Planta

, Volume 231, Issue 4, pp 887–899

Coordinated regulation of anthocyanin biosynthesis in Chinese bayberry (Myrica rubra) fruit by a R2R3 MYB transcription factor

  • Shan-Shan Niu
  • Chang-Jie Xu
  • Wang-Shu Zhang
  • Bo Zhang
  • Xian Li
  • Kui Lin-Wang
  • Ian B. Ferguson
  • Andrew C. Allan
  • Kun-Song Chen
Original Article

Abstract

Chinese bayberry (Myrica rubra) is a fruit crop with cultivars producing fruit ranging from white (Shuijing, SJ) to red (Dongkui, DK) and dark red-purple (Biqi, BQ), as a result of different levels of anthocyanin accumulation. Genes encoding the anthocyanin biosynthesis enzymes chalcone synthase, chalcone isomerase, flavanone 3-hydroxylase (F3H), flavonoid 3′-hydroxylase (F3′H), dihydroflavonol 4-reductase (DFR), anthocyanidin synthase (ANS) and UDP-glucose:flavonoid 3-O-glucosyltransferase (UFGT), as well as MrMYB1, a R2R3 MYB transcription factor homologous to known activators of anthocyanin biosynthesis, were isolated from ripe fruit of BQ. Differences in mRNA abundance of MrF3H, MrF3′H, MrDFR1, MrANS and MrUFGT were highly correlated with differential accumulation of anthocyanins between cultivars, suggesting coordinated regulation by transcription factors. The transcript level of MrMYB1 was strongly associated with the anthocyanin content in ripe fruit of the three cultivars, as well as different anthocyanin containing tissues of BQ fruit. Fruit bagging strongly inhibited anthocyanin accumulation in fruit as well as the expression of all anthocyanin biosynthetic genes and MrMYB1. Over-expression of MrMYB1 stimulated both anthocyanin accumulation and activated an Arabidopsis-DFR promoter in tobacco (Nicotiana tabacum). MrMYB1d, an allele with a 1 bp deletion at nucleotide 30 of coding sequence, was observed in SJ and DK fruit, suggesting that a nonsense mutation of the MYB1 protein may be responsible for no or low expression of MYB1 in the white and red fruit. These results show that coordinated expression of multiple biosynthetic genes is involved in anthocyanin accumulation in Chinese bayberry fruit, and this is regulated by MrMYB1.

Keywords

Anthocyanin biosynthesis Chinese bayberry Coordinated gene expression Nonsense mutation MYB Transcription factor 

