Western corn rootworm (Diabrotica virgifera virgifera LeConte) population dynamics

1 The western corn rootworm Diabrotica virgifera virgifera LeConte is a major insect pest of field maize, Zea mays L. Larvae can cause substantial injury by feeding on maize roots. Larval feeding may destroy individual roots or root nodes, and reduce plant growth, stability, and yield. Costs associated with managing corn rootworms in continuous maize are annually one of the largest expenditures for insect management in the United States Corn Belt.
2 Even though D. virgifera virgifera has been studied intensively for over 50 years, there is renewed interest in the biology, ecology, and genetics of this species because of its ability to rapidly adapt to management tactics, and its aggressive invasive nature.
3 This article provides a comprehensive review of D. virgifera virgifera population dynamics, specifically: diapause, larval and adult development, seasonality, spatial and temporal dynamics at local and landscape scales, invasiveness in North America and Europe, and non-trophic interactions with other arthropods.
4 Gaps in current knowledge are identified and discussed especially within the context of challenges that scientists in North America and Europe are currently facing regarding pest dynamics and the need to develop appropriate management strategies for each geographic area.     

Purification and kinetic analysis of acetylcholinesterase from western corn rootworm,Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae)

Acetylcholinesterase (AchE, EC 3.1.1.7) was purified from western corn rootworm (WCR, Diabrotica virgifera virgifera) beetles by affinity chromatography. The purification factor reached over 20,000-fold with a specific activity of 169.5 μmol/min/mg and a yield of 23%. The V_max values for hydrolyzing acetylthiocholine (ATC), acetyl-(β-methyl)thiocholine (AβMTC), propionylthiocholine (PTC), and S-butyrylthiocholine (BTC) were 184.8, 140.5, 150.2, and 18.8 μmol/min/mg, respectively, and K_m values were 19.7, 18.5, 14.1, and 11.0 μM, respectively. The first three substrates showed significant inhibition to the AchE at higher concentrations, whereas BTC showed inhibition at the concentrations of 0.25–2 mM but activation at >4 mM. AchE activity was almost completely inhibited by 1 μM eserine and BW284C15, respectively, but only 12% of AchE activity were inhibited by ethopropazine at the same concentration. These results suggested that the purified AchE from WCR was a typical insect AchE. Insecticides or their oxidative metabolites, chlorpyrifos-methyl oxon, carbofuran, carbaryl, malaoxon, and paraoxon, used in in vitro kinetic study exhibited high inhibition to AchE purified from WCR. However, chlorpyrifos-methyl oxon and carbofuran showed at least 36- and 4-fold, respectively, higher inhibitory potency than the remaining insecticides examined. Results from our in vitro inhibition of AchE agreed quite well with the previously published in vivo bioassay data. Arch. Insect Biochem. Physiol. 39:118–125, 1998. © 1998 Wiley-Liss, Inc.     

Behaviour and ecology of the western corn rootworm (Diabrotica virgifera virgifera LeConte)

1 The western corn rootworm (WCR) is a historic pest with a legacy of resistance and behavioural plasticity. Its behaviour and nutritional ecology are important to rootworm management. The success of the most effective and environmentally benign rootworm management method, annual crop rotation, was based on an understanding of rootworm behaviour and host–plant relationships. Enthusiastic adoption of crop rotation, provided excellent rootworm management, but also selected for behavioural resistance to this cultural control.
2 Though well-studied, significant gaps in WCR biology remain. Understanding the topics reviewed here (mating behaviour, nutritional ecology, larval and adult movement, oviposition, alternate host use, and chemical ecology) is a starting point for adapting integrated pest management and insect resistance management (IRM) to an expanding WCR threat. A presentation of significant questions and areas in need of further study follow each topic.
3 The expansion of WCR populations into Europe exposes this pest to new environmental and regulatory conditions that may influence its behaviour and ecology. Reviewing the state of current knowledge provides a starting point of reference for researchers and pest management decision-makers in North America and Europe.
4 The trend toward increasing adoption of transgenic maize will place an increasing premium on understanding WCR behaviour. IRM plans designed to promote sustainable deployment of transgenic hybrids are grounded on assumptions about WCR movement, mating and ovipositional behaviour. Preserving the utility of new and old management options will continue to depend on a thorough understanding of WCR biology, even as the ecological circumstances and geography of WCR problems become more complex.     

