Polymorphic microsatellite loci from the western corn rootworm (Insecta: Coleoptera: Chrysomelidae) and cross‐amplification with other Diabrotica spp

Corn rootworms (Diabrotica spp.) make up the major insect pest complex of corn in the US and Europe, and there is a need for molecular markers for genetics studies. We used an enrichment strategy to develop microsatellite markers from the western corn rootworm (Diabrotica virgifera virgifera). Of 54 loci isolated, 25 were polymorphic, and of these, 17 were surveyed for variability in 59 wild individuals. In addition, the potential for cross‐amplification of these microsatellites was surveyed for Mexican, northern, and southern corn rootworms. Nine microsatellite loci showed Mendelian inheritance and are likely to be useful in population genetics studies.     

Corn rootworm survey in North Dakota and a comparison of two sticky traps

Northern, Diabrotica barberi Smith & Lawrence, and western, D. virgifera virgifera LeConte, corn rootworms are major economic pests of corn, Zea mays L., in the United States. This research was conducted to determine the geographic distribution, abundance, and species composition of Diabrotica species in North Dakota, and to compare effectiveness of unbaited green Scentry™ Multigard and unbaited yellow Pherocon^® AM/NB sticky traps for monitoring. Fifty-one corn fields were monitored using traps from July through October of the 2013, 2014, and 2015 growing seasons for rootworm beetle activity. The overall species composition was 61% D. barberi and 39% D. v. virgifera. Both species were frequently captured, and the highest densities (i.e. >10 beetles per trap per week) were found in southeastern North Dakota. Low densities (i.e. <0.1 beetles per trap per week) of D. barberi were found in areas further north, but no D. v. virgifera were captured in those fields. The two different coloured sticky traps were not significantly different across 38 sites for D. barberi and across 21 sites for D. v. virgifera. However, green Scentry™ Multigard traps captured more D. barberi beetles than yellow Pherocon^® AM/NB traps at 68% of the 38 fields. In contrast, the yellow Pherocon^® AM/NB traps captured more D. v. virgifera beetles at 57% of the 21 fields. Findings also indicated that, although D. barberi was the predominant species in surveyed fields, populations rarely reached the economic threshold. Our study observed that economic populations of corn rootworms were infrequent among the field sites trapped in North Dakota. As a result, producers should scout fields regularly for corn rootworm populations levels to make sound pest management decisions. This knowledge can enable producers to effectively protect their crop when control is economically justified, and the information can also provide input cost savings when populations do not warrant control efforts.     

Volatile semiochemical impact on trapping and distribution in maize of northern and western corn rootworm beetles (Coleoptera: Chrysomelidae)

Abstract 1 Field studies evaluated plant attractants and analogues as tools to move corn rootworm beetles (Diabrotica spp.) into areas to be treated with toxic baits for population suppression via mass removal/annihilation of reproductive adults. 2 When dispensed from sticky traps in maize, 2‐phenyl‐1‐ethylamine and 2‐phenyl‐1‐ethanol captured more northern corn rootworm, Diabrotica barberi, than did 4‐methoxyphenethanol. Only 2‐phenyl‐1‐ethanol attracted the western corn rootworm, Diabrotica virgifera virgifera, but not until maize matured beyond milk stage. 3 Attraction of D. barberi to the amine, alone or blended with 2‐phenyl‐1‐ethanol, occurred before and after maize flowered but not during intervening silk or blister stages. Attraction recurred during early milk stage at or before 50% emergence of adult female D. barberi or D. v. virgifera, respectively, and before populations declined for the season. 4 Synergistic interaction of 2‐phenyl‐1‐ethylamine with 2‐phenyl‐1‐ethanol in attracting D. barberi females did not occur until maize matured to late milk stage. 5 The amine‐alcohol blend (0.44 point sources m^?2) doubled the density of D. barberi but not D. v. virgifera when applied to small plots within mostly milk‐stage or younger maize. Traps without bait within attractant‐treated plots captured more female, but not male, D. barberi than did traps in untreated control plots, hinting that females accounted for most of the observed increase in beetle density. 6 The results suggest that attractants can be used despite phenological limitations to concentrate preovipositional females within field areas and thus to complement a variety of corn rootworm control strategies.     