Abbreviations

ANS

Anthocyanidin synthase

bHLH

Basic-helix-loop-helix

BQ

Biqi

CDS

Coding sequence

CHI

Chalcone isomerase

CHS

Chalcone synthase

DAFB

Days after full bloom

DFR

Dihydroflavonol 4-reductase

DK

Dongkui

EBGs

Early biosynthetic genes

F3H

Flavanone 3-hydroxylase

F3′H

Flavonoid 3′-hydroxylase

LBGs

Late biosynthetic genes

LUC

Firefly luciferase

Q-PCR

Real-time quantitative PCR

REN

Renilla luciferase

SJ

Shuijing

TFs

Transcription factors

UFGT

UDP-glucose:flavonoid 3-O-glucosyltransferase

References

  1. Aharoni A, De Vos CHR, Wein M, Sun ZK, Greco R, Kroon A, Mol JNM, O’Connell AP (2001) The strawberry FaMYB1 transcription factor suppresses anthocyanin and flavonol accumulation in transgenic tobacco. Plant J 28:319–332CrossRefPubMedGoogle Scholar
  2. Ahmed N, Maekawa M, Noda K (2009) Anthocyanin accumulation and expression pattern of anthocyanin biosynthesis genes in developing wheat coleoptiles. Biol Plant 53:223–228CrossRefGoogle Scholar
  3. Albert NW, Lewis DH, Zhang H-B, Irving LJ, Jameson PE, Davies KM (2009) Light-induced vegetative anthocyanin pigmentation in Petunia. J Exp Bot 60:2191–2202CrossRefPubMedGoogle Scholar
  4. Allan AC, Hellens RP, Laing WA (2008) MYB transcription factors that colour our fruit. Trends Plant Sci 13:99–102CrossRefPubMedGoogle Scholar
  5. Ban Y, Honda C, Hatsuyama Y, Igarashi M, Bessho H, Moriguchi T (2007) Isolation and functional analysis of a MYB transcription factor gene that is a key regulator for the development of red coloration in apple skin. Plant Cell Physiol 48:958–970CrossRefPubMedGoogle Scholar
  6. Borevitz JO, Xia Y, Blount J, Dixon RA, Lamb C (2000) Activation tagging identifies a conserved MYB regulator of phenylpropanoid biosynthesis. Plant Cell 12:2383–2393CrossRefPubMedGoogle Scholar
  7. Boss PK, Davies C, Robinson SP (1996) Analysis of the expression of anthocyanin pathway genes in developing Vitis vinifera L cv Shiraz grape berries and the implications for pathway regulation. Plant Physiol 111:1059–1066PubMedGoogle Scholar
  8. Broun P (2005) Transcriptional control of flavonoid biosynthesis: a complex network of conserved regulators involved in multiple aspects of differentiation in Arabidopsis. Curr Opin Plant Biol 8:272–279CrossRefPubMedGoogle Scholar
  9. Chagne D, Carlisle CM, Blond C, Volz RK, Whitworth CJ, Oraguzie NC, Crowhurst RN, Allan AC, Espley RV, Hellens RP, Gardiner SE (2007) Mapping a candidate gene (MdMYB10) for red flesh and foliage colour in apple. BMC Genomics 8:212CrossRefPubMedGoogle Scholar
  10. Chen KS, Xu CJ, Zhang B, Ferguson IB (2004) Red bayberry: botany and horticulture. Hortic Rev 30:83–114Google Scholar
  11. Cominelli E, Gusmaroli G, Allegra D, Galbiati M, Wade HK, Jenkins GI, Tonelli C (2008) Expression analyssis of anthocyanin regulatory genes in response to different light qualities in Arabidopsis thaliana. Plant Physiol 165:886–894CrossRefGoogle Scholar
  12. Davies KM, Schwinn KE (2003) Transcriptional regulation of secondary metabolism. Funct Plant Biol 30:913–925CrossRefGoogle Scholar
  13. Dubos C, Le Gourrierec J, Baudry A, Huep G, Lanet E, Debeaujon I, Routaboul JM, Alboresi A, Weisshaar B, Lepiniec L (2008) MYBL2 is a new regulator of flavonoid biosynthesis in Arabidopsis thaliana. Plant J 55:940–953CrossRefPubMedGoogle Scholar
  14. Espley RV, Hellens RP, Putterill J, Stevenson DE, Kutty-Amma S, Allan AC (2007) Red colouration in apple fruit is due to the activity of the MYB transcription factor, MdMYB10. Plant J 49:414–427CrossRefPubMedGoogle Scholar
  15. Espley RV, Brendolise C, Chagné D, Kutty-Amma S, Green S, Volz R, Putterill J, Schouten HJ, Gardiner SE, Hellens RP, Allan AC (2009) Multiple repeats of a promoter segment causes transcription factor autoregulation in red apples. Plant Cell 21:168–183CrossRefPubMedGoogle Scholar
  16. Fang ZX, Zhang YH, Lü Y, Ma GP, Chen JC, Liu DH, Ye XQ (2009) Phenolic compounds and antioxidant capacities of bayberry juices. Food Chem 113:884–888CrossRefGoogle Scholar
  17. Galego L, Almeida J (2007) Molecular genetic basis of flower colour variegation in Linaria. Genet Res 89:129–134CrossRefPubMedGoogle Scholar
