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.     

Nutritional resources used by the invasive maize pest Diabrotica virgifera virgifera in its new South-east-European distribution range

Food utilization by adults of the invasive maize (Zea mays L.) (Poaceae) pest western corn rootworm (WCR; Diabrotica virgifera virgifera LeConte) (Coleoptera: Chrysomelidae) was investigated in the south‐eastern part of its new European distribution range. At weekly intervals over a 10‐week period, 10 beetles per field were collected from six fields that had a high abundance of flowering weeds and six fields with a low abundance of flowering weeds, with the aim of understanding adult feeding behaviour in Europe and comparing this behaviour with North American WCR. Gut content analysis was performed to determine the use of maize tissue and weed pollen with regard to maize phenology. Furthermore, all pollen found within the gut was quantified and identified to plant species level. The use of maize tissue by adult WCR changed over time according to maize phenology. Pollen originating from plants other than maize was used more frequently as the maize matured. Adults fed on pollen from 19 of 25 different plant species found in maize fields and showed a preference for the pollen of specific weeds. Pollen from weed species was found more often in beetles from fields with a high abundance of weeds compared to beetles from fields with a low abundance of weeds. Females consumed greater amounts of pollen than males, the latter feeding on a wider diversity of host plants. The pollen resources used by adult WCR in Hungary were more diverse compared to WCR in the USA, which may contribute to the invasion success of WCR in Europe.     

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.     

Genes, gene flow and adaptation of Diabrotica virgifera virgifera

1  Diabrotica virgifera virgifera has emerged as a major pest of cultivated maize, due to a combination of its high capacity to inflict economic damage, adaptability to pest management techniques and invasiveness.
2 This review presents a survey of the current state of knowledge about the genetics of D.   v.   virgifera . In addition, the tools and resources currently available to Diabrotica geneticists are identified, as are areas where knowledge is lacking and research should be prioritized.
3 A substantial amount of information has been published concerning the molecular phylogenetic relationships of D.   v.   virgifera to other chrysomelids.
4 There is a growing literature focused on the population genetics and evolution of the species. Several adaptations to anthropogenic selection pressure have been studied, with resistance to synthetic insecticides providing some particularly well-characterized examples.
5 A notable deficiency is a lack of studies directed toward the formal genetics of D.   v.   virgifera .     

Population density of Diabrotica virgifera virgifera LeConte beetles in Serbian first year and continuous maize fields

A 5‐year field survey examined western corn rootworm (WCR) (Diabrotica virgifera virgifera LeConte) beetle density in Serbia from 2002 to 2006. First‐, second‐, third‐, fourth‐ and fifth‐year maize fields were sampled; they represented 64.61%, 21.66%, 9.45%, 3.53% and 0.75% of all sampled fields respectively. Results showed that the mean WCR beetle population density from 794 maize fields differed depending on cropping history. Minimum mean WCR/trap/day was 0.0 in the first‐year maize fields in 2002 and 2006. Maximum mean WCR/trap/day was registered in the fourth‐year and the fifth‐year maize fields (27.8 and 21.2 respectively). Mean population density of WCR adults increased with the number of years of continuous maize from 1.17, 4.61, 6.41, 10.30 up to 13.53 WCR/trap/day for first‐fifth‐year maize fields respectively. Mean WCR/trap/day ± SE exceeded the economic population threshold of >6 WCR/trap/day in third‐year continuous maize fields. Out of 794 maize fields, 697 (87.78%) registered a mean population density below the <6 beetles/trap/day threshold. In only 97 fields was WCR population density >6 beetles/trap/day, a finding that predicts a risk of economic damage to a subsequent maize planting. These data are representative of the Serbian situation from 2002 to 2006; they indicate that WCR are well dispersed across commercial maize fields in Serbia. These results provide new insight into the current low WCR population densities in maize fields managed by crop rotation, a finding that can help in creating long‐term management strategy.     

