Skip to main content
SpringerLink
Log in

Cloning and expression analyses of the anthocyanin biosynthetic genes in mulberry plants

  • Original Paper
  • Published:
Molecular Genetics and Genomics Aims and scope Submit manuscript

Abstract

Anthocyanins are natural food colorants produced by plants that play important roles in their growth and development. Mulberry fruits are rich in anthocyanins, which are the most important active components of mulberry and have many potentially beneficial effects on human health. The study of anthocyanin biosynthesis will bring benefits for quality improvement and industrial exploration of mulberry fruits. In the present study, nine putative genes involved in anthocyanin biosynthesis in mulberry plants were identified and cloned. Sequence analysis revealed that the mulberry anthocyanin biosynthetic genes were conserved and had counterparts in other plants. Spatial transcriptional analysis showed detectable expression of eight of these genes in different tissues. The results of expression and UPLC analyses in two mulberry cultivars with differently colored fruit indicated that anthocyanin concentrations correlated with the expression levels of genes associated with anthocyanin biosynthesis including CHS1, CHI, F3H1, F3′H1, and ANS during the fruit ripening process. The present studies provide insight into anthocyanin biosynthesis in mulberry plants and may facilitate genetic engineering for improvement of the anthocyanin content in mulberry fruit.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

CHS:

Chalcone synthase

CHI:

Chalcone isomerase

F3H:

Flavanone 3-hydroxylase

F3′H:

Flavonoid 3′-hydroxylase

F3′5′H:

Flavonoid 3′,5′-hydroxylase

DFR:

Dihydroflavonol reductase

ANS:

Anthocyanin synthase

DAA:

Days after anthesis

qRT-PCR:

Quantitative reverse transcription-polymerase chain reaction

UPLC:

Ultra performance liquid chromatography

References

  • Almeida JR, D’Amico E, Preuss A, Carbone F, de Vos CH, Deiml B, Mourgues F, Perrotta G, Fischer TC, Bovy AG, Martens S, Rosati C (2007) Characterization of major enzymes and genes involved in flavonoid and proanthocyanidin biosynthesis during fruit development in strawberry (Fragaria × ananassa). Arch Biochem Biophys 465:61–71. doi:10.1016/j.abb.2007.04.040

    Article  PubMed  CAS  Google Scholar 

  • Boss PK, Davies C, Robinson SP (1996) Expression of anthocyanin biosynthesis pathway genes in red and white grapes. Plant Mol Biol 32:565–569

    Article  PubMed  CAS  Google Scholar 

  • de Pascual-Teresa S, Sanchez-Ballesta MT (2008) Anthocyanins: from plant to health. Phytochem Rev 7:281–299

    Article  Google Scholar 

  • de Vetten N, Ter Horst J, van Schaik H-P, de Boer A, Mol J, Koes R (1999) A cytochrome b5 is required for full activity of flavonoid 3′, 5′-hydroxylase, a cytochrome P450 involved in the formation of blue flower colors. Proc Natl Acad Sci USA 96:778–783

    Article  PubMed  PubMed Central  Google Scholar 

  • Du Q, Zheng J, Xu Y (2008) Composition of anthocyanins in mulberry and their antioxidant activity. J Food Compos Anal 21:390–395

    Article  CAS  Google Scholar 

  • Dugo P, Mondello L, Errante G, Zappia G, Dugo G (2001) Identification of anthocyanins in berries by narrow-bore high-performance liquid chromatography with electrospray ionization detection. J Agr Food Chem 49:3987–3992

    Article  CAS  Google Scholar 

  • Ferrer J-L, Jez JM, Bowman ME, Dixon RA, Noel JP (1999) Structure of chalcone synthase and the molecular basis of plant polyketide biosynthesis. Nat Struct Mol Biol 6:775–784