Comparative efficacy assessment of fungi,nematodes and insecticides to control western corn rootworm larvae in maize

All three larval instars of Diabrotica virgifera virgifera LeConte (western corn rootworm, Coleoptera: Chrysomelidae) feed on the roots of maize, Zea mays (L.). We assessed the efficacies of the following four agents in controlling these larvae: (1) the entomopathogenic fungus Metarhizium anisopliae (Metsch.) Sorokin (Hypocreales: Clavicipitaceae), (2) the nematode Heterorhabditis bacteriophora Poinar (Nematoda: Rhabditida), (3) a tefluthrin-based soil insecticide and (4) clothianidin-coated seeds. The agents were applied in field plot experiments in southern Hungary in 2006 and 2007. Efficacy was assessed by comparing the number of emerging D. v. virgifera adults and corresponding root damage among treatments and untreated controls. All agents significantly reduced D. v. virgifera numbers and root damage, but the relative success of each treatment was variable. On average across fields and years, the nematode and the two insecticides reduced D. v. virgifera by 65 ± 34% SD, while the fungus reduced D. v. virgifera by 31 ± 7%. According to the node injury scale, the agents prevented 23–95% of potential root damage. Large-scale commercialisation of these biological agents could offer viable and practical control options against D. v. virgifera.     

Evaluating Tripsacum‐introgressed maize germplasm after infestation with western corn rootworms (Coleoptera: Chrysomelidae)

Maize (Zea mays L.) is a valuable commodity throughout the world, but corn rootworms (Chrysomelidae: Diabrotica spp.) often cause economic damage and increase production costs. Current rootworm management strategies have limitations, and in order to create viable management alternatives, researchers have been developing novel maize lines using Eastern gamagrass (Tripsacum dactyloides L.) germplasm, a wild relative of maize that is resistant to rootworms. Ten maize Tripsacum‐introgressed inbred lines derived from recurrent selection of crosses with gamagrass and teosinte (Zea diploperennis Iltis) recombinants and two public inbred lines were assessed for susceptibility to western corn rootworm (Diabrotica virgifera virgifera LeConte) and yield in a two‐year field study. Two experimental maize inbred lines, SDG11 and SDG20, had mean root damage ratings that were significantly lower than the susceptible public line B73. Two other experimental maize inbred lines, SDG12 and SDG6, appeared tolerant to rootworm damage because they exhibited yield increases after rootworm infestation in both years. In the majority of cases, mean yield per plant of experimental maize lines used in yield analyses was equal to or exceeded that of the public inbred lines B73 and W64A. Our study indicates that there is potential to use Tripsacum‐introgressed maize germplasm in breeding programs to enhance plant resistance and/or tolerance to corn rootworms, although further research on insect resistance and agronomic potential of this germplasm needs to be conducted in F₁ hybrids.     

A review of the natural enemies of beetles in the subtribe Diabroticina (Coleoptera: Chrysomelidae): implications for sustainable pest management

Diabroticina is a speciose subtribe of New World Chrysomelidae (Subfamily Galerucinae: Tribe Luperini) that includes pests such as corn rootworms, cucumber beetles and bean leaf beetles (e.g. Diabrotica, Acalymma, Cerotoma species). The evolution and spread of pesticide resistance, the European invasion of Diabrotica v. virgifera LeConte, and possible development of resistance due to the large-scale deployment of Diabrotica-active Bt maize in North America have generated a sense of urgency in developing biological control options against Diabroticina pests. In the present study, we review available knowledge on biological control options, including 290 publications on natural enemy–Diabroticina associations in the New World. Several natural enemy species or groups appear to be promising candidates for control strategies with different ecological rationales. We propose that future research should pursue: (1) development of inundative biological control products, particularly mass-produced entomopathogenic nematodes and fungi, (2) understanding of specific natural enemies of Diabroticina larvae throughout the Americas and of adults particularly in higher altitudes of Central America or northern South America including potential classical biological control agents against D. v. virgifera; (3) enhancement of natural enemies through cultural practices, i.e., reduced tillage, reduced weed control, cover crops, diversified crop rotations or soil amendments. Research and action must be coordinated to accelerate the exploration of biological control options.     

