrosses to male sterile (Ogura cms) turnip rape line and TaqMan based real-time qPCR method were used simultaneously. The TaqMan qPCR detection system was good when used for selecting homozygous plants out of heterozygous ones before flowering. This enabled interpollination instead of inbreeding, which is a benefit with cross-pollinating crop species like turnip rape. According to testcross results of the offspring between male sterile line and homozygous (Rfk1/Rfk1) turnip rape plants, 100% fertility was reached. It was a good indication, that turnip rape with Ogura cms can be restored with the Kosena Rfk1 gene. However, during the subsequent selection of homozygous (Rfk1/Rfk1) plants, it was observed that the Rfk1 gene was still unstable in the genome of turnip rape, and thus the Rfk1 gene needs to be stabilized before its full exploitation in commercial hybrid production is possible.

Keywords

Turnip rape Brassica rapa L. Kosena Rfk1 gene Ogura cms TaqMan qPCR 

Notes

Acknowledgments

We thank for Dr. Takako Sakai and Dr. Jun Imamura for providing the breeding lines RfA4 and RfA12, and their technical and scientific support in PCR and qPCR analysis. The study was supported by National Emergency Supply Agency, Finnish Ministry of Agriculture and Forestry, Mildola Oy, Maatalouskesko Oy and RaisioYhtymä Oy.

