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Protocols for Microapplicator-assisted Infection of Lepidopteran Larvae with Baculovirus
Published on: August 23, 2008
Baculovirus insecticide production in insect larvae
Nikolai van Beek1, David C Davis
1Chesapeake PERL Inc, Savage, MD, USA.
Methods in Molecular Biology (Clifton, N.J.)
|October 24, 2007
Summary
Baculovirus insecticides are widely used, with recombinant viruses showing promise for faster pest control. This study details benchtop production methods for wild-type and genetically modified baculoviruses, addressing challenges in large-scale manufacturing.
Area of Science:
- Entomology
- Biotechnology
- Insect Pathology
Background:
- Baculovirus-based insecticides are globally utilized, with significant commercial success in crops like soybean and cotton.
- Current commercial products rely on wild-type baculoviruses, but recombinant viruses encoding insecticidal proteins offer enhanced efficacy, particularly for crops sensitive to feeding damage.
Purpose of the Study:
- To describe benchtop-scale production methods for two distinct baculoviruses: wild-type Helicoverpa zea nucleopolyhedrovirus and a recombinant Autographa californica multiple nucleopolyhedrovirus.
- To address the challenges associated with low-cost in vivo production of recombinant baculoviruses, specifically concerning host survival time.
Main Methods:
- Production of wild-type Helicoverpa zea nucleopolyhedrovirus in Helicoverpa zea larvae, requiring isolated rearing due to larval aggression.
- Production of recombinant Autographa californica multiple nucleopolyhedrovirus in Trichoplusia ni larvae, utilizing a tetracycline transactivator system to control expression of an insect-specific scorpion toxin (LqhIT2).
Main Results:
- Successful benchtop-scale production of wild-type H. zea nucleopolyhedrovirus was achieved.
- Recombinant AcMNPV production in T. ni yielded wild-type quantity and quality by repressing toxin expression, despite the toxin's potential to limit progeny virus production.
Conclusions:
- Benchtop-scale production protocols for both wild-type and recombinant baculoviruses are feasible.
- The tetracycline transactivator system provides a viable method for producing recombinant baculoviruses with controlled expression of insecticidal genes, overcoming limitations of host survival.
