Plant Molecular Biology

, Volume 50, Issue 2, pp 187–196 | Cite as

A single-base deletion in soybean flavonoid 3′-hydroxylase gene is associated with gray pubescence color

  • Kyoko Toda
  • Daijun Yang
  • Naoki Yamanaka
  • Satoshi Watanabe
  • Kyuya Harada
  • Ryoji Takahashi
Article

Abstract

The T locus of soybean (Glycine max (L.) Merr.) controls pubescence and seed coat color and is presumed to encode flavonoid 3′-hydroxylase (F3′H). The dominant T and the recessive t allele of the locus produce brown and gray pubescence, respectively. PCR primers were constructed based on the sequence of a soybean EST clone homologous to the F3′H gene. A putative full-length cDNA, sf3′h1 was isolated by 3′ and 5′ RACE. Sequence analysis revealed that sf3′h1 consists of 1690 nucleotides encoding 513 amino acids. It had 68% and 66% homology with corresponding F3′H protein sequences of petunia and Arabidopsis, respectively. A conserved amino acid sequence of F3′H proteins, GGEK, was found in the deduced polypeptide. Sequence analysis of the gene from a pair of near-isogenic lines for T, To7B (TT, brown) and To7G (tt, gray) revealed that they differed by a single C deletion in the coding region of To7G. The deletion changed the subsequent reading frame resulting in a truncated polypeptide lacking the GGEK consensus sequence and the heme-binding domain. Genomic Southern analysis probed by sf3′h1 revealed restriction fragment length polymorphisms between cultivars with different pubescence color. Further, sf3′h1 was mapped at the same position with T locus on LG3(c2). PCR-RFLP analysis was performed to detect the single-base deletion. To7B and three cultivars with brown pubescence exhibited shorter fragments, while To7G and three cultivars with gray pubescence had longer fragments due to the single-base deletion. The PCR-RFLP marker co-segregated with genotypes at the Tlocus in a F2 population segregating for the T locus. The above results strongly suggest that sf3′h1 represents the T gene of soybean responsible for pubescence color and that the single-base deletion may be responsible for gray pubescence color.

deletion F3′H flavonoid flavonoid 3′-hydroxylase P450 pubescence color soybean 

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Copyright information

© Kluwer Academic Publishers 2002

Authors and Affiliations

  • Kyoko Toda
    • 1
  • Daijun Yang
    • 1
  • Naoki Yamanaka
    • 2
  • Satoshi Watanabe
    • 2
  • Kyuya Harada
    • 2
  • Ryoji Takahashi
    • 1
  1. 1.National Institute of Crop ScienceTsukuba, IbarakiJapan
  2. 2.Graduate School of Science and TechnologyChiba UniversityMatsudo, ChibaJapan

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