    Article  CAS  Google Scholar 

  • He NJ, Zhang C, Qi XW, Zhao SC, Tao Y, Yang GJ, Lee TH, Wang XY, Cai QL, Li D, Lu MZ, Liao ST, Luo GQ, He RJ, Tan X, Xu YM, Li T, Zhao AC, Jia L, Fu Q, Zeng QW, Gao C, Ma B, Liang JB, Wang XL, Shang JZ, Song PH, Wu HY, Fan L, Wang Q, Shuai Q, Zhu JJ, Wei CJ, Zhu-Salzman KY, Jin DC, Wang JP, Liu T, Yu MD, Tang CM, Wang ZJ, Dai FW, Chen JF, Liu Y, Zhao ST, Lin TB, Zhang SG, Wang JY, Wang J, Yang HM, Yang GW, Wang J, Paterson AH, Xia QY, Ji DF, Xiang ZH (2013) Draft genome sequence of the mulberry tree Morus notabilis. Nat Commun 4:2445

    Article  PubMed  PubMed Central  Google Scholar 

  • Holton TA, Cornish EC (1995) Genetics and biochemistry of anthocyanin biosynthesis. Plant Cell 7:1071

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Holton TA, Brugliera F, Lester DR, Tanaka Y, Hyland CD, Menting JG, Lu C-Y, Farcy E, Stevenson TW, Cornish EC (1993) Cloning and expression of cytochrome P450 genes controlling flower colour. Nature 366:276–279

    Article  PubMed  CAS  Google Scholar 

  • Honda C, Kotoda N, Wada M, Kondo S, Kobayashi S, Soejima J, Zhang Z, Tsuda T, Moriguchi T (2002) Anthocyanin biosynthetic genes are coordinately expressed during red coloration in apple skin. Plant Physiol Biochem 40:955–962

    Article  CAS  Google Scholar 

  • Jeong S, Goto-Yamamoto N, Hashizume K, Esaka M (2006) Expression of the flavonoid 3′-hydroxylase and flavonoid 3′, 5′-hydroxylase genes and flavonoid composition in grape (Vitis vinifera). Plant Sci 170:61–69

    Article  CAS  Google Scholar 

  • Johnson ET, Yi H, Shin B, Oh B-J, Cheong H, Choi G (1999) Cymbidium hybrida dihydroflavonol 4-reductase does not efficiently reduce dihydrokaempferol to produce orange pelargonidin-type anthocyanins. Plant J 19:81–85

    Article  PubMed  CAS  Google Scholar 

  • Johnson ET, Ryu S, Yi H, Shin B, Cheong H, Choi G (2001) Alteration of a single amino acid changes the substrate specificity of dihydroflavonol 4-reductase. Plant J 25:325–333

    Article  PubMed  CAS  Google Scholar 

  • Koes R, Verweij W, Quattrocchio F (2005) Flavonoids: a colorful model for the regulation and evolution of biochemical pathways. Trends Plant Sci 10:236–242

    Article  PubMed  CAS  Google Scholar 

  • Lee JY, Moon SO, Kwon YJ, Rhee SJ, Park HR, Choi SW (2004) Identification and quantification of anthocyanins and flavonoids in mulberry (Morus sp.) cultivars. Food Sci Biotechnol 13:176–184

    CAS  Google Scholar 

  • Lev-Yadun S, Gould KS (2009) Role of anthocyanins in plant defence. Anthocyanins. Springer, New York, pp 22–28

    Google Scholar 

  • Liu XM, Xiao GS, Chen WD, Xu YJ, Wu JJ (2004) Quantification and purification of mulberry anthocyanins with macroporous resins. J Biomed Biotechnol 5:326–331

    Article  Google Scholar 

  • Lodhi MA, Ye G-N, Weeden NF, Reisch BI (1994) A simple and efficient method for DNA extraction from grapevine cultivars and Vitis species. Plant Mol Biol Rep 12:6–13

    Article  CAS  Google Scholar 

  • Lukačin R, Britsch L (1997) Identification of strictly conserved histidine and arginine residues as part of the active site in Petunia hybrida flavanone 3β-hydroxylase. Eur J Biochem 249:748–757