Morphological integration and modularity in Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) hind wings

The morphological integration of the hind wings of the western corn rootworm Diabrotica virgifera virgifera LeConte was investigated to get a better insight of the undergone by this invasive species. Geometric morphometric methods were used to test two modularity hypotheses associated with the wing development and function (hypothesis H1: anterior/posterior or H2: distal/proximal wing parts). Both hypotheses were rejected and the results showed the integrated behavior of the hind wings of D. v. virgifera. The hypothesized modules do not represent separate units of variation, so in a similar fashion as exhibited by the model species Drosophila melanogaster, the hind wings of D. v. virgifera act as a single functional unit. The moderate covariation strength found between anterior and posterior and distal and proximal parts of the hind wing of D. v. virgifera confirms its integrated behavior. We conclude that the wing shape shows internal integration, which could enable flexibility and thus enhance flight maneuverability. This study contributes to the understanding of morphological integration and modularity on a non-model organism. Additionally, these findings lay the groundwork for future flight performance and biogeographical studies on how wing shape and size vary across the endemic and expanded/invaded range in the USA and Europe infested with D. v. virgifera.     

A review of resistance breeding options targeting western corn rootworm (Diabrotica virgifera virgifera LeConte)

1 This review presents the latest research regarding maize resistance breeding against western corn rootworm (WCR) in the U.S.A. and Europe.
2 Investigations in Europe on the development of maize cultivars possessing resistant mechanisms against WCR are just beginning. In 2003, the European Commission implemented measures aimed at slowing down the spread of the WCR in Europe. Nevertheless, this pest has already been found in 20 countries of the European region. To establish a sustainable production system, the evaluation of native (nontransgenic) resistance in maize cultivars is essential.
3 This review emphasizes the future challenges involved in the research of native resistance breeding in maize against the insect.     

Screening of entomopathogenic nematodes for virulence against the invasive western corn rootworm, Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae) in Europe

Entomopathogenic nematode species available in Europe were screened for their efficacy against both the root-feeding larvae and silk-feeding adults of the western corn rootworm, Diabrotica virgifera virgifera LeConte. Laboratory screening tests were aimed at the selection of candidate biological control agents for the management of this invasive alien pest in Europe. Steinernema glaseri, S. arenarium, S. abassi, S. bicornutum, S. feltiae, S. kraussei, S. carpocapsae and Heterorhabditis bacteriophora were studied to determine their virulence against third instar larvae and adults of D. v. virgifera in small-volume arenas (using nematode concentrations of 0.5, 0.8, 7.9 and 15.9 infective juveniles cm-2). All nematode species were able to invade and propagate in D. v. virgifera larvae, but adults were rarely infected. At concentrations of 7.9 and 15.9 cm-2, S. glaseri, S. arenarium, S. abassi and H. bacteriophora caused the highest larval mortality of up to 77%. Steinernema bicornutum, S. abassi, S. carpocapsae and H. bacteriophora appeared to have a high propagation level, producing 5970+/-779, 5595+/-811, 5341+/-1177 and 4039+/-1025 infective juveniles per larva, respectively. Steinernema glaseri, S. arenarium, S. feltiae, S. kraussei and H. bacteriophora were further screened at a concentration of 16.7 nematodes cm-2 against third instar larvae in medium-volume arenas (sand-filled trays with maize plants). Heterorhabditis bacteriophora, S. arenarium and S. feltiae caused the highest larval mortality with 77+/-16.6%, 67+/-3.5%, and 57+/-17.1%, respectively. In a next step, criteria for rating the entomopathogenic nematode species were applied based on results obtained for virulence and propagation, and for current production costs and availability in Europe. These criteria were then rated to determine the potential of the nematodes for further field testing. Results showed the highest potential in H. bacteriophora, followed by S. arenarium and S. feltiae, for further testing as candidate biological control agents.     

