Uncovering tea-specific secondary metabolism using transcriptomic and metabolomic analyses in grafts of Camellia sinensis and C. oleifera
- 106 Downloads
Camellia sinensis (L.) Kuntze and Camellia oleifera C. Abel (Theaceae) are closely related perennial woody shrubs, but the accumulation of metabolites and gene expression patterns are quite different between these two species. In order to understand the mechanisms behind the accumulation and biosynthesis of tea-specific secondary metabolites and the key genes that regulate their target pathways, 1-year-old clone cuttings of C. sinensis and C. oleifera were grafted in both directions, and self-grafted C. sinensis were used as controls. The transcriptomes and metabolomes of leaves and roots from the grafts were analyzed. We found that 1375 unigenes were up-regulated in the leaves of the CS-CO grafts (C. sinensis scion, C. oleifera stock), while 2437 unigenes were down-regulated. OPLS-DA models established for 7230 and 3223 mass spectra peaks were obtained in the positive and negative modes by LC-MS detection. Association analysis of the secondary metabolism pathways was performed, and the relative gene expressions of 14 genes from the transcriptome screening were verified by qRT-PCR. Among the differential metabolites screened and identified, we found that the relative levels of theanine and caffeine decreased significantly, and that many of the genes in these metabolic pathways were also down-regulated. In contrast, the levels of flavonoids apparently increased, and the expression of related genes in the flavonoid biosynthetic pathway were mostly up-regulated.
KeywordsCamellia sinensis C. oleifera Grafts Metabolome Transcriptome Secondary metabolism
This study was supported by the Natural Science Foundation of Anhui Province (Grant No.1608085QC60), the National Natural Science Foundation of China (NSFC) (Grant No. 31300576), the Changjiang Scholars and Innovative Research Team in University (Grant No. IRT_15R01), and Tea Plant Germplasm Resources Innovation Team Project of Fujian Academy of Agricultural Science (STIT2017-3-12). We appreciated Chun Liu (Beijing Genome Institute at Shenzhen, China) for technical support and analysis. We were also grateful to the elixigen editing service for the language polishing.
WD, JH, and YF prepared the material for sequencing and analyzed the data.YT, BZ, and ML participated in data analysis. WD, JH, YF, and RW were responsible for drafting and revising the manuscript. ZZ and XW guided this research.
Compliance with ethical standards
The authors declare that they complied with ethical standards.
The authors declare that they have no competing interests.
Data achieving statement
The clean data of grafts of Camellia sinensis and C. oleifera will be available in the NCBI SRA (http://www.ncbi.nlm.nih.gov/Traces/sra_sub/sub.cgi) under project accession number PRJNA429946 if the manuscript is accepted for publication in the tree genetics and genomes prior to publication.
- Cheng S, Wang Y, Li J et al (2004) Study on the relationship between the endogenous hormones and flavonoids in Ginkgo biloba leaf. Scientia Silvae Sinicae 40:45–49Google Scholar
- Cookson SJ, Clemente Moreno MJ, Hevin C, Nyamba Mendome LZ, Delrot S, Trossat-Magnin C, Ollat N (2013) Graft union formation in grapevine induces transcriptional changes related to cell wall modification, wounding, hormone signalling, and secondary metabolism. J Exper Bot 64:2997–3008. https://doi.org/10.1093/jxb/ert144 CrossRefGoogle Scholar
- Deng WW, Fan YB, Gu CC et al (2017) Changes in morphological characters and secondary metabolite contents in leaves of grafting seedlings with Camellia sinensis as scions and C. oleifera as stocks. J Trop Subtrop Bot 25:35–42Google Scholar
- Grabherr MG, Haas BJ, Yassour M, Levin JZ, Thompson DA, Amit I, Adiconis X, Fan L, Raychowdhury R, Zeng Q, Chen Z, Mauceli E, Hacohen N, Gnirke A, Rhind N, di Palma F, Birren BW, Nusbaum C, Lindblad-Toh K, Friedman N, Regev A (2011) Trinity: reconstructing a full-length transcriptome without a genome from RNA-Seq data. Nat Biotechnol 29(7):644–652. https://doi.org/10.1038/nbt.1883 CrossRefPubMedPubMedCentralGoogle Scholar