References

  1. Brown GG, Formanova N, Jin H, Wargachuk R, Dendy C, Patil P, Laforest M, Zhang J, Cheung WY, Landry BS (2003) The radish Rfo restorer gene of Ogura cytoplasmic male sterility encodes a protein with multiple pentatricopeptide repeats. Plant J 35:262–272CrossRefPubMedGoogle Scholar
  2. Budahn H, Schrader O, Peterka H (2008) Development of a complete set of disomic rape-radish chromosome-addition lines. Euphytica 162:117–128CrossRefGoogle Scholar
  3. Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J, Kubista M, Mueller R, Nolan T, Pfaffl W, Shipley GR, Vandesompele J, Wittwer CT (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55:611–622CrossRefPubMedGoogle Scholar
  4. Chang SB, de Jong H (2005) Production of alien chromosome additions and their utility in plant genetics. Cytogenet Genome Res 109:335–343CrossRefPubMedGoogle Scholar
  5. Chevre AM, This P, Eber F, Deschamps M, Renard M, Delseny M, Quiros CF (1991) Characterization of disomic addition lines Brassica napus-Brassica nigra by isozyme, fatty acid, and RFLP markers. Theor Appl Genet 81:43–49CrossRefGoogle Scholar
  6. Fleiss JL, Levin B, Cho Paik M (2003) Statistical methods for rates and proportions, 3rd edn. Wiley-Interscience, Hoboken, NJGoogle Scholar
  7. Formanova N, Li XQ, Ferrie AMR, DePauw M, Keller WA, Landry B, Brown GG (2006) 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 61:269–281CrossRefPubMedGoogle Scholar
  8. Heyn FW (1978) Introgression of restorer genes from Raphanus sativus into cytoplasmic male sterile Brassica napus and the genetics of fertility restoration. Proceedings of 5th International Rapeseed Conference, vol 1. Malmö, Sweden, pp 82–83Google Scholar
  9. Hu XY, Sullivan-Gilbert M, Kubik T, Danielson J, Hnatiuk N, Marchione W, Greene T, Thompson SA (2008) Mapping of the Ogura fertility restorer gene Rfo and development of Rfo allele-specific markers in canola (Brassica napus L.). Mol Breed 22:663–674CrossRefGoogle Scholar
  10. Imai R, Koizuka N, Fujimoto H, Hayakawa T, Sakai T, Imamura J (2003) Delimitation of the fertility restorer locus Rfk1 to a 43-kb contig in Kosena radish (Raphanus sativus L.). Mol Gen Genom 269:388–394CrossRefGoogle Scholar
  11. Iwabuchi M, Koizuka N, Fujimoto H, Sakai T, Imamura J (1999) Identification and expression of the kosena radish (Raphanus sativus cv. Kosena) homologue of the ogura radish CMS-associated gene, orf138. Plant Mol Biol 39:183–188CrossRefPubMedGoogle Scholar
  12. Kaneko Y, Yano H, Bang SW, Matsuzawa Y (2003) Genetic stability and maintenance of Raphanus sativus lines with an added Brassica rapa chromosome. Plant Breed 122:239–243CrossRefGoogle Scholar
  13. Koizuka N, Imai R, Iwabuchi M, Sakai T, Imamura J (2000) Genetic analysis of fertility restoration and accumulation of ORF125 mitochondrial protein in the kosena radish (Raphanus sativus cv. Kosena) and a Brassica napus restorer line. Theor Appl Genet 100:949–955CrossRefGoogle Scholar
  14. Koizuka N, Imai R, Fujimoto H, Hayakawa T, Kimura Y, Kohno-Murase J, Sakai T, Kawasaki S, Imamura J (2003) Genetic characterization of a pentatricopeptide repeat protein gene, orf687, that restores fertility in the cytoplasmic male-sterile Kosena radish. Plant J 34:407–415CrossRefPubMedGoogle Scholar
  15. Lange W, Toxopeus H, Lubberts JH, Dolstra O, Harrewijn JL (1989) The development of Raparadish (x Brassicoraphanus, 2n = 38), a new crop in agriculture. Euphytica 40:1–14Google Scholar
  16. Lee KH, Namai H (1992) Pollen fertility and seed set percentage after backcrossing of sesquidiploids (AAC genomes) derived from interspecific hybrid between Brassica campestris L. (AA) and B. oleracea L. (CC) and frequency distribution of aneuploids in the progenies. Jpn J Breed 42:43–53Google Scholar
  17. Lee KH, Namai H (1994) Cytogenetic and morphological characteristics of new types of diploids (2n = 22, 24, 40) derived from consecutive selfing of aneuploids in brassica crops. Euphytica 72:15–22CrossRefGoogle Scholar
  18. Leflon M, Eber F, Letanneur JC, Chelysheva L, Coriton O, Huteau V, Ryder CD, Barker G, Jenczewski E, Chevre AM (2006) Pairing and recombination at meiosis of Brassica rapa (AA) × Brassica napus (AACC) hybrids. Theor Appl Genet 113:1467–1480CrossRefPubMedGoogle Scholar
  19. Metz PLJ, Jacobsen E, Nap JP, Pereira A, Stiekema WJ (1997) The impact on biosafety of the phosphinothricin-tolerance transgene in inter-specific B.rapa × B.napus hybrids and their successive backcrosses. Theor Appl Genet 95:442–450CrossRefGoogle Scholar
  20. Mikkelsen TR, Jensen J, Jørgensen RB (1996) Inheritance of oilseed rape (Brassica napus) RAPD markers in a backcross progeny with Brassica campestris. Theor Appl Genet 92:492–497CrossRefGoogle Scholar
  21. Mimida N, Sakamoto W, Murata M, Motoyoshi F (1999) TERMINAL FLOWER 1-like genes in Brassica species. Plant Sci 142:155–162CrossRefGoogle Scholar
  22. Muller PY, Janovjak H, Miserez AR, Doppie Z (2002) Processing of gene expression data generated by quantitative real-time RT-PCR. BioTechniques 32:1372–1379PubMedGoogle Scholar
  23. Niemela T, Seppanen M, Jauhiainen L, Tulisalo U (2006) Yield potential of spring turnip rape synthetics and composite hybrids compared with open-pollinated commercial cultivars. Can J Plant Sci 86:693–700Google Scholar
  24. Peterka H, Budahn H, Schrader O, Ahne R, Schutze W (2004) Transfer of resistance against the beet cyst nematode from radish (Raphanus sativus) to rape (Brassica napus) by monosomic chromosome addition. Theor Appl Genet 109:30–41CrossRefPubMedGoogle Scholar
  25. Primard-Brisset C, Poupard JP, Horvais R, Eber F, Pelletier G, Renard M, Delourme R (2005) A new recombined double low restorer line for the Ogu-INRA cms in rapeseed (Brassica napus L.). Theor Appl Genet 111:736–746CrossRefPubMedGoogle Scholar
  26. Quiros CF, Ochoa O, Kianian SF, Douches D (1987) Analysis of the Brassica oleracea genome by the generation of B. campestris-oleracea chromosome addition lines: characterization by isozymes and rDNA genes. Theor Appl Genet 74:758–766Google Scholar
  27. Sakai T, Liu HJ, Iwabuchi M, Kohno-Murase J, Imamura J (1996) Introduction of a gene from fertility restored radish (Raphanus sativus) into Brassica napus by fusion of X-irradiated protoplasts from a radish restorer line and iodacetoamide-treated protoplasts from a cytoplasmic male-sterile cybrid of B.napus. Theor Appl Genet 93:373–379CrossRefGoogle Scholar
  28. Sovero M (1987) Cytoplasmic male sterility in turnip-rape (Brassica campestris L.). Dissertation, University of ManitobaGoogle Scholar
  29. Struss D, Bellin U, Röbbelen G (1991) Development of B-genome chromosome addition lines of B.napus using different interspecific Brassica hybrids. Plant Breed 106:209–214CrossRefGoogle Scholar
  30. Yi CX, Zhang J, Chan KM, Liu XK, Hong Y (2008) Quantitative real-time PCR assay to detect transgene copy number in cotton (Gossypium hirsutum). Anal Biochem 375:150–152CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Tarja Niemelä
    • 1
  • Mervi Seppänen
    • 1
  • Lauri Jauhiainen
    • 2
  • Unto Tulisalo
    • 1
  1. 1.Department of AgricultureUniversity of HelsinkiHelsinkiFinland
  2. 2.MTT Agrifood Research Finland, Research ServicesJokioinenFinland

Personalised recommendations