Assessing larval rootworm behaviour after contacting maize roots: impact of germplasm,rootworm species and diapause status

Current methods of screening maize (Zea mays L.) germplasm for susceptibility or resistance to corn rootworms (Coleoptera: Chrysomelidae) rely primarily on information from large‐scale field experiments. Due to labour and cost constraints associated with field trials, alternative evaluation methods are desirable. We used a previously developed behavioural bioassay to: (1) investigate the host search behaviour of rootworm larvae after contact with 14 maize genotypes, (2) compare the behaviour of non‐diapausing Diabrotica virgifera virgifera LeConte, diapausing D. v. virgifera, and diapausing D. barberi Smith & Lawrence and (3) determine if this technique can be used to separate susceptible vs. resistant maize genotypes. The majority of rootworm larvae engaged in intensive (local search) behaviour after exposure to maize roots, whereas larvae continued to exhibit extensive (ranging) behaviour after contact with negative controls. Even though a transgenic hybrid with resistance to D. v. virgifera was included in analyses, quantitative path measurements were similar among genotypes and only differed between specific maize lines and controls. Notably, there were differences in host search behaviour among rootworm groups, with non‐diapausing D. v. virgifera having more convoluted paths and engaging in intensive search more frequently than diapausing rootworms. Correlations between larval path measurements and historic root damage ratings were not significant, although there were weak positive correlations between historic adult emergence densities and measures of path linearity. However, due to the lack of significant behavioural differences among maize lines with a range of susceptibility levels, we concluded that this bioassay is not useful in screening maize germplasm for rootworm resistance.     

Supercooling and cold-hardiness in eggs of western and northern corn rootworms

Oviposition by northern corn rootworms, Diabrotica barberi Smith and Lawrence, and western corn rootworms, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), key pests of corn in the Great Plains of the USA, occurs in the soil during late summer. Overwintering eggs are exposed to variable soil moisture and temperatures below ?5 °C. The winter mortality of eggs in the soil is a primary factor that determines the potential for larval injury to corn the following spring. Our studies aimed to determine the comparative supercooling capacities of northern and western corn rootworm eggs and to assess egg mortality following brief exposure to extreme low temperature, ranging from ?12.0 to ?21.5 °C, under three moisture regimes. Eggs of northern corn rootworm were supercooled to a temperature as low as ?27 °C, and survived supercooling to a greater extent than did western corn rootworm eggs. Moisture treatment prior to supercooling had little effect on northern corn rootworm eggs. Western corn rootworm eggs were more resistant than northern corn rootworm eggs to the effects of desiccation followed by supercooling. The survival of northern corn rootworm eggs was better than western corn rootworms under dry conditions, followed by exposure to temperatures of ?12.0 and ?17.5 °C, but was very low at ?21.5 °C, regardless of the moisture regime. The results suggest that moisture and temperature may interact in the soil environment to determine the overwintering survival of corn rootworms. It is evident from these studies that both rootworm species experience mortality at temperatures well above the supercooling points of the eggs, but that differences exist in the effects of substrate moisture treatments on the cold‐hardiness of eggs from the two species.     

Comparative susceptibility of larval instars and pupae of the western corn rootworm to infection by three entomopathogenic nematodes

