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Molecular Biology Reports

, Volume 39, Issue 2, pp 1721–1729 | Cite as

Effects of different elicitors on yield of tropane alkaloids in hairy roots of Anisodus acutangulus

  • Guoyin KaiEmail author
  • Sheng Yang
  • Yan Zhang
  • Xiuqin Luo
  • Xueqing Fu
  • Ang Zhang
  • Jianbo Xiao
Article

Abstract

The four tropane alkaloids have played a pivotal role in controlling diseases such as the toxic and septic shock, the organophosphorus poison and the acute lung injury. Here, the elicitation effect of different elicitors on the production of tropane alkaloids and the molecular mechanism of enzyme genes in the pathway was firstly demonstrated in hairy roots of Anisodus acutangulus. The results showed ethanol, methyl jasmonate and Ag+ could improve the accumulation of tropane alkaloids up to 1.51, 1.13 and 1.08 times after 24 h treatment, respectively (P < 0.05), whereas salicylic acid decreased the average content of tropane alkaloids. Furthermore, expression profile analysis results revealed that up-regulation of hyoscyamine-6b-hydroxylase (AaH6H) and little regulation of tropinone reducase II (AaTR2) elicited by ethanol, increased expression of putrescine N-methyltransferase I (AaPMT1) elicited by Ag+, elevated expression of tropinone reducase I (AaTR1) elicited by methyl jasmonate, respectively, resulted in tropane alkaloids improvement. Our results showed that hairy root culture of A. acutangulus in combination with elicitors was a promising way for production of tropane alkaloids in the future.

Keywords

Anisodus acutangulus Hairy roots Tropane alkaloids Elicitors Biosynthetic genes 

Abbreviations

A. acutangulus

Anisodus acutangulus

TA

Tropane alkaloids

MeJA

Methyl jasmonate

SA

Salicylic acid

EtOH

Ethanol

SA/EtOH

Salicylic acid dissolved in ethanol

RT-PCR

Reverse transcriptase-polymerase chain reaction

DW

Dry weight

FW

Fresh weight

Notes

Acknowledgments

This work was supported by National Natural Science Fund (30900110), Shanghai Science and Technology Committee Project (10JC1412000, 09QH1401900, 06QA14038, 08391911800, 073158202, 075405117, 065458022, 05ZR14093), Shanghai Education Committee Fund (09ZZ138, 06DZ015, J50401), Zhejiang Provincial Natural Science Fund (Y2080621), Fujian Science and Technology Committee Key Special Project (2008NZ0001-4), National Transgenic Organism New Variety Culture Key Project (2009ZX08012-002B), Project from Shanghai Normal University (SK200830, CH030). Miss Pin Liu’s (Shanghai Jiaotong University, China) kind assistance with HPLC analysis is also acknowledged.

