Formation of volatiles in response to tea green leafhopper (Empoasca onukii Matsuda) herbivory in tea plants: a multi-omics study

Abstract

Key message

Combined transcriptome and metabolome analysis of fresh leaf infestation by tea green leafhoppers (Empoasca (Matsumurasca) onukii Matsuda) suggests roles for alternative pre-mRNA splicing and mRNAs in the regulation of aroma formation in tea plants.

Abstract

Oriental Beauty is a high-grade, oolong tea with a pronounced honey-like aroma and rich ripe fruit flavor that develops primarily as a result of the infestation of the fresh leaves by tea green leafhoppers (Empoasca (Matsumurasca) onukii Matsuda). Here, we used PacBio Iso-Seq and RNA-seq analyses to determine the full-length transcripts and gene expression profiles of fresh tea leaves in response to E. (M.) onukii herbivory. We investigated the relationship between RNA-seq, tea metabolites, and aroma response mechanisms in leaves infested by leafhoppers. We found 3644 differentially expressed genes, of which 2552 were up- and 1092 were down-regulated. A total of 49,913 alternative splicing events were predicted, including 324 differential AS events. Moreover, 3105 differentially expressed transcripts were also identified, of which 2295 were up- and 810 were down-regulated. The characterization of expression patterns of the key gene transcript isoforms involved in the aroma formation pathways identified 130 differentially expressed metabolites, 97 of which were up- and 33 were down-regulated. Two key aroma compounds (phenylacetaldehyde and 4-hydroxybenzaldehyde) were highly correlated with genes of the aroma formation pathways. Our results revealed that pre-mRNA AS plays a crucial role in the metabolic regulation surrounding aroma formation under leafhopper herbivory in tea plants.

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Data availability

The transcriptome sequence data have been deposited into the NCBI Sequence Read Archive (SRA) under the accession number of PRJNA683753.

References

  1. Achard P, Cheng H, Grauwe LD, Decat J, Schoutteten H, Moritz T, Straeten DVD, Peng J, Harberd NP (2006) Integration of plant responses to environmentally activated phytohormonal signals. Science 311:91–94

    CAS  PubMed  Article  Google Scholar 

  2. Ali GS, Reddy ASN (2006) ATP, phosphorylation and transcription regulate the mobility of plant splicing factors. J Cell Sci 119:3527–3538

    CAS  PubMed  Article  Google Scholar 

  3. Arimura G-I, Köpke S, Kunert M, Volpe V, David A, Brand P, Dabrowska P, Maffei M, Boland W (2008) Effects of feeding spodoptera littoralis on lima bean leaves: IV. diurnal and nocturnal damage differentially initiate plant volatile emission. Plant Physiol 146:965–973

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  4. Aubourg S, Lecharny A, Bohlmann J (2002) Genomic analysis of the terpenoid synthase (AtTPS) gene family of Arabidopsis thaliana. Mol Genet Genomics 267:730–745

    CAS  PubMed  Article  Google Scholar 

  5. Bor T, Aljaloud SO, Gyawali R, Ibrahim SA (2016) Chapter 26 – antimicrobials from herbs, spices, and plants. Fruits Vegetables & Herbs 551–578

  6. Cho JY, Mizutani M, Shimizu BI, Kinoshita T, Ogura M, Tokoro K, Lin ML, Sakata K (2007) Chemical profiling and gene expression profiling during the manufacturing process of Taiwan Oolong tea “Oriental Beauty.” Biosci Biotech Biochem 71:1476–1486

    CAS  Article  Google Scholar 

  7. Davison PA, Hunter CN, Horton P (2002) Overexpression of β-carotene hydroxylase enhances stress tolerance in Arabidopsis. Nature 418:203–206

    CAS  PubMed  Article  Google Scholar 

  8. Dicke M, Ian TB (2010) The evolutionary context for herbivore-induced plant volatiles: beyond the ‘cry for help.’ Trends Plant Sci 15:167–175

    CAS  PubMed  Article  Google Scholar 

  9. Dong F, Fu XM, Watanabe N, Su XG, Yang ZY (2016) Recent advances in the emission and functions of plant vegetative volatiles. Molecules 21:124–134

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  10. Dong F, Yang Z, Baldermann S, Sato Y, Asai T, Watanabe N (2011) Herbivore-induced volatiles from tea (Camellia sinensis) plants and their involvement in intraplant communication and changes in endogenous nonvolatile metabolites. J Agric Food Chem 59:13131–13135

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  11. Dudareva N, Klempien A, Muhlemann JK, Kaplan I (2013) Biosynthesis, function and metabolic engineering of plant volatile organic compounds. New Phytol 198:16–32

