Biologia plantarum 2010, 54:344-348 | DOI: 10.1007/s10535-010-0060-9

Efficient production of transgenic tomatoes via Agrobacterium-mediated transformation

P. Kaur1, K. C. Bansal1,*
1 National Research Centre on Plant Biotechnology, Indian Agricultural Research Institute, New Delhi, India

Cotyledonary leaves of 9-d-old tomato (Lycopersicon esculentum Mill.) were co-cultivated with Agrobacterium tumefaciens GV 3101 harboring binary vector pBI101 containing kanamycin resistance gene (npt II) as selection marker. Murashige and Skoog (MS) inorganic salts with Gamborg's B5 vitamins supplemented with optimized concentrations of zeatin riboside and indole-acetic acid resulted in enhanced regeneration efficiency. Under optimized conditions of plant regeneration, transformation frequency in cvs. Pusa Ruby, Pusa Uphar and DT-39 was greater than 37 %. Transformed shoots were selected on kanamycin medium and the presence of the transgene in the primary transformants was confirmed by PCR. Integration of the npt II gene in the tomato genome was further confirmed by Southern blot analysis. RT-PCR analysis using neomycin phospho-transferase (npt II) gene-specific primers confirmed the expression of the transgene in transgenic plants. Transformed plants were successfully transferred to phytotron, where these plants grew to maturity and produced flowers and fruits.

Keywords: cotyledonary leaf explant; genetic transformation; Lycopersicon esculentum; zeatin riboside
Subjects: Agrobacterium tumefaciens; auxins; cytokinins; in vitro culture, direct organogenesis; Lycopersicon esculentum; polymerase chain reaction (PCR); Southern blot analysis; tomato; transgenic plants

Received: June 17, 2008; Accepted: January 10, 2009; Published: June 1, 2010Show citation

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Kaur, P., & Bansal, K.C. (2010). Efficient production of transgenic tomatoes via Agrobacterium-mediated transformation. Biologia plantarum, 54(2), 344-348. doi: 10.1007/s10535-010-0060-9.
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    References

    1. Agharbaoui, Z., Greer, A.F., Tabaeizadeh, Z.: Transformation of the wild tomato Lycopersicon chilense Dun. by Agrobacterium tumefaciens. - Plant Cell Rep. 15: 102-105, 1995. Go to original source...
    2. Bříza, J., Pavingerová, D., Přikrylová, P., Gazdová, J., Vlasák, J., Niedermeierová, H.: Use of phosphomannose isomerasebased selection system for Agrobacterium-mediated transformation of tomato and potato. - Biol. Plant. 52: 453-461, 2008.
    3. Chandel, G., Katiyar, S.K.: Organogenesis and somatic embryogenesis in tomato (Lycopersicon esculentum Mill.). - Adv. Plant Sci. 13: 11-17, 2000.
    4. Cho, M.A., Moon, C.Y., Liu, J.R., Choi, P.S.: Agrobacterium-mediated transformation in Citrullus lanatus. - Biol. Plant. 52: 365-369, 2008. Go to original source...
    5. Cortina, C., Culianez-Macia, F.A.: Tomato transformation and transgenic plant production. - Plant Cell Tissue Organ Cult. 76: 269-275, 2004. Go to original source...
    6. Ellul, P., Garcia-Sogo, B., Pineda, B., Rios, G., Roig, L.A., Moreno, V.: The ploidy level of transgenic plants in Agrobacterium-mediated transformation of tomato cotyledons (Lycopersicon esculentum L. Mill.) is genotype and procedure dependent. - Theor. appl. Genet. 106: 231-238, 2003. Go to original source...
    7. Frary, A., Earle, E.D.: An examination of factors affecting the efficiency of Agrobacterium-mediated transformation of tomato. - Plant Cell Rep. 16: 235-240, 1996. Go to original source...
    8. Gamborg, O.L., Miller, R.A., Ojima, K.: Nutrient requirements of suspension cultures of soybean root cells. - Exp. Cell Res. 50: 151-158, 1968. Go to original source...
    9. Gunay, A.L., Rao, P.S.: In vitro propagation of hybrid tomato plants (Lycopersicon esculentum L.) using hypocotyl and cotyledon explants. - Ann. Bot. 45: 205-207, 1980.
    10. Hamza, S., Chupeau, Y.: Re-evaluation of conditions for plant regeneration and Agrobacterium-mediated transformation from tomato (Lycopersicon esculentum). - J. exp. Bot. 44: 1837-1845, 1993.
    11. Hu, W., Phillips, G.C.: A combination of over growth-control and antibiotics improves Agrobacterium tumefaciens-mediated transformation efficiency for cultivated tomato (L. esculentum). - In Vitro cell. dev. Biol. Plant 37: 12-18, 2001. Go to original source...
    12. Katia, H., Lipp, J., Brown, T.A.: Enhanced transformation of tomato co-cultivated with Agrobacterium tumefaciens C58C1Rifr::pGSFR1161 in the presence of acetosyringone. - Plant Cell Rep. 12: 422-425, 1993.
    13. McCormick, S., Niedermeyer, J., Fry, J., Barnason, A., Horsch, R., Fraley, R.: Leaf disc transformation of cultivated tomato (L. esculentum) using Agrobacterium tumefaciens. - Plant Cell Rep. 5: 81-84, 1986. Go to original source...
    14. Murashige, T., Skoog, F.: A revised medium for rapid growth and bioassays with tobacco tissue cultures. - Physiol. Plant. 15: 473-497, 1962. Go to original source...
    15. Park, J., Yi, B., Lee, C.: Effects of plant growth regulators, bud length, donor plant age, low temperature treatment and glucose concentration on callus induction and plant regeneration in anther culture of cherry tomato 'Mini-Carol'. - J. korean Soc. hort. Sci. 42: 32-37, 2001.
    16. Park, S.H., Morris, Jay. L., Park, J., Hirschi, K.D., Smith, R.H.: Efficient and genotype-independent Agrobacterium-mediated tomato transformation. - J. Plant Physiol. 160:1253-1257, 2003. Go to original source...
    17. Raj, S.K., Singh, R., Pandey, S.K., Singh, B.P.: Agrobacterium-mediated tomato transformation and regeneration of transgenic lines expressing tomato leaf curl virus coat protein gene for resistance against TLCV infection. - Curr. Sci. 88: 1674-1679, 2005.
    18. Sambrook, J., Fristsch, E.F., Maniatis, T.: Molecular Cloning: A Laboratory Manual. 2nd Ed. - Cold Spring Harbor Laboratory Press, Cold Spring Harbor - New York 1989.
    19. Van Roekel, J.S.C., Damm, B., Melchers, L.S., Hoekema, A.: Factors influencing transformation frequency of tomato (Lycopersicon esculentum). - Plant Cell Rep. 12: 644-647, 1993. Go to original source...

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