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Transformation of Saussurea medusa for hairy roots and jaceosidin production

  • Genetic Transformation and Hybridization
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Abstract

Axenically grown Saussurea medusa plantlets were inoculated with four Agrobacterium rhizogenes strains, and hairy root lines were established with A. rhizogenes strain R1601 in N6 medium. PCR and Southern hybridization confirmed integration of the T-DNA fragment of the Ri plasmid from A. rhizogenes into the genome of S. medusa hairy roots. In N6 medium, maximum biomass of the hairy root cultures was achieved [8 g (dry weight) per liter; growth ratio 35-fold] after 21 days of culture. The amount of jaceosidin extracted from the hairy root cultures was 46 mg/l (production ratio of 37-fold) after 27 days of culture. The maximum jaceosidin content obtained using N6 medium was higher than that obtained with Modified White, MS or B5 medium. In N6 medium, the tip segments were more efficient for hairy root growth and jaceosidin production than the middle and basal regions of the root.

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Abbreviations

AS:

Acetosyringone

BA:

Benzyladenine

cef:

Cefotaxime sodium

DW:

Dry weight

FW:

Fresh weight

HPLC:

High-performance liquid chromatography

IAA:

Indole-3-acetic acid

km:

Kanamycin

NAA:

α-Naphthaleneacetic acid

SDS:

Sodium dodecyl sulfate

References

  • Aird ELH, Hamill JD, Rhodes MJC (1988) Cytogenetic analysis of hairy root cultures from a number of plant species transformed by Agrobacterium rhizogenes. Plant Cell Tissue Organ Cult 15:47–57

    Google Scholar 

  • Binns AN, Tomashow JV (1988) Cell biology of Agrobacterium infection and transformation of plants. Annu Rev Microbiol 42:575–606

    Article  CAS  Google Scholar 

  • Chu C, Wang C, Sun C, Hsu C, Yin C, Chu C, Bi F (1975) Establishment of an efficient medium for anther culture of rice through comparative experiments on the nitrogen sources. Sci Sin 18:223–231

    PubMed  Google Scholar 

  • David C, Chilton MD, Tempe J (1984) Conservation of T-DNA in plants regenerated from hairy root cultures. Biotechnology 2:73–76

    CAS  Google Scholar 

  • Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15

    Google Scholar 

  • Fan CQ, Yue JM (2003) Biologically active phenols from Saussurea medusa. Bioorg Med Chem 11:703–708

    Article  CAS  PubMed  Google Scholar 

  • Flores HE, Vivanco JM, Loyola-Vargas VM (1999) Radicle biochemistry: the biology of root-specific metabolism. Trends Plant Sci 4:220–226

    Article  PubMed  Google Scholar 

  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158

    CAS  PubMed  Google Scholar 

  • Jouanin L (1984) Restriction map of an agropine-type Ri plasmid and its homologies with Ti plasmids. Plasmid 12:91–102

    CAS  PubMed  Google Scholar 

  • Kamada H, Okamura N, Satake M, Harada H, Shimomura K (1986) Alkaloid production by hairy root cultures in Atropa belladonna. Plant Cell Rep 5:239–242

    CAS  Google Scholar 

  • Krolicka A, Staniszewska I, Bielawski K, Malinski E, Szafranek J, Lojkowska E (2001) Establishment of hairy root cultures of Ammi majus. Plant Sci 160:259–264

    Article  CAS  PubMed  Google Scholar 

  • Lee KT, Suzuki T, Yamakawa T, Kodama T (1999) Production of tropane alkaloids by transformed root cultures of Atropa belladonna in stirred bioreactors with a stainless steel net. Plant Cell Rep 18:567–571

    Article  CAS  Google Scholar 

  • Liu L, Xiao X, Zhang L (1985) Effect of the flavonoids from Saussurea involucrate on DNA synthesis of cancer cells. J Lanzhou Univ Nat Sci 21:80–83

    CAS  Google Scholar 

  • Mano Y, Nabeshima S, Matsui C, Ohkawa H (1986) Production of tropane alkaloids by hairy root cultures of Scopolia japonica. Agric Biol Chem 50:2715–2722

    CAS  Google Scholar 

  • Moyano E, Fornale S, Palazon J, Cusido RM, Bonfill M, Morles C, Pinol MT (1999) Effect of Agrobacterium rhizogenes T-DNA on alkaloid production in Solanaceae plants. Phytochemistry 52:1287–1292

    Article  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    CAS  Google Scholar 

  • Owen HR, Miller AR (1992) An examination and correction of plant tissue culture basal medium formulations. Plant Cell Tissue Organ Cult 28:147–150

    CAS  Google Scholar 

  • Pythoud F, Sinkar VP, Nester EW, Gordon MP (1987) Increased virulence of Agrobacterium rhizogenes conferred by the vir region of pTiBo542: application to genetic engineering of poplar. Biotechnology 5:1323–1327