Quick scouting of eggs of western corn rootworm (Diabrotica virgifera virgifera LeConte, 1868) from soil

Main method to control the American Corn Rootworm is crop rotation (Camprag et. al., 1994) but we don't know how to determine the possible number of larvae under fall so we cannot use autumn cereals to change the row of cultivated plants. The pest spends almost 10 months in soil in egg and larval state (Chiang, 1973). There are two methods for scouting Diabrotica eggs and larval instars from soil over the winter. One of the two most important methods is holding soil samples on fixed temperature (Fromm et al., 1999). This method takes more than one and a half month but its result is highly reliable. The conventional egg-washing technique takes fewer days to count the number of Diabrotica eggs in soil but it has lower effectiveness than the other one because the eggs in a sample cannot be counted correctly. Our results show that the effectiveness of egg washing with high concentrated salty water (NaCl) is high and the method is quick enough to help planning the crop rotation even under the autumn period (Takács et al., 2004).     

Behavioral and electrophysiological dose-response relationships in adult western corn rootworm (Diabrotica virgifera virgifera LeConte) for host pollen amino acids

A strong correlation is shown between taste cell inputs and phagostimulatory outputs with predominant dietary pollen amino acids for western corn rootworm, Diabrotica virgifera virgifera. Behavioral and electrophysiological dose-response profiles in adult beetles are presented for five major free amino acids in host pollens. Differential responses were found with strongest phagostimulation and sensory response elicited by L-alanine and L-serine, followed in order by L-proline and beta-alanine. gamma-Aminobutyric acid gave the weakest and most sporadic response. ED(50) values for phagostimulation and chemosensory input were 28.3nmol/disk and 13mM, respectively, for L-alanine and 17nmol/disk and 11mM, respectively, for serine. Threshold values for the responses were approximately 1-2mM. These behavioral and chemosensory dose-response ranges correspond closely to levels of free amino acids present in host plant pollens. Use of these response values in development of a pollen chemosensory code for western corn rootworm feeding is discussed.     

Reproductive Biology of Celatoria compressa (Diptera: Tachinidae), a Parasitoid of Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae)

The tachinid Celatoria compressa, a parasitoid of adult Diabrotica species in North America has been studied as a candidate classical biological control agent for the western corn rootworm, Diabrotica virgifera virgifera, in Europe. Prior to its potential importation, a thorough understanding of the parasitoid's reproductive biology is essential, and is an important component in the evaluation of a species as a biological control agent. In this study it has been clarified that C. compressa belongs to a group of a few tachinid species characterised by having eggs that contain fully developed larvae which are laid directly into the host. After mating, the egg load of females increased steadily from day 1 to a maximum egg load on day 4. Thereafter eggs containing fully developed first instar larvae reached a maximum of 31 in 69 eggs. At the first day of larviposition, females laid on average only five eggs into multiple hosts, which is in contrast to the availability of 18 eggs containing fully developed first instars in the uterus per female at that time. During a mean female's larviposition period of 23 days, a total of 33 first instars were larviposited into the hosts, which is only half of the female's egg load. Lifetime fecundity of C. compressa was significantly correlated with longevity. However no relationship was found between body size and either lifetime fecundity or longevity. In this study, an inverse host density-dependent pattern of percent parasitism was shown for C. compressa under 24-h fixed-time laboratory conditions, reflecting a Holling type II response. The number of host parasitized per C. compressa female reached an upper limit of 10 hosts with an increasing host density, which can be explained by the long host handling time of C. compressa.     

Mating success and spermatophore composition in Western corn rootworm (Coleoptera: Chrysomelidae)

Western corn rootworm (Diabrotica virgifera virgifera LeConte) resistance management for transgenic (or Bt) corn hinges on understanding the mating behavior and biology of this adaptable insect pest. During mating, the male transfers sperm and additional, previously uncharacterized material, to the female in the form of a spermatophore. We investigated the composition of rootworm spermatophores. Proteins were found to be a major component, and the stable isotope (15)N was used to assess the fate of spermatophore nitrogen in mated female beetles and their eggs. We also performed longevity studies on mated and virgin females under three different diet treatments and investigated the relationships between morphometric characteristics and spermatophore volume of mating pairs of beetles. The stable isotope analysis determined that nitrogen provided to the female in the spermatophore was incorporated into the eggs. We found that virgin female beetles on a corn diet lived significantly longer than mated female beetles on the same diet. There were significant positive relationships between male size parameters (head capsule width, pronotum width, and elytral length) and spermatophore volume, and ampulla and spermatophylax volume.     