References

  1. 1.
    Wang H, Zhang SH, Huang LS, Zhao RH, Wei HP (1999) Protective effects of anisodine and anisodamine on acute lung injury induced by oleic acid in rats. Chin J Crit Care Med 19(11):663–664Google Scholar
  2. 2.
    Wu GL, Zheng QY, Tian N (2002) Protective effects of anisodine, buscopan, anisodamine on acute renal failure with damages of renal microvessels in rat. Chin J Microcirc 12(1):6–8Google Scholar
  3. 3.
    Zayed R, Wink M (2004) Induction of tropane alkaloid formation in transformed root cultures of Brugmansia suaveolens (Solanaceae). Z Naturforsch C 59(11–12):863–867PubMedGoogle Scholar
  4. 4.
    Zhang L, Ding R, Chai YR, Bonfill M, Piñol MT, Xu TF, Pi Y, Wang ZN, Zhang HM, Kai GY, Liao ZH, Sun XF, Tang KX (2004) Engineering tropane biosynthetic pathway in Hyoscyamus niger hairy root cultures. Proc Natl Acad Sci USA 101:6786–6791PubMedCrossRefGoogle Scholar
  5. 5.
    Hashimoto T, Yamada Y (1987) Purification and characterization of hyoscyamine 6 beta-hydroxylase from root cultures of Hyoscyamus niger L. Hydroxylase and epoxidase activities in the enzyme preparation. Eur J Biochem 164:277–285PubMedCrossRefGoogle Scholar
  6. 6.
    Oksman-Caldentey KM, Kivel O, Hiltunen R (1991) Spontaneousshoot organogenesis and plant regeneration from hairy root cultures of Hyoscyamus muticus. Plant Sci 78:129–136CrossRefGoogle Scholar
  7. 7.
    Li L, Wang J, Wang W, Lu Y, Wang YL, Zhou GY, Kai GY (2008) Optimization of induction and culture conditions and tropane alkaloid production in hairy roots of Anisodus acutangulus. Biotechnol Bioprocess Eng 13:606–612CrossRefGoogle Scholar
  8. 8.
    Boitel-Conti M, Laberche JC, Lanoue A, Ducrocq C, Sangwan-Norreel BS (2000) Influence of feeding precursors on tropane alkaloid production during an abiotic stress in Datura innoxia transformed roots. Plant Cell Tissue Organ Cult 60:131–137CrossRefGoogle Scholar
  9. 9.
    Pitta-Alvarez SI, Spollansky TC, Giulietti AM (2000) The influence of different biotic and abiotic elicitors on the production and profile of tropane alkaloids in hairy root cultures of Brugmansia candida. Enzyme Microb Technol 26(2–4):252–258PubMedCrossRefGoogle Scholar
  10. 10.
    Zhao J, Davis LC, Verpoorte R (2005) Elicitor signal transduction leading to production of plant secondary metabolites. Biotechnol Adv 23:283–333PubMedCrossRefGoogle Scholar
  11. 11.
    El Jaber-Vazdekis N, Barres ML, Ravelo AG, Zarate R (2008) Effects of elicitors on tropane alkaloids and gene expression in Atropa baetica transgenic hairy roots. J Nat Prod 71(12):2026–2031PubMedCrossRefGoogle Scholar
  12. 12.
    Kai GY, Zhang Y, Chen JF, Li L, Yan XM, Zhang R, Liao P, Lu X, Wang W, Zhou GY (2009) Molecular characterization and expression analysis of two distinct putrescine N-methyltransferases from roots of Anisodus acutangulus. Physiol Plant 135:121–129PubMedCrossRefGoogle Scholar
  13. 13.
    Kai GY, Li L, Jiang YX, Yan XM, Zhang Y, Lu X, Liao P, Chen JB (2009) Molecular cloning and characterization of two tropinone reductases in Anisodus acutangulus and enhancement of tropane alkaloids production in AaTRI-transformed hairy roots. Biotechnol Appl Biochem 54(3):177–186PubMedCrossRefGoogle Scholar
  14. 14.
    Kai GY, Chen JF, Li L, Zhou GY, Zhou LM, Zhang L, Chen YH, Zhao LX (2007) Molecular cloning and characterization of a new cDNA encoding hyoscyamine 6β-hydroxylase from roots of Anisodus acutangulus. J Biochem Mol Biol 40(5):715–722PubMedCrossRefGoogle Scholar
  15. 15.
    Cardillo AB, María Otalvaro Alvarez A, Calabró Lopez A, Enrique Velásquez Lozano M, Rodríguez Talou J, María Giulietti A (2009) Anisodamine production from natural sources: seedlings and hairy root cultures of Argentinean and Colombian Brugmansia candida plants. Planta Med 76(4):402–405PubMedCrossRefGoogle Scholar
  16. 16.
    Chen H, Chen F (2000) Effect of yeast elicitor on the secondary metabolism of Ti-transformed Salvia miltiorrhiza cell suspension cultures. Plant Cell Rep 19:710–717CrossRefGoogle Scholar
  17. 17.
    Durrant WE, Dong X (2004) Systemic acquired resistance. Annu Rev Phytopathol 42:185–209PubMedCrossRefGoogle Scholar
  18. 18.
    Hirano H, Shimomura K, Yamakawa T (2003) Responses of transformed roots of Atropa belladonna to treatment with salicylic acid and other phenolic compounds. Plant Biotechnol 20(1):97–100CrossRefGoogle Scholar
  19. 19.
