Advertisement

Plant Molecular Biology Reporter

, Volume 33, Issue 2, pp 253–263 | Cite as

Molecular Cloning and Characterization of a Short-Chain Dehydrogenase Showing Activity with Volatile Compounds Isolated from Camellia sinensis

  • Ying Zhou
  • Ling Zhang
  • Jiadong Gui
  • Fang Dong
  • Sihua Cheng
  • Xin Mei
  • Linyun Zhang
  • Yongqing Li
  • Xinguo Su
  • Susanne Baldermann
  • Naoharu Watanabe
  • Ziyin Yang
Original Paper

Abstract

Camellia sinensis synthesizes and emits a large variety of volatile phenylpropanoids and benzenoids (VPB). To investigate the enzymes involved in the formation of these VPB compounds, a new C. sinensis short-chain dehydrogenase/reductase (CsSDR) was isolated, cloned, sequenced, and functionally characterized. The complete open reading frame of CsSDR contains 996 nucleotides with a calculated protein molecular mass of 34.5 kDa. The CsSDR recombinant protein produced in Escherichia coli exhibited dehydrogenase-reductase activity towards several major VPB compounds in C. sinensis flowers with a strong preference for NADP/NADPH co-factors, and showed affinity for (R)/(S)-1-phenylethanol (1PE), phenylacetaldehyde, benzaldehyde, and benzyl alcohol, and no affinity for acetophenone (AP) and 2-phenylethanol. CsSDR showed the highest catalytic efficiency towards (R)/(S)-1PE. Furthermore, the transient expression analysis in Nicotiana benthamiana plants validated that CsSDR could convert 1PE to AP in plants. CsSDR transcript level was not significantly affected by floral development and some jasmonic acid-related environmental stress, and CsSDR transcript accumulation was detected in most floral tissues such as receptacle and anther, which were main storage locations of VPB compounds. Our results indicate that CsSDR is expressed in C. sinensis flowers and is likely to contribute to a number of floral VPB compounds including the 1PE derivative AP.

Keywords

Camellia sinensis 1-Phenylethanol Phenylpropanoids Short chain dehydrogenase Volatile compound 

Abbreviations

ADH

Alcohol dehydrogenases

AP

Acetophenone

CsSDR

C. sinensis short chain dehydrogenase/reductase

CsSDRL

CsSDR-like

GGPP

Geranylgeranyl pyrophosphate

GPP

Geranyl pyrophosphate

ORF

Open reading frame

1PE

1-Phenylethanol

SDR

Short-chain dehydrogenase/reductase

VPB

Volatile phenylpropanoids and benzenoids

Notes

Acknowledgements

This work was supported by the “100 Talents Programme of the Chinese Academy of Sciences” (Y321011001 and 201209), the National Natural Science Foundation for Young Scholar of China (30900997), and the Foundation of Key Laboratory of Plant Resources Conservation and Sustainable Utilization, South China Botanical Garden, Chinese Academy of Sciences.

Supplementary material

11105_2014_751_MOESM1_ESM.doc (1.8 mb)
Figure S1 SDS-PAGE and Western blot analyses of CsSDR and CsSDRL expressed in Escherichia coli (DOC 1845 kb)
11105_2014_751_MOESM2_ESM.doc (116 kb)
Figure S2 (DOC 116 kb)
11105_2014_751_MOESM3_ESM.doc (53 kb)
Figure S3 (DOC 53 kb)
11105_2014_751_MOESM4_ESM.doc (56 kb)
Figure S4 (DOC 56 kb)