Protease activities in the midgut of Western corn rootworm (Diabrotica virgifera virgifera LeConte)

The Western corn rootworm is one of the most economically important pests in corn. One possibility for controlling this pest is the cultivation of transgenic corn expressing Bacillus thuringiensis (Bt) toxins, such as Cry3A, Cry34Ab1/Cry35Ab1, and Cry3Bb1. However, widespread cultivation of the resulting Bt corn may result in the development of resistant pest populations. The Bt toxins are processed by proteases in the midgut of susceptible insects. Thus, protease activity studies were conducted using the midgut juice (pH 5.75) from third instars larvae of the susceptible Western corn rootworm. As a result, the activities of the serine endopeptidases trypsin, chymotrypsin, elastase, cathepsin G, plasmin, and thrombin; the cysteine endopeptidases cathepsin L, papain, cathepsin B, and cathepsin H; the aspartic endopeptidase pepsin; the metallo endopeptidase saccharolysin; the exopeptidase aminopeptidase, and the omegapeptidase acylaminoacylpeptidase were detected. These results are of basic interest but also lead to reference systems for the identification of protease-mediated resistance mechanisms in potentially resistant individuals.     

Adult movements of newly introduced alien Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae) from non-host habitats

Mark-release-recapture experiments were undertaken in order to investigate the movement of adult Diabrotica virgifera virgifera LeConte from accidental introduction points towards suitable habitats, such as its host plant, Zea mays L. In Hungary in 2003 and 2004, nine mark-release-recapture experiments were carried out in a grass steppe area and lucerne field, in which two small maize fields (10x10 m) had been planted 300 m distant from the central release point. After each release of 5500 to 6000 marked D. v. virgifera, beetle recaptures were recorded three times using non-baited yellow sticky traps placed on 30, 105, 205 and 305 m radii around the release point. In seven out of 15 recapture periods (47%), beetle populations showed no directional movements, and their movements towards any particular habitat cannot be predicted. During five recapture periods (33%), beetle populations showed a uni-directional movement, and in three cases (20%) a bi-directional movement was observed. In 10 out of 15 recapture periods (67%), the released populations moved in a direction that was comparable with the mean wind direction during these periods; thus, beetle movements were slightly correlated with wind direction. On average over sites and years, beetles were not preferentially moving towards the two small maize fields (located 300 m from the release point) compared to other directions. However, beetles moved significantly more frequently in the direction of naturally-occurring maize fields within a radius of 1500 m than towards other habitats. Beetles stayed more frequently within flowering lucerne fields out to a radius of 300 and 600 m than in non-flowering lucerne or other habitats. On average, 2.8% (SD 3.2) of all recaptured beetles arrived in one of the two small maize fields located 300 m from their release point indicating that there is a high risk of a founder population establishing. Habitat management cannot be suggested as a means of preventing the beetle's initial dispersal because movement was usually non-directional, and alternative food plants were used prior to reaching maize.     

The selection of a non-diapause strain of Diabrotica virgifera (Coleoptera: Chrysomelidae)

A non-diapause strain of Diabrotica virgifera LeConte was selected after nine generations by the technique of mass selection. The passive consequence of chilling the eggs of this species is a synchronization of hatch; the chilling does not break diapause. A hypothetical model (and its mathematical expression) explaining diapause in this species was derived from the data. Initial hatch in the non-diapause strain occurred in 24 days at 20°. In an unchilled-unselected population, the mean minimum period of development to initial hatch (both pre-diapause and post-diapause development) was ca. 24 days, and the duration of the mean dormant period was ca. 78 days. The duration of the dormant period is probably quantitatively inherited since this character appears to have enormous genetic plasticity.