    Suresh B, Thimmaraju R, Bhagyalakshmi N, Ravishankar GA (2004) Polyamine and methyl jasmonate-influenced enhancement of betalaine production in hairy root cultures of Beta vulgaris grown in a bubble column reactor and studies on efflux of pigments. Process Biochem 39:2091–2096CrossRefGoogle Scholar
  20. 20.
    Wang YD, Wu JC, Yuan YJ (2007) Salicylic acid-induced taxol production and isopentenyl pyrophosphate biosynthesis in suspension cultures of Taxus chinensis var. mairei. Cell Biol Int 31:1179–1183PubMedCrossRefGoogle Scholar
  21. 21.
    Rischer H, Oresic M, Seppanen-Laakso T, Katajamaa M, Lammertyn F, Ardiles-Diaz W, Van Montagu MC, Inze D, Oksman-Caldentey KM, Goossens A (2006) Gene-tometabolite networks for terpenoid indole alkaloid biosynthesis in Catharanthus roseus cells. Proc Natl Acad Sci USA 103:5614–5619PubMedCrossRefGoogle Scholar
  22. 22.
    Steczko J, Donoho GA, Dixon JE, Sugimoto T, Axelrod B (1991) Effect of ethanol and low-temperature culture on expression of soybean lipoxygenase L-1 in Escherichia coli. Protein Expr Purif 2(2–3):221–227PubMedCrossRefGoogle Scholar
  23. 23.
    Kuo HF, Tsai YF, Young LS, Lin CY (2000) Ethanol treatment triggers a heat shock-like response but no thermotolerance in soybean (Glycine max cv. Kaohsiung No. 8) seedlings. Plant Cell Environ 23:1099–1108CrossRefGoogle Scholar
  24. 24.
    Gundlach H, Mfiller MJ, Kutchan TM, Zenk MH (1992) Jasmonic acid is a signal transducer in elicitor-induced plant cell cultures. Proc Natl Acad Sci USA 89:2389–2393PubMedCrossRefGoogle Scholar
  25. 25.
    Guo BH, Kai GY, Gong YF, Jin HB, Wang YC, Miao ZQ, Sun XF, Tang KX (2007) Molecular cloning and heterologous expression of a 10-deacetylbaccatin III-10-O-acetyl transferase cDNA from Taxus x media. Mol Biol Rep 34:89–95PubMedCrossRefGoogle Scholar
  26. 26.
    Kang SM, Jung HY, Kang YM, Yun DJ, Bahk JD, Yang JK, Choi MS (2004) Effects of methyl jasmonate and salicylic acid on the production of tropane alkaloids and the expression of PMT and H6H in adventitious root cultures of Scopolia parviflora. Plant Sci 166:745–751CrossRefGoogle Scholar
  27. 27.
    Caretto S, Quarta A, Durante M, Nisi R, De Paolis A, Blando F, Mita G (2009) Methyl jasmonate and miconazole differently affect arteminisin production and gene expression in Artemisia annua suspension cultures. Plant Biol 13:51–58CrossRefGoogle Scholar
  28. 28.
    Xiao Y, Gao S, Di P, Chen J, Chen W, Zhang L (2009) Lithospermic acid B is more responsive to silver ions (Ag+) than rosmarinic acid in Salvia miltiorrhiza hairy root cultures. Biosci Rep 30(1):33–40PubMedCrossRefGoogle Scholar
  29. 29.
    Xiao Y, Di P, Chen J, Liu Y, Chen W, Zhang L (2009) Characterization and expression profiling of 4-hydroxyphenylpyruvate dioxygenase gene (Smhppd) from Salvia miltiorrhiza hairy root cultures. Mol Biol Rep 36(7):2019–2029PubMedCrossRefGoogle Scholar
  30. 30.
    Sato F, Hashimoto T, Haciya A, Tamura K, Choi K-B, Morishige T, Fujimoto H, Yamada Y (2001) Metabolic engineering of plant alkaloid biosynthesis. Proc Natl Acad Sci USA 98:367–372PubMedCrossRefGoogle Scholar
  31. 31.
    Moyano E, Fornalé S, Palazón J, Cusidó RM, Bagni N, Piñol MT (2002) Alkaloid production in Duboisia hybrid hairy root cultures overexpressing the pmt gene. Phytochemistry 59:697–702PubMedCrossRefGoogle Scholar
  32. 32.
    Lee KT, Hirano H, Yamakawa T, Kodama T, Igarashi Y, Shimomura K (2001) Responses of transformed root culture of Atropa belladonna to salicylic acid stress. J Biosci Bioeng 91:586–589PubMedCrossRefGoogle Scholar
  33. 33.
    Xu YH, Wang JW, Wang S, Wang JY, Chen XY (2004) Characterization of GaWRKY1, a cotton transcription factor that regulates the sesquiterpene synthase gene (+)-δ-cadinene synthase-A. Plant Physiol 135:507–515PubMedCrossRefGoogle Scholar
  34. 34.
    Frankfater CR, Dowd MK, Triplett BA (2009) Effect of elicitors on the production of gossypol and methylated gossypol in cotton hairy roots. Plant Cell Tissue Organ Cult 98:341–349CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Guoyin Kai
    • 1
    Email author
  • Sheng Yang
    • 1
  • Yan Zhang
    • 1
  • Xiuqin Luo
    • 1
  • Xueqing Fu
    • 1
  • Ang Zhang
    • 1
  • Jianbo Xiao
    • 1
  1. 1.Laboratory of Plant Biotechnology, College of Life and Environment SciencesShanghai Normal UniversityShanghaiPeople’s Republic of China

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