The ineffectiveness of insecticide seed coatings and planting-time soil insecticides as Diabrotica virgifera virgifera LeConte population suppressors

Abstract:  The effect of any management strategy on pest population levels must be researched and determinations need to be made as to how that strategy might work based on the control objectives. In certain areas of Europe, the objective is to contain or eradicate the western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, population. In order to evaluate the impact of insecticide seed coatings and/or planting-time applications of insecticides as WCR population suppressors, plot trials and large field observations were carried out in Italy over a 5-year period. Larval, pupal and adult densities, along with root damage ratings, were estimated at different locations. Data from these studies revealed that the number of WCR adults emerging from untreated plots did not differ from the number of beetles emerging from those treated with insecticides, whether as seed coating or in-furrow applications. Both seed insecticide coatings (imidacloprid, fipronil, thiamethoxam, tefluthrin) and soil insecticides applied in-furrow (chlorpyrifos, diazinon, tefluthrin) did not reduce the number of beetles emerging from monoculture fields, either in plot trials or large field observations. Observations in the USA had previously shown that soil insecticides applied at planting time partially protected basal roots from economic damage, but did not reduce corn rootworm populations. Similarly, in Europe, it has been demonstrated that not only the application of soil insecticides at planting time but also insecticide seed coatings have no role in the containment and/or eradication of WCR. Although insecticide seed coatings and soil insecticides applied in-furrow may provide protection against economic damage to roots, these management strategies do not have an impact on WCR populations and therefore are useless in WCR containment and eradication programmes.     

Miscanthus,a host for larvae of a European population of Diabrotica v. virgifera

The EU policies for decreasing reliance on fossil fuels and reducing greenhouse gas emissions present challenging targets for bioenergy production. Field trials in Europe of Miscanthus×giganteus (Poales: Poaceae) over the past 20 years have shown that this plant is suitable for the production of biomass but the economic models promoting its use for this purpose often assume that controlling its pests will cost little or nothing. Laboratory experiments were conducted to determine whether this plant is suitable for the development of larvae of the western corn rootworm (WCR), Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae), which is an important pest of maize. Larvae of WCR, originating from a Central and South Eastern European population, can develop on Miscanthus.     

International cooperation on western corn rootworm ecology research: state-of-the-art and future research

1 Invasive pest species are challenging partly because the invasion process may be highly dynamic and because of the lack of knowledge of many researchers, professionals and farmers in the newly-invaded regions. The chrysomelid Diabrotica virgifera virgifera LeConte is such an invasive pest. It has been the main pest of continuous maize in the U.S.A. for more than 60 years and is currently spreading throughout Europe.
2 In the area with a long history of this pest (Central and North America), scientific knowledge concerning the ecology of this pest has accumulated over the last decades. This resource is of great importance to both America and Europe and has to be gathered, shared and adapted to new situations. We therefore examined, both qualitatively and quantitatively, the scientific literature relating to D.   virgifera virgifera ecology.
3 The quantitative analysis suggests that research on D.   virgifera virgifera ecology is still in its infancy in Europe and suffers from geographical barriers (between Europe and North America and between linguistic areas within Europe) and that scientific communication should be strengthened both between North America and Europe and within Europe.
4 As a first solution to this problem, we introduce three companion review articles that constitute a landmark for D.   virgifera virgifera research, enabling European and American scientists and decision-makers to orient themselves and discover new opportunities for research. We also stress that international research cooperation is the most important key to successfully manage invasive species.     

Biology of the European large raspberry aphid (Amphorophora idaei): its role in virus transmission and resistance breakdown in red raspberry

1 The European large raspberry aphid Amphorophora idaei Börner is the most important vector of viral diseases afflicting commercially grown red raspberry ( Rubus idaeus L.) in Northern Europe, with European raspberry production amounting to 416 000 tonnes per annum. This review synthesizes existing knowledge on its biology and interactions with other organisms, including its host plant and the viral pathogens it vectors.
2 Information about trophic interactions with other insect herbivores and natural enemies is reviewed. Vine weevils Otiorhynchus sulcatus compromise aphid resistance in some raspberry cultivars, increasing A.   idaei abundance by 80%. Parasitoids show mixed success in parasitizing A.   idaei , although Aphidius ervi attack rates more than doubled when A.   idaei fed on a partially susceptible raspberry cultivar, compared with a resistant variety. These findings are discussed in the context of potential biological control as part of an integrated pest and disease management framework.
3  Amphorophora idaei transmits four known viruses: Black raspberry necrosis virus, Raspberry leaf mottle virus, Raspberry leaf spot virus and Rubus yellow net virus , with A.   idaei taking as little as 2 min to transmit some viruses.
4 Existing control strategies, including resistant cultivars, insecticides and eradication of disease from parent plants, are described. In particular, strong selection pressures have resulted in A .  idaei overcoming genetic resistance in many raspberry cultivars and most insecticides are now ineffective.
5 Future directions for the sustained control of A.   idaei are suggested, taking into consideration the possible effects of climate change and also changes in agronomic practices in U.K. agriculture.     