    CAS  Google Scholar 

  12. Dunn WB, Broadhurst D, Begley P, Zelena E, Francis-McIntyre S, Anderson N, Brown M, Knowles JD, Halsall A, Haselden JN (2011) Procedures for large-scale metabolic profiling of serum and plasma using gas chromatography and liquid chromatography coupled to mass spectrometry. Nat Protoc 6:1060–1083

    CAS  PubMed  Article  Google Scholar 

  13. Foissac S, Sammeth M (2007) ASTALAVISTA: dynamic and flexible analysis of alternative splicing events in custom gene datasets. Nucleic Acids Res 35:297–299

    Article  Google Scholar 

  14. Gonda I, Bar E, Portnoy V, Lev S, Burger J, Schaffer AA, Ya T, Gepstein S, Giovannoni JJ, Katzir N, Lewinsohn E (2010) Branched-chain and aromatic amino acid catabolism into aroma volatiles in Cucumis melo L. fruit. J Exp Bot 61:1111–1123

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  15. Hackl T, Hedrich R, Jo S, F¨orster F (2014) Proovread: large-scale high-accuracy PacBio correction through iterative short read consensus. Bioinformatics 21:3004–3022

    Article  CAS  Google Scholar 

  16. Ho CT, Zheng X, Li SM (2015) Tea aroma formation. Food Sci Hum Well 4:9–27

    Article  Google Scholar 

  17. Jozefczuk S, Klie S, Catchpole G, Szymanski J, Cuadros-Inostroza A, Steinhauser D, Selbig J, Willmitzer L (2010) Metabolomic and transcriptomic stress response of Escherichia coli. Mol syst biol 6:364–380

    PubMed  PubMed Central  Article  Google Scholar 

  18. Kachroo A, Robin GP (2013) Systemic signaling during plant defense. Curr Opin Plant Biol 16:527–533

    CAS  PubMed  Article  Google Scholar 

  19. Kaminaga Y, Schnepp J, Peel G, Kish CM, Ben-Nissan G, Weiss D, Orlova I, Lavie O, Rhodes D, Wood K (2006) Plant phenylacetaldehyde synthase is a bifunctional homotetrameric enzyme that catalyzes phenylalanine decarboxylation and oxidation. J Biol Chem 281:23357–23366

    CAS  PubMed  Article  Google Scholar 

  20. Kim D, Langmead B, Salzberg SL (2015) HISAT: a fast spliced aligner with low memory requirements. Nat Methods 12:357–360

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  21. Kind T, Wohlgemuth G, Lee DY, Lu Y, Palazoglu M, Shahbaz S, Fiehn O (2009) FiehnLib: mass spectral and retention index libraries for metabolomics based on quadrupole and time-of-flight Gas Chromatography/Mass Spectrometry. Anal Chem 81:10038–10048

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  22. Laloum T, Martín G, Duque P (2017) Alternative splicing control of abiotic stress responses. Trends Plant Sci 23:140–150

    PubMed  Article  CAS  PubMed Central  Google Scholar 

  23. Lei B, Zhao XH, Zhang K, Zhang J, Ren W, Zhu R (2013) Comparative transcriptome analysis of tobacco (Nicotiana tabacum) leaves to identify aroma compound-related genes expressed in different cultivated regions. Mol Biol Rep 40:345–357

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  24. Li CF, Xu YX, Ma JQ, Jin JQ, Chen L (2016) Biochemical and transcriptomic analyses reveal different metabolite biosynthesis profiles among three color and developmental stages in ‘Anji Baicha’ (Camellia sinensis). BMC Plant Biol 16:195–212

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  25. Li M, Li L, Dunwell JM, Qiao X, Liu X, Zhang S (2014) Characterization of the lipoxygenase (LOX) gene family in the Chinese white pear (Pyrus bretschneideri) and comparison with other members of the Rosaceae. BMC Genom 15:1–12

    Google Scholar 

  26. Li Y, Mi X, Zhao S, Zhu J, Wei C (2020) Comprehensive profiling of alternative splicing landscape during cold acclimation in tea plant. BMC Genom 21:1–6

    Article  Google Scholar 

  27. Ma CY, Li JX, Chen W, Wang WW, Qi DD, Pang S, Miao AQ (2018) Study of the aroma formation and transformation during the manufacturing process of oolong tea by solid-phase micro-extraction and gas chromatography–mass spectrometry combined with chemometrics. Food Res Int 108:413–422

    CAS  PubMed  Article  Google Scholar 

  28. Maffei M, Mithöfer A, Boland W (2007) Before gene expression: Early events in plant-insect interaction. Trends Plant Sci 12:310–316

