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Production of transgenic Podophyllum peltatum via Agrobacterium tumefaciens-mediated transformation

  • Original Papers
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Biologia Plantarum

Abstract

Transgenic Podophyllum peltatum plants were successfully produced by Agrobacterium tumefaciens-mediated transformation. Embryogenic callus was co-cultivated with Agrobacterium tumefaciens harboring a binary vector pBI 121 carrying β-glucuronidase (GUS) and neomycinphosphotransferase (NPT II) gene. GUS-histochemical analysis revealed that, 50 µM acetosyringone treatments during Agrobacterium infection and 3 d co-cultivation with Agrobacterium showed enhanced transformation efficiency. Percentage of GUS positive callus increased rapidly as the subculture time proceeded on selection medium containing 100 mg dm−3 kanamycin. Kanamycin resistant somatic embryos were formed from embryogenic callus after cultivation with 11.35 µM abscisic acid (ABA) for 3 weeks and then on hormone-free selection medium. Somatic embryos were germinated and converted into plantlets on medium containing 2.89 µM gibberellic acid (GA3). The integration of GUS and NPT II gene into transgenic plants was confirmed by polymerase chain reaction and Southern analysis.

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Abbreviations

ABA:

abscisic acid

CaMV:

cauliflower mosaic virus

2,4-D:

2,4-dichlorophenoxyacetic acid

GA3 :

gibberellic acid

GUS:

β-glucuronidase

MS:

Murashige and Skoog

NOS:

nopaline synthase

NPT II:

neomycinphosphotransferase II

PCR:

polymerase chain reaction

PTOX:

podophyllotoxin

References

  • Ayres, N.M., Park, W.D.: Genetic transformation of rice. — Crit. Rev. Plant Sci. 13: 219–239, 1994.

    Article  CAS  Google Scholar 

  • Cervera, M., Pina, J.A., Juarez, J., Navarvo, L., Pena, L.: Agrobacterium-mediated transformation of citrange: factors affecting transformation and regeneration. — Plant Cell Rep. 18: 271–278, 1998.

    Article  CAS  Google Scholar 

  • Chilton, M.D., Currier, T.C., Farrand, S.K., Bendich, A.J., Gordon, M.P., Nester, E.W.: Agrobacterium tumefaciens DNA and PS8 bacteriophage DNA not detected in crown gall tumors. — Proc. nat. Acad. Sci. USA 71: 3672–3676, 1974.

    Article  CAS  PubMed  Google Scholar 

  • Damayanthi, Y., Lown, J.W.: Podophyllotoxins: current status and recent developments. — Curr. Med. Chem. 5: 205–52, 1998.

    CAS  PubMed  Google Scholar 

  • Franklin, G., Lakshmi Sita, G.: Agrobacterium tumefaciensmediated transformation of egg plant (Solanum melongena L.) using root explants. — Plant Cell Rep. 21: 549–554, 2003.

    CAS  PubMed  Google Scholar 

  • Giri, A., Giri, C.C., Dhingra, V., Narasu, M.L.: Enhanced podophyllotoxin production from Agrobacterium rhizogenes transformed cultures of P. hexandrum. — Nat. Prod. Lett. 15: 229–235, 2001.

    CAS  PubMed  Google Scholar 

  • Godwin, I., Todd, G., Ford-Lloyd, B., Newbury, H.J.: The effects of acetosyringone and pH on Agrobacteriummediated transformation vary according to plant species. — Plant Cell Rep. 9: 671–675, 1991.

    Article  CAS  Google Scholar 

  • Han, K.H., Meilan, R., Ma, C., Strauss, S.H.: An Agrobacterium tumefaciens transformation protocol effective on a variety of cottonwood hybrids (genus Populus). — Plant Cell Rep. 19: 315–320, 2000.

    Article  CAS  Google Scholar 

  • Hiei, Y., Komari, T., Kubo, T.: Transformation of rice mediated by Agrobacterium tumefaciens. — Plant mol. Biol. 35: 205–218, 1997.

    Article  CAS  PubMed  Google Scholar 

  • Howe, A., Sato, S., Dweikat, I., Fromm, M., Clemente, T.: Rapid and reproducible Agrobacterium-mediated transformation of Sorghum. — Plant Cell Rep. 25: 784–791, 2006.

    Article  CAS  PubMed  Google Scholar 

  • Kim, Y.S., Lim, S., Choi, Y.E., Anbazhagan, V.R.: High frequency plant regeneration via somatic embryogenesis in Podophyllum peltatum L., an important medicinal plant for source of anticancer drug. — Curr. Sci. 92: 662–666, 2007.