    Google Scholar 

  • Rhodes MJC, Robins RJ, Hamill JD, Parr AJ, Hilton MG, Walton NJ (1990) Properties of transformed root cultures. In: Charlwood BV, Rhodes MJC (eds) Proc Phytochem Soc Eur Secondary Prod Plant Tissue Cult. Clarendon, Oxford, pp 201–225

    Google Scholar 

  • Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, pp 35–37

    Google Scholar 

  • Schinella GR, Giner RM, Recio MC, de Buschiazzo PM, Rios JL, Manez S (1998) Anti-inflammatory effects of South American Tanacetum vulgare. J Pharm Pharmacol 50:1069–1074

    CAS  PubMed  Google Scholar 

  • Sevon N, Oksman-Caldentey KM (2002) Agrobacterium rhizogenes-mediated transformation: root cultures as a source of alkaloids. Planta Med 68:859–868

    Article  CAS  PubMed  Google Scholar 

  • Slightom JL, Durand-Tardif M, Jouanin L, Tepfer D (1986) Nucleotide sequence analysis of TL-DNA of Agrobacterium rhizogenes agropine type plasmid: identification of open reading frames. J Biol Chem 261:108–121

    CAS  PubMed  Google Scholar 

  • Tepfer DA (1984) Transformation of several species of higher plants by Agrobacterium rhizogenes: sexual transmission of the transformed genotype and phenotype. Cell 37:959–967

    Article  CAS  PubMed  Google Scholar 

  • Vervliet G, Holsters M, Teuchy H, Van MM, Schell J (1975) Characterization of different plaque-forming and defective temperate phages in Agrobacterium. J Gen Virol 26:33–48

    CAS  PubMed  Google Scholar 

  • Wang W-Z (1997) Determination of the content of flavonoids in tissue culture of S. medusa by RP-HPLC. Chin Trad Herb Drugs 28:654–656

    CAS  Google Scholar 

  • White PR (1963) The cultivation of plant and animal cells, 2nd edn. Ronald, New York

    Google Scholar 

  • White FF, Nester EW (1980) Hairy root: plasmid encodes virulence in Agrobacterium rhizogenes. J Bacteriol 141:1134–1141

    CAS  PubMed  Google Scholar 

  • Woerdenbag HJ, Merfort I, Passreiter CM, Schmidt TJ, Willuhn G, Van Uden W, Pras N, Kampinga HH, Konings WT (1994) Cytotoxicity of flavonoids and sesquiterpene lactones from Arnica species against the GLC4 and the COLO 320 cell lines. Planta Med 60:434–437

    CAS  PubMed  Google Scholar 

  • Wysokinska H, Chmiel A (1997) Transformed root cultures for biotechnology. Acta Biotechnol 17:131–159

    CAS  Google Scholar 

  • Yoshikawa T, Furuya T (1987) Saponin production by cultures of Panax ginseng transformed with Agrobacterium rhizogenes. Plant Cell Rep 6:449–453

    CAS  Google Scholar 

  • Zhao D-X, Qiao C-L, Wang Y (1998) Cell culture and selection of high flavonoids-producing cell lines in Saussurea medusa. Acta Bot Sin 40:515–520

    CAS  Google Scholar 

  • Zhao D-X, Xing J-M, Li M-Y, Lu D-P, Zhao G (2001) Optimization of growth and jaceosidin production in callus and cell suspension cultures of Saussurea medusa. Plant Cell Tissue Organ Cult 67:227–234

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Prof. Zhong-ping Lin, National Laboratory of Protein Engineering and Genetic Engineering of Plants, Peking University, Beijing, and Prof. Min Sun, Center for Agricultural Biotechnology, Southwest Agricultural University, Chongqing, for providing the Agrobacterium rhizogenes strains. We thank Prof. Zhong-Jian Jia, Department of Chemistry, Lanzhou University, Lanzhou, and Dr. Jing-yuan Song, Institute of Medicinal Plants, Chinese Academy of Medical Sciences, Beijing, for the generous gifts of standard samples of jaceosidin and mannopine. This investigation was supported by the National Science Foundation of China (No. 39970896).

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Authors and Affiliations

  1. Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, 20 Nanxincun, Xiangshan, Beijing, 100093, China

    Dexiu Zhao, Chunxiang Fu & Yaqiong Chen

  2. Department of Biology, University of Waterloo, 200 University Avenue West, Waterloo, ON, Canada, N2L 3G1

    Fengshan Ma

  3. Southern Crop Protection and Food Research Center, Agriculture and Agri-Food Canada, 1391 Sandford Street, London, ON, Canada, N5V 4T3

    Fengshan Ma

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  1. Dexiu Zhao
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  2. Chunxiang Fu
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Correspondence to Dexiu Zhao.

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Communicated by K.K. Kamo

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Zhao, D., Fu, C., Chen, Y. et al. Transformation of Saussurea medusa for hairy roots and jaceosidin production. Plant Cell Rep 23, 468–474 (2004). https://doi.org/10.1007/s00299-004-0840-9

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  • DOI: https://doi.org/10.1007/s00299-004-0840-9

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