Biologia plantarum 2014, 58:618-626 | DOI: 10.1007/s10535-014-0450-5

Isolation and expression analysis of anthocyanin biosynthetic genes in Morus alba L.

J. Li1,2, R. -H. Lü1, A. -C. Zhao1, X. -L. Wang1, C. -Y. Liu1, Q. -Y. Zhang1, X. -H. Wang1, D. Umuhoza1, X. -Y. Jin3, C. Lu1, Z. -G. Li1, M. -D. Yu1,*
1 State Key Laboratory of Silkworm Genome Biology, College of Biotechnology, Southwest University, Chongqing, P.R. China
2 Guiyang College of Traditional Chinese Medicine, Guiyang, P.R. China
3 College of Agronomy and Biotechnology, Southwest University, Chongqing, P.R. China

Anthocyanins from mulberry fruits are used in medicine. However, little anthocyanin can be detected in other tissues and sometimes also mulberry fruits are colorless. The aim of this study was to investigate which gene or genes have the strongest correlation with the anthocyanin biosynthesis. The expression of several anthocyanin synthesis genes were determined in different tissues of two white and two purple fruit cultivars. Genes encoding dihydroflavonol reductase (MaDFR) and anthocyanidin synthase (MaANS) showed a high expression only in fruit tissue of purple-fruit cultivars. During the development of mulberry fruits, the anthocyanin content was well correlated with the transcripts abundance of MaDFR, MaANS, and MaCHS (encoding chalcone synthase). The skin of female mulberry flowers turns red under irradiance because of up-regulated expressions of MaCHS, MaDFR, and MaANS. These three genes may control the anthocyanin biosynthesis in mulberry and up-regulation of them may greatly increase the anthocyanin content.

Keywords: anthocyanidin synthase; chalcone isomerase; chalcone synthase; dihydroflavonol reductase; flavanone-3-hydroxylase; phenylalanine ammonia lyase
Subjects: anthocyanin biosynthesis-related genes; anthocyanin accumulation; flavonoids; phylogenetic tree; mulberry
Species: Morus alba

Received: October 21, 2013; Revised: May 14, 2014; Accepted: May 19, 2014; Published: December 1, 2014Show citation

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Li, J., Lü, R.-H., Zhao, A.-C., Wang, X.-L., Liu, C.-Y., Zhang, Q.-Y., ... Yu, M.-D. (2014). Isolation and expression analysis of anthocyanin biosynthetic genes in Morus alba L. Biologia plantarum, 58(4), 618-626. doi: 10.1007/s10535-014-0450-5.
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    References

