Administering and Detecting Protein Marks on Arthropods for Dispersal Research

Monitoring arthropod movement is often required to better understand associated population dynamics, dispersal patterns, host plant preferences, and other ...

Arid-Land Agricultural Research Center

The adult dustywing, Semidalis flinti Meinander (Neuroptera: Coniopterygidae), begins producing circular-shaped waxy particles after eclosion. The waxy material, which forms the particles, is extruded from individual pores found in clusters on the abdomen. Pores also are present in two rows of three pores on the frontalis and two pores on the first segment of each antennae. The pores have a rosette-like appearance and each pore extrudes dual waxy ribbons. As each ribbon extends a short distance out of the pore, it begins to curl back on itself until the end makes contact with the ribbon. The curled end then breaks free from the extruding ribbon to form the circular waxy particles with fluted edges approximately 2.75-microm diameter. The adults use the particles to cover all parts of their body except for their eyes and appear to lightly coat their antennae. The lipid portion of the particles consists largely of free fatty acids, almost exclusively the 24-carbon fatty acid, tetracosanoic acid. Minor lipid classes are hydrocarbons, fatty alcohols and unidentified material.

Characterization of the cuticular surface wax pores and the waxy particles of the dustywing, Semidalis flinti (Neuroptera: Coniopterygidae).

The sweetpotato whitefly, Bemisia tabaci (Gennadius) is fed on by a wide variety of generalist predators, but there is little information on these predator-prey interactions. A laboratory investigation was conducted to quantify the foraging behavior of the adults of five common whitefly predators presented with a surfeit of whitefly eggs, nymphs, and adults. The beetles, Hippodamia convergens Guerin-Meneville and Collops vittatus (Say) fed mostly on whitefly eggs, but readily and rapidly preyed on all of the whitefly lifestages. The true bugs, Geocoris punctipes (Say) and Orius tristicolor (Say) preyed almost exclusively on adult whiteflies, while Lygus hesperus Knight preyed almost exclusively on nymphs. The true bugs had much longer prey handling times than the beetles and spent much more of their time feeding (35-42%) than the beetles (6-7%). These results indicate that generalist predators vary significantly in their interaction with this host, and that foraging behavior should be considered during development of a predator-based biological control program for B. tabaci.

Foraging behavior and prey interactions by a guild of predators on various lifestages of Bemisia tabaci.

Four continuous cell lines were established from the embryos of the glassy-winged sharpshooter, Homalodisca coagulata (Say), an economically important insect vector of bacterial pathogens of grape, almond, citrus, oleander, and other agricultural and ornamental plantings. The cell lines were designated GWSS-Z10, GWSS-Z15, GWSS-G3, and GWSS-LH. The GWSS-Z10, GWSS-Z15, and GWSS-G3 lines were cultured in Ex-Cell 401 medium supplemented with 10% fetal bovine serum (FBS), whereas the GWSS-LH line was cultured in LH medium supplemented with 20% FBS. The cell lines were characterized in terms of their morphology, growth, protein composition, and polymerase chain reaction- amplification patterns of their chromosomal deoxyribonucleic acid. The population doubling times of GWSS-Z10, GWSS-Z15, GWSS-G3, and GWSS-LH were 46.2, 90.9, 100.3, and 60.2 h, respectively. These lines should be useful for the study of insect-pathogenic viruses of leafhoppers, aphids, treehoppers, and other related insects as well as plant-pathogenic viruses that are transmitted by these insects.

Characterization of cell lines developed from the glassy-winged sharpshooter, Homalodisca coagulata (Hemiptera: Cicadellidae).

A 2-yr study was conducted in a citrus orchard (Citrus sinensis L. Osbeck cultivar Valencia) to determine the influence of plant water stress on the population dynamics of glassy-winged sharpshooter, Homalodisca vitripennis (Germar). Experimental treatments included irrigation at 100% of the crop evapotranspiration rate (ET(c)) and continuous deficit-irrigation regimens at 80 and 60% ET(c). Microclimate and plant conditions monitored included temperature and humidity in the tree canopy, leaf surface temperature, water potential, and fruit quality and yield. Glassy-winged sharpshooter population densities and activity were monitored weekly by a combination of visual inspections, beat net sampling, and trapping. Glassy-winged sharpshooter populations were negatively affected by severe plant water stress; however, population densities were not linearly related to decreasing water availability in plants. Citrus trees irrigated at 60% ET(c) had significantly warmer leaves, lower xylem water potential, and consequently hosted fewer glassy-winged sharpshooter eggs, nymphs, and adults than trees irrigated at 80% ET(c). Citrus trees irrigated at 100% ET(c) hosted similar numbers of glassy-winged sharpshooter stages as trees irrigated at 60% ET(c) and a lower number of glassy-winged sharpshooter nymphs than the 80% ET(c) treatment, specifically during the nymphal density peak in mid-April to early July. Irrigation treatments did not affect populations of monitored natural enemies. Although the adult glassy-winged sharpshooter population was reduced, on average, by 50% in trees under severe water stress, the total number of fruit and number of fruit across several fruit grade categories were significantly lower in the 60% ET(c) than in the 80 and 100% ET(c) irrigation treatments.

