Plant Molecular Biology

, Volume 63, Issue 1, pp 125–135 | Cite as

A single-base deletion in soybean flavonol synthase gene is associated with magenta flower color

  • Ryoji Takahashi
  • Stephen M. Githiri
  • Kouta Hatayama
  • Emilyn G. Dubouzet
  • Norimoto Shimada
  • Toshio Aoki
  • Shin-ichi Ayabe
  • Tsukasa Iwashina
  • Kyoko Toda
  • Hisakazu Matsumura
Article

Abstract

The Wm locus of soybean [Glycine max (L.) Merr.] controls flower color. Dominant Wm and recessive wm allele of the locus produce purple and magenta flower, respectively. A putative full-length cDNA of flavonol synthase (FLS), gmfls1 was isolated by 5′ RACE and end-to-end PCR from a cultivar Harosoy with purple flower (WmWm). Sequence analysis revealed that gmfls1 consisted of 1,208 nucleotides encoding 334 amino acids. It had 59–72% homology with FLS proteins of other plant species. Conserved dioxygenase domains A and B were found in the deduced polypeptide. Sequence comparison between Harosoy and Harosoy-wm (magenta flower mutant of Harosoy; wmwm) revealed that they differed by a single G deletion in the coding region of Harosoy-wm. The deletion changed the subsequent reading frame resulting in a truncated polypeptide consisting of 37 amino acids that lacked the dioxygenase domains A and B. Extracts of E. coli cells expressing gmfls1 of Harosoy catalyzed the formation of quercetin from dihydroquercetin, whereas cell extracts expressing gmfls1 of Harosoy-wm had no FLS activity. Genomic Southern analysis suggested the existence of three to four copies of the FLS gene in the soybean genome. CAPS analysis was performed to detect the single-base deletion. Harosoy and Clark (WmWm) exhibited longer fragments, while Harosoy-wm had shorter fragments due to the single-base deletion. The CAPS marker co-segregated with genotypes at Wm locus in a F2 population segregating for the locus. Linkage mapping using SSR markers revealed that the Wm and gmfls1 were mapped at similar position in the molecular linkage group F. The above results strongly suggest that gmfls1 represents the Wm gene and that the single-base deletion may be responsible for magenta flower color.

Keywords

Deletion FLS Flavonol synthase Flower color Soybean Wm locus 

Abbreviations

FLS

Flavonol synthase

F3′H

Flavonoid 3′-hydroxylase

F3′5′H

Flavonoid 3′5′-hydroxylase

DDBJ accession number

AB246668 

Notes

Acknowledgments

We thank Dr. R.L. Nelson at USDA/ARS University of Illinois for providing the seeds of T235. We are grateful to Dr. Joseph G. Dubouzet (Natl. Inst. Agrobiol. Sci.) for critical reading of the manuscript, and Dr. Eduàrdo R. Benitez (Nat 1. Inst. Crop. Sci.) for taking photographs.

