Development of a monoclonal antibody to detect predation of the sweetpotato whitefly, Bemisia tabaci

Abstract

We have developed an enzyme-linked immunosorbent assay (ELISA) for detection of whitefly antigen in the guts of predaceous arthropods based on a monoclonal antibody (MAb) produced in response to Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae) egg antigen. We observed positive antigenantibody reactions with the egg and the adult female stages of certain whitefly species and not with others. This MAb did not cross-react with any of the whitefly nymphs or adult males we tested. Moreover, this MAb did not cross-react with the various life stages of other insect species we examined. This immunologically-based technique to identify whitefly egg facilitates examinations of predator gut contents in the field. Such a specific and sensitive whitefly egg immunoassay will expedite the characterization of the B. tabaci predator complex in the southwestern United States.

Use of a gut content ELISA to detect whitefly predator feeding activity after field exposure to different insecticide treatments

“…Since D. nr. divergens feeds only on adults, and since the whitefly egg-specific ELISA can't differentiate between an egg and an egg-carrying (i.e., gravid) female meal (Hagler et al 1993) a positive ELISA reaction can be attributed to predation on adult females. With the exception of perhaps Z. renardii , the other whitefly predators examined in this study readily prey on both whitefly eggs and adults .…”

“…Following incubation, the insect macerates were discarded from each plate and a 360-ml aliquot of 1.0% non-fat dry milk in distilled water was added to each well for 30 min at 278C to block any unoccupied antigenic sites in the wells. The non-fat milk was emptied from each plate and a 50-ml aliquot of anti-whitefly monoclonal antibody was added to each well of the ELISA plate (Hagler et al 1993). The ELISA plates were then incubated for 1 h at 278C.…”

“…We determined the proportion of predators that had recently fed on whitefly using an established whitefly-specific ELISA (Hagler et al 1993). Each predator was macerated in 500-ml of Tris-buffered saline (TBS) (pH 7.4).…”

“…Here, we present an assessment of the impact that each management regime had on predator feeding activity. We analyzed the gut contents of several key whitefly predators in cotton plots exposed to various insecticide regimes over two growing seasons using a whitefly specific enzymelinked immunosorbent assay (ELISA) (Hagler et al 1993). The ELISA provides a quick and accurate indirect method (i.e., a non-visual method) to assess predation (see Luck et al 1988).…”

Identifying the predator complex of Homalodisca vitripennis (Hemiptera: Cicadellidae): a comparative study of the efficacy of an ELISA and PCR gut content assay

“…Contemporary assays of stomach content include monoclonal antibody (mAb)-based, enzyme-linked immunosorbent assays (ELISA), which detect species-speciWc proteins (and sometimes life-stage-speciWc proteins) (Greenstone and Morgan 1989;Hagler et al 1991Hagler et al , 1993Hagler et al , 1994Symondson and Liddell 1996;Greenstone 1996;Fournier et al 2006;Harwood et al 2007a), and polymerase chain reaction (PCR)-based assays, which detect species-speciWc DNA (all life stages) (Agustí et al 1999(Agustí et al , 2003Harper et al 2005;de León et al 2006;Harwood et al 2007b). …”