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Genetic engineering of plant cells

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  1. Introduction .
  2. Agrobacterium tumefaciens- mediated transformation.
  3. Genetic engineering.
  4. Direct gene transfer into plants.
  5. Viruses as gene vectors.
  6. Conclusion.

In this text, we shall see how selection data have been used to assist the design of experiments on the modification of plants using recombinant DNA (genetic engineering). It is evident that in some situations the potential of in vitro selection is limited by a number of difficulties. It is partly because of these difficulties that genetic engineering methods are assuming an increasingly important role in the modification of plants. We now consider some of these methods.The plant pathogen Agrobacterium tumefaciens causes crown gall disease in higher plants. Infection by the bacterium commonly results in the formation of timorous outgrowths, the morphology of which depends on the plant species. The bacterium is now known to possess a large (95-160 Md) tumour-inducing (Ti) plasmid, part of which (the T-DNA) is transferred and incorporated into the host genome during infection.

[...] Protoplasts are incubated with the DNA at 22-24 C for about 30 min in the presence of PEG. Alternatively, protoplasts are subjected to electroporation in the presence of the DNA to be inserted. During these treatments the recombinant DNA construct is taken up into the protoplasts which are then washed and cultured. In this way it has been possible to transfer genes directly into protoplasts of graminaceous plants such as Trilicum monococcum and Lolium multiflorum. The presence of the genes in the transformed cells can be determined by assessing levels of antibiotic resistance. [...]


[...] Clearly therefore, CaMV has potential for the transfer of agriculturally desirable traits into crops. Conclusion Attempts are being made to circumvent this problem through the use of binary vectors, i.e. one viral genome carrying foreign genes, and the second carrying viral genes essential for replication, etc. However, there are also general problems of disease symptoms which may be produced after expression of viral genes in the host plant. Clearly, much more research will be required before viruses acquire a major role in plant genetic engineering. SUMMARY It is [...]


[...] Such progress has shown that the genetic engineering of monocotyledons is a viable proposition. One of the major remaining problems withcereals, however, is our lack of knowledge of reliable systems to regenerate whole plants from culture. Another method of inserting genes directly into plant cells is by microinjection. This work requires the provision of some finely adjustable equipment for the micromanipulation and injection of cells. In Japan, researchers have demonstrated the microinjection of berberine-labelled DNA into protoplasts. Berberine shows up yellow under ultraviolet light and was easily visualized in the cells using a microscope fitted with epifluorescence optics. [...]

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