Advertisement

Establishment of Persicaria minor hairy roots and analysis of secreted β-caryophyllene in medium broth

  • Mehdi Farshad Ashraf
  • Che Radziah Che Mohd Zain
  • Zamri Zainal
  • Normah Mohd Noor
  • Nurina Anuar
  • Masturah Markom
  • Ismanizan Ismail
Original Paper

Abstract

A method for hairy root induction of Persicaria minor, was developed and investigated, and a qualitative evaluation of the sesquiterpenes in the culture medium was performed. The transgenic status was confirmed by PCR analysis using rolB and virD specific primers. The efficiency with which four different strains of Agrobacterium rhizogenes (A4, ATCC43056, ATCC15834 and ATCC13333) and different concentrations of acetosyringone induced hairy roots from in vitro leaf segments of P. minor was evaluated. Strains A4 and ATCC13333 were able to induce hairy root in an average of 6 days of co-cultivation and exhibited significantly higher transformation efficiencies, 20 and 21.3 %, respectively, compared to the other strains in the presence of 100 µM acetosyringone. Half-strength Murashige–Skoog basal medium supplemented with Gamborg vitamins supported the greatest increase (35-fold) of growth index of hairy roots relative to the initial weight after 8 weeks of culture. GC–MS results showed that β-caryophyllene was the main sesquiterpenes detected and was quantified with standards, with total peak areas of 7.934. Results suggested that hairy roots secreted sesquiterpenes into the medium while preserving viable hairy roots as a source of specialized metabolites.

Keywords

Persicaria minor Sesquiterpene Hairy root culture β-Caryophyllene 

Abbreviations

AS

Acetosyringone

GI

Growth index

HR (s)

Hairy root (s)

MS

Murashige–Skoog

TF

Transformation frequency

Notes

Acknowledgments

The authors gratefully thank the Ministry of Science, Technology and Innovation (MOSTI) of Malaysia through the Science Fund research grant (02-01-02-SF0909) and UKM Research University grant (DLP-2013-008) for funding this research.

