biologia plantarum

International journal on Plant Life established by Bohumil Nìmec in 1959

Biologia plantarum 63:765-775, 2019 | DOI: 10.32615/bp.2019.146

Transcriptome sequencing flower petals reveals insights into regulation of flavonoid biosynthesis in Osmanthus fragrans

Y.J. HAN*, M.F. DONG, H.Y. WANG, X.D. WANG, K. LI, F.D. SHANG*
School of Life Sciences, State Key Laboratory of Cotton Biology, State Key Laboratory of Crop Stress Adaptation and Improvement, Key Laboratory of Plant Stress Biology, Laboratory of Plant Germplasm and Genetic Engineering, Henan University, Kaifeng 475004, P.R. China

Osmanthus fragrans Lour., one of the top 10 most popular flowers in China, is known for both its beauty and fragrance. It is rich in flavonoids, a class of secondary metabolites with significant neuroprotective, free-radical scavenging, and anti-oxidant activity. To understand the mechanisms regulating flavonoid biosynthesis, we conducted transcriptome sequencing O. fragrans flowers to analyze gene expressions during the full flowering stage. The RNA was isolated separately from petals of cvs. Yingui and Dangui, which were treated or not with jasmonic acid, salicylic acid, or abscisic acid. A total of 142 029 unigenes were denovo assembled, and 50 918 unigenes were annotated. The differentially expressed genes were identified, annotated, and classified. The results of transcriptome sequencing and real-time PCR revealed higher expressions of phenylalanine ammonia-lyase (PAL), PAL1, chalcone synthase (CHS), flavanone-3-hydroxylase (F3H), flavonol synthase (FLS), and lower expressions of dihydroflavonol-4-reductase (DFR), anthocyanidin synthase (ANS) in 'Yingui' than in 'Dangui'. Such an expression pattern facilitated the higher accumulation of flavonoids in 'Yingui'. Several genes of the flavonoid biosynthesis pathway were upregulated by jasmonic acid and salicylic acid in both the cultivars leading to flavonoid accumulation in their petals. In the v-myb avian myeloblastosis viral oncogene homolog 1(MYB1)-overexpressing petals, the expressions of PAL, PAL1, CHI, and FLS increased. The results suggest that MYB1 may participate in the flavonoid biosynthesis pathway and regulate the expression of some upstream genes in O. fragrans.

Keywords: abscisic acid, DEGs, jasmonic acid, MYB transcription factor, salicylic acid.

Received: December 12, 2018; Revised: October 28, 2019; Accepted: November 28, 2019; Published online: December 18, 2019  Show citation

ACS AIP APA ASA Harvard Chicago Chicago Notes IEEE ISO690 MLA NLM Turabian Vancouver
HAN, Y.J., DONG, M.F., WANG, H.Y., WANG, X.D., LI, K., & SHANG, F.D. (2019). Transcriptome sequencing flower petals reveals insights into regulation of flavonoid biosynthesis in Osmanthus fragrans. Biologia plantarum, 63, Article 765-775. https://doi.org/10.32615/bp.2019.146
Share...
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    Supplementary files