  18. Gonzalez A, Zhao M, Leavitt JM, Lloyd AM (2008) Regulation of the anthocyanin biosynthetic pathway by the TTG1/bHLH/Myb transcriptional complex in Arabidopsis seedlings. Plant J 53:814–827CrossRefPubMedGoogle Scholar
  19. Grotewold E (2006) The genetics and biochemistry of floral pigments. Annu Rev Plant Biol 57:761–780CrossRefPubMedGoogle Scholar
  20. Hellens R, Allan A, Friel E, Bolitho K, Grafton K, Templeton M, Karunairetnam S, Gleave A, Laing W (2005) Transient expression vectors for functional genomics, quantification of promoter activity and RNA silencing in plants. Plant Methods 1:13CrossRefPubMedGoogle Scholar
  21. Huang C-H, Yu B, Teng Y-W, Su J, Shu Q, Cheng Z-Q, Zeng L-Q (2009) Effects of fruit bagging on coloring and related physiology, and qualities of red Chinese sand pears during fruit maturation. Sci Hortic 121:149–158CrossRefGoogle Scholar
  22. Jia H-J, Araki A, Okamoto G (2004) Influence of fruit bagging on aroma volatiles and skin coloration of ‘Hakuho’ peach (Prunus persica Batsch). Postharvest Biol Technol 35:61–68CrossRefGoogle Scholar
  23. Ju Z-G, Liu C-L, Yuan Y-B (1995) Activities of chalcone synthase and UDPGal: flavonoid-3-o-glycosyltransferase in relation to anthocyanin synthesis in apple. Sci Hortic 63:175–185CrossRefGoogle Scholar
  24. Kim S, Binzel ML, Park S, Yoo K-S, Pike LM (2004) Inactivation of DFR (dihydroflavonol 4-reductase) gene transcription results in blockage of anthocyanin production in yellow onions (Allium cepa). Mol Breed 14:253–263CrossRefGoogle Scholar
  25. Kobayashi S, Ishimaru M, Hiraoka K, Honda C (2002) Myb-related genes of the Kyoho grape (Vitis labruscana) regulate anthocyanin biosynthesis. Planta 215:924–933CrossRefPubMedGoogle Scholar
  26. Kobayashi S, Goto-Yamamoto N, Hirochika H (2004) Retrotransposon-induced mutations in grape skin color. Science 304:982CrossRefPubMedGoogle Scholar
  27. Koes R, Verweij W, Quattrocchio F (2005) Flavonoids: a colorful model for the regulation and evolution of biochemical pathways. Trends Plant Sci 10:236–242CrossRefPubMedGoogle Scholar
  28. Kumar S, Tamura K, Nei M (2004) MEGA3:integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5:150–163CrossRefPubMedGoogle Scholar
  29. Lo Piero AR, Consoli A, Puglisi I, Orestano G, Recupero GR, Petrone G (2005) Anthocyaninless cultivars of sweet orange lack to express the UDP-glucose flavonoid 3-O-glucosyl transferase. J Plant Biochem Biotechnol 14:9–14Google Scholar
  30. Matsui K, Umemura Y, Ohme-Takagi M (2008) AtMYBL2, a protein with a single MYB domain, acts as a negative regulator of anthocyanin biosynthesis in Arabidopsis. Plant J 55:954–967CrossRefPubMedGoogle Scholar
  31. Matus JT, Aquea F, Arce-Johnson P (2008) Analysis of the grape MYB R2R3 subfamily reveals expanded wine quality-related clades and conserved gene structure organization across Vitis and Arabidopsis genomes. BMC Plant Biol 8:83CrossRefPubMedGoogle Scholar
  32. Matus JT, Loyola R, Vega A, Peña-Neira A, Bordeu E, Arce-Johnson P, Alcalde JA (2009) Post-veraison sunlight exposure induces MYB-mediated transcriptional regulation of anthocyanin and flavonol synthesis in berry skins of Vitis vinifera. J Exp Bot 60:853–867CrossRefPubMedGoogle Scholar
  33. Nakatsuka T, Haruta KS, Pitaksutheepong C, Abe Y, Kakizaki Y, Yamamoto K, Shimada N, Yamamura S, Nishihara M (2008) Identification and characterization of R2R3-MYB and bHLH transcription factors regulating anthocyanin biosynthesis in gentian flowers. Plant Cell Physiol 49:1818–1829CrossRefPubMedGoogle Scholar
  34. Palapol Y, Ketsa S, Lin-Wang K, Ferguson IB, Allan AC (2009) A MYB transcription factor regulates anthocyanin biosynthesis in mangosteen (Garcinia mangostana L.) fruit during ripening. Planta 229:1323–1334CrossRefPubMedGoogle Scholar
  35. Park J-S, Kim J-B, Cho K-J, Cheon C-I, Sung M-K, Choung M-G, Roh K-H (2008) Arabidopsis R2R3-MYB transcription factor AtMYB60 functions as a transcriptional repressor of anthocyanin biosynthesis in lettuce (Lactuca sativa). Plant Cell Rep 27:985–994CrossRefPubMedGoogle Scholar
  36. Paz-Ares J, Ghosal D, Wienand U, Peterson PA, Saedler H (1987) The regulatory c1 locus of Zea mays encodes a protein with homology to myb proto-oncogene products and with structural similarities to transcriptional activators. EMBO J 6:3553–3558PubMedGoogle Scholar
