Development of Gene-based Markers for the Allelic Selection of the Restorer-of-fertility Gene, Rfo, in Radish (Raphanus sativus)

  • Kim, Sunggil (Department of Plant Biotechnology, Biotechnology Research Institute, Chonnam National University) ;
  • Lim, Heerae (Biotech Application Team, Dongbu Advanced Research Institute, Dongbu HiTek Co., Ltd.) ;
  • Cho, Kang-Hee (National Institute of Horticultural & Herbal Science, RDA) ;
  • Park, Pue Hee (National Institute of Horticultural & Herbal Science, RDA) ;
  • Park, Suhyung (National Institute of Horticultural & Herbal Science, RDA) ;
  • Sung, Soon-Kee (Biotech Application Team, Dongbu Advanced Research Institute, Dongbu HiTek Co., Ltd.) ;
  • Oh, Daegeun (National Institute of Horticultural & Herbal Science, RDA) ;
  • Kim, Ki-Taek (National Institute of Horticultural & Herbal Science, RDA)
  • Received : 2009.08.12
  • Published : 20090900

Abstract

Cytoplasmic male sterility (CMS) and fertility restoration have been utilized as valuable tools for $F_1$-hybrid seed production in many crops despite laborious breeding processes. Molecular markers for the selection of CMS-related genes help reduce the expenses and breeding times. A previously reported genomic region containing the Ppr-B gene, which is responsible for restoration of fertility and corresponds to the Rfo locus, was used to develop gene-based or so-called "functional" markers for allelic selection of the restorer-of-fertility gene (Rfo) in $F_1$-hybrid breeding of radish (Raphanus sativus L.) Polymorphic sequences among Rfo alleles of diverse breeding lines of radish were examined by sequencing the Ppr-B alleles. However, presence of Ppr-B homolog, designated as Ppr-D, interferes on specific PCR amplification of Ppr-B in certain breeding lines. The organization of Ppr-D, resolved by genome walking, revealed extended homology with Ppr-B even in the promoter region. Interestingly, PCR amplification of Ppr-D was repeatedly unsuccessful in certain breeding lines implying the lack of Ppr-D in these radishes. Ppr-B could only be successfully amplified for analysis through designing primers based on the sequences unique to Ppr-B that exclude interference from Ppr-D gene. Four variants of Rfo alleles were identified from 20 breeding lines. A combination of three molecular markers was developed in order to genotype the Rfo locus based on polymorphisms among four different variants. These markers will be useful in facilitating $F_1$-hybrid cultivar development in radish.

Keywords

Acknowledgement

Supported by : Rural Development Administration

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