Production of insect resistant, transgenic turfgrass plants with an insecticidal protein gene.

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In our project, we obtained the following research results.1.A higher frequency of embryogenic callus induction from seeds was obtained using a revised callus induction medium, indicating the establishment of an efficient cell-to-plant regeneration system of turfgrass plants in vitro. 2.In electroporation, modification of buffer components and electric conditions improved gene transfer efficiency to protoplasts. 3.In particle bombardment, adjustment of several mechanical conditions improved gene transfer efficiency to intact culture cells. 4.Transgenic plants with a marker gene were obtained by electroporation, and the transformation was confirmed by PCR-southrn analysis. Difference in gene expression level between transformants with different types of promoters of transgenes was analyzed. 5.Among various types of bt genes, we found that the CryIA(c) produced the protein which gave the strongest insecticidal activity to Jananese lawn grass cutworm and Bluegrass webworm, the severest harmful insects of turfgrass. 6.The CryIA(c) gene (wild-type) was introduced into turgrass plants. Insect resistance of the transgenic plants was examined by bioassay with Bluegrass webworm. However, none of the transgenic plants showed resistance to the insect. 7.To enhance the insecticidal activity of the CryIA(c) gene, we intend 1)to produce a CryIA(c) gene with a stronger promoter, and 2)to modify the sequence of the coding region of the gene. We hope that the use of the gene with a stronger promoter or the modified gene may lead to the production of transgenic turfgrass plants with substantial insect resistance.