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Euphytica

, Volume 188, Issue 1, pp 141–151 | Cite as

Ant28 gene for proanthocyanidin synthesis encoding the R2R3 MYB domain protein (Hvmyb10) highly affects grain dormancy in barley

  • Eiko HimiEmail author
  • Yuko Yamashita
  • Naoto Haruyama
  • Takashi Yanagisawa
  • Masahiko Maekawa
  • Shin Taketa
Article

Abstract

A number of anthocyanin- and proanthocyanidin-free mutants (ant mutants) in barley were induced and selected because of breeding interest to reduce proanthocyanidins, which could cause haze and degrade the quality of beer. Ant loci, known as anthocyanin or proanthocyanidin synthesis genes, are classified into Ant1 to Ant30 through allelism tests. However, only the Ant18 gene has been molecularly shown to encode dihydroflavonol 4-reductase (DFR), which is involved in both anthocyanin and proanthocyanidin synthesis. In this study, an R2R3 MYB gene of barley was isolated by PCR and named Hvmyb10 due to its similarity to Tamyb10 of wheat, which is a candidate for the R-1 gene grain color regulator. The predicted amino acid sequences of Hvmyb10 showed high similarity not only to Tamyb10 but also to TT2, the proanthocyanidin regulator of Arabidopsis. Non-synonymous nucleotide substitutions in the Hvmyb10 gene were found in all six ant28 mutants tested. Mapping showed that a polymorphism in Hvmyb10 perfectly cosegregated with the ant 28 phenotype on the distal region of the long arm of chromosome 3H. These results demonstrate that ant28 encodes Hvmyb10, the R2R3 MYB domain protein that regulates proanthocyanidin accumulation in developing grains. The reduced grain dormancy of ant28 mutants compared with those of the respective wild types indicates that Hvmyb10 is a key factor in grain dormancy in barley.

Keywords

Barley Proanthocyanidin Grain dormancy Ant mutants 

Abbreviations

CHI

Chalcone isomerase

CHS

Chalcone synthase

DAP

Days after pollination

DFR

Dihydroflavonol 4-reductase

F3H

Flavanone 3-hydroxylase

GI

Germination index

LAR

Leucoanthocyandin reductase

Notes

Acknowledgments

We thank Dr. Udda Lundqvist, Nordic Genetic Resource Center, and Dr. Fredric Ottosson, Department of Crop Science, Swedish University of Agricultural Sciences, Alnarp, Sweden, for kindly providing the grains of the ant28 mutants. This work was supported by Ofu-Kai for the Promotion of Education and Culture at Japan Women’s University, the Elizabeth Arnold Fuji Foundation, and a grant from the Ministry of Agriculture, Forestry, and Fisheries of Japan. (Development of crop production technology for all year round multi-utilization of paddy fields).

Supplementary material

10681_2011_552_MOESM1_ESM.docx (98 kb)
Supplementary material 1 (DOCX 97 kb)