    Download fileHan6041 Suppl.pdf

    File size: 1011.61 kB

    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. Bairoch, A., Apweiler, R.: The SWISS-PROT protein sequence data bank and its supplement TrEMBL in 1999. - Nucl. Acids Res. 28: 49-54, 1999. Go to original source...
    3. Baudry, A., Heim, M.A., Dubreucq, B., Caboche, M., Weisshaar, B., Lepiniec, L.: TT2, TT8, and TTG1 synergistically specify the expression of BANYULs and proanthocyanidin biosynthesis in Arabidopsis thaliana. - Plant J. 39: 366-380, 2010. Go to original source...
    4. Bedon, F., Bomal, C., Caron, S., Levasseur, C., Boyle, B., Mansfield, S.D., Gershenzon, J., Grima-Pettenati, J., Séguin, A., MacKay, J.: Subgroup 4 R2R3-MYBs in conifer trees: gene family expansion and contribution to the isoprenoid- and flavonoid-oriented responses. - J. exp. Bot. 61: 3847-3864, 2010. Go to original source...
    5. Bovy, A.G., De Vos, R., Kemper, M., Schijen, E., Pertejo, M.A., Muir, S., Collins, G., Robinson, S., Verhoeyen, M., Hughes, S.: High-flavonol tomatoes resulting from the heterologous expression of the maize transcription factor genes LC and C1. - Plant Cell 14: 2509-2526, 2002. Go to original source...
    6. Broun, P.: Transcriptional control of flavonoid biosynthesis: a complex network of conserved regulators involved in multiple aspects of differentiation in Arabidopsis. - Curr. Opin. Plant Biol. 8: 272-279, 2005. Go to original source...
    7. Czemmel, S., Stracke, R., Weisshaar, B., Cordon, N., Harris, N.N., Walker, A.R., Robinson, S.P., Bogs, J.: The grapevine R2R3-MYB transcription factor VvMYBF1 regulate flavonol synthesis in developing grape berries. - Plant Physiol. 151: 1513-1530, 2009. Go to original source...
    8. Debeaujon, I., Peeters, A.J., Léon-Kloosterziel, K.M., Koornneef, M.: 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: 853-871, 2001. Go to original source...
    9. Espley, R.V., Hellens, R.P., Putterill, J.,Stevenson, D.E.,Kutty-Amma, S.,Allan, A.C.: Red colouration in apple fruit is due to the activity of the MYB transcription factor, MdMYB10. - Plant J. 49: 414-427, 2007. Go to original source...
    10. Finn, R.D., Tate, J., Mistry, J., Coggill, P.C., Sammut, S.J., Hotz, H.R., Ceric, G., Forslund, K., Eddy, S.R., Sonnhammer, E.L., Bateman, A.: The Pfam protein families database. - Nucl. Acids Res. 36: D281-D288, 2007. Go to original source...
    11. Guan, C., Song, X., Ji, J., Li, X.Z., Jin, C., Li, J., Wang, G.: Salicylic acid treatment enhances expression of chalcone isomerase gene and accumulation of corresponding flavonoids during fruit maturation of Lycium chinense. - Eur. Food Res. Technol. 239: 857-865, 2014. Go to original source...
    12. Gonzalez, A., Zhao, M., Leavitt, J.M., Lloyd, A.M.: Regulation of the anthocyanin biosynthetic pathway by the TTG1/bHLH/Myb transcriptional complex in Arabidopsis seedlings. - Plant J. 53: 814-827, 2010. Go to original source...
    13. Grabherr, M.G., Haas, B.J., Yassour, M., Levin, J.Z., Thompson, D.A., Amit, I., Adiconis, X., Fan, L., Raychowdhury, R., Zeng, Q., Chen, Z., Mauceli, E., Hacohen, N., Gnirke, A., Rhind, N., Palma, F.D., Birren, B.W., Nusbaum, C., Lindblad-Toh, K., Friedman, N., Regev, A.: Trinity: reconstructing a full-length transcriptome without a genome from RNA-Seq data[J]. - Nat. Biotechnol. 29: 644-652, 2011. Go to original source...
    14. Han, Y.J., Wang, X.H., Chen, W.C., Dong, M.F., Yuan, W.J., Shang, F.D.: Differential expression of carotenoid-related genes determines diversified carotenoid coloration in flower petal of Osmanthus fragrans. - Tree Genet. Genomes 10: 329-338, 2014a. Go to original source...