As a first step towards the development of an ecologically rational control strategy against western corn rootworm (WCR; Diabrotica virgifera virgifera LeConte, Coleoptera: Chrysomelidae) in Europe, we compared the susceptibility of the soil living larvae and pupae of this maize pest to infection by three entomopathogenic nematode (EPN) species. In laboratory assays using sand-filled trays, Heterorhabditis bacteriophora Poinar and H. megidis Poinar, Jackson & Klein (both Rhabditida: Heterorhabditidae) caused comparable mortality among all three larval instars and pupae of D. v. virgifera. In soil-filled trays, H. bacteriophora was slightly more effective against third larval instars and pupae, and H. megidis against third larval instars, compared to other developmental stages. In both sand and soil, Steinernema feltiae (Filipjev) (Rh.: Steinernematidae) was least effective against second instars. In conclusion, all larval instars of D. v. virgifera show susceptibility to infection by all three nematodes tested. It is predicted that early application against young larval instars would be most effective at preventing root feeding damage by D. v. virgifera. Applications of nematodes just before or during the time period when third instars are predominant in the field are likely to increase control efficacy. According to our laboratory assays, H. bacteriophora and H. megidis appear to be the most promising candidates for testing in the field. I. Hiltpold similarly contributed to this paper as the first author.     

A selective insecticidal protein from Pseudomonas mosselii for corn rootworm control

The coleopteran insect western corn rootworm (WCR, Diabrotica virgifera virgifera) is an economically important pest in North America and Europe. Transgenic corn plants producing Bacillus thuringiensis (Bt) insecticidal proteins have been useful against this devastating pest, but evolution of resistance has reduced their efficacy. Here, we report the discovery of a novel insecticidal protein, PIP‐47Aa, from an isolate of Pseudomonas mosselii. PIP‐47Aa sequence shows no shared motifs, domains or signatures with other known proteins. Recombinant PIP‐47Aa kills WCR, two other corn rootworm pests (Diabrotica barberi and Diabrotica undecimpunctata howardi) and two other beetle species (Diabrotica speciosa and Phyllotreta cruciferae), but it was not toxic to the spotted lady beetle (Coleomegilla maculata) or seven species of Lepidoptera and Hemiptera. Transgenic corn plants expressing PIP‐47Aa show significant protection from root damage by WCR. PIP‐47Aa kills a WCR strain resistant to mCry3A and does not share rootworm midgut binding sites with mCry3A or AfIP‐1A/1B from Alcaligenes that acts like Cry34Ab1/Cry35Ab1. Our results indicate that PIP‐47Aa is a novel insecticidal protein for controlling the corn rootworm pests.     

Natural occurrence of insect pathogenic fungi and insect parasitic nematodes in <Emphasis Type="Italic">Diabrotica virgifera virgifera</Emphasis> populations

The Western Corn Rootworm D. virgifera virgifera Le Conte (Coleoptera: Chrysomelidae), a serious pest of maize, has been recently introduced into Europe. Several approaches for its control are presently under investigation including microbial agents. In order to get information on the role of naturally occurring pathogens in the regulation of Diabrotica populations, we started an investigation in established populations in Hungary, Romania, Serbia, Austria, and Italy in 2005 and 2006. In infested maize fields in Hungary, plants and their root systems were grubbed out and larvae and pupae were collected. Adult D. v. virgifera were collected in Hungary, Austria, Romania, Serbia and Italy. Additionally, the occurrence of entomopathogenic fungi in soils of maize fields was determined using Galleria mellonella and Tenebrio molitor larvae as bait insects. The density of entomopathogenic fungi was obtained by plating soil suspension on selective medium. Metarhizium anisopliae and Beauveria spp. infections were found in 1.4% of field collected larvae, 0.2% of field collected pupae and 0.05% of field collected adults. Whereas natural infections of D. v. virgifera were rarely found, a high density of insect pathogenic fungi was recorded in Hungarian soils. M. anisopliae could be detected in every maize field either using the “bait method” or a “selective medium” method. This is the first report of a natural occurrence of entomoparasitic nematodes (Heterorhabditis sp., Steinernema sp.) in Diabrotica v. virgifera in Europe.     

