Agrobacterium-mediated transformation of polyploid cereals. The efficiency of selection and transgene expression in wheat

• Autorzy: Przetakiewicz A , Karas A. , Orczyk W. , Nadolska-Orczyk A.
• Języki publikacji: EN

Abstrakty

EN

Three combinations of Agrobacterium tumefaciens strains and vectors were used in the transformation of selected Polish wheat cultivars. The combinations were: two hypervirulent strains, AGL1, containing the pDM805 binary plasmid, and EHA101, containing pGAH; and the common Agro strain LBA4404, harboring the super-binary pTOK233 vector. pDM805 contained bar under the control of Ubi1 promoter, pGAH had nptII under nos, and pTOK233 had hpt under 35S. Additionally, pDM805 and pTOK233 carried the gus reporter gene under the Act1 promoter or 35S promoter, respectively. The highest selection rate was 12.6% and was obtained with EHA101(pGAH) on a kanamycin-containing medium. Sixty-five of the plants grown on that medium were PCR positive. The second best combination was LBA4404(pTOK233) and kanamycin selection, which gave an average transformation rate of 2.3%. Phosphinothricin selection gave 1.0% transformation efficiency, while hygromycin, depending on the strain/vector used, gave from 0.2 to 0.4%. PCR tests in T1 revealed that 67% of the lines showed a 3:1 segregation ratio, and 11% a 15:1 ratio, while in 22%, segregation was non-Mendelian. The high number of T0 transgenic plants containing one copy of the transgene was confirmed via Southern blot analysis. Kanamycin resistance in the T1 generation was very low; in some lines, all the progeny were kanamycin sensitive. GUS expression, only tested in young T1 plants, was in agreement with Mendelian segregation in three out of the twelve tested. The factors influencing the efficiency of selection and transgene expression are discussed in this paper.

• Wydawca: -
• Czasopismo: Cellular and Molecular Biology Letters
• Rocznik: 2004
• Tom: 09
• Numer: 4B
• Opis fizyczny: p.903-917,fig.,ref.

Twórcy

autor

Przetakiewicz A

• Plant Breeding and Acclimatization Institute, Radzikow, 05-870 Blonie, Poland

autor

Karas A.

autor

Orczyk W.

autor

Nadolska-Orczyk A.