Western corn rootworm adult movement and possible egg laying in fields bordering maize

Abstract:  Western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, larval damage in maize following soybean was observed in Croatia in 2003 along the edges and within soybean fields which bordered continuous maize fields in previous year. The explanation was that WCR adults moved from the continuous maize to the neighbouring soybean fields to lay eggs. This study was designed to measure how far WCR adults will enter into neighbouring fields to lay eggs. The WCR adult population was monitored in continuous maize fields in 2003 and 2005 by using Pherocon^® AM non-baited yellow sticky traps in the middle and on the borders of the maize field and at different distances and directions into neighbouring fields planted by wheat and soybean. Larval presence and root damage ratings (Iowa State University 1–6 ) were recorded at different locations within the maize field in following years. Approximately, the same concentration of WCR adults was recorded along the edges of the maize fields as recorded in the centres of those fields. A significant number of WCR adults was recorded up to a distance of 50 m into neighbouring fields. Regression analysis showed medium negative correlation between distance from previous maize field and root damage in the following year. Findings indicate that WCR egg lying can reach approximately 20 m into fields neighbouring maize fields and that significant root damage caused by WCR larvae in first-year maize following soybean and wheat can happen up to a distance of 20 m into those fields. Most farmers's fields in Croatia are up to approximately 50 m wide. As an edge effect for WCR egg laying can reach approximately 20 m into fields neighbouring maize fields, our research results indicate that it is possible to see WCR larval damage in rotated fields without those WCR's being the variant form.     

Systemic RNAi in western corn rootworm,Diabrotica virgifera virgifera,does not involve transitive pathways

Western corn rootworm (WCR, Diabrotica virgifera virgifera LeConte) is highly sensitive to orally delivered double‐stranded RNA (dsRNA). RNAi in WCR is systemic and spreads throughout the insect body. This raises the question whether transitive RNAi is a mechanism that functions in WCR to amplify the RNAi response via production of secondary siRNA. Secondary siRNA production is achieved through RNA‐dependent RNA polymerase (RdRP) activity in other eukaryotic organisms, but RdRP has not been identified in WCR and any other insects. This study visualized the spread of the RNAi‐mediated knockdown of Dv v‐ATPase C mRNA throughout the WCR gut and other tissues using high‐sensitivity branched DNA in situ hybridization. Furthermore, we did not detect either secondary siRNA production or transitive RNAi in WCR through siRNA sequence profile analysis. Nucleotide mismatched sequences introduced into either the sense or antisense strand of v‐ATPase C dsRNAs were maintained in siRNAs derived from WCR fed with the mismatched dsRNAs in a strand specific manner. The distribution of all siRNAs was restricted to within the original target sequence regions, which may indicate the lack of new dsRNA synthesis leading to production of secondary siRNA. Thus, the systemic spread of RNAi in WCR may be derived from the original dsRNA molecules taken up from the gut lumen. These results indicate that the initial dsRNA dose is important for a lethal systemic RNAi response in WCR and have implications in developing effective dsRNA traits to control WCR and in resistance management to prolong the durability of RNAi trait technology.     

Characterization of the spectrum of insecticidal activity of a double-stranded RNA with targeted activity against Western Corn Rootworm (Diabrotica virgifera virgifera LeConte)

The sequence specificity of the endogenous RNA interference pathway allows targeted suppression of genes essential for insect survival and enables the development of durable and efficacious insecticidal products having a low likelihood to adversely impact non-target organisms. The spectrum of insecticidal activity of a 240 nucleotide (nt) dsRNA targeting the Snf7 ortholog in Western Corn Rootworm (WCR; Diabrotica virgifera virgifera) was characterized by selecting and testing insects based upon their phylogenetic relatedness to WCR. Insect species, representing 10 families and 4 Orders, were evaluated in subchronic or chronic diet bioassays that measured potential lethal and sublethal effects. When a specific species could not be tested in diet bioassays, the ortholog to the WCR Snf7 gene (DvSnf7) was cloned and corresponding dsRNAs were tested against WCR and Colorado potato beetle (Leptinotarsa decemlineata); model systems known to be sensitive to ingested dsRNA. Bioassay results demonstrate that the spectrum of activity for DvSnf7 is narrow and activity is only evident in a subset of beetles within the Galerucinae subfamily of Chrysomelidae (>90 % identity with WCR Snf7 240 nt). This approach allowed for evaluating the relationship between minimum shared nt sequence length and activity. A shared sequence length of ≥21 nt was required for efficacy against WCR (containing 221 potential 21-nt matches) and all active orthologs contained at least three 21 nt matches. These results also suggest that WCR resistance to DvSnf7 dsRNA due to single nucleotide polymorphisms in the target sequence of 240 nt is highly unlikely.     