    Article  PubMed  Google Scholar 

  • Pang Y, Peel GJ, Wright E, Wang Z, Dixon RA (2007) Early steps in proanthocyanidin biosynthesis in the model legume Medicago truncatula. Plant Physiol 145:601–615. doi:10.1104/pp.107.107326

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Ravindra P, Narayan M (2003) Antioxidant activity of the anthocyanin from carrot (Daucus carota) callus culture. Int J Food Sci Nutr 54:349–355

    Article  PubMed  CAS  Google Scholar 

  • Sadilova E, Stintzing FC, Carle R (2006) Anthocyanins, colour and antioxidant properties of eggplant (Solanum melongena L.) and violet pepper (Capsicum annuum L.) peel extracts. Z Naturforsch C 61:527–535

    PubMed  CAS  Google Scholar 

  • Salvatierra A, Pimentel P, Moya-Leon MA, Caligari PD, Herrera R (2010) Comparison of transcriptional profiles of flavonoid genes and anthocyanin contents during fruit development of two botanical forms of Fragaria chiloensis ssp. chiloensis. Phytochemistry 71:1839–1847

    Article  PubMed  CAS  Google Scholar 

  • Sánchez MD (2000) World distribution and utilization of mulberry, potential for animal feeding. FAO electronic conference on mulberry for animal production (Morus1-L). pp 1–11

  • Shih C-H, Chu IK, Yip WK, Lo C (2006) Differential expression of two flavonoid 3′-hydroxylase cDNAs involved in biosynthesis of anthocyanin pigments and 3-deoxyanthocyanidin phytoalexins in sorghum. Plant Cell Physiol 47:1412–1419

    Article  PubMed  CAS  Google Scholar 

  • Shimada N, Aoki T, Sato S, Nakamura Y, Tabata S, Ayabe S (2003) A cluster of genes encodes the two types of chalcone isomerase involved in the biosynthesis of general flavonoids and legume-specific 5-deoxy (iso) flavonoids in Lotus japonicus. Plant Physiol 131:941–951

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Singhal BK, Khan MA, Dhar A, Baqual FM, Bindroo BB (2010) Approaches to industrial exploitation of mulberry (mulberry sp.) fruits. J Fruit Ornam Plant Res 18:83–99

    Google Scholar 

  • Stintzing FC, Stintzing AS, Carle R, Frei B, Wrolstad RE (2002) Color and antioxidant properties of cyanidin-based anthocyanin pigments. J Agr Food Chem 50:6172–6181

    Article  CAS  Google Scholar 

  • Wu T, Tang Q, Gao ZC, Yu ZP, Song HZ, Zheng XD, Chen W (2013) Blueberry and mulberry juice prevent obesity development in C57BL/6 mice. PLoS One 8:e77585

    Article  PubMed  CAS  PubMed Central  Google Scholar 

Download references

Acknowledgments

This project was funded by the research grants from the National Hi-Tech Research and Development Program of China (No. 2013AA100605-3), the “111” Project (B12006), the Science Fund for Distinguished Young Scholars of Chongqing (Grant No. cstc2011jjjq0010).

Author information

Authors and Affiliations

  1. State Key Laboratory of Silkworm Genome Biology, Southwest University, Beibei, Chongqing, 400715, China

    Xiwu Qi, Qin Shuai, Hu Chen, Li Fan, Qiwei Zeng & Ningjia He

Authors
  1. Xiwu Qi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Qin Shuai
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hu Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Li Fan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Qiwei Zeng
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ningjia He
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ningjia He.

Additional information

Communicated by S. Hohmann.

Nucleotide sequence data reported are available in the GenBank databases under the Accession Numbers KF438041–KF438048.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 48 kb)

Supplementary material 2 (PDF 1689 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Qi, X., Shuai, Q., Chen, H. et al. Cloning and expression analyses of the anthocyanin biosynthetic genes in mulberry plants. Mol Genet Genomics 289, 783–793 (2014). https://doi.org/10.1007/s00438-014-0851-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00438-014-0851-3

Keywords

Search

Navigation