References

  1. Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402CrossRefPubMedGoogle Scholar
  2. Baker KE, Parker R (2004) Nonsense-mediated mRNA decay: terminating erroneous gene expression. Curr Opin Cell Biol 16:293–299CrossRefPubMedGoogle Scholar
  3. Buttery BR, Buzzell RI (1973) Varietal differences in leaf flavonoids of soybeans. Crop Sci 13:103–106Google Scholar
  4. Buttery BR, Buzzell RI (1976) Flavonol glycoside genes and photosynthesis in soybean. Crop Sci 16:547–550CrossRefGoogle Scholar
  5. Buzzell RI, Buttery BR (1974) Flavonol glycoside genes in soybeans. Can J Genet Cytol 16:897–899Google Scholar
  6. Buzzell RI, Buttery BR (1977) Inheritance and linkage of a magenta flower gene in soybeans. Can J Genet Cytol 19:749–751Google Scholar
  7. Buzzell RI, Buttery BR (1982) Genetics of black pigmentation of soybean seed coats/hila. Soybean Genet Newsl 9:26–29Google Scholar
  8. Buzzell RI, Buttery BR (1998) Genetics of leaf waviness in soybean. Soybean Genet Newsl 25:23–24Google Scholar
  9. Conti E, Izaurralde E (2005) Nonsense-mediated mRNA decay: molecular insights and mechanistic variations across species. Curr Opin Cell Biol 17:316–325CrossRefPubMedGoogle Scholar
  10. Cregan PB, Jarvic T, Bush AL, Shoemaker RC, Lark KG, Kahler AL, Kaya N, VanToai TT, Lohnes DG, Chung J, Specht JE (1999) An integrated genetic map of the soybean genome. Crop Sci. 39:1464–1490CrossRefGoogle Scholar
  11. Dixon RA, Steele CL (1999) Flavonoids and isoflavonoids—a gold mine for metabolic engineering. Trends Plant Sci 4:394–400CrossRefPubMedGoogle Scholar
  12. Fasoula DA, Stephens PA, Nickell CD, Vodkin LO (1995) Cosegregation of purple-throat flower color with dihydroflavonol reductase polymorphism in soybean. Crop Sci 35:1028–1031CrossRefGoogle Scholar
  13. Fehr WR, Caviness CE, Burmood DT, Pennington JS (1971) Stage of development descriptions for soybeans, GLYCINE MAX (L.) Merrill. Crop Sci 11:929–931CrossRefGoogle Scholar
  14. Forkmann G (1991) Flavonoids as flower pigments: the formation of the natural spectrum and its extension by genetic engineering. Plant Breed 106:1–26CrossRefGoogle Scholar
  15. Holton TA, Brugliera F, Tanaka Y (1993) Cloning and expression of flavonol synthase from Petunia hybrida. Plant J 4:1003–1010CrossRefPubMedGoogle Scholar
  16. Holton TA, Cornish EC (1995) Genetics and biochemistry of anthocyanin biosynthesis. Plant Cell 7:1071–1083CrossRefPubMedGoogle Scholar
  17. Lander ES, Green P, Abrahamson J, Barlow A, Day MJ, Lincoln SE, Newberg L (1987) Mapmaker: an interactive computer package for constructing primary genetic linkage map of experimental and natural populations. Genomics 1:174–181CrossRefPubMedGoogle Scholar
  18. Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucl Acid Res 8:4321–4325Google Scholar
  19. Palmer RG, Pfeiffer TW, Buss GR, Kilen TC (2004) Qualitative genetics. In: Boerma HR, Specht JE (eds) Soybeans: improvement, production, and uses, 3rd edn. Agron Monogr 16. ASA, CSSA, and SSSA, Madison, WI, pp. 137–233Google Scholar
  20. Rode MW, Bernard RL (1975) Inheritance of wavy leaf. Soybean Genet Newsl 2:42–44Google Scholar
  21. Scott-Moncrieff R (1936) A biochemical survey of some Mendelian factors for flower color. J Genet 32:117–170CrossRefGoogle Scholar
  22. Shah DM, Hightower RC, Meagher RB (1983) Genes encoding actin in higher plants: intron positions are highly conserved but the coding sequences are not. J Mol Appl Genet 2:111–126PubMedGoogle Scholar
  23. Shirley BW (1996) Flavonoid biosynthesis: new functions for an old pathway. Trends Plant Sci 1:377–382CrossRefGoogle Scholar
  24. Shirley BW (2001) Flavonoid biosynthesis. A colorful model for genetics, biochemistry, cell biology, and biotechnology. Plant Physiol 126:485–493CrossRefGoogle Scholar
  25. Stafford HA, (1990) Flavonoid metabolism. CRC Press, Boca Raton, FL, pp. 1–298Google Scholar
  26. Toda K, Yang D, Yamanaka N, Watanabe S, Harada K, Takahashi R (2002) A single-base deletion in soybean flavonoid 3′-hydroxylase gene is associated with gray pubescence color. Plant Mol Biol 50:187–196CrossRefPubMedGoogle Scholar
  27. Todd JJ, Vodkin LO (1996) Duplications that suppress and deletions that restore expression from a chalcone synthase multigene family. Plant Cell 8:687–699CrossRefPubMedGoogle Scholar
  28. Zabala G, Vodkin LO (2005) The wp mutation of Glycine max carries a gene-fragment-rich transposon of the CACTA superfamily. Plant Cell 17:2619–2632CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  • Ryoji Takahashi
    • 1
  • Stephen M. Githiri
    • 1
    • 2
  • Kouta Hatayama
    • 1
    • 3
  • Emilyn G. Dubouzet
    • 1
    • 4
  • Norimoto Shimada
    • 5
  • Toshio Aoki
    • 5
  • Shin-ichi Ayabe
    • 5
  • Tsukasa Iwashina
    • 6
  • Kyoko Toda
    • 1
  • Hisakazu Matsumura
    • 1
  1. 1.National Institute of Crop Science and University of TsukubaTsukubaJapan
  2. 2.Department of Crop ScienceUniversity of NairobiNairobiKenya
  3. 3.National Institute of Advanced Industrial Science and TechnologyBiomass Technology Research CenterHiroshimaJapan
  4. 4.National Institute of Advanced Industrial Science and TechnologyTsukubaJapan
  5. 5.Nihon UniversityFujisawaJapan
  6. 6.Tsukuba Botanical GardenNational Science MuseumTsukubaJapan

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