Diabrotica virgifera virgifera LeConte: inconspicuous leaf beetle--formidable challenges to agriculture

In the universe of entomology with its close to one million described and an estimated ten million undescribed species, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysometidae), the Western corn rootworm (WCR), is one of the economically most important pests. It causes annual treatment costs and damages of a billion dollars in the US (Metcalf 1986). Similar costs are predicted for Europe which the beetle invaded 15 years ago. Due to lack of natural enemies it is now expanding its territory at a rapid rate. With prior experience gained in Illinois, USA, and subsequent largely unsuccessful efforts to stop the pest in Europe, eradication efforts, although attempted in France and in the Veneto region of northern Italy, are not a viable sustainable strategy for the future nor a long term solution. Crop rotation, so far one of the best cultural management options of practicing entomologists, is beginning to show weaknesses specificly on Glycine max (Shaw et al. 1978, Levine et al. 2002, Spencer et al. 2005, Kiss et al. 2005, Tollefson and Prasifka 2006). WCR also was discovered on alternative hosts such as Cucurbita pepo in Slovenia (Hummel et al. 2007a, 2007b), C.maxima in Hungary (Moeser and Vidal 2001), and Serbia (Baca 1993, Baca and Berger 1994, Baca unpublished 1998, Baca 2007). WCR thus turns out to remain a challenge to general entomolog'sts, agricultural and ecological entomologists, geneticists, epidemiologists, crop protection engineers, phytosanitary services and economists alike. WCR and H. sapiens today are reaching a labile ecological equilibrium of coexistence, with new surprises on both sides in the "arms race" just waiting around the corner. Most experts will agree that WCR is in Europe to stay and is readily expanding from here to Asia and Africa if given the slightest chance.- The really unsolved question remains how to manage WCR sensibly, economically, and sustainably, and how and where to train a new generation of capable, reliable, all round entomological pest managers needed for the future.     

Characterizing the Mechanism of Action of Double-Stranded RNA Activity against Western Corn Rootworm (Diabrotica virgifera virgifera LeConte)

RNA interference (RNAi) has previously been shown to be effective in western corn rootworm (WCR, Diabrotica virgifera virgifera LeConte) larvae via oral delivery of synthetic double-stranded RNA (dsRNA) in an artificial diet bioassay, as well as by ingestion of transgenic corn plant tissues engineered to express dsRNA. Although the RNAi machinery components appear to be conserved in Coleopteran insects, the key steps in this process have not been reported for WCR. Here we characterized the sequence of events that result in mortality after ingestion of a dsRNA designed against WCR larvae. We selected the Snf7 ortholog (DvSnf7) as the target mRNA, which encodes an essential protein involved in intracellular trafficking. Our results showed that dsRNAs greater than or equal to approximately 60 base-pairs (bp) are required for biological activity in artificial diet bioassays. Additionally, 240 bp dsRNAs containing a single 21 bp match to the target sequence were also efficacious, whereas 21 bp short interfering (si) RNAs matching the target sequence were not. This result was further investigated in WCR midgut tissues: uptake of 240 bp dsRNA was evident in WCR midgut cells while a 21 bp siRNA was not, supporting the size-activity relationship established in diet bioassays. DvSnf7 suppression was observed in a time-dependent manner with suppression at the mRNA level preceding suppression at the protein level when a 240 bp dsRNA was fed to WCR larvae. DvSnf7 suppression was shown to spread to tissues beyond the midgut within 24 h after dsRNA ingestion. These events (dsRNA uptake, target mRNA and protein suppression, systemic spreading, growth inhibition and eventual mortality) comprise the overall mechanism of action by which DvSnf7 dsRNA affects WCR via oral delivery and provides insights as to how targeted dsRNAs in general are active against insects.     

Refuge or Reservoir? The Potential Impacts of the Biofuel Crop Miscanthus x giganteus on a Major Pest of Maize

Background

Interest in the cultivation of biomass crops like the C4 grass Miscanthus x giganteus (Miscanthus) is increasing as global demand for biofuel grows. In the US, Miscanthus is promoted as a crop well-suited to the Corn Belt where it could be cultivated on marginal land interposed with maize and soybean. Interactions (direct and indirect) of Miscanthus, maize, and the major Corn Belt pest of maize, the western corn rootworm, (Diabrotica virgifera virgifera LeConte, WCR) are unknown. Adding a perennial grass/biomass crop to this system is concerning since WCR is adapted to the continuous availability of its grass host, maize (Zea mays).