References

  1. Abokitse K, Hummel W (2003) Cloning, sequence analysis, and heterologous expression of the gene encoding a (S)-specific alcohol dehydrogenase from Rhodococcus erythropolis DSM 43297. Appl Microbiol Biotechnol 62:380–386CrossRefPubMedGoogle Scholar
  2. Arimura G, Köpke S, Kunert M, Volpe V, David A, Brand P, Dabrowska P, Maffei ME, Boland W (2008) Effects of feeding Spodoptera littoralis on lima bean leaves: IV. Diurnal and nocturnal damage differentially initiate plant volatile emission. Plant Physiol 146:965–973CrossRefPubMedCentralPubMedGoogle Scholar
  3. Brosche M, Strid A (1999) Cloning, expression, and molecular characterization of a small pea gene family regulated by low levels of ultraviolet B radiation and other stresses. Plant Physiol 121:479–487CrossRefPubMedCentralPubMedGoogle Scholar
  4. de Kraker JW, Franssen MC, Dalm MC, de Groot A, Bouwmeester HJ (2001) Biosynthesis of germacrene A carboxylic acid in chicory roots. Demonstration of a cytochrome P450 (+)-germacrene a hydroxylase and NADP + −dependent sesquiterpenoid dehydrogenase(s) involved in sesquiterpene lactone biosynthesis. Plant Physiol 125:1930–1940CrossRefPubMedCentralPubMedGoogle Scholar
  5. Dong F, Yang ZY, 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 59:13131–13135CrossRefPubMedGoogle Scholar
  6. Dong F, Yang ZY, Baldermann S, Kajitani Y, Ota S, Kasuga H, Imazeki Y, Ohnishi T, Watanabe N (2012) Characterization of l-phenylalanine metabolism to acetophenone and 1-phenylethanol in the flowers of Camellia sinensis using stable isotope labeling. J Plant Physiol 169:217–225CrossRefPubMedGoogle Scholar
  7. Effmert U, Dinse C, Piechulla B (2008) Influence of green leaf herbivory by Manduca sexta on floral volatile emission by Nicotiana suaveolens. Plant Physiol 146:1996–2007CrossRefPubMedCentralPubMedGoogle Scholar
  8. Filling C, Berndt KD, Benach J, Knapp S, Prozorovski T, Nordling E, Ladenstein R, Jörnvall H, Oppermann U (2002) Critical residues for structure and catalysis in short-chain dehydrogenases/reductases. J Biol Chem 277:25677–25684CrossRefPubMedGoogle Scholar
  9. Finsterbusch A, Lindemann P, Grimm R, Eckerskorn C, Luckner M (1999) Delta(5)-3beta-hydroxysteroid dehydrogenase from Digitalis lanata Ehrh. — a multifunctional enzyme in steroid metabolism? Planta 209:478–486CrossRefPubMedGoogle Scholar
  10. Höffken HW, Duong M, Friedrich T, Breuer M, Hauer B, Reinhardt R, Rabus R, Heider J (2006) Crystal structure and enzyme kinetics of the (S)-specific 1-phenylethanol dehydrogenase of the denitrifying bacterium Strain EbN1. Biochemistry 45:82–93CrossRefPubMedGoogle Scholar
  11. Itoh N, Morihama R, Wang J, Okada K, Mizuguchi N (1997) Purification and characterization of phenylacetaldehyde reductase from a styrene-assimilating Corynebacterium strain, ST-10. Appl Environ Microbiol 63:3783–3788PubMedCentralPubMedGoogle Scholar
  12. Joshi R, Poonam SR, Guleria S, Babu GD, Kumari M, Gulati A (2011) Characterization of volatile components of tea flowers (Camellia sinensis) growing in Kangra by GC/MS. Nat Prod Commun 6:1155–1158PubMedGoogle Scholar
  13. Jörnvall H, Persson B, Krook M, Atrian S, Gonzàlez-Duarte R, Jeffery J, Ghosh D (1995) Short-chain dehydrogenases/reductases (SDR). Biochemistry 34:6003–6013CrossRefPubMedGoogle Scholar
  14. Kallberg Y, Oppermann U, Jornvall H, Persson B (2002) Short-chain dehydrogenases/reductases (SDRs). Eur J Biochem 269:4409–4417CrossRefPubMedGoogle Scholar
  15. Kallberg Y, Persson B (2006) Prediction of coenzyme specificity in dehydrogenases/reductases. A hidden Markov model-based method and its application on complete genomes. FEBS J 273:1177–1184CrossRefPubMedGoogle Scholar
  16. Kavanagh KL, Jörnvall H, Persson B, Oppermann U (2008) Medium- and short-chain dehydrogenase/reductase gene and protein families: the SDR superfamily: functional and structural diversity within a family of metabolic and regulatory enzymes. Cell Mol Life Sci 65:3895–3906CrossRefPubMedCentralPubMedGoogle Scholar
  17. Kessler D, Diezel C, Baldwin IT (2010) Changing pollinators as a means of escaping herbivores. Curr Biol 20:237–242CrossRefPubMedGoogle Scholar
  18. Kniemeyer O, Heider J (2001) (S)-1-Phenylethanol dehydrogenase of Azoarcus sp. strain EbN1, an enzyme of anaerobic ethylbenzene catabolism. Arch Microbiol 176:129–135CrossRefPubMedGoogle Scholar
  19. Liu ZZ, Wang JL, Huang X, Xu WH, Liu ZM, Fang RX (2003) The promoter of a rice glycine-rich protein gene, Osgrp-2, confers vascular-specific expression in transgenic plants. Planta 216:824–833PubMedGoogle Scholar
  20. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real time quantitative PCR and 2−∆∆ct method. Methods 25:402–408CrossRefPubMedGoogle Scholar
