Regeneration of phenotypically normal English elm (Ulmus procera) plantlets following transformation with an Agrobacterium tumefaciens binary vector.
A transformation system was developed for English elm (Ulmus procera Salisbury) using Agrobacterium tumefaciens C58 pMP90 p35SGUS/INTRON, allowing for the transfer of foreign genes and regeneration of phenotypically normal elm plantlets. The PCR analysis indicated that both nptII and uidA genes were stably inserted in the plant genome. beta-Glucuronidase histochemical and fluorimetric assays revealed expression of the uidA gene in the shoots, leaves, stems and roots of regenerated transgenic plants. The DNA-DNA hybridizations confirmed the presence of the uidA gene in regenerant plants. Factors influencing successful transformation and regeneration of elms included: identifying gene-transfer-proficient Agrobacterium strains for use with elms; developing an infection protocol allowing T-DNA transfer while retaining the ability to remove inciting bacteria; and identifying selection conditions to eliminate non-transformed material and choice of regeneration medium to allow shoot production. The potential utility of an effective elm transformation and regeneration system in the control of Dutch elm disease is discussed.
Gartland
J S
JS
Plant Biotechnology Research Group, Molecular & Life Sciences, University of Abertay Dundee, Scotland, UK.
McHugh
A T
AT
Brasier
C M
CM
Irvine
R J
RJ
Fenning
T M
TM
Gartland
K M
KM
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D015139:Blotting, Southern; D010641:Phenotype; D010935:Plant Diseases / Q000235:genetics* / Q000382:microbiology; D018520:Plant Shoots / Q000254:growth & development; D018547:Plant Stems / Q000254:growth & development; D030821:Plants, Genetically Modified / Q000235:genetics* / Q000254:growth & development / Q000382:microbiology; D016133:Polymerase Chain Reaction; D014170:Transformation, Genetic / Q000235:genetics*; D014197:Trees / Q000235:genetics* / Q000254:growth & development