Methodology/Principal Findings

In a greenhouse and field study, we investigated WCR development and oviposition on Miscanthus. The suitability of Miscanthus for WCR development varied across different WCR populations. Data trends indicate that WCR populations that express behavioural resistance to crop rotation performed as well on Miscanthus as on maize. Over the entire study, total adult WCR emergence from Miscanthus (212 WCR) was 29.6% of that from maize (717 WCR). Adult dry weight was 75–80% that of WCR from maize; female emergence patterns on Miscanthus were similar to females developing on maize. There was no difference in the mean no. of WCR eggs laid at the base of Miscanthus and maize in the field.

Conclusions/Significance

Field oviposition and significant WCR emergence from Miscanthus raises many questions about the nature of likely interactions between Miscanthus, maize and WCR and the potential for Miscanthus to act as a refuge or reservoir for Corn Belt WCR. Responsible consideration of the benefits and risks associated with Corn Belt Miscanthus are critical to protecting an agroecosystem that we depend on for food, feed, and increasingly, fuel. Implications for European agroecosystems in which Miscanthus is being proposed are also discussed in light of the WCR''s recent invasion into Europe.     

Evidence of Field-Evolved Resistance to Bifenthrin in Western Corn Rootworm (Diabrotica virgifera virgifera LeConte) Populations in Western Nebraska and Kansas

Pyrethroid insecticides have been used to control larvae or adults of the western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, a key pest of field corn in the United States. In response to reports of reduced efficacy of pyrethroids in WCR management programs in southwestern areas of Nebraska and Kansas the present research was designed to establish a baseline of susceptibility to the pyrethroid insecticide, bifenthrin, using susceptible laboratory populations and to compare this baseline with susceptibility of field populations. Concentration-response bioassays were performed to estimate the baseline susceptibility. From the baseline data, a diagnostic concentration (LC₉₉) was determined and used to test adults of both laboratory and field populations. Larval susceptibility was also tested using both laboratory and field populations. Significant differences were recorded in adult and larval susceptibility among WCR field and laboratory populations. The highest LC₅₀ for WCR adults was observed in populations from Keith 2 and Chase Counties, NE, with LC₅₀s of 2.2 and 1.38 μg/vial, respectively, and Finney County 1, KS, with 1.43 μg/vial, as compared to a laboratory non-diapause population (0.24 μg/vial). For larvae, significant differences between WCR field and laboratory populations were also recorded. Significant differences in mortalities at the diagnostic bifenthrin concentration (LC₉₉) were observed among WCR adult populations with western Corn Belt populations exhibiting lower susceptibility to bifenthrin, especially in southwestern Nebraska and southwestern Kansas. This study provides evidence that resistance to bifenthrin is evolving in field populations that have been exposed for multiple years to pyrethroid insecticides. Implications to sustainable rootworm management are discussed.     

Selection of entomopathogenic fungi for the control of the western corn rootworm Diabrotica virgifera virgifera

Abstract:  The western corn rootworm Diabrotica virgifera virgifera Le Conte (Col., Chrysomelidae), a serious pest of maize, has been recently introduced into Europe. Several approaches for its control are presently under investigation including microbial agents. During a field survey in Hungary in 2005, naturally occurring entomopathogenic fungi were found to attack this pest. These novel isolates together with standard isolates were tested for virulence against D. v. virgifera larvae and adults. Twenty strains of Metarhizium anisopliae , Beauveria bassiana and Beauveria brongniartii were used in bioassays in the laboratory. Larvae and adults were dipped into a spore suspension with a concentration of 1 × 10⁷ conidia (con.)/ml. They were kept for 14 days at 22°C (±2°C) and 70% relative humidity. The number of infected larvae and adults were counted and infection rates were calculated. Adults were significantly more susceptible to entomopathogenic fungi than larvae. The most virulent isolate infected about 47% of larvae ( M. anisopliae Ma2277), whereas the infection rate in adults was up to 97% ( M. anisopliae Ma2275). Isolates of M. anisopliae caused significantly higher mortalities than isolates of B. brongniartii and B. bassiana . Most of the adult beetles were killed within 12 days. Isolates from D. v. virgifera were more virulent than those from other hosts.