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Zusammenfassung Ein Nicht-Diapause-Stamm von Diabrotica virgifera wurde durch Massenselektion nach neun Generationen selektiert. Die passive Folge des Kühlens der Eier dieser Art (was bei normaler Zucht des normalen Stamms geübt wird) ist eine Synchronisation des Schlüpfens; das Kühlen bricht nicht die Diapause. Ein hypothetisches Modell (und sein mathematischer Ausdruck), das die Diapause dieser Art erklärt, wurde von den gewonnenen Daten abgeleitet. Erstes Schlüpfen im Nicht-Diapause-Stamm fand nach 24 d bei 20° statt. In einer nicht gekühlten, nicht selektierten Population war die mittlere Minimalperiode der Entwicklung bis zum ersten Schlüpfen (Prä- und Postdiapause-Entwicklung) ca 24 d, die Dauer der mittleren Dormanzperiode ca 78 d (nach der Formel berechnet). Die Dauer der Dormanzperiode wird wahrscheinlich quantitativ vererbt; denn diese Eigenschaft scheint eine sehr grosse genetische Plastizität zu haben.

     

Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae) larval feeding behavior on transgenic maize (MON 863) and its isoline

Diabrotica species (Coleoptera: Chrysomelidae) larval behavior studies have posed a challenge to researchers because of the subterranean life cycle of this pest. To fully understand how the western corn rootworm, Diabrotica virgifera virgifera LeConte, injures the maize, Zea mays L., root system, its behavior must be studied. For example, larvae that can detect an area of the root that has a lower amount of toxin, whether from an insecticide or a transgenic maize plant, have an increased chance of survival. This study assessed D. v. virgifera larval feeding behavior on rootworm-susceptible maize and maize containing a biotechnology-derived trait (MON 863) with resistance to D. v. virgifera first instar feeding. Maize plants were grown in a medium that allowed for direct observation and measurements during feeding of larval stadia. Neonates were placed on maize seedlings, and data were taken at 3, 6, 9, and 12 d postinfestation on resistant and susceptible maize. On rootworm-susceptible maize, neonate larvae aggregated at the root tips and began actively feeding, and then they moved to older root tissue. Conversely, some larvae that ingested Cry 3Bb1 from the resistant maize exhibited no movement. Other larvae on the resistant maize moved continuously, sampling root hairs or root tissue but not actively feeding. The continuously moving larvae had visibly empty guts, suggesting possible nonpreference for the resistant root. This study contributes to our understanding of D. v. virgifera larval behavior and provides insight into questions surrounding the potential evolution of behavioral and biochemical resistance to Cry3Bb1.     

Western corn rootworm (Coleoptera: Chrysomelidae) beetle emergence from weedy Cry3Bbl rootworm-resistant transgenic corn

Three greenhouse experiments were conducted to evaluate western corn rootworm, Diabrotica virgifera virgifera LeConte, beetle emergence from individual pots containing glyphosate-tolerant transgenic corn, Zea mays L., expressing the Cry3Bbl endotoxin from the soil bacterium Bacillus thuringiensis Berliner (MON863), nontransgenic glyphosate-tolerant isoline corn, grassy weeds (giant foxtail, Setariafaberi R.A.W. Herrm; and large crabgrass, Digitaria sanguinalis (L.) Scop.), and combinations thereof infested with 40 neonate larvae. In the first two experiments, pots with corn and weed combinations were treated with glyphosate 5 d after larval infestation to kill the weeds. The third experiment was similar to the first two except that untreated corn-weed combinations were added. In all three experiments beetle emergence varied significantly. Beetle recovery generally did not vary between the nontransgenic, nontransgenic + weeds, and MON863 + weeds. Significantly more beetles were recovered from MON863 + weeds than MON863 alone or weeds alone. Beetle emergence from MON863 + weeds was likely enhanced by larvae that initially survived on weeds before application of glyphosate. Preliminary data indicated that fecundity was highest from beetles reared on nontransgenic isoline corn and fewer eggs were laid by beetles reared on MON863 alone. Egg viability was generally lowest from beetles reared on MON863. The implications of these results in relation to insect resistant management are discussed.     