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  29. Mao XZ, Cai T, Olyarchuk JG, Wei LP (2005) Automated genome annotation and pathway identification using the KEGG Orthology (KO) as a controlled vocabulary. Bioinformatics 21:3787–3793

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  30. Marquez Y, Brown JWS, Simpson C, Barta A, Kalyna M (2012) Transcriptome survey reveals increased complexity of the alternative splicing landscape in Arabidopsis. Genome Res 22:1184–1195

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  31. Mei X, Liu X, Zhou Y, Wang XQ, Zeng LT, Fu XM, Li JL, Tang JC, Dong F, Yang ZY (2017) Formation and emission of linalool in tea (Camellia sinensis) leaves infested by tea green leafhopper (Empoasca (Matsumurasca) onukii Matsuda). Food Chem 237:356–363

    CAS  PubMed  Article  Google Scholar 

  32. Ogura M, Kinoshita T, Shimizu BI, Shirai F, Sakata K (2008) Identification of aroma components during processing of the famous formosa Oolong tea “Oriental Beauty.” ACS Sym Ser 988:87–97

    CAS  Article  Google Scholar 

  33. Pérez AG, Olías R, Luaces P, Sanz C (2002) Biosynthesis of strawberry aroma compounds through amino acid metabolism. J Agric Food Chem 50:4037–4042

    PubMed  Article  CAS  PubMed Central  Google Scholar 

  34. Podstolski A, Havkin-Frenkel D, Malinowski J, Blount JW, Dixon RA (2002) Unusual 4–hydroxybenzaldehyde synthase activity from tissue cultures of vanilla orchid Vanilla planifolia. Phytochemistry 61:611–620

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  35. Qiao D, Yang C, Chen J, Guo Y, Li Y, Niu S, Cao K, Chen Z (2019) Comprehensive identification of the full-length transcripts and alternative splicing related to the secondary metabolism pathways in the tea plant ( Camellia sinensis ). Sci Rep 9:2709–2812

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  36. Schwender J, Gemünden C, Lichtenthaler HK (2001) Chlorophyta exclusively use the 1-deoxyxylulose 5-phosphate/2-C-methylerythritol 4-phosphate pathway for the biosynthesis of isoprenoids. Planta 212:416–423

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  37. Scott ER, Li X, Wei JP, Kfoury N, Friedman DR (2020) Changes in tea plant secondary metabolite profiles as a function of leafhopper density and damage. Front Plant Sci 11:636–651

    PubMed  PubMed Central  Article  Google Scholar 

  38. Shen S, Park JW, Lu Z, Lin L, Henry MD, Wu YN, Zhou Q, Xing Y (2014) rMATS: Robust and flexible detection of differential alternative splicing from replicate RNA-Seq data. Proc Natl Acad 111:5593–5601

    Article  CAS  Google Scholar 

  39. Simão FA, Waterhouse RM, Panagiotis I, Kriventseva EV, Zdobnov EM (2015) BUSCO: assessing genome assembly and annotation completeness with single-copy orthologs. Bioinformatics 31:3210–3212

    PubMed  Article  CAS  PubMed Central  Google Scholar 

  40. Takeo T, Tsushida T (1980) Changes in lipoxygenase activity in relation to lipid degradation in plucked tea shoots. Phytochemistry 19:2521–2522

    CAS  Article  Google Scholar 

  41. Thatcher SR, Zhou W, Leonard A, Wang BB, Beatty M, Zastrow-Hayes G, Zhao X, Baumgarten A, Li B (2014) Genome-wide analysis of alternative splicing in zea mays: landscape and genetic regulation. Plant Cell 26:3472–3487

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  42. Vasconsuelo A, Boland R (2007) Molecular aspects of the early stages of elicitation of secondary metabolites in plants. Plant Ence 172:861–875

    CAS  Google Scholar 

  43. Vranová E (2013) Systems understanding of isoprenoid pathway regulation in arabidopsis. Isoprenoid Synth Plants Microorg 33:475–491

    Google Scholar 

  44. Walling LL (2000) The myriad plant responses to herbivores. J Plant Growth Regul 19:195–216

    CAS  PubMed  Article  Google Scholar 

  45. Wang C, Lv S, Wu Y, Gao X, Li J, Zhang W, Meng Q (2016a) Oolong tea made from tea plants from different locations in Yunnan and Fujian, China showed similar aroma but different taste characteristics. Springerplus 5:576

