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Camptothecin and 10-hydroxycamptothecin from Camptotheca acuminata hairy roots

  • Physiology and Biochemistry
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Abstract

Camptothecin (CPT) is an anticancer and antiviral alkaloid produced by the Chinese tree Camptotheca acuminata (Nyssaceae) and some other species belonging to the families Apocynaceae, Olacaceae, and Rubiaceae. Bark and seeds are currently used as sources for the drug. Several attempts have been made to produce CPT from cell suspensions; however, the low yields obtained limit this approach. Cultures of differentiated cell types may be an alternative source of alkaloid production. Hairy root cultures of C. acuminata were established from tissue transformed with Agrobacterium rhizogenes strains ATCC 15834 and R-1000. Integration of the genes responsible for the hairy-root phenotype (rol genes) into the plant genome was verified by DNA gel blot analysis. The hairy roots produce and secrete CPT as well as the more potent and less toxic natural derivative, 10-hydroxycamptothecin (HCPT), into the medium. Remarkably, the cultures were able to synthesize the alkaloids at levels equal to, and sometimes greater than, the roots in planta, i.e., 1.0 and 0.15 mg/g dry weight for CPT and the HCPT, respectively.

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Fig. 1a–c.
Fig. 2.

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Abbreviations

CPT :

Camptothecin

DR :

Dry weight

FW :

Fresh weight

HCPT :

10-Hydroxycamptothecin

HPLC :

High-performance liquid chromatography

kb :

Kilobases

NAA :

1-Napththalenacetic acid

TLC :

Thin-layer chromatography

References

  • Arvy MP, Imbault N, Naudascher F, Thiersault M, Doireae P (1994) 2,4-D and alkaloid accumulation in periwinkle cell suspensions. Biochimie 76:410–416

    Article  CAS  Google Scholar 

  • Bodley AL, Shapiro TA (1995) Molecular and cytotoxic effects of camptothecin, a topoisomerase I inhibitor, on trypanosomes and Leishmania. Proc Natl Acad Sci USA 92:3726–3730

    CAS  Google Scholar 

  • Bonhomme V, Laurain-Mattar D, Lacoux J, Fliniaux MA, Jacquin-Dubreuil A (2000) Tropane alkaloid production by hairy roots of Atropa belladonna obtained after transformation with Agrobacterium rhizogenes 15834 and Agrobacterium tumefaciens containing rol A, B, C genes only. J Biotechnol 81:151–158

    Article  CAS  Google Scholar 

  • Ciddi V, Shuler ML (2000) Camptothecin from callus cultures of Nothapodytes foetida. Biotechnol Lett 22:129–132

    Article  CAS  Google Scholar 

  • De Luca V, St. Pierre B (2000) The cell and developmental biology of alkaloid biosynthesis. Trends Plant Sci 5:168–173

    Article  PubMed  Google Scholar 

  • Flores HE, Medina-Bolivar F (1995) Root culture and plant natural products: "unearthing" the hidden half of plant metabolism. Plant Tissue Cult Biotechnol 1:59–74

    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 

  • Fulzele DP, Satdive RK, Pol BB (2001) Growth and production of camptothecin by cell suspension cultures of Nothapodytes foetida. Planta Med 67:150–152

    Article  CAS  Google Scholar 

  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:104–106

    PubMed  Google Scholar 

  • Gunasekera SP, Badawi MM, Cordell GA, Farnsworth NR, Chitnis M (1979) Plant anticancer agents X. Isolation of camptothecin and 9-methoxycamptothecin from Ervatamia heyneana. J Nat Prod 42:475–477

    CAS  Google Scholar 

  • Hengel AJ van, Harkes MP, Wichers HJ, Hessleink PGM, Buitelaar RM (1992) Characterization of callus formation and camptothecin production by cell lines of Camptotheca acuminata. Plant Cell Tissue Org Cult 28:11–18

    Google Scholar 

  • Imanishi S, Hashizume K, Nakakita M, Kojima H, Matsubayashi Y, Hashimoto T, Sakagami Y, Yamada Y, Nakamura K (1998) Differential induction by methyl jasmonate of genes encoding ornithine decarboxylase and other enzymes involved in nicotine biosynthesis in tobacco cell cultures. Plant Mol Biol 38:1101–1111

    Article  CAS  PubMed  Google Scholar 

  • Jain AK, Nessler CL (1996) Clonal propagation of Camptotheca acuminata through shoot bud culture. Plant Cell Tissue Org Cult 44:229–233

    Google Scholar 

  • Kitajima M, Fischer U, Nakamura M, Ohsawa M, Ueno M, Takayama H, Unger M, Stöckigt J, Aimi N (1998) Anthraquinones from Ophiorrhiza pumila tissue and cell cultures. Phytochemistry 48:107–111

    Article  CAS  Google Scholar 

  • Kjeldsen E, Svejstrup JQ, Gromova II, Alsner J, Westergaard O (1992) Camptothecin inhibits both the cleavage and religation reactions of eukaryotic DNA topoisomerase I. J Mol Biol 228:1025–1030