    15. Han, S., Wang, C.W., Wang, W.L., Jiang, J.: Mitogen-activated protein kinase 6 controls root growth in Arabidopsis by modulating Ca2+-based Na+ flux in root cell under salt stress. - J. Plant Physiol. 171: 26-34, 2014b. Go to original source...
    16. Han, Y.J., Chen, W.C., Yang, F.B., Wang, X.H., Dong, M.F., Zhou, P., Shang, F.D.: cDNA-AFLP analysis on 2 Osmanthus fragrans cultivars with different flower colour and molecular characteristics of OfMYB1 gene. - Trees 29: 931-940, 2015. Go to original source...
    17. Han, Y.J., Liu, L.X., Dong, M.F., Yuan, W.J., Shang, F.D.: cDNA cloning of the phytoene synthase (PSY) and expression analysis of PSY and carotenoid cleavage dioxygenase genes in Osmanthus fragrans. - Biologia 68: 258-263, 2013. Go to original source...
    18. Han, Y.J., Wang, H.Y., Wang, X.D., Li, K., Dong, M.F., Li, Y., Zhu, Q., Shang, F.D.: Mechanism of floral scent production in Osmanthus fragrans and the production and regulation of its key floral constituents, β-ionone and linalool. - Hort. Res. 6: 106, 2019. Go to original source...
    19. Han, Y.J., Wu, M., Cao, L.Y. Shang, F.D.: Characterization of OfWRKY3, a transcription factor that positively regulates the carotenoid cleavage dioxygenase gene OfCCD4 in Osmanthus fragrans. - Plant Mol. Biol. 91: 485-496, 2016. Go to original source...
    20. Hao, G., Du, X., Zhao, F., Ji, H.: Fungal endophytes-induced abscisic acid is required for flavonoid accumulation in suspension cells of Ginkgo biloba. -Biotechnol. Lett. 32: 305-314, 2010. Go to original source...
    21. Harris, M.A., Clark, J., Ireland, A., Foulger, R.: The Gene Ontology (GO) database and informatics resource. - Nucl. Acids Res. 32: D258-D261, 2004. Go to original source...
    22. He, Y.X., Yuan, W.J., Dong, M.F., Han, Y.J., Shang, F.D.: The first genetic map in sweet osmanthus (Osmanthus fragrans Lour.) using specific locus amplified fragment sequencing. -Front. Plant Sci. 8: 1621, 2017. Go to original source...
    23. Hernandez, J.M., Heine, G.F., Irani, N.G., Feller, A., Kim, M.G., Matulnik, T., Chandler, V.L., Grotewold, E.: Different mechanisms participate in the R-dependent activity of the R2R3 MYB transcription factor C1. - J. biol. Chem. 279: 48205-48213, 2004. Go to original source...
    24. Huse, S.M., Huber, J.A., Morrison, H.G., Sogin, M.L., Welch, D.M.: Accuracy and quality of massively parallel DNA pyrosequencing. - Genome Biol. 8: R143, 2007. Go to original source...
    25. Kanehisa, M., Goto, S., Kawashima, S., Okuno, Y., Hattori, M.: The KEGG resource for deciphering the genome. - Nucleic Acids Res. 32: D277-D280, 2004. Go to original source...
    26. Koes, R., Verweij, W., Quattrocchio, F.: Flavonoids: a colorful model for the regulation and evolution of biochemical pathways. - Trends Plant Sci. 10: 236-242, 2005. Go to original source...
    27. Livak, K.J., Schmittgen, T.D.: Analysis of relative gene expression data using real-time quantitative PCR and the 2 (-Delta Delta C(T)) method. - Methods 25: 402-408, 2001. Go to original source...
    28. Love, M.I., Huber, W., Anders, S.: Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. - Genome Biol. 15: 550, 2014. Go to original source...
    29. Margulies, M., Egholm, M., Altman, W.E., Attiya, S., Bader, J.S.: Genome sequencing in microfabricated high-density picolitre reactors. - Nature 437: 376-380, 2005. Go to original source...
    30. Mathews, H., Clendennen, S.K., Caldwell, C.G., Liu, X.L., Connors, K., Matheis, N., Schuster, D.K., Menasco, D.J., Wagoner, W., Lightner, J.: Activation tagging in tomato identifies a transcriptional regulator of anthocyanin biosynthesis, modification, and transport. - Plant Cell 15: 1689-1703, 2003. Go to original source...
    31. Mendhulkar, V.D., Vakil, M.M.A.: Elicitation of flavonoids by salicylic acid and Penicillium expansum in Andrographis paniculata (Burm. f.) Nees. cell culture. - Res. Biotechnol. 2:1-9, 2013.
