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Philip J Dale

Publications and source records attributed to Philip J Dale.

5 recordsLinked to original sources
PubMed2006

Brassica oleracea.

A better understanding of the genetic basis underlying the genotype dependence of Brassica oleracea transformation is enabling researchers to distinguish between recalcitrant and successful candidate genotypes for routine transformation. In this chapter we outline an A. tumefaciens-mediated transformation method for B. oleracea using 4-d-old cotyledonary explants and a model B. oleracea doubled haploid genotype, AG DH1012. After only 3 wk on kanamycin selection, the first transgenic shoots can be isolated. Transformation efficiencies in the region of 10 to 25% (based on 50-125 PCR-positive independent shoots from 500 infected explants) are typically achieved. For researchers wishing to use their own plant genotype, we highlight the tissue culture phenotypes that are conducive to efficient transformation.

Agrobacterium tumefaciens↗
PubMed2004

Mapping of transposable element Dissociation inserts in Brassica oleracea following plant regeneration from streptomycin selection of callus.

To investigate the potential of heterologous transposons as a gene-tagging system in broccoli (Brassica oleracea var. italica), we have introduced a Dissociation ( Ds)-based two-element transposon system. Ds has been cloned into a 35S-SPT excision-marker system, with transposition being driven by an independent 35S-transposase gene construct. In three successive selfed generations of plants, there was no evidence of germinal-excision events. In a previous study, we overcame this apparent inability to produce B. oleracea plants with germinal excisions by performing a novel tissue-culture technique to select for fully green shoots from seed with somatic excision events. The results showed a very high efficiency of regeneration of fully green plants (up to 65%), and molecular analysis showed that the plants contained the equivalent of a germinal-excision event. In this study, we followed the previous work by using inverse and nested PCR to generate probes of flanking genomic DNA adjacent to independently reinserted Ds elements, and these were hybridised to DNA from a double-haploid mapping population of B. oleracea. Seventeen Ds insertions and the original Ds T-DNA site have been localised, and these are spread over six (out of nine) linkage groups. Distribution of inserts show that 15 were found on a different linkage group to the original 'launch' site, and of these 11 were found to be clustered on two separate groups. Previous studies in other plant species have found that germinal excision of Ds predominantly moves to sites linked close to the donor site. However, this study shows a potential to produce plants with Ds insertion scattered over many unlinked sites.

Blotting, Southern↗
PubMed2002

Herbicide sensitivity of transgenic multiple herbicide-tolerant oilseed rape.

Glyphosate and glufosinate-ammonium herbicide tolerance traits were combined into both winter and spring lines of Brassica napus L. This allowed the study of possible interactions between these transgenes in two genetic backgrounds when treated with a variety of herbicides. Selective herbicides that are commonly used within Brassica crops showed no adverse effects on the transgenic plants or their null controls. Lines containing both glyphosate and glufosinate transgenes remained tolerant to their respective herbicides, regardless of the presence of the second tolerance transgene. Lines containing only a single transgene retained tolerance to the encoded trait and did not show cross-tolerance to the second. Null lines were killed by either herbicide. All plant lines, regardless of their transgene content, were found to be equally susceptible to three herbicides (paraquat, metsulfuronmethyl and mecoprop), commonly used to remove volunteer B napus from succeeding crops and set-a-side land.

2-Methyl-4-chlorophenoxyacetic Acid↗
PubMed2002

Potential for the environmental impact of transgenic crops.

In recent years, there has been increasing interest in how changes in agricultural practice associated with the introduction of particular genetically modified (GM) crops might indirectly impact the environment. There is also interest in any effects that might be associated with recombinant and novel combinations of DNA passing into the environment, and the possibility that they may be taken up by microorganisms or other live biological material. From the current state of knowledge, the impact of free DNA of transgenic origin is likely to be negligible compared with the large amount of total free DNA. We can find no compelling scientific arguments to demonstrate that GM crops are innately different from non-GM crops. The kinds of potential impacts of GM crops fall into classes familiar from the cultivation of non-GM crops (e.g., invasiveness, weediness, toxicity, or biodiversity). It is likely, however, that the novelty of some of the products of GM crop improvement will present new challenges and perhaps opportunities to manage particular crops in creative ways.

Consumer Product Safety↗