  21. Maffei ME, Gertsch J, Appendino G (2011) Plant volatiles: production, function and pharmacology. Nat Prod Rep 28:1359–1380CrossRefPubMedGoogle Scholar
  22. Manriquez D, El-Sharkawy I, Flores FB, El-Yahyaoui F, Regad F, Bouzayen M, Latché A, Pech JC (2006) Two highly divergent alcohol dehydrogenases of melon exhibit fruit ripening-specific expression and distinct biochemical characteristics. Plant Mol Biol 61:675–685CrossRefPubMedGoogle Scholar
  23. Moummou H, Tonfack LB, Chervin C, Benichou M, Youmbi E, Ginies C, Latché A, Pech JC, van der Rest B (2012) Functional characterization of SlscADH1, a fruit-ripening-associated short-chain alcohol dehydrogenase of tomato. J Plant Physiol 169:1435–1444CrossRefPubMedGoogle Scholar
  24. Negre-Zakharov F, Long MC, Dudareva N (2009) Floral scents and fruit aromas inspired by nature. In: Osbourn AE, Lanzotti V (eds) Plant-derived natural products, synthesis, function, and application. Springer, Berlin, pp 405–431CrossRefGoogle Scholar
  25. Okamoto S, Yu F, Harada H, Okajima T, Hattan J, Misawa N, Utsumi R (2011) A short-chain dehydrogenase involved in terpene metabolism from Zingiber zerumbet. FEBS J 278:2892–2900CrossRefPubMedGoogle Scholar
  26. Oppermann U, Filling C, Hult M, Shafqat N, Wu X, Lindh M, Shafqat J, Nordling E, Kallberg Y, Persson B, Jörnvall H (2003) Short-chain dehydrogenases/reductases (SDR): the 2002 update. Chem Biol Interact 143–144:247–253CrossRefPubMedGoogle Scholar
  27. Pennacchio A, Pucci B, Secundo F, La Cara F, Rossi M, Raia CA (2008) Purification and characterization of a novel recombinant highly enantioselective short-chain NAD(H)-dependent alcohol dehydrogenase from Thermus thermophilus. Appl Environ Microbiol 74:3949–3958CrossRefPubMedCentralPubMedGoogle Scholar
  28. Pichersky E, Noel JP, Dudareva N (2006) Biosynthesis of plant volatiles: nature's diversity and ingenuity. Science 331:808–811CrossRefGoogle Scholar
  29. Rasch B, Büchel C, Gais S, Born J (2007) Odor cues during slow-wave sleep prompt declarative memory consolidation. Science 315:1426–1429CrossRefPubMedGoogle Scholar
  30. Sakai M, Hirata H, Sayama H, Sekiguchi K, Itano H, Asai T, Dohra H, Hara M, Watanabe N (2007) Production of 2-phenylethanol in roses as the dominant floral scent compound from l-phenylalanine by two key enzymes, a PLP-dependent decarboxylase and a phenylacetaldehyde reductase. Biosci Biotechnol Biochem 71:2408–2419CrossRefPubMedGoogle Scholar
  31. Schiestl FP (2010) The evolution of floral scent and insect chemical communication. Ecol Lett 13:643–656CrossRefPubMedGoogle Scholar
  32. Schwab W, Davidovich-Rikanati R, Lewinsohn E (2008) Biosynthesis of plant-derived flavor compounds. Plant J 54:712–732CrossRefPubMedGoogle Scholar
  33. Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599CrossRefPubMedGoogle Scholar
  34. Theis N, Kesler K, Adler LS (2009) Leaf herbivory increases floral fragrance in male but not female Cucurbita pepo subsp texana (Cucurbitaceae) flowers. Am J Bot 96:897–903CrossRefPubMedGoogle Scholar
  35. Tieman D, Taylor M, Schauer N, Fernie AR, Hanson AD, Klee HJ (2006) Tomato aromatic amino acid decarboxylases participate in synthesis of the flavor volatiles 2-phenylethanol and 2-phenylacetaldehyde. Proc Natl Acad Sci U S A 103:8287–8292CrossRefPubMedCentralPubMedGoogle Scholar
  36. Tieman DM, Loucas HM, Kim JY, Clark DG, Klee HJ (2007) Tomato phenylacetaldehyde reductases catalyze the last step in the synthesis of the aroma volatile 2-phenylethanol. Phytochemistry 68:2660–2669CrossRefPubMedGoogle Scholar
  37. Wu JQ, Hettenhausen C, Meldau S, Baldwin IT (2007) Herbivory rapidly activates MAPK signaling in attacked and unattacked leaf regions but not between leaves of Nicotiana attenuate. Plant Cell 19:1096–1122CrossRefPubMedCentralPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Ying Zhou
    • 1
  • Ling Zhang
    • 1
  • Jiadong Gui
    • 1
    • 2
  • Fang Dong
    • 3
  • Sihua Cheng
    • 4
  • Xin Mei
    • 1
  • Linyun Zhang
    • 4
  • Yongqing Li
    • 1
  • Xinguo Su
    • 3
  • Susanne Baldermann
    • 5
  • Naoharu Watanabe
    • 6
  • Ziyin Yang
    • 1
    • 2
  1. 1.Key Laboratory of Plant Resource Conservation and Sustainable Utilization, South China Botanical GardenChinese Academy of SciencesGuangzhouChina
  2. 2.University of Chinese Academy of SciencesBeijingChina
  3. 3.Guangdong Food and Drug Vocational CollegeGuangzhouChina
  4. 4.College of Horticultural ScienceSouth China Agricultural UniversityGuangzhouChina
  5. 5.Institute of Nutritional ScienceUniversity of PotsdamNuthetalGermany
  6. 6.Graduate School of Science and TechnologyShizuoka UniversityHamamatsuJapan

Personalised recommendations