    32. Mehrtens, F., Kranz, H., Bednarek, P., Weisshaar, B.: The Arabidopsis transcription factor MYB12 is a flavonol-specific regulator of phenylpropanoid biosynthesis. -J. Plant Physiol. 138: 1083-1096, 2005. Go to original source...
    33. Mondal, S.K., Roy, S.: Genome-wide sequential, evolutionary, organizational and expression analyses of phenylpropanoid biosynthesis associated MYB domain transcription factors in Arabidopsis. - J. Biomol. Struct. Dynamics 36: 1577-1601, 2017. Go to original source...
    34. Morita, Y., Saitoh, M., Hoshino, A., Nitasaka, E., Iida, S.: Isolation of cDNAs for R2R3-MYB, bHLH and WDR transcriptional regulators and identification of c and ca mutations conferring white flowers in the Japanese morning glory. - Plant Cell Physiol. 47: 457-470, 2006. Go to original source...
    35. Nakamura, N., Masako, F.M., Miyazaki, K., Tanaka, Y.: RNAi suppression of the anthocyanidin synthase gene in Torenia hybrida yields white flowers with higher frequency and better stability than antisense and sense suppression. - Plant Biotechnol. 23: 13-17, 2006. Go to original source...
    36. Nakatsuka, T., Abe, Y., Kakizaki,Y., Yamamura, S., Nishihara, M.: Production of red-flowered plants by genetic engineering of multiple flavonoid biosynthetic genes. - Plant Cell Rep. 26: 1951-1959, 2007. Go to original source...
    37. Nesi, N., Jond, C., Debeaujon, I., Caboche, M., Lepiniec, L.: 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: 2099-2114, 2001. Go to original source...
    38. Nishihara, M., Nakatsuka, T., Yamamura, S.: Flavonoid components and flower color change in transgenic tobacco plants by suppression of chalcone isomerase gene. - FEBS Lett. 579: 6074-6078, 2005. Go to original source...
    39. Novaes, E., Drost, D.R., Farmerie, W.G., Pappas, G.J.Jr., Grattapaglia, D., Sederoff, R.R., Kirst, M.: High-throughput gene and SNP discovery in Eucalyptus grandis, an uncharacterized genome. - BMC Genomics 9: 312, 2008. Go to original source...
    40. Peters, D.J., Constabel, C.P.: Molecular analysis of herbivore-induced condensed tannin synthesis: cloning and expression of dihydroflavonol reductase from trembling aspen (Populus tremuloides). - Plant J. 32: 701-712, 2002. Go to original source...
    41. Qi, T., Xie, D.: The jasmonate-ZIM-domain proteins interact with the WD-Repeat/bHLH/MYB complexes to regulate jasmonate-mediated anthocyanin accumulation and trichome initiation in Arabidopsis thaliana. - Plant Cell 23: 1795-1814, 2011. Go to original source...
    42. Quattrocchio, F., Verweij, W., Kroon, A., Spelt, C., Mol, J., Koes, R.: PH4 of Petunia is an R2R3 MYB protein that activates vacuolar acidification through interactions with basic-helix-loop-helix transcription factors of the anthocyanin pathway. - Plant Cell 18: 1274-1291, 2006. Go to original source...
    43. Quattrocchio, F., Wing, J., Van der Woude, K., Souer, E., De Vetten, N., Mol, J., Koes, R.: Molecular analysis of the anthocyanin 2 gene of petunia and its role in the evolution of flower colour. - Plant Cell 11: 1433-1444, 1999. Go to original source...
    44. Ramsay, N.A., Glover, B.J.: MYB-bHLH-WD40 protein complex and the evolution of cellular diversity. - Trends Plant Sci. 10: 63-70, 2005. Go to original source...
    45. Schwinn, K., Venail, J., Shang, Y., Mackay, S., Alm, V., Butelli, E., Oyama, R., Bailey, P., Davies, K., 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...
    46. Seitz, C., Vitten, M., Steinbach, P., Hartl, S., Hirsche, J., Rhthje, W., Treutter, D., Forkmann, G.: Redirection of anthocyanin synthesis in Osteospermum hybrida by a two-enzyme manipulation strategy. - Phytochemistry 68: 824-833, 2007. Go to original source...