    1. Altschul, S.F., Gish, W., Miller, W., Myers, E.W., Lipman, D.J.: Basic local alignment search tool. - J. mol. Biol. 215: 403-410, 1990. Go to original source...
    2. Balzergue, S., Dubreucq, B., Chauvin, S., Le-Clainche, I., Le Boulaire, F., De Rose, R., Lepiniec, L.: Improved PCR-walking for large-scale isolation of plant T-DNA borders. - Biotechniques 30: 496-498, 502, 504, 2001. Go to original source...
    3. Boddu, J., Svabek, C., Sekhon, R., Gevens, A., Nicholson, R.L., Jones, A.D., Chopra, S.: Expression of a putative flavonoid 3'-hydroxylase in Sorghum mesocotyls synthesizing 3-deoxyanthocyanidin phytoalexins. - Physiol. mol. Plant Pathol. 65: 101-113, 2004. Go to original source...
    4. Boss, P.K., Davies, C., Robinson, S.P.: 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: 1059-1066, 1996. Go to original source...
    5. Britsch, L., Dedio, J., Saedler, H., Forkmann, G.: Molecular characterization of flavanone 3-beta-hydroxylases. Consensus sequence, comparison with related enzymes and the role of conserved histidine residues. - Eur. J. Biochem. 217: 745-754, 1993. Go to original source...
    6. Chan, K.C., Ho, H.H., Huang, C.N., Lin, M.C., Chen, H.M., Wang, C.J.: Mulberry leaf extract inhibits vascular smooth muscle cell migration involving a block of small GTPase and Akt/NF-kappaB signals. - J. Agr. Food Chem. 57: 9147-9153, 2009. Go to original source...
    7. Chang, C.C., Yang, M.H., Wen, H.M., Chern, J.C.: Estimation of total flavonoid content in propolis by two complementary colorimetric methods. - J. Food Drug Anal. 10: 178-182, 2002.
    8. Chapple, C.: Molecular-genetic analysis of plant cytochrome P450-dependent monooxygenases. - Ann. Rev. Plant Physiol. Plant mol. Biol. 49: 311-343, 1998. Go to original source...
    9. Chiu, L.W., Zhou, X., Burke, S., Wu, X., Prior, R.L., Li, L.: The purple cauliflower arises from activation of a MYB transcription factor. - Plant Physiol. 154: 1470-1480, 2010. Go to original source...
    10. Debes, M.A., Arias, M.E., Grellet-Bournonville, C.F., Wulff, A.F., Martinez-Zamora, M.G,, Castagnaro, A.P., Diaz-Ricci, J.C.: White-fruited Duchesnea indica (Rosaceae) is impaired in ANS gene expression. - Amer. J. Bot. 98: 2077-2083, 2011. Go to original source...
    11. Deroles, S.: Anthocyanin biosynthesis in plant cell cultures: a potential source of natural colourants. - In: Kevin, G., Kevin, D., Chris, W. (ed.): Anthocyanins: Biosynthesis, Functions and Applications. Pp. 107-117, Springer, New York 2009.
    12. Goldsbrough, A., Belzile, F., Yoder, J.I.: Complementation of the tomato anthocyanin without (aw) mutant using the dihydroflavonol 4-reductase gene. - Plant Physiol. 105: 491-496, 1994. Go to original source...
    13. Grayer, R.J., Harborne, J.B.: A survey of antifungal compounds from higher plants 1982-1993. - Phytochemistry 37: 19-42, 1994. Go to original source...
    14. Grotewold, E.: The genetics and biochemistry of floral pigments. - Annu. Rev. Plant Biol. 57: 761-780, 2006. Go to original source...
    15. Hasemann, C.A., Kurumbail, R.G., Boddupalli, S.S., Peterson, J.A., Deisenhofer, J.: Structure and function of cytochromes P450: a comparative analysis of three crystal structures. - Structure 3: 41-62, 1995. Go to original source...
    16. Holton, T.A., Cornish, E.C.: Genetics and biochemistry of anthocyanin biosynthesis. - Plant Cell 7: 1071-1083, 1995. Go to original source...
    17. Honda, C., Kotoda, N., Wada, M., Kondo, S., Kobayashi, S., Soejima, J., Moriguchi, T.: Anthocyanin biosynthetic genes are coordinately expressed during red coloration in apple skin. - Plant Physiol. Biochem. 40: 955-962, 2002. Go to original source...