References

  1. Amiel E, Ofir R, Dudai N, Soloway E, Rabinsky T, Rachmilevitch Sh (2012) β-Caryophyllene, a compound isolated from the biblical balm of gilead (Commiphora gileadensis), is a selective apoptosis inducer for tumor cell lines. Evidence-Based Complementary and Alternative Medicine. doi: 10.1155/2012/872394 PubMedCentralPubMedGoogle Scholar
  2. Amoah BK, Wu H, Sparks C, Jones HD (2001) Factors influencing Agrobacterium-mediated transient expression of uidA in wheat inflorescence tissue. J Exp Bot 52(358):1135–1142. doi: 10.1093/jexbot/52.358.1135 CrossRefPubMedGoogle Scholar
  3. Asadollahi MA, Maury J, Moller K, Nielsen KF, Schalk M, Clark A, Nielsen J (2008) Production of plant sesquiterpenes in Saccharomyces cerevisiae: effect of ERG9 repression on sesquiterpene biosynthesis. Biotechnol Bioeng 99(3):666–677. doi: 10.1002/bit.21581 CrossRefPubMedGoogle Scholar
  4. Ashour ML, El-Readi M, Youns M, Mulyaningsih S, Sporer F, Efferth T, Wink M (2009) Chemical composition and biological activity of the essential oil obtained from Bupleurum marginatum (Apiaceae). J Pharm Pharmacol 61(8):1079–1087. doi: 10.1211/jpp.61.08.0012 CrossRefPubMedGoogle Scholar
  5. Ashraf MF, Maheran AA, Mihdzar AK, Stanslas J, Farokhian E (2013) In vitro tuberization of Chlorophytum Borivilianum Sant & Fern (Safed musli) as influenced by sucrose, CCC and culture systems. Plant Cell Physiol 54(8):1356–1364. doi: 10.1093/pcp/pct083 CrossRefGoogle Scholar
  6. Ayadi R, Trémouillaux-Guiller J (2003) Root formation from transgenic calli of Ginkgo biloba. Tree Physiol 23(10):713–718. doi: 10.1093/treephys/23.10.713 CrossRefPubMedGoogle Scholar
  7. Banerjee S, Singh S, Rahman LU (2012) Biotransformation studies using hairy root cultures—a review. Biotechnol Adv 30:461–468. doi: 10.1016/j.biotechadv.2011.08.010 CrossRefPubMedGoogle Scholar
  8. Chae YA, Yu HS, Song JS, Chun HK, Park SU (2010) Indigo production in hairy root cultures of Polygonum tinctorium Lour. Biotechnol Lett 22:1527–1530. doi: 10.1023/A:1005668625822 Google Scholar
  9. Chandra Sh, Chandra R (2011) Engineering secondary metabolite production in hairy roots. Phytochem Rev 10:371–395. doi: 10.1007/s11101-011-9210-8 CrossRefGoogle Scholar
  10. Cheng AX, Xiang CY, Li JX, Yang CQ, Hu WL, Wang LJ, Lou YG, Chen XY (2007) The rice (E)-beta-caryophyllene synthase (OsTPS3) accounts for the major inducible volatile sesquiterpenes. Phytochemistry 68:1632–1641. doi: 10.1016/j.phytochem.2007.04.008 CrossRefPubMedGoogle Scholar
  11. Chiang L, Abdullah MA (2007) Enhanced anthraquinones production from adsorbent-treated Morinda elliptica cell suspension cultures in production medium strategy. Process Biochem 42:757–763. doi: 10.1016/j.procbio.2007.01.005 CrossRefGoogle Scholar
  12. Danphitsanuparn P, Boonsnongcheep P, Boriboonkaset T, Chintapakorn Y, Prathanturarug S (2012) Effects of Agrobacterium rhizogenes strains and other parameters on production of isoflavonoids in hairy roots of Pueraria candollei Grah. ex Benth.var, candollei. Plant Cell, Tissue Organ Cult 111:315–322CrossRefGoogle Scholar
  13. Di Cola A, Constantino P, Spano L (1996) Cell commitment and rolB gene expression in the induction of root differentiation. Plant Cell, Tissue Organ Cult 46:203–209. doi: 10.1007/BF02307096 CrossRefGoogle Scholar
  14. Fernandes ES, Passos GF, Medeiros R et al (2007) Anti-inflammatory effects of compounds alpha-humulene and (−)-trans-caryophyllene isolated from the essential oil of Cordia verbenacea. Eur J Pharmacol 569(3):228–236. doi: 10.1016/j.ejphar.2007.04.059 CrossRefPubMedGoogle Scholar
  15. Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50(1):151–158. doi: 10.1016/0014-4827(68)90403-5 CrossRefPubMedGoogle Scholar
  16. Gertsch J, Leonti M, Raduner S, Racz I, Chen J, Xie X, Altmann K, Karsak M, Zimmer A (2008) β-Caryophyllene is a dietary cannabinoid. Proceedings of the National Acadamey of Sciences. 105:9099–9104. doi: 10.1073/pnas.0803601105 CrossRefGoogle Scholar