    47. Sonnhammer, E.L.L., Koonin, E.V.: Orthology, paralogy and proposed classification for paralog subtypes. - Trends Genet. 18: 619-620, 2003. Go to original source...
    48. Spelt, C., Quattrocchio, F., Mol, J.N.M., Koes. R.: Anthocyanin1 of Petunia encodes a basic helix-loop-helix protein that directly activates transcription of structural anthocyanin genes. - Plant Cell 12: 1619-1632, 2000. Go to original source...
    49. Stracke, R., Ishihara, H., Huep, G., Barsch, A., Mehrtens, F., Niehaus, K., Weisshaar, B.: Differential regulation of closely related R2R3-MYB transcription factors controls flavonol accumulation in different parts of the Arabidopsis thaliana seedling. - Plant J. 50: 660-677, 2007. Go to original source...
    50. Takos, A.M., Jaffé, F.W., Jacob, S.R., Bogs, J., Robinson, S.P., Walker, A.R.: Light-induced expression of a MYB gene regulates anthocyanin biosynthesis in red apples. - Plant Physiol. 142: 1216-1232, 2006. Go to original source...
    51. Tanaka, Y., Ohmiya, A.: Seeing is believing: engineering anthocyanin and carotenoid biosynthetic pathways. - Curr. Opin. Biotechnol. 19: 190-197, 2008. Go to original source...
    52. Tanaka, Y., Sasaki, N., Ohmiya, A.: Biosynthesis of plant pigments: anthocyanins, betalains and carotenoids. - Plant J. 54: 733-749, 2010. Go to original source...
    53. Tatusov, R.L., Fedorova, N.D., Jackson, J.D., Jacobs, A.R., Kiryutin, B., Koonin, E.V., Krylov, D.M., Mazumder, R., Mekhedov, S.L., Nikolskaya, A.N., Rao, B.S., Smirnov, S., Sverdlov, A.V., Vasudevan, S., Wolf, Y.I., Yin, J.J., Natale, D.A.: The COG database: an updated version includes eukaryotes. - BMC Bioinformatics 4: 41, 2003. Go to original source...
    54. Tsuda, S., Fukui, Y., Nakamra, N., Katsumoto, Y., Sakakibara, K., Fukuchi-Mizutani, M., Ohira, K., Ueyama, Y., Ohkawa, H., Holton, T.A., Kusumi, T., Tanaka, Y.: Flower colour modification of Petunia hybrida commercial varieties by metabolic engineering. - Plant Biotechnol. 21: 377-386, 2004. Go to original source...
    55. Vimolmangkang, S., Han, Y., Wei, G.C., Korban, S.S.: An apple MYB transcription factor, MdMYB3, is involved in regulation of anthocyanin biosynthesis and flower development. - BMC Plant Biol. 13: 176, 2013. Go to original source...
    56. Wang, Z., Gerstein, M., Snyder, M.: RNA-Seq: a revolutionary tool for transcriptomics. - Nat. Rev. Genet. 10: 57-63, 2009. Go to original source...
    57. Wei, W.L., Qi, X.Q., Wang, L.H., Zhang, Y.X., Hua, W., Li, D.H., Zhang, X.R.: Characterization of the sesame (Sesamum indicum L.) global transcriptome using Illumina paired-end sequencing and development of EST-SSR markers. - BMC Genomics 12: 451, 2011. Go to original source...
    58. Winkel-Shirley, B.: Flavonoid biosynthesis. A colourful model for genetics, biochemistry, cell biology, and biotechnology. - Plant Physiol. 126: 485-493, 2001. Go to original source...
    59. Xiang, Q.B. Liu, Y.L. An Illustrated Monograph of the Sweet Osmanthus Varieties in China. - Zhejiang Science & Technology Press, Hangzhou 2007.
    60. Zifkin, M., Jin, A., Ozga, J.A., Zaharia, L.I., Irina, L., Schernthaner, J.P., Gesell, A., Abrams, S.R., Kennedy, J.A., Constabel, C.P.: Gene expression and metabolite profiling of developing highbush blueberry fruit indicates transcriptional regulation of flavonoid metabolism and activation of abscisic acid metabolism. - Plant Physiol. 158: 200-224, 2012. Go to original source...
    61. Zuker, A., Tzfira, T., Ben-Meir, H., Ovadis, M., Shklarman, E., Itzhaki, H., Forkamnn, G., Martens, S., Nata-Sharir, I., Weiss, D., Vainstein, A.: Modification of flower colour and fragrance by antisense suppression of the flavanone 3-hydroxylase gene. - Mol Breed. 9: 33-41, 2002. Go to original source...

    Return to the content