  37. Romero I, Sanchez-Ballesta MT, Maldonado R, Escribano MI, Merodio C (2008) Anthocyanin, antioxidant activity and stress-induced gene expression in high CO2-treated table grapes stored at low temperature. Plant Physiol 165:522–530CrossRefGoogle Scholar
  38. Rowen DD, Cao MS, Lin-Wang K, Cooney JM, Jensen DJ, Austin PT, Hunt MB, Norling C, Hellens RP, Schaffer RJ, Allan AC (2009) Environmental regulation of leaf colour in red 35S:PAP1 Arabidopsis thaliana. New Phytol 182:102–115CrossRefGoogle Scholar
  39. Saitoh K, Onishi K, Mikami I, Thidar K, Sano Y (2004) Allelic diversification at the C (OsC1) locus of wild and cultivated rice: nucleotide changes associated with phenotypes. Genetics 168:997–1007CrossRefPubMedGoogle Scholar
  40. Schwinn K, Venail J, Shang Y, Mackay S, Alm V, Butelli E, Oyama R, Bailey P, Davies K, Martin C (2006) A small family of MYB-regulatory genes controls floral pigmentation intensity and patterning in the genus Antirrhinum. Plant Cell 18:831–851CrossRefPubMedGoogle Scholar
  41. Shan LL, Li X, Wang P, Cai C, Zhang B, Sun CD, Zhang WS, Xu CJ, Ferguson IB, Chen KS (2008) Characterization of cDNAs associated with lignification and their expression profiles in loquat fruit with different lignin accumulation. Planta 227:1243–1254CrossRefPubMedGoogle Scholar
  42. Spelt C, Quattrocchio F, Mol JN, Koes R (2000) anthocyanin1 of petunia encodes a basic helix-loop-helix protein that directly activates transcription of structural anthocyanin genes. Plant Cell 12:1619–1632CrossRefPubMedGoogle Scholar
  43. Stracke R, Werber M, Weisshaar B (2001) The R2R3-MYB gene family in Arabidopsis thaliana. Curr Opin Plant Biol 4:447–456CrossRefPubMedGoogle Scholar
  44. Streisfeld MA, Rausher MD (2009) Genetic changes contributing to the parallel evolution of red floral pigmentation among Ipomoea species. New Phytol 183:751–763CrossRefPubMedGoogle Scholar
  45. Takos AM, Jaffé FW, Jacob SR, Bogs J, Robinson SP, Walker AR (2006) Light-induced expression of a MYB gene regulates anthocyanin biosynthesis in red apples. Plant Physiol 142:1216–1232CrossRefPubMedGoogle Scholar
  46. Ubi BE, Honda C, Bessho H, Kondo S, Wada M, Kobayashi S, Moriguchi T (2006) Expression analysis of anthocyanin biosynthetic genes in apple skin: effect of UV-B and temperature. Plant Sci 170:571–578CrossRefGoogle Scholar
  47. Walker AR, Lee E, Bogs J, McDavid DAJ, Thomas MR, Robinson SP (2007) White grapes arose through the mutation of two similar and adjacent regulatory genes. Plant J 49:772–785CrossRefPubMedGoogle Scholar
  48. Wrolstand RE, Culbertson JD, Cornwell CJ, Mattick L (1982) Detection of adulteration in blackberry juice concentrates and wines. J Assoc Off Anal Chem 6:1417–1423Google Scholar
  49. Yin XR, Chen KS, Allan AC, Wu RM, Zhang B, Lallu N, Ferguson IB (2008) Ethylene-induced modulation of genes associated with the ethylene signalling pathway in ripening kiwifruit. J Exp Bot 59:2097–2108CrossRefPubMedGoogle Scholar
  50. Yuan Y, Chiu L-W, Li L (2009) Transcriptional regulation of anthocyanin biosynthesis in red cabbage. Planta 230:1141–1153CrossRefPubMedGoogle Scholar
  51. Zhang WS, Li X, Zheng JT, Wang GY, Sun CD, Ferguson IB, Chen KS (2008) Bioactive components and antioxidant capacity of Chinese bayberry (Myrica rubra Sieb. and Zucc.) fruit in relation to fruit maturity and postharvest storage. Eur Food Res Technol 227:1091–1097CrossRefGoogle Scholar
  52. Zhang SM, Gao ZS, Xu CJ, Chen KS, Wang GY, Zheng JT, Lu T (2009) Genetic diversity of chinese bayberry (Myrica rubra Sieb. et Zucc.) accessions revealed by amplified fragment length polymorphism. HortScience 44:487–491Google Scholar
  53. 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 40:22–34CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Shan-Shan Niu
    • 1
  • Chang-Jie Xu
    • 1
  • Wang-Shu Zhang
    • 1
  • Bo Zhang
    • 1
  • Xian Li
    • 1
  • Kui Lin-Wang
    • 2
  • Ian B. Ferguson
    • 2
  • Andrew C. Allan
    • 2
  • Kun-Song Chen
    • 1
  1. 1.Laboratory of Fruit Quality Biology/The State Agriculture Ministry Laboratory of Horticultural Plant Growth, Development and Quality ImprovementZhejiang UniversityHangzhouPeople’s Republic of China
  2. 2.The New Zealand Institute of Plant and Food ResearchAucklandNew Zealand

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