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Theoretical and Applied Genetics

, Volume 118, Issue 3, pp 433–441 | Cite as

Identification of a novel chimeric gene, orf725, and its use in development of a molecular marker for distinguishing among three cytoplasm types in onion (Allium cepa L.)

  • Sunggil KimEmail author
  • Eul-Tai Lee
  • Dong Youn Cho
  • Taeho Han
  • Haejeen Bang
  • Bhimanagouda S. Patil
  • Yul Kyun Ahn
  • Moo-Kyoung Yoon
Original Paper

Abstract

A novel chimeric gene with a 5′ end containing the nearly complete sequence of the coxI gene and a 3′ end showing homology with chive orfA501 was isolated by genome walking from two cytoplasm types: CMS-S and CMS-T, both of which induce male-sterility in onion (Allium cepa L.). In addition, the normal active and variant inactive coxI genes were also isolated from onions containing the normal and CMS-S cytoplasms, respectively. The chimeric gene, designated as orf725, was nearly undetectable in normal cytoplasm, and the copy number of the normal coxI gene was significantly reduced in CMS-S cytoplasm. RT-PCR results showed that orf725 was not transcribed in normal cytoplasm. Meanwhile, the normal coxI gene, which is essential for normal mitochondrial function, was not expressed in CMS-S cytoplasm. However, both orf725 and coxI were transcribed in CMS-T cytoplasm. The expression of orf725, a putative male-sterility-inducing gene, was not affected by the presence of nuclear restorer-of-fertility gene(s) in male-fertility segregating populations originating from the cross between a male-sterile plant containing either CMS-T or CMS-S and a male-fertile plant whose genotypes of nuclear restorer gene(s) might be heterozygous. The specific stoichiometry of orf725 and coxI in the mtDNA of the three cytoplasm types was consistent among diverse germplasm. Therefore, a molecular marker based on the relative copy numbers of orf725 and coxI was designed for distinguishing among the three cytoplasm types by one simple PCR. The reliability and applicability of the molecular marker was shown by testing diverse onion germplasm.

Keywords

Mitochondrial Genome coxI Gene Chimeric Gene Restorer Gene Orf725 Gene 
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

Acknowledgments

This study was supported by the “Specific Joint Agricultural Research-promoting Projects (Project No. 200803A01081219)”, RDA, Republic of Korea, and partially supported by the Biotechnology Research Institute, Chonnam National University, Republic of Korea. We thank Seung-Hee Kim and Seon Chong for their dedicated technical help.

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Sunggil Kim
    • 1
    Email author
  • Eul-Tai Lee
    • 2
  • Dong Youn Cho
    • 3
  • Taeho Han
    • 1
  • Haejeen Bang
    • 4
  • Bhimanagouda S. Patil
    • 4
  • Yul Kyun Ahn
    • 5
  • Moo-Kyoung Yoon
    • 5
  1. 1.Department of Plant Biotechnology, Biotechnology Research InstituteChonnam National UniversityGwangjuSouth Korea
  2. 2.Mokpo Experiment StationNational Institute of Crop Science, RDAMuanSouth Korea
  3. 3.ONBREETECH Corp.HaenamSouth Korea
  4. 4.Department of Horticultural Sciences, Vegetable and Fruit Improvement CenterTexas A&M UniversityCollege StationUSA
  5. 5.National Horticultural Research Institute, RDASuwonSouth Korea

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