Two MYB transcription factors (CsMYB2 and CsMYB26) are involved in flavonoid biosynthesis in tea plant [Camellia sinensis (L.) O. Kuntze]
Tóm tắt
Flavonoids are secondary metabolites that play important roles in the entire tea plant life cycle and have potential health-promoting properties. MYB transcription factors (TFs) are considered potentially important regulators of flavonoid biosynthesis in plants. However, the molecular mechanisms by which MYB TFs regulate the flavonoid pathway in tea plant remain unknown. In this study, two R2R3-MYB TFs (CsMYB2 and CsMYB26) involved in flavonoid biosynthesis in tea plant were investigated. The genes encoding CsMYB2 and CsMYB26 were cloned from the tea plant cultivar ‘Longjing 43’. Phylogenetic analysis showed that CsMYB2 and CsMYB26 were grouped into the proanthocyanidin biosynthesis-related MYB clade. Multiple sequence alignment revealed that conserved motif 1 in the two MYB factors was related to the bHLH TF. Subcellular localization assays suggested that CsMYB2 localized in the nucleus. Promoter analysis indicated that CsMYB2, CsMYB26 and the related structural genes contain MYB recognition elements. The expression levels of the CsMYB2 and CsMYB26 genes and the structural genes in the flavonoid biosynthesis pathway were determined in leaves from various sites in the two tea plant cultivars ‘Longjing 43’ and ‘Baiye 1 hao’. The expression levels of these genes were correlated with the accumulated flavonoid content. The results demonstrated that the expression level of CsF3’H may be regulated by CsMYB2 and that CsMYB26 expression is positively correlated with CsLAR expression. The relative transcriptional level of CsMYB26 may be the main reason for the different epigallocatechin contents between the tea plant cultivars ‘Longjing 43’ and ‘Baiye 1 hao’. Our results will serve as a reference for the potential regulatory roles of CsMYB2 and CsMYB26 in flavonoid biosynthesis in tea plant and may also assist biologists in improving tea quality.
Tài liệu tham khảo
Chen Y, Yu M, Xu J, Chen X, Shi J. Differentiation of eight tea (Camellia sinensis) cultivars in China by elemental fingerprint of their leaves. J Sci Food Agr. 2009;89(14):2350–5.
Wang Y, Gao L, Wang Z, Liu Y, Sun M, Yang D, Wei C, Shan Y, Xia T. Light-induced expression of genes involved in phenylpropanoid biosynthetic pathways in callus of tea (Camellia sinensis (L.) O. Kuntze). Sci Hortic. 2012;133:72–83.
Wu ZJ, Li XH, Liu ZW, Xu ZS, Zhuang J. De novo assembly and transcriptome characterization: novel insights into catechins biosynthesis in Camellia sinensis. BMC Plant Bio. 2014;14(1):1471–2229.
Jiang XL, Li Z, Li YJ, Liu L, Wu YH, Qian YM, Gao LP, Tan HY, Xia T, Meng F, Wang YS, Li MZ. Analysis of accumulation patterns and preliminary study on the condensation mechanism of proanthocyanidins in the tea plant [Camellia sinensis]. Sci Rep. 2015;5:8742.
Feng K, Xu ZS, Que F, Liu JX, Wang F, Xiong AS. An R2R3-MYB transcription factor, OjMYB1, functions in anthocyanin biosynthesis in Oenanthe javanica. Planta. 2017;247(4):1–15.
Xu ZS, Feng K, Que F, Wang F, Xiong AS. A MYB transcription factor, DcMYB6, is involved in regulating anthocyanin biosynthesis in purple carrot taproots. Sci Rep. 2017;7:45324.
Ramsay NA, Beverley JG. MYB-bHLH-WD40 protein complex and the evolution of cellular diversity. Trends Plant Sci. 2005;10(2):63–70.
Kranz HD, D M GR, Jin HL, Leyva A, Meissner RC, Petroni K, UA BM, Martin C, Smeekens S, Tonelli C, PA J. Towards functional characterisation of the members of the R2R3-MYB gene family from Arabidopsis thaliana. Plant J. 1998;16(2):263–76.
Stracke R, W M, Weisshaar B. The R2R3-MYB gene family in Arabidopsis thaliana. Curr Opin Plant Biol. 2001;4(5):447–56.
Nathalie N, C J, Isabelle D, Michel C, 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. 2001;13(9):2099–114.
Czemmel S, Stracke R, Weisshaar B, Cordon N, Harris NN, Walker AR, Robinson SP, Bogs J. The grapevine R2R3-MYB transcription factor VvMYBF1 regulates flavonol synthesis in developing grape berries. Plant Physiol. 2009;151(3):1513–30.
Cathie M, Javier PA. MYB transcription factors in plant. Trends Genet. 1997;13(2):67–73.
Sun B, Zhu Z, Cao P, Chen H, Chen C, Zhou X, Mao Y, Lei J, Jiang Y, Meng W, Wang Y, Liu S. Purple foliage coloration in tea (Camellia sinensis L.) arises from activation of the R2R3-MYB transcription factor CsAN1. Sci Rep. 2016;6:32534.
Takos AM, Jaffe FW, Jacob SR, Bogs J, Robinson SP, Walker AR. Light-induced expression of a MYB gene regulates anthocyanin biosynthesis in red apples. Plant Physiol. 2006;142(3):1216–32.
Terrier N, Torregrosa L, Ageorges A, Vialet S, Verries C, Cheynier V, Romieu C. Ectopic expression of VvMybPA2 promotes proanthocyanidin biosynthesis in grapevine and suggests additional targets in the pathway. Plant Physiol. 2009;149(2):1028–41.
