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Stable transformation of Populus and incorporation of pest resistance by electric discharge particle acceleration

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Abstract

Three different target tissues (protoplast-derived cells, nodules, and stems) and two unrelated hybrid genotypes of Populus (P. alba x P. grandidentata ‘Crandon’ and P. nigra ‘Betulifolia’ x P. trichocarpa) have been stably transformed by electric discharge particle acceleration using a 18.7 kb plasmid containing NOS-NPT, CaMV 35S-GUS, and CaMV 35S-BT. Four transformed plants of one hybrid genotype, NC5339, containing all 3 genes were recovered and analyzed. Two expressed GUS and one was highly resistant to feeding by 2 lepidopteran pests (the forest tent caterpillar, Malacosoma disstria, and the gypsy moth, Lymantria dispar.) Pretreatment of the target tissues, fine-tuning of the bombardment parameters, and the use of a selection technique employing flooding of the target tissues were important for reliable recovery of transformed plants.

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Abbreviations

BA:

benzyladenine

NAA:

1-naphthaleneacetic acid

TDZ:

thidiazuron

DB:

2,6-dichlorobenzonitrile

WPM:

woody plant medium

Kan:

kanamycin

Carb:

carbenicillin

Ben:

benylate

gus:

β-glucuronidase (EC 3.2.1.31)

NPT:

neomycin phosphoransferase (EC 2.7.1.95) gene

GUS:

betaglucuronidase gene

BT:

modified endotoxin gene originally from Bacillus thuringiensis (B.t.)

PCR:

polymerase chain reaction

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Communicated by J.M. Widholm

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McCown, B.H., McCabe, D.E., Russell, D.R. et al. Stable transformation of Populus and incorporation of pest resistance by electric discharge particle acceleration. Plant Cell Reports 9, 590–594 (1991). https://doi.org/10.1007/BF00232339

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  • DOI: https://doi.org/10.1007/BF00232339

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