    18. Jaakola, L.: New insights into the regulation of anthocyanin biosynthesis in fruits. - Trends Plant Sci. 18: 477-83, 2013. Go to original source...
    19. Jaakola, L., Pirttila, A.M., Halonen, M., Hohtola, A.: Isolation of high quality RNA from bilberry Vaccinium myrtillus L. fruit. - Mol. Biotechnol. 19: 201-203, 2001. Go to original source...
    20. Jayaprakasam, B., Vareed, S.K., Olson, L.K., Nair, M.G.: Insulin secretion by bioactive anthocyanins and anthocyanidins present in fruits. - J. Agr Food Chem. 53: 28-31, 2005. Go to original source...
    21. Jin, H.: Multi-functionality and diversity within the plant MYBgene family. - Plant mol. Biol. 41: 577-585, 1997.
    22. Johnson, E.T., Yi, H.K., Shin, B.C., Oh, B.J., Cheong, H.S., Choi, G.: Cymbidium hybrida dihydroflavonol 4-reductase does not efficiently reduce dihydrokaempferol to produce orange pelargonidin-type anthocyanins. - Plant J. 19: 81-85, 1999. Go to original source...
    23. Kim, S., Binzel, M.L., Yoo, K.S., Park, S., Pike, L.M.: Pink P, a new locus responsible for a pink trait in onions Allium cepa resulting from natural mutations of anthocyanidin synthase. - Mol. Genet. Genomics 272: 18-27, 2004. Go to original source...
    24. Kim, S., Yoo, K.S., Pike, L.M.: Development of a codominant PCR-based marker for allelic selection of the pink trait in onion Allium cepa, based on the insertion mutation in the promoter of the anthocyanidin synthase gene. - Theor. appl. Genet. 110: 573-578, 2005. Go to original source...
    25. Kim, S.H., Lee, J.R., Hong, S.T., Yoo, Y.K., An, G., Kim, S.R.: Molecular cloning and analysis of anthocyanin biosynthesis genes preferentially expressed in apple skin. - Plant Sci. 165: 403-413, 2003. Go to original source...
    26. Kobayashi, S., Ishimaru, M., Ding, C.K., Yakushiji, H., Goto, N.: Comparison of UDP-glucose: flavonoid 3-O-glucosyltransferase UFGT gene sequences between white grapes Vitis vinifera and their spots with red skin. - Plant Sci. 160: 543-550, 2001. Go to original source...
    27. Koes, R.E.F., Quattrocchio, M., Mol, J.N.: The flavonoid biosynthetic pathway in plants: function and evolution. - Bioessays 16: 123-132, 1994. Go to original source...
    28. Kotepong, P., Ketsa, S., Van Doorn, W.G.: A white mutant of Malay apple fruit Syzygium malaccense lacks transcript expression and activity for the last enzyme of anthocyanin synthesis, and the normal expression of a MYB transcription factor. - Funct. Plant Biol. 38: 75-86, 2011. Go to original source...
    29. Kubo, H., Nawa, N., Lupsea, S.A.: Anthocyaninless1 gene of Arabidopsis thaliana encodes a UDP-glucose:flavonoid-3-O-glucosyltransferase. - J. Plant Res. 120: 445-449, 2007. Go to original source...
    30. Lacombe, E., Hawkins, S., Van Doorsselaere, J., Piquemal, J., Goffner, D., Poeydomenge, O., Grima Pettenati, J.: Cinnamoyl CoA reductase, the first committed enzyme of the lignin branch biosynthetic pathway: cloning, expression and phylogenetic relationships. - Plant J. 11: 429-441, 1997. Go to original source...
    31. Lukacin, R., Britsch, L.: Identification of strictly conserved histidine and arginine residues as part of the active site in Petunia hybrida flavanone 3-beta-hydroxylase. - Eur. J. Biochem. 249: 748-757, 1997. Go to original source...
    32. Macheix, J.J., Fleuriet, A., Billot, J.: Fruit Phenolics. - CRC Press, Boca Raton 1990.
    33. Martin, C., Carpenter, R., Sommer, H., Saedler, H., Coen, E.S,: Molecular analysis of instability in flower pigmentation of Antirrhinum majus, following isolation of the pallida locus by transposon tagging. - EMBO J. 4: 1625-1630, 1985. Go to original source...
