Efficiency of transformation of Polish cultivars of pea [Pisum sativum L.] with various regeneration capacity by using hypervirulent Agrobacterium tumefaciens strains

• Autorzy: Pniewski T , Kapusta J
• Języki publikacji: EN

Abstrakty

EN

An Agrobacterium-mediated transformation method of pea has been developed for several edible and fodder cultivars of pea (Pisum sativum L.), characterized previously in their potential for regeneration via organogenesis) The most appropriate explant, which was susceptible to Agrobacterium infection and capable of regenerating transgenic plants, turned out to be a slice of an immature embryo, including the embryo axis and the basal part of a cotyledon. Three hypervirulent strains of A. tumefaciens were tested: AgL0, AgL1 and EHA105. Each carried the binary vector pP35SGIB containing the uid gene, with an intron under control of the 35S promoter, and the bar gene conferring resistance to phosphinotricin. Strain AgL0 was found to be efficient for the majority of cultivars, followed by AgL1 and EHA105. Transformation efficiency varied from 0.7 to 4.1%, depending on cultivar and Agrobacterium strain. The transformation efficiency of particular pea cultivars did not clearly correspond to their regeneration capacity, which - although indispensable - was not a critical parameter of successful transformation. The presence of integrated genes in pea genomic DNA was detected by the PCR. T-DNA was stably transmitted to the progeny, as it was confirmed by Southern hybridization. The activity of introduced genes was analysed by the histochemical GUS assay and by painting leaves or by spraying transgenic plants with the herbicide Basta.

Słowa kluczowe

EN

transformation; Agrobacterium tumefaciens; transgenic plant; regeneration; plant genetics; Polish cultivar; organogenesis; pea; Pisum sativum

• Wydawca: -
• Czasopismo: Journal of Applied Genetics
• Rocznik: 2005
• Tom: 46
• Numer: 2
• Opis fizyczny: p.139-147,fig.,ref.

Twórcy

autor

Pniewski T

• Institute of Plant Genetics, Polish Academy of Sciences, Strzeszynska 34, 60-479 Poznan, Poland

autor

Kapusta J

• Institue of Biotechnology and Antibiotics, Warszawa, Poland

Bibliografia

• Bean SJ, Gooding PS, Mullineaux PM, Davies DR, 1997. A simple system for pea transformation. Plant Cell Rep 16: 513-519.
• Casey R, Davies DR, 1993. Peas: Genetics, Molecular Biology and Biotechnology. CAB International, Wallingford, UK.
• Chee PP, Slightom JL, 1994. Transformation of soybean (Glycine max) via Agrobacterium tumefaciens and analysis of transformed plants. In: Gartland KMA, Davey MR, eds. Methods in
• Molecular Biology. Humana Press, Totowa, NJ 44: 101-119.
• Davies DR, Hamilton J, Mullineaux PM, 1993. Transformation of peas. Plant Cell Rep 12: 180-183.
• De Kathen A, Jacobsen HJ, 1990. Agrobacterium tumefaciens-mediated transformation of Pisum sativum L. using binary and cointegrate vectors. Plant Cell Rep 9: 276-279.
• Gamborg OL, Miller RA, Ojima K, 1968. Nutrient requirements for suspension cultures of soybean root cells. Exp Cell Res 50: 151-158.
• Garfinkel DJ, Nester EW, 1980. Agrobacterium tumefaciens mutants affected in crown gall tumorigenesis and octopine catabolism. J Bacteriol 144: 732-743.
• Grant JE, Cooper PA, Mcara AE, Frew TJ, 1995. Transformation of peas (Pisum sativum L.) using immature cotyledons. Plant Cell Rep 15: 254-258.
• Grant JE, Cooper PA, Gilpin BJ, Hoglund SJ, Reader JK, Pither-Joyce MD, Timmerman- Vaughan GM, 1998. Kanamycin is effective for selecting transformed peas. Plant Sci 139:159-164.
• Griga M, Novák FJ, 1990. Pea (Pisum sativum L.). In: Bajaj YPS, ed. Biotechnology in Agriculture and Forestry 10. Legumes and Oilseed Crops I. Springer-Verlag, Berlin, II.1-65 - II- 1-94.
• Jasińska Z, Kotecki A, 1993. Pea. In: Legume crops. Wydawnictwo Naukowe PWN, Warszawa, III—61 - III—88 (in Polish).
• Jefferson RA, 1987. Assaying chimeric genes in plants: The GUS gene fusion system. Plant Mol Biol Rep 5: 387-405.
• Li H, Wylie SJ, Jones MGK, 2000. Transgenic yellow lupin (Lupinus luteus). Plant Cell Rep 19: 634—637.
• Lulsdorf MM, Rempel H, Jackson JA, Baliski DS, Hobbs SLA, 1991. Optimizing the production of transformed pea (Pisum sativum L.) callus using disarmed Agrobacterium tumefaciens strains. Plant Cell Rep 9: 479-483.
• McGarvey P, Kaper JM, 1991. A simple and rapid method for screening transgenic plants using the PCR. BioFeedback 4: 428-432.
• Murashige T, Skoog F, 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plantarum 15: 473-479.
• Nadolska-Orczyk A, Orczyk W, 2000. Study of the factors influencing Agrobacterium- mediated transformation of pea (Pisum sativum L.). Mol Breeding 6: 185-194.
• Perrin Y, Vaquera C, Gerrard I, Sack M, Drossard J, Stöger E, et al, 2000. Transgenic pea seeds as bioreactors for the production of a single-chain Fv fragment (scFV) antibody used in cancer diagnosis and therapy. Mol Breeding 6: 345-352.
• Pigeaire A, Abernethy D, Smith PM, Simpson K, Fletcher N, Lu C, et ah, 1997. Transformation of grain legume (Lupinus angustifolius L.) via Agrobacterium tumefaciens-mediated gene transfer to shoot apices. Mol Breeding 3: 341-349.
• Pniewski T, Wachowiak J, Kapusta J, Legocki AB, 2003. Organogenesis and long-term micropropagation of Polish pea cultivars. Acta Soc Bot Pol 72: 295-302.
• Polowick PL, Quandt J, Mahon JD, 2000. The ability of pea transformation technology to transfer genes into peas adapted to western Canadian growing conditions. Plant Sci 153: 161-170.
• Puonti-Kaerlas J, Eriksson T, Engström P, 1990. Production of transgenic pea (Pisum sativum L.) plants by Agrobacterium tumefaciens - mediated gene transfer. Theor Appl Genet 80: 246-252.
• Puonti-Kaerlas J, Eriksson T, Engström P, 1992. Inheritance of a bacterial hygromycin phosphotransferase gene in the progeny of primary transgenic pea plants. Theor Appl Genet 84: 443-450.
• Schroeder HE, Schotz AH, Wardley-Richardson T, Spencer D, Higgins TJV, 1993. Transformation and regeneration of two cultivars of pea (Pisum sativum L.). Plant Physiol 101: 751-757.
• Vervliet G, Holsters M, Teuchy H, van Montagu M, Schell J, 1975. Characterisation of different plaque-forming and defective temperate phages in Agrobacterium strains. J Gen Virol 26: 33-48.