Advertisement

Theoretical and Applied Genetics

, Volume 111, Issue 6, pp 1191–1200 | Cite as

Development of a molecular marker specific to a novel CMS line in radish (Raphanus sativus L.)

  • Seok-Hyeon NahmEmail author
  • Hee-Jeong Lee
  • Shi-Woo Lee
  • Gyun-Young Joo
  • Chee-Hark Harn
  • Seung-Gyun Yang
  • Byung-Whan Min
Original Paper

Abstract

In this study, we have investigated the cytoplasmic male sterility (CMS) of a novel male sterile radish line, designated NWB CMS. The NWB CMS was crossed with 16 fertile breeding lines, and all the progenies were completely male sterile. The degree of male sterility exhibited by NWB CMS is more than Ogura CMS from the Cruciferae family. The NWB CMS was found to induce 100% male sterility when crossed with all the tested breeding lines, whereas the Ogura CMS did not induce male sterility with any of the breeding lines. PCR analysis revealed that the molecular factor that influenced Ogura CMS, the orf138 gene, was absent in the NWB CMS line, and that the orf138 gene was not also expressed in this CMS line. In order to identify the cytoplasmic factors that confer male sterility in the NWB CMS line, we carried out RFLP analyses with 32 mitochondrial genes, all of which were used as probes. Fourteen genes exhibited polymorphisms between the NWB CMS line and other radish cultivars. Based on these RFLP data, intergenic primers were developed in order to amplify the intergenic regions between the polymorphic genes. Among these, a primer pair at the 3′ region of the atp6 gene (5′-cgcttggactatgctatgtatga-3′) and the 5′ region of the nad3 gene (5′-tcatagagaaatccaatcgtcaa-3′) produced a 2 kbp DNA fragment as a result of PCR. This DNA fragment was found to be specific to NWB CMS and was not present in other CMS types. It appears that this fragment could be used as a DNA marker to select NWB CMS line in a radish-breeding program.

Keywords

Mitochondrial Genome Male Sterility Cytoplasmic Male Sterility Breeding Line RFLP Analysis 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

This work was supported, in part, by the Crop Functional Genomics Center of the 21st Century Frontier Research Program, funded by the Ministry of Science and Technology, and by a grant of the Biogreen21 Research Program funded by the Rural Development Administration of the Republic of Korea. We also would like to thank Mr. Soo Young Chung, for his technical help with the NWB CMS line.