- Haas BJ, Papanicolaou A, Yassour M, Grabherr M, Blood PD, Bowden J, Couger MB, Eccles D, Li B, Lieber M, MacManes MD, Ott M, Orvis J, Pochet N, Strozzi F, Weeks N, Westerman R, William T, Dewey CN, Henschel R, LeDuc RD, Friedman N, Regev A (2013) De novo transcript sequence reconstruction from RNA-Seq: reference generation and analysis with trinity. Nat Protocol 8(8):1494–1512. https://doi.org/10.1038/nprot.2013.084 CrossRefGoogle Scholar
- Hu Y, Wang HF, Dong SQ et al (2016) Effect of sucrose treatment on flavonoid content and antioxidant activity of Sedum aizoon leaves. Modern Food Sci Tech 32:250–255Google Scholar
- Jiang X, Liu Y, Li W, Zhao L, Meng F, Wang Y, Tan H, Yang H, Wei C, Wan X, Gao L, Xia T (2013) Tissue-specific, development-dependent phenolic compounds accumulation profile and gene expression pattern in tea plant [Camellia sinensis]. PLoS One 8(4):e62315. https://doi.org/10.1371/journal.pone.0062315 CrossRefPubMedPubMedCentralGoogle Scholar
- Li CF, Zhu Y, Yu Y, Zhao QY, Wang SJ, Wang XC, Yao MZ, Luo D, Li X, Chen L, Yang YJ (2015) Global transcriptome and gene regulation network for secondary metabolite biosynthesis of tea plant (Camellia sinensis). BMC Genomics 16:560. https://doi.org/10.1186/s12864-015-1773-0 CrossRefPubMedPubMedCentralGoogle Scholar
- Li M, Li Y, Guo L, Gong N, Pang Y, Jiang W, Liu Y, Jiang X, Zhao L, Wang Y, Xie DY, Gao L, Xia T (2017) Functional characterization of tea (Camellia sinensis) MYB4a transcription factor using an integrative approach. Front Plant Sci 8:943. https://doi.org/10.3389/fpls.2017.00943 CrossRefPubMedPubMedCentralGoogle Scholar
- McCombie G, Browning LM, Titman CM, Song M, Shockcor J, Jebb SA, Griffin JL (2009) ω-3 oil intake during weight loss in obese women results in remodelling of plasma triglyceride and fatty acids. Metabolomics 5:363–374. https://doi.org/10.1007/s11306-009-0161-7 CrossRefPubMedPubMedCentralGoogle Scholar
- McKenna A, Hanna M, Banks E, Sivachenko A, Cibulskis K, Kernytsky A, Garimella K, Altshuler D, Gabriel S, Daly M, DePristo MA (2010) The genome analysis toolkit: a MapReduce framework for analyzing next-generation DNA sequencing data. Genome Res 20:1297–1303. https://doi.org/10.1101/gr.107524.110 CrossRefPubMedPubMedCentralGoogle Scholar
- Pertea G, Huang X, Liang F, Antonescu V, Sultana R, Karamycheva S, Lee Y, White J, Cheung F, Parvizi B, Tsai J, Quackenbush J (2003) TIGR gene indices clustering tools (TGICL): a software system for fast clustering of large EST datasets. Bioinformatics 19(5):651–652. https://doi.org/10.1093/bioinformatics/btg034 CrossRefPubMedGoogle Scholar
- Savoi S, Wong DC, Arapitsas P et al (2016) Transcriptome and metabolite profiling reveals that prolonged drought modulates the phenylpropanoid and terpenoid pathway in white grapes (Vitis vinifera L.) BMC Plant Biol 16:67. https://doi.org/10.1186/s12870-016-0760-1 CrossRefPubMedPubMedCentralGoogle Scholar
- Stulen I (1986) Interactions between nitrogen and carbon metabolism in a whole plant context. Developments in Plant and Soil Sciences, vol 19. Springer, Dordrecht, pp 261-278Google Scholar
- Sun B, Zhu Z, Cao P, Chen H, Chen C, Zhou X, Mao Y, Lei J, Jiang Y, Meng W, Wang Y, Liu S (2016) Purple foliage coloration in tea (Camellia sinensis L.) arises from activation of the R2R3-MYB transcription factor CsAN1. Sci Rep 6(32534). https://doi.org/10.1038/srep32534
- Tai Y, Wei C, Yang H, Zhang L, Chen Q, Deng W, Wei S, Zhang J, Fang C, Ho C, Wan X (2015) Transcriptomic and phytochemical analysis of the biosynthesis of characteristic constituents in tea (Camellia sinensis) compared with oil tea (Camellia oleifera). BMC Plant Biol 15:190. https://doi.org/10.1186/s12870-015-0574-6 CrossRefPubMedPubMedCentralGoogle Scholar
- Wiklund S, Johansson E, Sjöström L, Mellerowicz EJ, Edlund U, Shockcor JP, Gottfries J, Moritz T, Trygg J (2008) Visualization of GC/TOF-MS-based metabolomics data for identification of biochemically interesting compounds using OPLS class models. Anal Chem 80:115–122. https://doi.org/10.1021/ac0713510 CrossRefPubMedGoogle Scholar
- Wu ZJ, Tian C, Jiang Q, Li XH, Zhuang J (2016) Selection of suitable reference genes for qRT-PCR normalization during leaf development and hormonal stimuli in tea plant (Camellia sinensis). Sci Rep 6(19748). https://doi.org/10.1038/srep19748