References

  1. Aastrup S, Outtrup H, Erdal K (1984) Location of the proanthocyanidins in the barley grain. Carlsberg Res Commun 49:105–109CrossRefGoogle Scholar
  2. Baskin JM, Baskin CC (2004) A classification system for seed dormancy. Seed Science Research 14:1–16Google Scholar
  3. Debeaujon I, Leon-Kloosterziel KM, Koornneef M (2000) Influence of the testa on seed dormancy, germination, and longevity in Arabidopsis. Plant Physiol 122(2):403–414PubMedCrossRefGoogle Scholar
  4. Debeaujon I, Peeters AJ, Leon-Kloosterziel KM, Koornneef M (2001) The TRANSPARENT TESTA12 gene of Arabidopsis encodes a multidrug secondary transporter-like protein required for flavonoid sequestration in vacuoles of the seed coat endothelium. Plant Cell 13(4):853–871PubMedGoogle Scholar
  5. Flintham J, Adlam R, Bassoi M, Holdsworth M, Gale M (2002) Mapping genes for resistance to sprouting damage in wheat. Euphytica 126:39–45CrossRefGoogle Scholar
  6. Flintham JE (2000) Different genetic components control coat-imposed and embryo-imposed dormancy in wheat. Seed Sci Res 10:43–50CrossRefGoogle Scholar
  7. Gale MD, Atkinson MD, Chinoy CN, Harcourt RL, Jia J, Li QY, Devos KM (1995) Genetic maps of hexaplid wheat. Proceedings of the eighth international wheat genetics symposium. China Agricultural Scienteh Press, BeijingGoogle Scholar
  8. Garvin DF, Miller-Garvin JE, Viccars EA, Jacobsen JV, Brown AHD (1998) Identification of molecular markers linked to ant28–484, a mutation that eliminates proanthocyanidin production in barley seeds. Crop Sci 38:1250–1255CrossRefGoogle Scholar
  9. Haruyama N, Oozeki M, Sotome T, Takayama T, Watanabe H, Okiyama T (2011) Development of caps marker linked to the proanthocyanidin-free gene ant28 in barley, and efficient usage for breeding paf cultivars. Bull Tochigi Agr Exp Stn 66:37–42Google Scholar
  10. Himi E, Maekawa M, Miura H, Noda K (2011a) Development of PCR markers for Tamyb10 related to R-1, red grain color gene in wheat. Theor Appl Genet 122(8):1561–1576PubMedCrossRefGoogle Scholar
  11. Himi E, Maekawa M, Noda K (2011b) Differential expression of three flavanone 3-hydroxylase genes in grains and coleoptiles of wheat. Int J Plant Genomics (2011:369460)Google Scholar
  12. Himi E, Mares DJ, Yanagisawa A, Noda K (2002) Effect of grain colour gene (R) on grain dormancy and sensitivity of the embryo to abscisic acid (ABA) in wheat. J Exp Bot 53(374):1569–1574PubMedCrossRefGoogle Scholar
  13. Himi E, Noda K (2004) Isolation and location of three homoeologous dihydroflavonol-4-reductase (DFR) genes of wheat and their tissue-dependent expression. J Exp Bot 55(396):365–375PubMedCrossRefGoogle Scholar
  14. Jende-Strid B (1978) Mutation frequencies obtained after sodium azide treatment in different barley varieties. Barley Genetics Newsl 8:55–57Google Scholar
  15. Jende-Strid B (1993) Genetic control of flavonoid biosynthesis in barley. Hereditas 119:187–204CrossRefGoogle Scholar
  16. Jende-Strid B (1994) Co-ordinator’s report: anthocyanin genes. Barley Genetics Newsl 24:162–165Google Scholar
  17. Jende-Strid B (1998) Proanthocyanidin-free 30, ant30. Barley Genetics Newsl 29:99Google Scholar
  18. Kristiansen K (1984) Biosynthesis of proanthocyanidins in barley: genetic control of the conversion of dihydroquercetin to catechin and procyanidins. Carlsberg Res Commun 49:503–524CrossRefGoogle Scholar
  19. Kristiansen K, Rohde W (1991) Structure of the Hordeum vulgare gene encoding dihydroflavonol-4-reductase and molecular analysis of ant18 mutants blocked in flavonoid synthesis. Mol Gen Genet 230(1–2):49–59PubMedCrossRefGoogle Scholar
  20. Lepiniec L, Debeaujon I, Routaboul JM, Baudry A, Pourcel L, Nesi N, Caboche M (2006) Genetics and biochemistry of seed flavonoids. Annu Rev Plant Biol 57:405–430PubMedCrossRefGoogle Scholar
  21. Mares D, Mrva K, Cheong J, Williams K, Watson B, Storlie E, Sutherland M, Zou Y (2005) A QTL located on chromosome 4A associated with dormancy in white- and red-grained wheats of diverse origin. Theor Appl Genet 111(7):1357–1364PubMedCrossRefGoogle Scholar
  22. Marles MA, Ray H, Gruber MY (2003) New perspectives on proanthocyanidin biochemistry and molecular regulation. Phytochemistry 64:367–383PubMedCrossRefGoogle Scholar
