Abstract
The use of two Agrobacterium tumefaciens strains for transformation of Triticum aestivum L. cv. Vesna was studied. Immature embryos, isolated 15 d after pollination, were co-cultivated with the super-binary LBA4404/pTOK233 and the binary AGL1/pDM805 vectors. While the transient GUS-intron expression was high (69.9 and 80.0 %), the number of plants regenerated on selective media containing hygromycin or phosphinotricin did not exceed 0.4 and 0.13 %, respectively. Nevertheless, the regenerated plants were fertile and produced seeds. The T0 plants, as well as the T1 seedlings, displayed the activity in the β-glucuronidase histochemical assay and a positive signal in PCR analysis for the presence of uidA gene sequences in their genomes. The data suggest that the transformation of wheat cv. Vesna with both Agrobacterium strains is feasible.
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Banerjee, A.K., Agrawal, D.C., Nalawade, S.M., Krishnamurthy, K.V.: Transient expression of β-glucuronidase in embryo axes of cotton by Agrobacterium and particle bombardment methods.-Biol. Plant. 45: 359-365, 2002.
Chan, M.T., Chang, H.H., Ho, S.L., Tong, W.F., Yu, S.M.: Agrobacterium-mediated production of transgenic rice plants expressing chimeric α-amylase promotor/β-glucuronidase gene.-Plant mol. Biol. 22: 491-406, 1993.
Cheng, M., Fry, J.E., Pang, S., Zhou, H., Hironaka, C.M., Duncan, D.R., Conner, T.W., Wan, Y.: Genetic transformation of wheat mediated by Agrobacterium tumefaciens.-Plant Physiol. 115: 971-980, 1997.
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.
Enríquez-Obregón, G.A., Vázquez-Padrón, R.I., Prieto-Samsónov, D.L., De la Riva, G.A., Selman-Housein, G.: Herbicide-resistant sugarcane (Saccharum officinarum L.) plants by Agrobacterium-mediated transformation.-Planta 206: 20-27, 1998.
Garfinkel, M., Nester, E.W.: Agrobacterium tumefaciens mutants affected in crown gall tumourigenesis and octopine catabolism.-J. Bacteriol. 144: 732-743, 1980.
Hiei, Y., Ohta, S., Komari, T., Kumashiro, T.: Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA.-Plant J. 6: 271-282, 1994.
Ishida, Y., Saito, H., Ohta, S., Hiei, Y., Komari, T., Kumashiro, T.: High efficiency transformation of maize (Zea mays L.) mediated by Agrobacterium tumefaciens.-Natur. Biotechnol. 14: 745-750, 1996.
Jefferson, R.A., Kavanagh, T.A., Bevan, M.W.: GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants.-EMBO J. 6: 3901-3907, 1987.
Khanna, H.K., Daggard, G.E.: Agrobacterium tumefaciens-mediated transformation of wheat using a superbinary vector and a polyamine-supplemented regeneration medium.-Plant Cell Rep. 21: 429-436, 2003.
McCormac, A.C., Wu, H., Bao, M., Wang, Y., Xu, R., Elliott, M.C., Chen, D.F.: The use of visual marker genes as cell-specific reporters of Agrobacterium-mediated T-DNA delivery to wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.).-Euphytica 99: 17-25, 1998.
Mitić, N., Nikolić, R., Nešković, M.: Somatic embryogenesis and plant regeneration from immature embryos of five wheat cultivars and their reciprocal hybrids.-Arch. biol. Sci. Belgrade 51: 99-104, 1999.
Murashige, T., Skoog, F.: A revised medium for rapid growth and bioassays with tobacco tissue cultures.-Physiol. Plant. 15: 473-497, 1962.
Tingay, S., McElroy, D., Kalla, R., Fieg, S., Wang, M., Thornton, S., Brettell, R.: Agrobacterium tumefaciens-mediated barley transformation.-Plant J. 11: 1369-1376, 1997.
Vasil, I.K.: Molecular improvement of cereals.-Plant mol. Biol. 25: 925-937, 1994.
Vasil, V., Castillo, A.M., Fromm, M.E., Vasil, I.K.: Herbicide resistant fertile transgenic wheat plants obtained by microprojectile bombardment of regenerable embryogenic callus.-Bio/Technology 10: 667-674, 1992.
Wu, H., McCormac, A.C., Elliott, M.C., Chen, D.F.: Agrobacterium-mediated stable transformation of cell suspension cultures of barley (Hordeum vulgare L.).-Plant Cell Tissue Organ Cult. 54: 161-171, 1998.
Wu, H., Sparks, C., Amoah, A., Jones, H.D.: Factors influencing successful Agrobacterium-mediated genetic transformation of wheat.-Plant Cell Rep. 21: 659-668, 2003.
Xia, G., Li, Z., He, C., Chen, H., Brettell, R.: Transgenic plant regeneration from wheat (Triticum aestivum L.) mediated by Agrobacterium tumefaciens.-Acta phytophysiol. sin. 25: 22-28, 1999.
Xiaomei, Z., Guangcheng, C., Rufa, L., Yongru, S., Wenbin, L.A.: A rapid and efficient DNA extraction method of genus Fagopyrum for RAPD analysis.-In: Javornik, B., Bohanec, B., Kreft, I. (ed.): Proceedings of the International Colloquium on Impact of Plant Biotechnology on Agriculture. Vol. 5–7. Pp. 171-175. Univ. Ljubljana, Ljubljana 1994.
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Mitić, N., Nikolić, R., Ninković, S. et al. Agrobacterium-Mediated Transformation and Plant Regeneration of Triticum aestivum L.. Biologia Plantarum 48, 179–184 (2004). https://doi.org/10.1023/B:BIOP.0000033442.15611.7d
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DOI: https://doi.org/10.1023/B:BIOP.0000033442.15611.7d