Bibliografia

• 1. Hiei, Y., Komari, T. and Kubo, T. Transformation of rice mediated by Agrobacterium tumefaciens. Plant Mol. Biol. 35 (1997) 205-218.
• 2. Nadolska-Orczyk, A. and Orczyk, W. Agrobacterium-mediated transformation of cereals. in: Plant Genetic Engineering. Vol 2. Improvement of Food Crops. (Jaiwal, P.K. and Singh, R.P. Eds.), Sci Tech Publishing LLC USA, 2003, 1-25.
• 3. De Framond, A.J., Back, E.W., Chilton, W.S., Kayes, L. and Chilton, M. Two unlinked T-DNAs can transform the same tobacco plant cell and segregate in the F1 generation. Mol. Gen. Genet. 202 (1986) 125-131.
• 4. Matthews, P.R., Wang, M.-B., Waterhouse, P.M., Thornton, S., Fieg, S.J., Gubler, F. and Jacobsen, J.V. Marker gene elimination from transgenic barley, using co-transformation with adjacent 'twin T-DNAs' on a standard Agrobacterium transformation vector. Mol. Breed. 7 (2001) 195-202.
• 5. Cheng, M., Fry, J.E., Pang, S., Zhou, H., Hironaka, C.M., Duncan, D.R. Conner, T.W. and Wan, Y. Genetic transformation of wheat mediated by Agrobacterium tumefaciens. Plant Physiol. 115 (1997) 971-980.
• 6. Hu, T., Metz, S., Chay, C., Zhou, H.P., Biest, N., Chen, G., Cheng, M., Feng, X., Radionenko, M., Lu, F. and Fry, J. Agrobacterium-mediated large-scale transformation of wheat (Triticum aestivum L.) using glyphosate selection. Plant Cell Rep. 21 (2003) 1010-1019.
• 7. Khanna, H.K. and Daggard, G.E. Agrobacterium tumefaciens-mediated transformation of wheat using a superbinary vector and a polyamine-supplemented regeneration medium. Plant Cell Rep. 21 (2003) 429-436.
• 8. Wu, H., Sparks, C., Amoah, B. and Jones, H.D. Factors influencing successful Agrobacterium-mediated genetic transformation of wheat. Plant Cell Rep. 21 (2003) 659-668.
• 9. Przetakiewicz, A., Orczyk, W. and Nadolska-Orczyk, A. The effect of auxin on plant regeneration of wheat, barley and triticale. Plant Cell, Tissue Org. Cult. 73 (2003) 245-256.
• 10. Murashige, T. and Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15 (1962) 473-497.
• 11. Gamborg, O.L., Miller, R.A. and Ojima, K. Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell. Res. 50 (1968) 151-158.
• 12. Hiei, Y., Ohta, S., Komari, T. and 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 (1994) 271-282.
• 13. Tingay, S., McElroy, D., Kalla, R., Fieg, S., Wang, M., Thornton, S. and Brettell, R. Agrobacterium tumefaciens-mediated barley transformation. Plant J. 11 (1997) 1369-1376.
• 14. Hayashi, H., Alia, Mustardy, L., Deshnium, P., Ida, M. and Murata, N. Transformation of Arabidopsis thaliana with the codA gene for choline oxidase; accumulation of glycinebetaine and enhanced tolerance to salt and cold stress. Plant J. 12 (1997) 133-142.
• 15. Garfinkel, M. and Nester, E.W. Agrobacterium tumefaciens mutants affected in crown gall tumourigenesis and octopine catabolism. J. Bacteriol. 144 (1980) 732-743.
• 16. Chilton, M.-D., Currier, T.C., Farrand, S.K., Bendich, A.J., Gordon, M.P. and Nester, E.W. Agrobacterium tumefaciens DNA and PS8 bacteriofage DNA not detectedin crown gall tumors. Proc. Natl. Acad. Sci. USA 71 (1974) 3672-3676.
• 17. Jefferson, R.A., Kavanagh, A. and Bevan, M.W. GUS fusions: ß-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6 (1987) 3901-3907.
• 18. Shaw, K.J., Rather, P.N., Hare R.S. and Miller G.H. Molecular genetics of aminoglycoside resistance genes and familial relationships of the aminoglycoside-modifying enzymes. Microbiol. Rev. 57 (1993) 138-163.
• 19. Hood, E.E., Gelvin, S.B., Melchers, L.S. and Hoekema, A. New Agrobacterium helper plasmids for gene transfer to plants. Transgenic Res. 2 (1993) 208-218.
• 20. Nadolska-Orczyk, A., Orczyk, W. and Przetakiewicz, A. Agrobacterium-mediated transformation of cereals - from technique development to its application. Acta Physiol. Plant. 22 (2000) 77-88.
• 21. Moore, G. Cereal chromosome structure, evolution and pairing. Annu. Rev. Plant Physiol. Plant Mol. Biol. 51 (2000) 195-222.
• 22. Patnaik, D., Khurana, P. Wheat biotechnology: A minireview. Electronic J. Biotechnol. 4 (2001) 74-102.
• 23. Przetakiewicz, A. Agrobacterium-mediated transformation of wheat and barley cultivars. Ph.D. thesis (2003) (in Polish).
• 24. Matzke, M., Aufsatz, W., Kanno, T., Daxinger, L., Papp, I., Mette, M.F. and Matzke, A.J.M. Genetic analysis of RNA-mediated transcriptional gene silencing. Biochim. Biophys. Acta 1677 (2004) 129-141.