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Biomedical subjects

K M Gartland

Publications and source records attributed to K M Gartland.

7 recordsLinked to original sources

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.

Agrobacterium tumefaciens↗

Analysis of genetically modified plant gene expression using GUS fluorimetry.

A fluorimetric assay method for the analysis of beta-glucuronidase (GUS) reporter gene expression in genetically modified plants is described. Optimization of this method for woody plants and a statistical approach suitable for comparisons of gene expression in different transformants or tissues of the same plant is described. Example data from elm (Ulmus procera) SR4 regenerant plants, shown to be genetically modified by PCR and DNA-DNA hybridizations, in which higher GUS expression levels are found in stems than in leaves demonstrates the utility of this approach.

DNA, Plant↗

Transformation of the genomic expression of plant cells.

Agrobacterium-induced transformation of plant cells results from integration of T-DNA of the Ti or Ri plasmids into the genome of susceptible plants. Expression of T-DNA genes induces physiological changes in transformed cells which modify normal plant development to produce proliferations characteristic of crown gall and hairy root diseases. Understanding of the molecular basis of the transformation events associated with these examples of naturally occurring genetic engineering of plant cells, has stimulated efforts to construct vectors for transferring specific genes into plants. Vector construction has progressed from the use of wild-type Ti plasmids, giving phenotypically abnormal regenerated plants, to non-oncogenic plasmids. The range of vectors now available should enable useful foreign genes to be inserted into a range of dicotyledons and monocotyledons without impairing normal plant development.

Arginine↗