Comparison of adult corn rootworm (Coleoptera: Chrysomelidae) sampling methods

Studies were conducted in Kansas corn and soybean fields during 1997 to compare various sampling methods, traps, and trap components for capturing three species of adult corn rootworms: western (Diabrotica virgifera virgifera Leconte), southern (D. undecimpunctata howardi Barber), and northern (D. barberi Smith & Lawrence). Lure constituents affected the species of beetle attracted to the trap. Traps with a lure containing 4-methoxycinnamaldehyde attracted more western corn rootworms, those with a lure containing eugenol were more attractive to northern corn rootworms, and those containing trans-cinnamaldehyde were most attractive to southern corn rootworms. Multigard sticky traps caught more beetles than did Pherocon AM sticky traps. In corn, a newly designed lure trap caught more beetles than did sticky traps on most occasions. Also, lure-baited sticky traps caught more beetles than did nonbaited sticky traps. Varying the color of the lure trap bottom did not affect the number caught. In soybeans, the new lure traps captured more beetles than did the nonbaited Multigard or Pherocon AM sticky traps. Results of this study suggest the new lure trap may provide a more accurate assessment of corn rootworm populations than traditional monitoring techniques and may be more esthetically pleasing to growers and consultants.     

Field-Cage Studies of Beauveria bassiana (Hyphomycetes: Moniliaceae) for the Suppression of Adult Western Corn Rootworm, Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae)

The efficacy of the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin, was tested as a control agent for adult western corn rootworm, Diabrotica virgifera virgifera LeConte, in walk-in field cages. Suspensions of B. bassiana conidia were applied to corn plants in cages into which laboratory-reared beetles had been released. Beetles were collected at 3 and 5 days post-application and evaluated in the laboratory for mortality. Mortality was 10, 29 and 50%, at rate equivalents of 7 X 10 12 , 2 X 10 13 (two applications), and 5 X 10 13 conidia/ ha, respectively. There was no significant difference in mortality of beetles collected at 3 days compared with 5 days post-application. Mortality due to B. bassiana was 24% when beetles were released into field cages 24 h post-application (5 X 10 13 conidia/ha) compared with 50% when beetles were present during the application. Beetle mortality declined significantly with increasing time from application in feeding assays carried out with leaf samples removed from plants at 0, 12, 24 and 72 h post-application. Mortality of beetles collected from treated plants within cages and maintained in the laboratory was found to overestimate the population decline by 10% when compared with beetle estimates from treated plants within field cages.     

Predator responses to novel haemolymph defences of western corn rootworm (Diabrotica virgifera) larvae

Many herbivorous arthropods use defensive chemistry to discourage predators from attacking. This chemistry relies on the ability of predators to rapidly learn to recognize and avoid offensive stimuli. Western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), employs multifaceted chemical defences in its haemolymph, which may contribute significantly to its success as a major economic pest. Here, we test the hypothesis that agrobiont predators can rapidly learn to recognize and avoid WCR larvae, and will thereby reduce their contribution to WCR suppression. In controlled feeding assays, the effectiveness of WCR haemolymph defences varied across three predator taxa (crickets, centipedes, and ants). Centipedes (Chilopoda: Lithobiidae) were minimally affected by WCR defences, but crickets [Gryllus pennsylvanicus Burmeister (Orthoptera: Gryllidae)] spent less time feeding on WCR than on an undefended control prey, house fly maggots. However, we uncovered no evidence indicating that experienced crickets rapidly learn to avoid WCR larvae, indicating that haemolymph defences offer few, if any, survival benefits for WCR. Colonies of ants [Lasius neoniger Emery (Hymenoptera: Formicidae)] switched from low worker participation in initial attacks on WCR to higher worker participation in subsequent attacks, indicating an attempt to overcome, rather than avoid, WCR haemolymph defences. These results suggest that a diverse assemblage of natural enemies will show a diverse array of behavioural responses to toxic pest prey, and highlight the importance of behavioural diversity in driving the function of natural enemy assemblages.