  17. Ismail I, Gor MCh, Hussein ZAM, Zainal Z, Noor NM (2011) Alteration of abiotic stress responsive genes in Polygonum minor roots by jasmonic acid elicitation. In: Vasanthaiah H (ed) Plants and environment. InTech., pp 49–88, ISBN: 978-953-307-779-6Google Scholar
  18. Jiang J (2005) Volatile composition of the laksa plant (Polygonum hydropiper L.), a potential source of green note aroma compounds. Flavour and Fragrance Journal. 20:455–459. doi: 10.1002/ffj.1363 CrossRefGoogle Scholar
  19. Khanuja SPS, Shasany AK, Darokar MP, Kumar S (1999) Rapid isolation of DNA from dry and fresh samples of plants producing large amounts of secondary metabolites and essential oils. Plant Molecular Biology Reporter. 17:1–7. doi: 10.1023/A:1007528101452 CrossRefGoogle Scholar
  20. Köllner T, Held M, Lenk C, Hiltpold I, Turlings TCJ, Gersgenzon J, Degenhardt J (2008) A maize (E)-β-caryophyllene synthase implicated in indirect defense responses against herbivores is not expressed in most American maize varieties. Plant Cell 20:482–494. doi: 10.1105/tpc.107.051672 CrossRefPubMedCentralPubMedGoogle Scholar
  21. Kubo I, Chaudhuri SK, Kubo Y, Sanchez Y, Ogura T, Saito T, Ishikawa H, Haraguchi H (1996) Cytotoxic and antioxidative sesquiterpenoids from Heterotheca inuloides. Planta Med 62(5):427–430. doi: 10.1055/s-2006-957932 CrossRefPubMedGoogle Scholar
  22. Kumar V, Sharma A, Prasad BCN, Gururaj HB, Ravishankar GA (2006) Agrobacterium rhizogenes mediated genetic transformation resulting in hairy root formation is enhanced by ultrasonication and acetosyringone treatment. Electron J Biotechnol 9(4):349–357. doi: 10.2225/vol9-issue4-fulltext-4 CrossRefGoogle Scholar
  23. Lee KT, Yamakawa T, Kodama T, Shimomura K (1998) Effects of chemicals on alkaloid production by transformed roots of Belladonna. Phytochemistry 49:2343–2347. doi: 10.1016/S0031-9422(97)01074-1 CrossRefGoogle Scholar
  24. Mehmetoglu U, Curtis WR (1997) Effects of abiotic inducers on sesquiterpene synthesis in hairy root and cell-suspension cultures of Hyoscyamus muticus. Appl Biochem Biotechnol 67:71–77. doi: 10.1007/BF02787842 CrossRefGoogle Scholar
  25. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497. doi: 10.1111/j.1399-3054.1962.tb08052.x CrossRefGoogle Scholar
  26. Sabulal B, Dan M, John A, Kurup R, Pradeep NS, Valsamma RK, George V (2006) Caryophyllene-rich rhizome oil of Zingiber nimmonii from South India: chemical characterization and antimicrobial activity. Phytochemistry 67(22):2469–2473. doi: 10.1016/j.phytochem.2006.08.003 CrossRefPubMedGoogle Scholar
  27. Sánchez-Muñoz BA, Aguilar MI, King-Díaz B, Rivero JF, Lotina-Hennsen B (2012) The sesquiterpenes β-caryophyllene and caryophyllene oxide isolated from Senecio salignus act as phytogrowth and photosynthesis inhibitors. Molecules 17:1437–1447. doi: 10.3390/molecules17021437 CrossRefPubMedGoogle Scholar
  28. Scholtmeijer K, Cankar K, Beekeilder J, Wosten HAB, Lugones LG, Bosch D (2014) Production of (+)-valencene in the mushroom-forming fungus S. commune. Appl Microbiol Biotechnol 98:5059–5068. doi: 10.1007/s00253-014-5581-2 CrossRefPubMedGoogle Scholar
  29. Sharafi A, Sohi HH, Mousavi A, Azadi P, Razavi Kh, Ntui VO (2013) A reliable and efficient protocol for inducing hairy roots in Papaver bacteatum. Plant Cell, Tissue Organ Cult 113:1–9. doi: 10.1007/s11240-012-0246-2 CrossRefGoogle Scholar
  30. Sheng J, Citovsky V (1996) Agrobacterium-plant cell DNA transport: have virulence proteins, will travel. Plant Cell 8(10):1699–1710. doi: 10.1105/tpc.8.10.1699 CrossRefPubMedCentralPubMedGoogle Scholar
  31. Sneden AT (2000) Protein kinasen C inhibitory phenylpropanoid glycosides from Polygonum species. In: Cutler SJ, Cutler HG (eds) Biologically active natural products: pharmaceuticals. CRC Press, Boca Raton, pp 172–180Google Scholar
