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Abstract
The physical barrier imposed by the thick microspore wall constitutes an obstacle for an efficient Agrobacterium-mediated transformation of vacuolate microspores prior to androgenic induction and haploid embryogenic commitment. It is thus necessary to implement additional methods to overcome this drawback. In this study, we focused on the optimization of a protocol to allow for the exogenous DNA to enter the microspore in an efficient manner. We tested different options, based on microprojectile bombardment, to be applied prior to agroinfiltration. From them, the best results were obtained through co-transformation by microspore bombardment with DNA-coated microprojectile particles, followed by Agrobacterium tumefaciens infection. This method provides an efficient means to integrate extraneous DNA into rapeseed microspores prior to androgenesis induction.
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