Generation of a recombinant Anticarsia gemmatalis MNPV bearing the itch mite toxin gene

ELOÍSA I. ARANA; CHRISTINA B. MCCARTHY; ALINA V. GOLDBERG; ALICIA SCIOCCO-CAP; DAVID O´REILLY; VICTOR ROMANOWSKI

Noordwijkerhout

Congreso; 34th Annual Meeting of the Society for Invertebrate Pathology; 2001

AgMNPV is today the most widely used biopesticide in more than one million hectares of soybean. It kills the velvetbean caterpillar Anticarsia genmatalis, but depending on the age and susceptibility of the host, this virus may take up to two weeks to kill the insect. During that time larvae may continue to feed. Taking into account these facts, our aim is to improve its pesticidal properties. The first altemative chosen to achieve this objective was the insertion of an insect-toxic gene in the viral genome. Using a system developed in our laboratory for the modification of fte AgMMV genome via homologous recombination, we inserted the itch mite toxin gene in the polh locus. This foreign gene was excised with Eco RI from the pEV-Tox34 plasmid and subcloned in pBSSK in order to add lhe appropiate restriction sites for its insertion in the transfer vector pBS-AgDU. UFLAG-286 cells were cotransfected with the parental virus DNA (rAgMMV polEllacZ) and the transfer vertor containing the candidate gene. Recombinants were detected by its white plaque phenotype and purified. The purification steps were screened by PCR and the genome structure was confirmed by restriction pattern. Bioassays are being carried out to evaluate the biological effects of this genetic modification.