  32. Sujatha G, Zdravković-Korać S, Ćalić D, Flamini G, Ranjitha Kumari BD (2013) High-efficiency Agrobacterium rhizogenes-mediated genetic transformation in Artemisia vulgaris: hairy root production and essential oil analysis. Ind Crops Prod 44:643–652. doi: 10.1016/j.indcrop.2012.09.007 CrossRefGoogle Scholar
  33. Tamogami Sh, Narita Y, Suzuki Sh, Nishizawa T, Hanai H, Noma M (2007) Volatile sesquiterpenes emitted from leaves of Polygonum longisetum treated with jasmonic acid and its amide conjugates. Journal of Pesticide Science. 32(3):264–269CrossRefGoogle Scholar
  34. Tao J, Li L (2006) Genetic transformation of Torenia fournieri L. mediated by Agrobacterium rhizogenes. South African Journal of Botany. 72:211–216. doi: 10.1016/j.sajb.2005.07.010 CrossRefGoogle Scholar
  35. Topal U, Sasaki M, Goto M, Otles S (2008) Chemical compositions and antioxidant properties of essential oils from nine species of Turkish plants obtained by supercritical carbon dioxide extraction and steam distillation. Int J Food Sci Nutr 59:619–634. doi: 10.1080/09637480701553816 CrossRefPubMedGoogle Scholar
  36. Wan Hassan WE (2007) Healing herbs of Malaysia (Cetak ulang). Federal Land Development Authority (FELDA), Kuala LumpurGoogle Scholar
  37. Wang R, Peng S, Zeng R, Ding LW, Zengfu X (2009) Cloning, expression and wounding induction of β-caryophyllene synthase gene from Mikania micrantha H.B.K. and allelopathic potential of β-caryophyllene. Allelopathy Journal. 24:35–44Google Scholar
  38. Whitehead IM, Ewing DF, Threlfall DR (1988) Sesquiterpenoids related to the phytoalexin debneyol from elicited cell suspension cultures of Nicotiana tabacum. Phytochemistry 27:1365–1370. doi: 10.1016/0031-9422(88)80195-X CrossRefGoogle Scholar
  39. Wiart Ch (2006) Medicinal plants of Asia and the pacific. CRC Press, Boca RatonGoogle Scholar
  40. Wu H, Sparks C, Amoah B, Jones HD (2003) Factors influencing successful Agrobacterium-mediated genetic transformation of wheat. Plant Cell Rep 21:659–668. doi: 10.1007/s00299-002-0564-7 PubMedGoogle Scholar
  41. Zhao JL, Zhou LG, Wu JY (2010) Promotion of Salvia miltiorrhiza hairy root growth and tanshinone production by polysaccharide–protein fractions of plant growth-promoting rhizobacterium Bacillus cereus. Process Biochem 45(9):1517–1522. doi: 10.1016/j.procbio.2010.05.034 CrossRefGoogle Scholar
  42. Zhou L, Tian T, Xue B, Song L, Liu L, Yu R (2012) Biosynthesis of coumarin glycosides by transgenic hairy roots of Polygonum multiflorum. Biosci Biotechnol Biochem 76(5):1008–1010. doi: 10.1271/bbb.110347 CrossRefPubMedGoogle Scholar
  43. Zhu LH, Holefors A, Ahlman A, Xue ZT, Welander M (2001) Transformation of the apple rootstock M.9/29 with the rolB gene and its influence on rooting and growth. Plant Sci 160(3):433–439. doi: 10.1016/S0168-9452(00)00401-5 CrossRefPubMedGoogle Scholar
  44. Zych M, Pietrosiuk A, Karasiewicz M, Bogacz A, Kujawski R, Mrozikiewicz PM, Patan AK, Furmanowa M (2008) Establishment of Rhodiola kirilowii hairy roots using Agrobacterium rhizogenes LBA 9402. Herba Polonica. 54(4):7–16Google Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Mehdi Farshad Ashraf
    • 1
    • 2
  • Che Radziah Che Mohd Zain
    • 2
  • Zamri Zainal
    • 1
    • 2
  • Normah Mohd Noor
    • 1
  • Nurina Anuar
    • 3
  • Masturah Markom
    • 3
  • Ismanizan Ismail
    • 1
    • 2
  1. 1.Plant Biotechnology Centre, Institute of Systems Biology (INBIOSIS)Universiti Kebangsaan MalaysiaBangiMalaysia
  2. 2.Faculty of Science and Technology, School of Biosciences and BiotechnologyUniversiti Kebangsaan MalaysiaBangiMalaysia
  3. 3.Department of Chemical and Process Engineering, Faculty of Engineering and Built EnvironmentUniversiti Kebangsaan MalaysiaBangiMalaysia

Personalised recommendations