References

  1. Albaum M, Luhrs R, Trautner J, Abel WO (1995) The Tokumasu radish mitochondrial genome contains two complete atp9 reading frame. Plant Mol Biol 29: 179–185CrossRefPubMedGoogle Scholar
  2. Bellaoui M, Pelletier G, Budar F (1997) The steady-state level of mRNA from the Ogura cytoplasmic male sterility locus in Brassica cybrids is determined post-transcriptionally by its 3′ region. EMBO J 16(16):5057–5068CrossRefPubMedGoogle Scholar
  3. Bentolila S, Alfonso AA, Hanson MR (2002) A pentatricopeptide repeat-containing gene restores fertility to cytoplasmic male-sterile plants. Proc Natl Acad Sci USA 99:10887–10892CrossRefPubMedGoogle Scholar
  4. Bonhomme S, Budar F, Lancelin D, Small I, Defrance MC, Pelletier G (1992) Sequence and transcript analysis of the Nco2.5 Ogura specific fragment correlated with cytoplasmic male-sterility in Brassica cybrids. Mol Gen Genet 235:340–348CrossRefPubMedGoogle Scholar
  5. Brown GG, Nataa F, Hua J, Richard W, Charles D, Prashant P, Martin L, Jinfa Z, Wing YC, Benoit SL (2003) The radish Rfo restorer gene of Ogura cytoplasmic male sterility encodes a protein with multiple pentatricopeptide repeats. Plant J 35(2):262–272CrossRefPubMedGoogle Scholar
  6. Cardi T, Earle ED (1997) Production of new CMS Brassica oleracea by transfer of ‘Anand’ cytoplasm from B. rapa through protoplast fusion. Theor Appl Genet 94:204–212CrossRefGoogle Scholar
  7. Grelon M, Budar F, Bonhomme S, Pelletier G (1994) Ogura cytoplasmic male-sterility (CMS)-associated orf138 is translated into a mitochondrial membrane polypeptide in male-sterile Brassica cybrids. Mol Gen Genet 243(5):540–547CrossRefPubMedGoogle Scholar
  8. Handa H, Gualberto JM, Grienenberger JM (1995) Characterization of the mitochondrial orfB gene and its derivative, orf224, a chimeric open reading frame specific to one mitochondrial genome of the “Polima” male-sterile cytoplasm in rapeseed (Brassica napus L.). Curr Genet 28(6):546–552CrossRefPubMedGoogle Scholar
  9. Hanson MR, Bentolia S (2004) Interactions of mitochondrial and nuclear genes that affect male gemetophyte development. Plant Cell 16:S154–S169CrossRefPubMedGoogle Scholar
  10. Homme YL, Brown GG (1993) Organizational differences between cytoplasmic male sterile and male fertile Brassica mitochondrial genomes are confined to a single transposed locus. Nucleic Acids Res 21(8):1903–1909PubMedGoogle Scholar
  11. Homme YL, Stahl RJ, Li X, Hameed A, Brown GG (1997) Male sterility induced by Brassica nap cytoplasm is correlated with expression of a chimeric gene homologous to the pol CMS-associated orf224 gene. Curr Genet 31:325–335CrossRefPubMedGoogle Scholar
  12. 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
  13. Jean M, Brown GG, Landry BS (1997) Genetic mapping of fertility restorer genes for the Polima cytoplasmic male sterility in canola (Brassica napus L.) using DNA markers. Theor Appl Genet 95:321–328CrossRefGoogle Scholar
  14. Kang BC, Nahm SH, Huh JH, Yoo HS, Yu JW, Lee MH, Kim BD (2001) An interspecific (Capsicum annuum x C. chinese) F2 linkage map in pepper using RFLP and AFLP markers. Theor Appl Genet 102(4):531–539CrossRefGoogle Scholar
  15. 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
  16. Krishnasamy S, Makaroff CA (1993) Characterization of the radish mitochondrial orfB locus: possible relationship with male sterility in Ogura radish. Curr Genet 24:156–163CrossRefPubMedGoogle Scholar
  17. Krishnasamy S, Makaroff CA (1994) Organ-specific reduction in the abundance of a mitochondrial protein accompanies fertility restoration in cytoplasmic male-sterile radish. Plant Mol Biol 26:935–946CrossRefPubMedGoogle Scholar
  18. Makaroff CA, Palmer JD (1988) Mitochondrial DNA rearrangements and transcriptional alterations in the male-sterile cytoplasm of Ogura radish. Mol Cell Biol 8:1474–1490PubMedGoogle Scholar
  19. Makaroff CA, Apel IJ, Palmer JD (1991) The role of cox2-associated repeated sequences in plant mitochondrial DNA rearrangements and radish cytoplasmic sterility. Curr Genet 19(3):183–190CrossRefPubMedGoogle Scholar
  20. Schnable PS, Wise RP (1998) The molecular basis of cytoplasmic male sterility. Trends Plant Sci 3:175–180CrossRefGoogle Scholar
  21. Singh M, Hamel N, Menassa R, Li XQ, Young B (1996) Nuclear genes associated with a single Brassica CMS restorer locus influence transcripts of three different mitochondrial gene regions. Genetics 143:505–516PubMedGoogle Scholar
  22. Yamagishi H, Terachi T (1996) Molecular and biological studies on male-sterile cytoplasm in the Cruciferae. III. Distribution of Ogura-type cytoplasm among Japanese wild radishes and Asian radish cultivars. Theor Appl Genet 93:325–332CrossRefGoogle Scholar
  23. Yamagishi H, Terachi T (2001) Intra- and inter-specific variations in the mitochondrial gene orf138 of Ogura-type male-sterile cytoplasm from Raphanus sativus and Raphanus raphanistrum. Theor Appl Genet 103:725–732CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Seok-Hyeon Nahm
    • 1
    Email author
  • Hee-Jeong Lee
    • 1
  • Shi-Woo Lee
    • 1
  • Gyun-Young Joo
    • 1
  • Chee-Hark Harn
    • 1
  • Seung-Gyun Yang
    • 1
  • Byung-Whan Min
    • 1
  1. 1.Biotechnology InstituteNong Woo Bio Co.YeojuSouth Korea

Personalised recommendations