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  46. Wang C, Lv SD, Wu YS, Lian M, Gao XM, Meng QX (2016b) Study of aroma formation and transformation during manufacturing process of Biluochun green tea in Yunnan Province by HS-SPME and GC-MS. J Sci Food Agric 96:4492–4498

    CAS  PubMed  Article  Google Scholar 

  47. Wang T, Wang H, Cai D, Gao Y, Gu L (2017) Comprehensive profiling of rhizome-associated alternative splicing and alternative polyadenylation in moso bamboo (Phyllostachys edulis). Plant J 91:684–699

    CAS  PubMed  Article  Google Scholar 

  48. Wang XQ, Zeng LT, Liao YY, Zhou Y, Xu XL (2018) An alternative pathway for the formation of aromatic aroma compounds derived from l-phenylalanine via phenylpyruvic acid in tea (Camellia sinensis (L.) O. Kuntze) leaves. Food Chem 270:17–24

    PubMed  Article  CAS  Google Scholar 

  49. Watanabe S, Hayashi K, Yagi K, Asai T, Mactavish H, Picone J, Turnbull C, Watanabe N (2002) Biogenesis of 2-phenylethanol in rose flowers: incorporation of [2H8]L-phenylalanine into 2-phenylethanol and its beta-D-glucopyranoside during the flower opening of Rosa “Hoh-Jun” and Rosa damascena Mill. Biosci Biotechnol Biochem 66:943–947

    CAS  PubMed  Article  Google Scholar 

  50. Wu TD, Watanabe CK (2005) GMAP: a genomic mapping and alignment program for mRNA and EST sequences. Bioinformatics 21:1859–1875

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  51. Xia E-H, Li FD, Tong W, Li PH, Wu Q, Zhao HJ (2019) Tea plant information archive: a comprehensive genomics and bioinformatics platform for tea plant. Plant Biotechnol J 17:1938–1953

    PubMed  PubMed Central  Article  Google Scholar 

  52. Xin Z, Ge L, Chen S, Sun X (2019) Enhanced transcriptome responses in herbivore-infested tea plants by the green leaf volatile (Z)-3-hexenol. J Plant Res 132:285–293

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  53. Xu QS, He YX, Yan XM, Zhao SQ, Zhu JY (2018) Unraveling a crosstalk regulatory network of temporal aroma accumulation in tea plant (Camellia sinensis) leaves by integration of metabolomics and transcriptomics. Environ Exp Bot 149:81–94

    Article  CAS  Google Scholar 

  54. Yang H, Xie S, Wang L, Jing S, Zhu X, Li X, Zeng W, Yuan H (2011) Identification of up-regulated genes in tea leaves under mild infestation of green leafhopper. Acta Hortic 130:476–481

    CAS  Article  Google Scholar 

  55. Zhao X, Chen S, Wang S, Shan W, Wang X, Lin Y, Su F, Yang Z, Yu X (2020) Defensive responses of tea plants (Camellia sinensis) against tea green leafhopper attack: a multi-omics study. Front Plant Sci 10:1705–1722

    PubMed  PubMed Central  Article  Google Scholar 

  56. Zhu CH, Li XF, Zheng JY (2018) Transcriptome profiling using Illumina- and SMRT-based RNA-seq of hot pepper for in-depth understanding of genes involved in CMV infection. Gene 666:123–133

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  57. Zou J, Chen J, Tang N, Gao YQ, Hong MS, Wei W, Cao HH, Jian W, Li N, Deng W (2018) Transcriptome analysis of aroma volatile metabolism change in tomato ( Solanum lycopersicum ) fruit under different storage temperatures and 1-MCP treatment. Postharvest Biol Tech 135:57–67

    CAS  Article  Google Scholar 

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Acknowledgements

This work was financially supported by the Targeted Poverty Alleviation and Rural Revitalization Project of the Agricultural Science and Technology Commissioner of Guangdong Province (KA1901003), and the Provincial Modern Agricultural Industrial Park Expert Service Team Docking Service of Xinfeng Country (D11920715).

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QW and G-SX were involved in funding acquisition and the conception, design, and supervision; S-FL contributed to data acquisition and analysis; and H-FL contributed to the intellectual contents, as well as the drafting, revision, and the final approval of the manuscript.

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Correspondence to Qin Wang.

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Liu, H., Li, S., Xiao, G. et al. Formation of volatiles in response to tea green leafhopper (Empoasca onukii Matsuda) herbivory in tea plants: a multi-omics study. Plant Cell Rep (2021). https://doi.org/10.1007/s00299-021-02674-9

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Keywords

  • Tea plant
  • Empoasca (matsumurasca) onukii matsuda
  • Transcriptome
  • Metabolome
  • Aroma