  23. Matus-Cadiz MA, Daskalchuk TE, Verma B, Puttick D, Chibbar RN, Gray GR, Perron CE, Tyler RT, Hucl P (2008) Phenolic compounds contribute to dark bran pigmentation in hard white wheat. J Agric Food Chem 56(5):1644–1653PubMedCrossRefGoogle Scholar
  24. McIntosh RA, Hart GE, Devos KM, Gale MD, Rogers WJ (1998) Catalogue of gene symbols for wheat. In: Proceedings of the ninth international wheat genetics symposium, 1998. University of Saskatchewan Extension Press, SaskatchewanGoogle Scholar
  25. Meldgaard M (1992) Expression of chalcone synthase, dihydroflavonol reductase, and flavanone-3-hydroxylase in mutants of barley deficient in anthocyanin and proanthocyanidin biosynthesis. Theor Appl Genet 83:695–706CrossRefGoogle Scholar
  26. Mori M, Uchino N, Chono M, Kato K, Miura H (2005) Mapping QTLs for grain dormancy on wheat chromosome 3A and the group 4 chromosomes, and their combined effect. Theor Appl Genet 110(7):1315–1323PubMedCrossRefGoogle Scholar
  27. Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8(19):4321–4325PubMedCrossRefGoogle Scholar
  28. Nakamura S, Abe F, Kawahigashi H, Nakazono K, Tagiri A, Matsumoto T, Utsugi S, Ogawa T, Handa H, Ishida H, Mori M, Kawaura K, Ogihara Y, Miura H (2011) A wheat homolog of MOTHER OF FT AND TFL1 acts in the regulation of germination. Plant Cell. doi: 10.1105/tpc.111.088492
  29. Nelson JC, Van Deynze AE, Autrique E, Sorrells ME, Lu YH, Negre S, Bernard M, Leroy P (1995) Molecular mapping of wheat. Homoeologous group 3. Genome 38:525–533PubMedCrossRefGoogle Scholar
  30. Nesi N, Jond C, Debeaujon I, Caboche M, Lepiniec L (2001) The Arabidopsis TT2 gene encodes an R2R3 myb domain protein that acts as a key determinant for proanthocyanidin accumulation in developing seed. Plant Cell 13(9):2099–2114PubMedGoogle Scholar
  31. Page RD (1996) Treeview: an application to display phylogenetic trees on personal computers. Comput Appl Biosci 12(4):357–358PubMedGoogle Scholar
  32. Pourcel L, Routaboul JM, Kerhoas L, Caboche M, Lepiniec L, Debeaujon I (2005) TRANSPARENT TESTA10 encodes a laccase-like enzyme involved in oxidative polymerization of flavonoids in Arabidopsis seed coat. Plant Cell 17(11):2966–2980PubMedCrossRefGoogle Scholar
  33. Price ML, van Scoyoc S, Butler LG (1978) A critical evaluation of the vanillin reaction as an assay for tannin in sorghum grain. J Agric Food Chem 26(5):1214–1218CrossRefGoogle Scholar
  34. Tonooka T, Kawada N, Yoshida M, Yoshioka T, Oda S, Hatta K, Hatano T, Fujita M, Kubo K (2010) Breeding of a new food barley cultivar “shiratae nijo” exhibiting no after-cooking discoloration. Breeding Sci 60:172–176CrossRefGoogle Scholar
  35. Varshney RK, Marcel TC, Ramsay L, Russell J, Roder MS, Stein N, Waugh R, Langridge P, Niks RE, Graner A (2007) A high density barley microsatellite consensus map with 775 SSR loci. Theor Appl Genet 114(6):1091–1103PubMedCrossRefGoogle Scholar
  36. von Wettstein D (2007) From analysis of mutants to genetic engineering. Annu Rev Plant Biol 58:1–19CrossRefGoogle Scholar
  37. Warner RL, Kudrna DA, Spaeth SC, Jones SS (2000) Dormancy in white-grain mutants of chinese spring wheat (Triticum aestivum L.). Seed Sci Res 10:51–60Google Scholar
  38. Zhao J, Dixon RA (2010) The ‘ins’ and ‘outs’ of flavonoid transport. Trends Plant Sci 15(2):72–80PubMedCrossRefGoogle Scholar
  39. Zimmermann IM, Heim MA, Weisshaar B, Uhrig JF (2004) Comprehensive identification of Arabidopsis thaliana MYB transcription factors interacting with R/B-like bHLH proteins. Plant J 40(1):22–34PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Eiko Himi
    • 1
    Email author
  • Yuko Yamashita
    • 1
  • Naoto Haruyama
    • 2
    • 3
  • Takashi Yanagisawa
    • 4
  • Masahiko Maekawa
    • 1
  • Shin Taketa
    • 1
  1. 1.Institute of Plant Science and ResourcesOkayama UniversityOkayamaJapan
  2. 2.Tochigi Prefectural Agricultural Experiment StationTochigiJapan
  3. 3.Tochigi Prefectural Sustainable Agriculture Extension CenterTochigiJapan
  4. 4.NARO Institute of Crop Science (NICS)TsukubaJapan

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