    34. Martin, C., Prescott, A., Mackay, S., Bartlett, J., Vrijlandt, E.: Control of anthocyanin biosynthesis in flowers of Antirrhinum majus. - Plant J. 1: 37-49, 1991. Go to original source...
    35. Montefiori, M., Espley, R.V., Stevenson, D., Cooney, J., Datson, P.M., Saiz, A., Allan, A.C.: Identification and characterisation of F3GT1 and F3GGT1, two lycosyltransferases responsible for anthocyanin biosynthesis in red-fleshed kiwifruit Actinidia chinensis. - Plant J. 65: 106-118, 2011. Go to original source...
    36. Murakami, K., Mihara, K., Omura, T.: The transmembrane region of microsomal cytochrome-P450 identified as the endoplasmic-reticulum retention signal. - J. Biochem. 116: 164-175, 1994. Go to original source...
    37. Nicoue, E.E., Savard, S., Belkacemi, K.: Anthocyanins in wild blueberries of Quebec: extraction and identification. - J. Agr. Food Chem. 55: 5626-5635, 2007. Go to original source...
    38. Niu, S.S., Xu, C.J., Zhang, W.S., Zhang, B., Li, X., Lin-Wang, K., Chen, K.S.: Coordinated regulation of anthocyanin biosynthesis in Chinese bayberry Myrica rubra fruit by a R2R3 MYB transcription factor. - Planta 231: 887-899, 2010. Go to original source...
    39. O'Neill, S.D., Tong, Y., Sporlein, B., Forkmann, G., Yoder, J.I.: Molecular genetic analysis of chalcone synthase in Lycopersicon esculentum and an anthocyanin-deficient mutant. - Mol. gen. Genet. 224: 279-288, 1990. Go to original source...
    40. Pawlowska, A.M., Oleszek, W., Braca, A. Quali-quantitative analyses of flavonoids of Morus nigra L. and Morus alba L., Moraceae fruits. - J. Agr. Food Chem. 56: 3377-3380, 2008. Go to original source...
    41. Qin, C.G., Li, N., Niu, W.Y., Ding, J., Zhang, R., Shang, X.Y.: Analysis and characterisation of anthocyanins in mulberry fruit. - Czech J. Food Sci. 2: 117-126, 2010. Go to original source...
    42. Santos, D.T., Cavalcanti, R.N., Rostagno, M.A., Queiroga, C.L., Eberlin, M.N., M. Meireles, A.A.: Extraction of polyphenols and anthocyanins from the jambul (Syzygium cumini) fruit jeels. - J. Food Public Health 3: 12-20, 2013.
    43. Schwinn, K., Venail, J., Shang, Y.J., Mackay, S., Alm, V., Butelli, E., Martin, C.: A small family of MYB-regulatory genes controls floral pigmentation intensity and patterning in the genus Antirrhinum. - Plant Cell 18: 831-851, 2006. Go to original source...
    44. Sommer, H., Saedler, H.: Structure of the chalcone synthase gene of Antirrhinum majus. - Mol. gen. Genet. 202: 429-434, 1986. Go to original source...
    45. Wang, X., Olsen, O., Knudsen, S.: Expression of the dihydroflavonol reductase gene in an anthocyanin-free barley mutant. - Hereditas 119: 67-75, 1993. Go to original source...
    46. Wei, Y.Z., Hu, F.C., Hu, G.B., Li, X.J., Huang, X.M., Wang, H.C.: Differential expression of anthocyanin biosynthetic genes in relation to anthocyanin accumulation in the pericarp of Litchi chinensis Sonn. - PLoS One 6: e19455, 2011. Go to original source...
    47. Yamazaki, S., Sato, K., Suhara, K., Sakaguchi, M., Mihara, K., Omura, T.: Importance of the proline-rich region following signal-anchor sequence in the formation of correct conformation of microsomal cytochrome P-450s. - J. Biochem. 114: 652-657, 1993. Go to original source...
    48. Zou, T.B., Wang, M., Gan, R.Y., Ling, W.H.: Optimization of ultrasound-assisted extraction of anthocyanins from mulberry, using response surface methodology. - Int. J. mol. Sci. 12: 3006-3017, 2011. Go to original source...

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