    CAS  Google Scholar 

  • Li, X.Q., Liu, C.N., Ritchie, S.W., Peng, J., Gelvin, S.B., Hodges, T.K.: Factors influencing Agrobacterium-mediated transient expression of gusA in rice. — Plant mol. Biol. 20: 1037–1048, 1992.

    Article  CAS  PubMed  Google Scholar 

  • Lichtenstein, C., Draper, J.: Genetic engineering of plants. — In: Glover, D.M. (ed.): DNA Cloning. Pp. 67–119. IRL Press, Oxford 1985.

    Google Scholar 

  • Manickavasagam, M., Ganapathi, A., Anbazhagan, V.R., Sudhakar, B., Selvaraj, N., Vasudevan, A., Kasthurirengan, S.: Agrobacterium-mediated genetic transformation and development of herbicide-resistant sugarcane (Saccharam species hybrids) using axillary buds. — Plant Cell Rep. 23: 134–143, 2004.

    Article  CAS  PubMed  Google Scholar 

  • May, G.D., Afza, R., Mason, H.A., Wiecko, A., Novak, F.J., Arntzen, C.J.: Generations of transgenic banana (Musa acuminata) plants via Agrobacterium-mediated transformation. — Biotechnology 13: 486–492, 1995.

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Nadeem, M., Palni, L.M.S., Purohit, A.N., Pandey, H., Nandi, S.K.: Propagation and conservation of Podophyllum hexandrum Royle: an important medicinal herb. — Biol. Conserv. 92: 121–129, 2000.

    Article  Google Scholar 

  • Saini, R., Jaiwal, P.K.: Efficient transformation of a recalcitrant grain legumes Vigna mungo L. Hepper via Agrobacteriummediated gene transfer into shoot apical meristem cultures. — Plant Cell Rep. 24: 164–171, 2005.

    Article  CAS  PubMed  Google Scholar 

  • Saini, R., Jaiwal, P.K.: Agrobacterium tumefaciens-mediated transformation of blackgram: an assessment of factors influencing the efficiency of uidA gene transfer. — Biol. Plant. 51: 69–74, 2007.

    Article  CAS  Google Scholar 

  • Schroeder, H.E., Schotz, A.H., Wardley-Richardson, T., Spencer, D., Higgins, T.J.V. Transformation and regeneration of two pea (Pisum sativum) cultivars. — Plant Physiol. 101: 751–757, 1993.

    Article  CAS  PubMed  Google Scholar 

  • Stachel, S.E., Messens, E., Montagu, M.V., Zambryski, P.: Identification of the signal molecules produced by wounded plant cells that activate T-DNA transfer in Agrobacterium tumefaciens. — Nature 318: 624–629, 1985.

    Article  Google Scholar 

  • Tiwari, R.K., Trivedi, M., Guang, Z.-C., Guo, G.-O., Zheng, G.- C.: Agrobacterium rhizogenes mediated transformation of Scutellaria baicalensis and production of flavonoids in hairy roots. — Biol. Plant. 52: 26–35, 2008.

    Article  CAS  Google Scholar 

  • Van Wordragen, M.F., Dons, H.J.M.: Agrobacterium tumefaciens mediated transformation of recalcitrant crops. — Plant mol. Biol. Rep. 10: 12–36, 1992.

    Article  Google Scholar 

  • Xiao, K., Zhang, C., Harrison, M., Wang, Z.Y.: Isolation and characterization of a novel plant promoter that directs strong constitutive expression of transgenes in plants. — Mol. Breed. 15: 221–231, 2005.

    Article  CAS  Google Scholar 

  • Yu, T.A., Yeh, S.D., Yang, J.S.: Comparison of the effects of kanamycin and geneticin on regeneration of papaya from root tissue. — Plant Cell Tissue Organ Cult. 74: 169–178, 2003.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Division of Forest Resources, College of Forest Sciences, Kangwon National University, Chunchon, 200-701, Kangwon-do, South Korea

    V. R. Anbazhagan & Y. -E. Choi

  2. Research and Education Center for Genetic Information, Nara Institute of Science and Technology, Nara, 630-0192, Japan

    Y. -S. Kim

Authors
  1. V. R. Anbazhagan
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  2. Y. -S. Kim
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  3. Y. -E. Choi
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Correspondence to Y. -E. Choi.

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Acknowledgements: This work was supported by 2007 Research Grant from Kangwon National University, and from the WCU project of the Ministry of Education, Science & Technology (MEST), Republic of Korea.

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Anbazhagan, V.R., Kim, Y.S. & Choi, Y.E. Production of transgenic Podophyllum peltatum via Agrobacterium tumefaciens-mediated transformation. Biol Plant 53, 637–642 (2009). https://doi.org/10.1007/s10535-009-0116-x

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  • DOI: https://doi.org/10.1007/s10535-009-0116-x

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