Seasonal population dynamics of Homalodisca vitripennis (Hemiptera: Cicadellidae) in sweet orange trees maintained under continuous deficit irrigation.

Lygus hesperus Knight and Lygus lineolaris (Palisot de Beauvois) (Heteroptera: Miridae) are economically important plant bugs on many crops worldwide. However, these omnivores are also facultative predators on a wide variety of insects. This study was conducted to quantify and compare herbivory and carnivory exhibited among different lifestages of these two insect pests. The feeding activity of a total of 422 individuals was observed for 1 h each in feeding arenas containing a cotton leaf disk and copious amounts of the sweetpotato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) eggs, nymphs and adults. The L. hesperus and L. lineolaris lifestages examined included adults and 3rd, 4th and 5th instar nymphs. Plant feeding occupied the majority of both species' time budget, regardless of the species or lifestage examined. There was a tendency for L. lineolaris lifestages to feed more often and for longer duration on plant tissue than L. hesperus. All lifestages of both species rarely fed on B. tabaci, but when they did, they preferred nymphs > adults > eggs. There were only a few cases where there were significant differences in predation rates and prey handling times exhibited among lifestages and between species, but juvenile L. hesperus tended to be more predaceous than juvenile L. lineolaris on whitefly nymphs and adults and 5th instar and adult L. lineolaris were significantly more herbaceous than their L. hesperus counterparts. In addition, the younger individuals of both species tended to have greater prey handling times than their older counterparts. The significance of these findings is discussed.

Diet selection exhibited by juvenile and adult lifestages of the omnivores Western tarnished plant bug, Lygus hesperus and tarnished plant bug, Lygus lineolaris.

A study was conducted in 2006 and 2007 designed to examine the foraging range of honey bees, Apis mellifera (Hymenoptera: Apidae), in a 15.2 km(2) area dominated by a 128.9 ha glyphosate-resistant Roundup Ready® alfalfa seed production field and several non-Roundup Ready alfalfa seed production fields (totaling 120.2 ha). Each year, honey bee self-marking devices were placed on 112 selected honey bee colonies originating from nine different apiary locations. The foraging bees exiting each apiary location were uniquely marked so that the apiary of origin and the distance traveled by the marked (field-collected) bees into each of the alfalfa fields could be pinpointed. Honey bee self-marking devices were installed on 14.4 and 11.2% of the total hives located within the research area in 2006 and 2007, respectively. The frequency of field-collected bees possessing a distinct mark was similar, averaging 14.0% in 2006 and 12.6% in 2007. A grand total of 12,266 bees were collected from the various alfalfa fields on seven sampling dates over the course of the study. The distances traveled by marked bees ranged from a minimum of 45 m to a maximum of 5983 m. On average, marked bees were recovered ~ 800 m from their apiary of origin and the recovery rate of marked bees decreased exponentially as the distance from the apiary of origin increased. Ultimately, these data will be used to identify the extent of pollen-mediated gene flow from Roundup Ready to conventional alfalfa.

Foraging range of honey bees, Apis mellifera, in alfalfa seed production fields.

Homalodisca vitripennis (Germar), a vector of Xylella fastidiosa, is associated with citrus plantings in California. Infested citrus orchards act as a source of vectors to adjacent vineyards where X. fastidiosa causes Pierce's disease. An analysis of the pattern and rate of movement of H. vitripennis and its egg parasitoid, Gonatocerus ashmeadi Girault, was conducted in a citrus orchard by using a protein mark-capture technique to quantify movement and net dispersal rates in the experimental areas. Treatments included irrigation at 100% of the crop evapotranspiration rate (ET(c)), 80, and 60% ET(c). Sex-specific net dispersal rates showed that H. vitripennis males and females moved consistently and contributed equally to the level of population change within treated areas. Trees irrigated at 60% ET(c) were the least preferred by H. vitripennis. Among all protein-marked individuals captured in the 60% ET(c) treatment, ≈ 75 and 88% in 2005 and 2006, respectively, were inflow individuals. Movement toward less preferable plants indicates that in agricultural landscapes dominated by perennial monocultures, there is a random component to H. vitripennis movement, which may result from the inability of H. vitripennis to use plant visual cues, olfactory cues, or both to make well-informed long-range decisions. The 80% ET(c) areas were a significant source of adult H. vitripennis and G. ashmeadi compared with the other treatments. Colonization rates by parasitoids were synchronized with the spatiotemporal distribution of H. vitripennis eggs. Results suggest that H. vitripennis movement from citrus into adjacent vineyards could be a result of random dispersal rather than oriented movement in response to host-plant characteristics.

