Transformation of pollen by particle bombardment
The development of pollen as a vector for direct gene transfer would be a significant advance in our ability to introduce genes into plants. Such methodology should be of general utility for many plant species, and in particular for the major monocotyledonous crop plants such as maize, wheat and barley that are recalcitrant to protoplast regeneration and that are not amenable to Agrobacterium based transformation techniques. A further advantage would be the avoidance of tissue culture steps that are time consuming and known to result in undesirable somaclonal variation. The potential of pollen as a vector for direct gene transfer has long been realized. For more than 10 years numerous investigators have attempted pollen-mediated transformation, several of which have claimed success [5, 8, 19, 23]. However the ultimate proof that transformation has taken place, that is, the demonstration of integration of foreign DNA into the nuclear genome at the molecular level and the genetic transmission of this DNA, is still lacking. This chapter presents a summary of research that has been directed towards pollen-mediated gene transfer, a detailed protocol for the delivery of DNA into pollen using particle bombardment and a discussion of factors that may be important for the successful application of this technique to obtain stably transformed plants.
KeywordsPollen Tube Pollen Germination Particle Bombardment Transient Expression Assay Direct Gene Transfer
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