Uematsu C, Katayama H, Makino I, Inagaki A, Arakawa O, Martin C. Peace, a MYB-like transcription factor, regulates petal pigmentation in flowering peach 'Genpei' bearing variegated and fully pigmented flowers. J Exp Bot. 2014;65(4):1081–94.
Zhou M, Liang YT, Zhang KX, Li FL, Yang PY, Tang YX, Wu YM, Shao JR. Identification of TT2 gene from floral transcriptome in Fagopyrum tataricum. Food Res Int. 2013;54(1):1331–3.
Passeri V, DF MS, Carvalho E, Paolocci F. The R2R3MYB VvMYBPA1 from grape reprograms the phenylpropanoid pathway in tobacco flowers. Planta. 2017;246(2):1–15.
Deluc L, Barrieu F, Marchive C, Lauvergeat V, Decendit A, Richard T, Carde JP, Merillon JM, Hamdi S. Characterization of a grapevine R2R3-MYB transcription factor that regulates the phenylpropanoid pathway. Plant Physiol. 2006;140(2):499–511.
Ravaglia D, E RV, Rebecca HK, Andreotti C, Ziosi V, Hellens RP, Costa G, Allan AC. Transcriptional regulation of flavonoid biosynthesis in nectarine (Prunus persica) by a set of R2R3 MYB transcription factors. BMC Plant Biol. 2013;13(1):1471–2229.
Feng K, Liu JX, Duan AQ, et al. AgMYB2 transcription factor is involved in the regulation of anthocyanin biosynthesis in purple celery (Apium graveolens L.)[J]. Planta. 2018;248(5):1249–61.
Ban Y, Honda C, Hatsuyama Y, Igarashi M, Bessho H, Moriguchi T. 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. 2007;48(7):958–70.
Li MZ, Li Y, Guo LL, Gong ND, Pang YZ, Jiang WB, Liu YJ, Jiang XL, Zhao L, Wang YS, Xie DY, Gao LP, Xia T. Functional characterization of tea (Camellia sinensis) MYB4a transcription factor using an integrative approach. Front Plant Sci. 2017;8:943.
Akagi T, Ikegami A, Tsujimoto T, Kobayashi S, Sato A, Kono A, Yonemori K. DkMyb4 is a Myb transcription factor involved in proanthocyanidin biosynthesis in persimmon fruit. Plant Physiol. 2009;151(4):2028–45.
Espley RV, Hellens RP, Putterill J, Stevenson DE, S KA, Allan AC. Red colouration in apple fruit is due to the activity of the MYB transcription factor, MdMYB10. The Plant J. 2007;49(3):414–27.
Wang KL, BK GK, Kortstee A, Karunairetnam S, Mcghie TK, Espley RV, Allan AC. An R2R3 MYB transcription factor associated with regulation of the anthocyanin biosynthetic pathway in Rosaceae. BMC Plant Biol. 2010;10(1):50.
Ashihara H, Deng WW, Mullen W, Crozier A. Distribution and biosynthesis of flavan-3-ols in Camellia sinensis seedlings and expression of genes encoding biosynthetic enzymes. Phytochemistry. 2010;71(5):559–66.
Mohammed KA, M J MAD, Bohm JBA, Towers J, HN G. Developmental changes in the composition of Proanthocyanidins from leaves of Sainfoin (Onobrychis viciifola Scop.) as determined by HPLC analysis. J Agr Food Chem. 1993;41(7):1066–70.
Nakatsuka T, S M, Yamada E, Fujita K, Kakizaki Y, Nishihara M. In Posidonia oceanica cadmium induces changes in DNA methylation and chromatin patternintion factors that regulate early flavonoid biosynthesis. in gentian flower J Exp Bot. 2012;63(2):695–709.
Yoshida K, Kume N, Nakaya Y, Yamagami A, Nakano T, Sakuta M. Comparative analysis of the triplicate proathocyanidin regulators in Lotus japonicus. Plant Cell Physiol. 2010;51(6):912–22.
Jin J, Tian F, Yang DC, Meng YQ, Kong L, Luo J, Gao G. PlantTFDB 4.0: toward a central hub for transcription factors and regulatory interactions in plants. Nucleic Acids Res. 2017;45(D1):D1040–5.
Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol. 2011;28(10):2731–9.
Wei C, Yang H, Wang S, et al. Draft genome sequence of Camellia sinensis var. sinensis provides insights into the evolution of the tea genome and tea quality. Proc Natl Acad Sci U S A. 2018;115(18):201719622.
Lescot M, Déhais P, Thijs G, et al. PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic Acids Res. 2002;30(1):325–7.
Li MZ, L Y, Guo LL, Gong ND, Pang YZ, Jiang WB, Liu YJ, Jiang XL, Zhao L, Wang YS, Xie DY, Gao LP, Xia T. Functional characterization of tea (Camellia sinensis) MYB4a transcription factor using an integrative approach. Front Plant Sci. 2017;8:943.
Michaelw P. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 2001;29(9):e45.
Pang Y, Peel GJ, Sharma SB, Tang Y, Dixon RA. A transcript profiling approach reveals an epicatechin-specific glucosyltransferase expressed in the seed coat of Medicago truncatula. Proc Natl Acad Sci. 2008;105(37):14210–5.
Xl J, Li Y, Li WW, Zhao L, Meng F, Wang YS, Tan HR, Yang H, W C WXC, Gao LP, Xia T. Tissue-specific, development-dependent phenolic compounds accumulation profile and gene expression pattern in tea plant (camellia sinensis). PLoS One. 2013;8(4):e62315.