Expression of the cry1Ac in `Arizona Common' Common Bermudagrass via Agrobacterium-mediated Transformation and Control of Black Cutworm

Abstract

Bermudagrass (Cynodon L.C. Rich.) is grown on more than 4 million ha in the southern United States. The black cutworm (Agrotis ipsilon Hufnagel) is the most commonly encountered pest of bermudagrass, especially on golf course greens. Developing insect-resistant cultivars is a very desirable substitute, both environmentally and economically, to using current synthetic pesticides. Here we report, for the first time, Agrobacterium-mediated transformation of `Arizona Common' common bermudagrass [Cynodon dactylon (L.) Pers.] with the Bacillus thuringiensis Berliner cry1Ac gene encoding an endotoxin active against black cutworm. Mature seeds were used for producing embryogenic callus, and calli were transformed with a plasmid containing a synthetic cry1Ac and the kanamycin resistance (nptII) genes. Putative transgenic calli and plantlets were selected on media containing 100 and 50 mg·L-1 G418, respectively. RNA-blot analysis of PCR-positive lines revealed the expression of the cry1Ac transgene in three out of five putative transgenic lines. The larvae fed on transgenic plant leaves experienced highly significant (over 80%) mortality.