Skip to main content
ARS Home » Southeast Area » Gainesville, Florida » Center for Medical, Agricultural and Veterinary Entomology » Insect Behavior and Biocontrol Research » Research » Publications at this Location » Publication #111364
Title: VITELLIN AND VITELLOGENIN IN PODISUS MACULIVENTRIS: IDENTIFICATION WITH MONOCLONAL ANTIBODIES AND REPRODUCTIVE RESPONSE TO DIET

Author
item Shapiro, Jeffrey
item WASSERMAN, HEATHER - UNIV. OF FL, GAINESVILLE
item GREANY, PATRICK - FORMER USDA, ARS EMPLOYEE
item NATION, JAMES - UNIV. OF FL, GAINESVILLE

Submitted to: Archives of Insect Biochemistry and Physiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/12/2000
Publication Date: 7/1/2000
Citation: Shapiro, J.P., Wasserman, H.A., Greany, P.D., Nation, J.L. 2000. Vitellin and vitellogenin in pthe Soldier Bug, Podisus maculiventris: Identification with monoclonal antibodies and reproductive response to diet. Archives of Insect Biochemistry and Physiology. 44:130-135.

Interpretive Summary: Improvements in the mass rearing of insect predators and parasites are crucial if augmentative biological control is to have major impact. The outcomes must include increased quantity, quality, and efficiency of arthropod production by commercial insectaries. Reproductive rate and quality are central to the enterprise. We are developing a tool, an ELISA or Enzyme-Linked Immunosorbent Assay, to assess and predict rates of reproduction in a predatory stinkbug, Podisus maculiventris. P. maculiventris is commonly produced and released to prey on caterpillar pests in field and greenhouse. This report first describes how females respond to feeding by producing a yolk protein precursor, which appears in their blood (hemolymph) within days after feeding. The protein is produced more rapidly and at higher levels in insects fed on caterpillars than in those fed on an artificial diet. A critical component of the ELISA was then ndeveloped, a monoclonal antibody that binds to and identifies a yolk protein and its precursor from eggs or hemolymph of reproducing females, but not males. The antibody bound to a single protein that is common to both egg yolk and hemolymph in the female. The ELISA will be initially directed at developing or improving artificial diets and at assessing product quality during mass rearing in insectaries. The concepts developed can apply to most insects, mites, or other arthropods that serve as natural enemies against pest species.

Technical Abstract: Reproductive responses were compared in adult Podisus maculiventris fed an artificial diet or a diet of larval Galleria mellonella. Adults were fed on day five after the adult molt, following four days on water only to allow complete previtellogenic development. Newly oviposited eggs contained major polypeptides of 171,000, 106,000, and 51,000 molecular weight. A major female-specific polypeptide of 171,000 molecular weight appeared in hemolymph only after feeding but preceeding follicular growth, and was tentatively identified as an apoprotein of vitellogenin, based on female specificity and electrophoretic mobility coincident with a major polypeptide of egg homogenates. Appearance of the putative apoprotein and follicular growth were delayed and reduced in females fed an artificial diet relative to those fed G. mellonella. Two monoclonal antibodies against egg proteins of P. maculiventris were selected by immuno-dot blot for strong reaction against egg and female hemolymph and null reaction against male hemolymph. On native Western blots, the antibodies reacted in egg homogenate against one tight fast-running protein band and in hemolymph against two slower-running bands, one tight and the other strong yet diffuse. On SDS blots, only one band reacted in both egg and hemolymph, running at the same Mr of 170k. This band corresponded to the female- specific hemolymph band that appeared within several days of feeding, further confirming its identity as a vitellogenin. These mAbs are being used to develop an enzyme-linked immunosorbent assay (ELISA) to quantitate reproductive response of females to diets of differing quality.