Abstract
A method for Agrobacterium-mediated transformation of hybrid poplar (Populus alba x P. grandidentata cv. ‘Crandon’) suspension cultures and regeneration of transformed plants is described. Transformants were recovered when suspension cultures were inoculated with Agrobacterium tumefaciens at a density of 107 colony-forming units ml-1, cocultivated for 48 h, and plated to cellulose acetate filters on Woody Plant Medium containing 4.5 μM 2,4-dichlorophenoxyacetic acid and 250 mg l-1 cefotaxime. Levels of cefotaxime greater than 250 mg l-1 were unnecessary for control of residual bacteria and inhibited callus growth. Transgenic plants were regenerated by culturing the transformed callus on media containing 0.11 to 27 μM thidiazuron. In contrast to thidiazuron, N6-benzyladenine had a negative effect on shoot regeneration; the callus became necrotic when we attempted to induce shoots with concentrations of 1.1 to 8.9 μM, and growth was inhibited when concentrations of 0.11 or 0.22 μM were used to regenerate callus from suspension cultures. Following cocultivation of poplar suspension cultures, we recovered transgenic plants containing the maize transposon Ac, and callus containing an insect toxin gene from Bacillus thuringiensis.
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Abbreviations
- BA:
-
N6-benzyladenine
- CIM:
-
callus initiation medium
- CaMV:
-
cauliflower mosaic virus
- cfu's:
-
colony-forming units
- HPT:
-
hygromycin phosphotransferase
- MS:
-
Murashige and Skoog medium (Murashige & Skoog 1962)
- NPT-II:
-
neomycin phosphotransferase-II
- PAR:
-
photosynthetically active radiation
- PCR:
-
polymerase-chain-reaction
- TDZ:
-
thidiazuron
- WPM:
-
Woody Plant Medium (Lloyd & McCown 1980)
- 2,4-d :
-
2,4-dichlorophenoxyacetic acid
References
An G (1985) High efficiency transformation of cultured tobacco cells. Plant Physiol. 79: 568–570
An G (1986) Development of plant promoter expression vectors and their use for analysis of differential activity of nopaline synthase promoter in transformed tobacco cells. Plant Physiol. 81: 86–91
An G, Ebert PR, Mitra A & Ha SB (1988) Binary vectors. In: Gelvin SB & Schilperoort RA (Eds) Plant Molecular Biology Manual (A3: pp 1–19). Kluwer Academic Publishers, Dordrecht
Baker B, Coupland G, Fedoroff N, Starlinger P & Schell J (1987) Phenotypic assay for excision of the maize controlling element Ac in tobacco. EMBO J. 6: 1547–1554
Barton KA, Whiteley HR & Yang N-S (1987) Bacillus thuringiensis ö-endotoxin expressed in transgenic Nicotiana tabacum provides resistance to lepidopteran insects. Plant Physiol. 85: 1103–1109
Coleman GD & Ernst SG (1989) In vitro shoot regeneration of Populus deltoides: effect of cytokinin and genotype. Plant Cell Rep. 8: 459–462
De Block M (1990) Factors influencing the tissue culture and the Agrobacterium tumefaciens-mediated transformation of hybrid aspen and poplar clones. Plant Physiol. 93: 1110–1116
Dekeyser R, Claes B, Marichal M, Van Montagu M & Caplan A (1989) Evaluation of selectable markers for rice transformation. Plant Physiol. 90: 217–223
Ellis DD, Lazaroff WR, Roberts DR, Flinn BS & Webb DT (1989) The effect of antibiotics on elongation and callus and bud formation from embryonic tissue of Picea glauca. Can. J. For. Res. 19: 1343–1346
Fillatti JJ, Sellmer J, McCown B, Haissig B & Comai L (1987) Agrobacterium mediated transformation and regeneration of Populus. Mol. Gen. Genet. 206: 192–199
Gheysen G, Van Montagu M & Zambryski P (1987) Integration of Agrobacterium tumefaciens transfer DNA (T-DNA) involves rearrangements of target plant DNA sequences. Proc. Natl. Acad. Sci. U.S.A. 84: 6169–6173