    CAS  Google Scholar 

  • López-Meyer M, Nessler CL (1997) Tryptophan decarboxylase is encoded by two autonomously regulated genes in Camptotheca acuminata which are differentially expressed during development and stress. Plant J 11:1167–1175

    Article  Google Scholar 

  • López-Meyer M, Nesler CL, McKnight TD (1994) Sites of accumulation of the antitumor alkaloid camptothecin in Camptotheca acuminata. Planta Med 60:558–560

    PubMed  Google Scholar 

  • Lu H, McKnight TD (1999) Tissue specific expression of the beta subunit of tryptophan synthase in Camptotheca acuminata, an indole alkaloid producing plant. Plant Physiol 120:43–52

    Article  CAS  Google Scholar 

  • Medina-Bolivar F, Flores HE (1995) Selection for hyoscyamine and cinnamoyl putrescine overproduction in cell and root cultures of Hyoscyamus muticus. Plant Physiol 108:1553–1560

    CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol Plant 15:473–479

    CAS  Google Scholar 

  • Priel E, Showalter SD, Blair DG (1991) Inhibition of human immunodeficiency virus (HIV-1) replication in vitro by non cytotoxic doses of camptothecin, a topoisomerase I inhibitor. AIDS Res Hum Retroviruses 7:65–72

    CAS  Google Scholar 

  • Roja M, Heble MR (1994) The quinoline alkaloids camptothecin and 9-methoxycamptothecin from tissue cultures and mature trees of Nothapodytes foetida. Phytochemistry 36:65–66

    Article  CAS  Google Scholar 

  • Saito K, Sudo H, Yamazaki M, Koseki-Nakamura M, Kitajima M, Takayama H, Aimi N (2001) Feasible production of camptothecin by hairy root culture of Ophiorrhiza pumila. Plant Cell Rep 20:267–271

    Article  CAS  Google Scholar 

  • Sakato K, Tanaka H, Mukai N, Misawa M (1974) Isolation and identification of camptothecin from cells of Camptotheca acuminata suspension cultures. Agric Biol Chem 38:217–218

    CAS  Google Scholar 

  • Tafur S, Nelson JD, DeLong DC, Svoboda GH (1976) Antiviral components of Ophiorrhiza mungos isolation of camptothecin and 10-methoxycamptothecin. Lloydia 39:261–262

    CAS  Google Scholar 

  • Wall ME, Wani MC, Cook CE, Palmer KH (1966) Plant anti-tumor agents I. The isolation and structure of camptothecin—a novel alkaloidal leukemia and tumor inhibitor from Camptotheca acuminata. J Am Chem Soc 88:3888–3890

    CAS  Google Scholar 

  • Whitmer S, Verpoorte R, Canel C (1999) Influence of auxins on alkaloid accumulation by a transgenic cell line S10 of Catharanthus roseus. In: Whitmer S (PhD Thesis) Aspects of terpenoid indole alkaloid formation by transgenic cell lines of Catharanthus roseus over-expressing tryptophan decarboxylase and strictosidine synthase. Leiden University, Leiden, The Netherlands, pp 51–61

  • Wiedenfeld H, Furmanowa M, Roeder E, Guzewska J, Gustowski H (1997) Camptothecin and 10-hydroxycamptothecin in callus and plantlets of Camptotheca acuminata. Plant Cell Tissue Org Cult 49:213–218

    Article  CAS  Google Scholar 

  • Zhang R, Li Y, Cai Q, Liu T, Sun H, Chambless B (1998) Preclinical pharmacology of the natural product anticancer agent 10-hydroxycamptothecin, an inhibitor of topoisomerase I. Cancer Chemother Pharmacol 41:257–267

    Article  CAS  Google Scholar 

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Acknowledgements

The authors are thankful to J. Fraga, K. Mitchell and A.L. Mercado for their technical assistance. Support for A.L. was provided through Universidad Autónoma del Estado de Morelos (UAEM). This work was also supported by the National Institutes of Health (CA57592) and Project J29064-B from the Consejo Nacional de Ciencia y Tecnología (CONACYT).

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Authors and Affiliations

  1. Department of Plant Pathology, Physiology and Weed Science, Virginia Tech, 413 Price Hall (0331), Blacksburg, VA 24060–0331, USA

    A. Lorence, F. Medina-Bolivar & C. L. Nessler

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  1. A. Lorence
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  2. F. Medina-Bolivar
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  3. C. L. Nessler
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Correspondence to C. L. Nessler.

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Communicated by K.K. Kamo

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Lorence, A., Medina-Bolivar, F. & Nessler, C.L. Camptothecin and 10-hydroxycamptothecin from Camptotheca acuminata hairy roots. Plant Cell Rep 22, 437–441 (2004). https://doi.org/10.1007/s00299-003-0708-4

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  • DOI: https://doi.org/10.1007/s00299-003-0708-4

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