Summary
A leaf-disk transformation experiment was performed with tobacco (Nicotiana tabacum L.) using a binary vector and a strain of Agrobacterium tumefaciens that carried a wild-type Ti-plasmid, pTiBo542. Although the majority of kanamycin-resistant, transgenic plants was morphologically normal, one of the plants was double-flowered and had a slightly wavy stem and leaves whose edges were bent slightly upwards. The abnormal morphology was controlled by a single, dominant Mendelian gene. Young plants that carried this gene were distinguishable from normal plants at the stage of cotyledons. The homozygotes, with respect to this gene, were more seriously deformed than the heterozygotes. DNA segments derived from the binary vector and from the TL-and TR-DNA of pTiBo542 were detected in the double-flowered plant, but the T-DNA genes involved in biosynthesis of phytohormones were absent from the plant. The abnormal morphology, the resistance to kanamycin, and the segments of foreign DNA were genetically linked, and the linkage was very tight, at least between the abnormal morphology and the resistance to kanamycin; the meiotic recombination frequency was less than 0.02%, if recombination occurred at all.
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References
An G, Watson BD, Stachel S, Gordon MP, Nester EW (1985) New cloning vehicles for transformation of higher plants. EMBO J 4:277–284
An G, Ebert PR, Mitra A, Ha SB (1988) Binary vectors. In: Gelvin SB, Schilperoort RA (eds) Plant molecular biology manual A3. Kluwer Academic, Dordrecht, pp 1–19
Bevan MW, Chilton M-D (1982) T-DNA of the Agrobacterium Ti-and Ri-plasmids. Annu Rev Genet 16:357–384
Chilton M-D, Currier TC, Farrand SK, Bendich AJ, Gordon MP, Nester EW (1974) Agrobacterium tumefaciens DNA and PS8 bacteriophage DNA not detected in crown gall tumors. Proc Natl Acad Sci USA 71:3672–3676
Depicker A, Van Montagu M, Schell J (1983) Plant cell transformation by Agrobacterium plasmids. In: Kosuge T, Meridith CP, Hollaender A (eds) Genetic engineering of plants: an agricultural perspective. Plenum Publishing, New York, pp 143–176
Fraley RT, Rogers SG, Horsh RB (1986) Genetic transformation in higher plants. CRC Crit Rev Plant Sci 4:1–46
Hitier H (1950) Étude génétique des caractères “énation” et “catacorolle” chez Nicotiana tabacum L. Ann Tabac Sect II 1:1–31
Hood EE, Jen G, Kayes L, Kramer J, Fraley RT, Chilton M-D (1984) Restriction endonuclease map of pTiBo542, a potential Ti-plasmid vector for genetic engineering of plants. Bio/ Technol 2:702–709
Hood EE, Chilton WS, Chilton M-D, Fraley RT (1986a) TDNA and opine synthetic loci in tumors incited by Agrobacterium tumefaciens A281 on soybean and alfalfa plants. J Bacteriol 168:1283–1290
Hood EE, Helmer GL, Fraley RT, Chilton M-D (1986b) The hypervirulence of Agrobacterium tumefaciens A281 is encoded in a region of pTiBo542 outside of T-DNA. J Bacteriol 168:1291–1301
Jin S, Komari T, Gordon MP, Nester EW (1987) Genes responsible for the supervirulence phenotype of Agrobacterium tumefaciens A281. J Bacteriol 169:4417–4425
Komari T, Halperin W, Nester EW (1986) Physical and functional map of supervirulent Agrobacterium tumefaciens tumorinducing plasmid pTiBo542. J Bacteriol 166:88–94
Komari T, Saito Y, Nakakido F, Kumashiro T (1989) Efficient selection of somatic hybrids in Nicotiana tabacum L. using a combination of drug resistance markers introduced by transformation. Theor Appl Genet 77:547–552
Linsmaier EM, Skoog F (1965) Organic growth factor requirements of tobacco tissue cultures. Physiol Plant 18:100–127
Nester EW, Gordon MP, Amasino RM, Yanofsky MF (1984) Crown gall: a molecular and physiological analysis. Annu Rev Plant Physiol 35:387–413
Schmulling T, Schell J, Spena A (1988) Single genes from Agrobacterium rhizogenes influence plant development. EMBO J 7:2621–2629
Strabala TJ, Bednarek SY, Bertoni G, Amasino RM (1989) Isolation and characterization of an ipt gene from the Ti-plasmid Bo542. Mol Gen Genet 216:388–394
Watson B, Currier TC, Gordon MP, Chilton M-D, Nester EW (1975) Plasmid required for virulence of Agrobacterium tumefaciens. J Bacteriol 123:255–264
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Communicated by K. Tsunewaki
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Komari, T. Genetic characterization of a double-flowered tobacco plant obtained in a transformation experiment. Theoret. Appl. Genetics 80, 167–171 (1990). https://doi.org/10.1007/BF00224381
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DOI: https://doi.org/10.1007/BF00224381