Hepburn AG, White J, Pearson L, Maunders MJ, Clarke LE, Prescott AG & Blundy KS (1985) The use of pNJ5000 as an intermediate vector for the genetic manipulation of Agrobacterium Ti-plasmids. J. Gen. Microbiol. 131: 2961–2969
Hood EE, Helmer GL, Fraley RT & Chilton M-D (1986) The hypervirulence of Agrobacterium tumefaciens A281 is encoded in a region of pTiBo542 outside of T-DNA. J. Bacteriol. 168: 1291–1301
Hooykaas PJJ (1988) Agrobacterium molecular genetics. In: Gelvin SB & Schilperoort RA (Eds) Plant Molecular Biology Manual (A4: pp 1–13). Kluwer Academic Publishers, Dordrecht
Howe GT (1991) Agrobacterium-mediated transformation of hybrid poplar (Populus alba x P. grandidentata) and analysis of foreign gene expression. Ph.D. dissertation, Oregon State Univ., Corvallis, OR, USA
Jorgensen R, Snyder C & Jones JDG (1987) T-DNA is organized predominantly in inverted repeat structures in plants transformed with Agrobacterium tumefaciens C58 derivatives. Mol. & Gen. Genet. 207: 471–477
Lloyd G & McCown B (1980) Commercially-feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot-tip culture. Comb. Proc. Int. Plant Prop. Soc. 30: 421–427
McCown BH, McCabe DE, Russell DR, Robison DJ, Barton KA & Raffa KF (1991) Stable transformation of Populus and incorporation of pest resistance by electric discharge particle acceleration. Plant Cell Rep. 9: 590–594
Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497
Okada K, Takebe I & Nagata T (1986) Expression and integration of genes introduced into highly synchronized plant protoplasts. Mol. Gen. Genet. 205: 398–403
Park YG & Son SH (1988) In vitro organogenesis and somatic embryogenesis from punctured leaf of Populus nigra x P. maximowiczii. Plant Cell Tiss. Org. Cult. 15: 95–105
Peerbolte R, Leenhouts K, Hooykaas-van Slogteren GMS, Hoge JHC, Wullems GJ, & Schilperoort RA (1986) Clones from a shooty tobacco crown gall tumor I: deletions, rearrangements and amplifications resulting in irregular T-DNA structures and organizations. Plant Mol. Biol. 7: 265–284
Pollock K, Barfield DG & Shields R (1983) The toxicity of antibiotics to plant cell cultures. Plant Cell Rep. 2: 36–39
Russell JA & McCown BH (1988) Recovery of plants from leaf protoplasts of hybrid-poplar and aspen clones. Plant Cell Rep. 7: 59–62
Sellmer JC & McCown BH (1989) Transformation in Populus spp. In: Bajaj YPS (Ed) Biotechnology in Agriculture and Forestry, Plant Protoplasts and Genetic Engineering II, Vol 9 (pp 155–172) Springer Verlag, Berlin/Heidelberg/New York
Sheikholeslam SN & Weeks DP (1987) Acetosyringone promotes high efficiency transformation of Arabidopsis thaliana explants by Agrobacterium tumefaciens. Plant Mol. Biol. 8: 291–298
Strauss SH, Howe GT & Goldfarb B (1991) Prospects for genetic engineering of insect resistance in forest trees. For. Ecol. Mgt. 43: 181–209
Tsang EWT, David H, David A & Dunstan DI (1989) Toxicity of antibiotics on zygotic embryos of white spruce (Picea glauca) cultured in vitro. Plant Cell Rep. 8: 214–216
van den Elzen PJM, Townsend J, Lee KY & Bedbrook JR (1985) A chimaeric hygromycin resistance gene as a selectable marker in plant cells. Plant Mol. Biol. 5: 299–302
Wagner DB, Furnier GR, Saghai-Maroof MA, Williams SM, Dancik BP & Allard RW (1987) Chloroplast DNA polymorphisms in lodgepole and jack pines and their hybrids. Proc. Natl. Acad. Sci. USA 84: 2097–2100
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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Howe, G.T., Goldfarb, B. & Strauss, S.H. Agrobacterium-mediated transformation of hybrid poplar suspension cultures and regeneration of transformed plants. Plant Cell Tiss Organ Cult 36, 59–71 (1994). https://doi.org/10.1007/BF00048316
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DOI: https://doi.org/10.1007/BF00048316