Plant water stress effects on the net dispersal rate of the insect vector Homalodisca vitripennis (Hemiptera: Cicadellidae) and movement of its egg parasitoid, Gonatocerus ashmeadi (Hymenoptera: Mymaridae).

Immunoproteins are markers that are useful for monitoring dispersal and/or pest consumption, but current application techniques are less effective for the large guild of piercing-sucking predators important in biocontrol. We quantified the use of protein immunomarks in tracking emigration of spined soldier bug, Podisus maculiventris Say (Hemiptera: Pentatomidae) and predation on the hornworm caterpillar, Manduca sexta L. (Lepidoptera: Sphingidae). An external protein mark was topically applied to adult P. maculiventris to assess persistence under field conditions for >2 wk. Internal marks were incorporated into the artificial diet of M. sexta to test retention of the internal mark in the prey and uptake of the mark by predators. External marks remained detectable in 100% of individuals after 3 d and >50% still tested positive at 12 d after application in the field. Internal diet-based marking was also effective in tracking feeding by P. maculiventris on M. sexta, especially using rabbit IgG that was far more persistent than chicken IgY. Nearly 90% of stink bugs fed caterpillars previously reared on protein-enriched diet retained their mark for 24 h. Surprisingly, diet concentration and time reared on diet had comparatively little impact on mark retention. Development on unmarked tomato leaves clearly diluted the initial diet mark, but plant-reared individuals that were marked were still successfully detected in 35 and 20% of the predators.

Employing immunomarkers to track dispersal and trophic relationships of a piercing-sucking predator, Podisus maculiventris (Hemiptera: Pentatomidae).

Alfalfa (Medicago sativa L.) is a highly attractive plant host to Lygus spp. and is used as a trap crop in California organic strawberries to influence the dispersion and dispersal of these pests, particularly Lygus hesperus Knight. The abundance and distribution of Lygus spp. nymphs between two trap crops separated by 50 strawberry rows was analyzed in 2008 and 2010. Nymphs demonstrated a bimodal distribution in strawberries between trap crops, where nymphs were most abundant and aggregated in alfalfa, when compared with interior strawberry rows, where nymphs were less abundant. The majority of nymphs were concentrated in trap crops and nymphal densities in interior strawberry rows were well below economic thresholds. The movement of Lygus spp. from a marked alfalfa trap crop into adjacent strawberry rows or trap crops was also studied in 2008 and 2009 using a chicken egg albumin enzyme-linked immunosorbent assay mark-capture technique. The majority of marked-captured L. hesperus adults and Lygus spp. nymphs remained in alfalfa trap crops, rather than dispersing out into strawberry rows at 24 h, 48 h, and 2 wk, postprotein application. The attenuation of Lygus spp. movement in alfalfa associated with organic strawberries is a key component of successful trap cropping. A small percentage of marked adults and nymphs were captured in neighboring alfalfa trap crops, located 62 m from the point of protein application, highlighting the dispersal capacity of this key pest.

Dispersion, distribution, and movement of Lygus spp. (Hemiptera: Miridae) in trap-cropped organic strawberries.

Alfalfa trap crops are currently used to manage Lygus spp. in organic strawberry fields on the California Central Coast. The retention of Lygus spp. in alfalfa creates aggregated distributions that provide improved opportunities for biological control by the introduced parasitoid Peristenus relictus (Ruthe). The abundance and distribution of P. relictus between two trap crops separated by 50 strawberry rows were analyzed in 2008 and 2010. Parasitism of Lygus spp. nymphs by P. relictus (measured by larval abundance and % parasitism) was greatest in alfalfa trap crops compared with strawberry rows. A significantly positive correlation between host nymphs and P. relictus larvae in and between trap crops was found. Movement of P. relictus adults from a marked alfalfa trap crop into adjacent strawberry rows or trap crops was also studied in 2008 and 2009 using a chicken egg-albumin enzyme-linked immunosorbent assay mark-capture technique. In 2008 and 2009, 85 and 49% of protein-marked wasps were captured from central trap crops, respectively, indicating that alfalfa trap crops act as a concentrated "host-density anchor" in organic strawberry fields.

Spatial density and movement of the Lygus spp. parasitoid Peristenus relictus (Hymenoptera: Braconidae) in organic strawberries with alfalfa trap crops.

James R. Hagler

USDA-ARS

Arid-Land Agricultural Research Center

Administering and Detecting Protein Marks on Arthropods for Dispersal Research

James R. Hagler

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Arid-Land Agricultural Research Center

Administering and Detecting Protein Marks on Arthropods for Dispersal Research

USDA-ARS