European populations of Diabrotica virgifera virgifera are resistant to aldrin,but not to methyl‐parathion

The western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is a major pest of cultivated corn in North America and has recently begun to invade Europe. In addition to crop rotation, chemical control is an important option for D. v. virgifera management. However, resistance to chemical insecticides has evolved repeatedly in the USA. In Europe, chemical control strategies have yet to be harmonized and no surveys of insecticide resistance have been carried out. We investigated the resistance to methyl‐parathion and aldrin of samples from nine D. v. virgifera field populations originating from two European outbreaks thought to have originated from two independent introductions from North America. Diagnostic concentration bioassays revealed that all nine D. v. virgifera field populations were resistant to aldrin but susceptible to methyl‐parathion. Aldrin resistance was probably introduced independently, at least twice, from North America into Europe, as there is no evident selection pressure to account for an increase of frequency of aldrin resistance in each of the invasive outbreaks in Europe. Our results suggest that organophosphates, such as methyl‐parathion, may still provide effective control of both larval and adult D. v. virgifera in the European invasive outbreaks studied.     

A critical evaluation of host ranges of parasitoids of the subtribe Diabroticina (Coleoptera: Chrysomelidae: Galerucinae: Luperini) using field and laboratory host records

The subtribe Diabroticina is a large group of New World Chrysomelidae that includes corn rootworms, cucumber beetles and other pests. Recent introductions of Diabrotica virgifera virgifera LeConte into Europe, and the development of resistances to current management practices of rootworms in the USA have increased interest in new sustainable options for managing those pests. The only parasitoids that have been shown to consistently target and develop inside the beetle adults are Centistes gasseni Shaw, Centistes diabroticae Gahan (both Hym.: Braconidae), and Celatoria diabroticae Shimer, Celatoria compressa (Wulp), Celatoria bosqi Blanchard, and Celatoria setosa Coquillett (all Diptera: Tachinidae). This review improves our understanding of the realised and potential host range of these known parasitoids by rectifying erroneous references in light of new host records and from laboratory host range tests. Based on this critical review, all tachinid and braconid species studied are considered to be specific at least to the level of subtribe, i.e. Diabroticina. Celatoria setosa, Celatoria diabroticae and C. bosqi, have a narrow realized and potential host range; the former is restricted to the genus Acalymma and the last two to the fucata and virgifera groups of the genus Diabrotica. The braconids Centistes gasseni and C. diabroticae are also specific. The realized host range of C. gasseni includes species in the Diabrotica fucata and virgifera groups; while its potential host range also includes Acalymma species. The realized and potential host range of Centistes diabroticae includes Acalymma species as well as species in the fucata and virgifera groups of Diabrotica. Celatoria compressa has the broadest realised range compared to the other species studied, since it was obtained from species in several genera of Diabroticina; and its potential host range may also include Old World Aulacophora species.     

Influence of adult diet on the reproductive biology and survival of the western corn rootworm,Diabrotica virgifera virgifera

Adult corn rootworm beetles,Diabrotica virgifera virgifera LeConte, were maintained on three dietary regimes which mimicked the progression of corn tissues that would be available in the field for beetles eclosing when corn was in the following growth stages: (A) after tassels had emerged but prior to silking and pollen shed; (B) while plants were silking and shedding pollen; and (C) after pollination was complete and silks had turned brown. A fourth regime (D) was established in which green corn silks, pollen, and leaves were provided throughout the study. The mean number of eggs laid per female over the 12-week duration of the study was 125, 235, 179, and 441 for regimes A, B, C, and D, respectively. Median length of life was 7.2, 7.2, 6.7, and 8.8 weeks for regimes A, B, C, and D respectively. Beetles in regime A laid a greater proportion of their eggs at an older age than did beetles from regimes B and C. To further investigate the influences on survival of changes in food quality of corn as plants and beetles aged, newly-eclosed beetles and beetles that had been maintained in caged plots of corn growing in a greenhouse for various lengths of time were caged on corn at different stages of growth, and the proportion of beetles surviving for 48 h was determined. Survival decreased as plants aged for both groups of beetles, but decreased at a faster rate for old than for young beetles.

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Influence de l'alimenation sur la reproduction et la survíe deDiabrotica virgifera virgifera

Résumé D. virgifera virgifera LeConte a été placé sur des régimes nutritifs mimant l'évolution des tissus du maïs disponsibles dans le champ lorsque les insectes éclosent est aux stades suivants du maïs: 1) après émergence de l'épi mâle, mais avant l'apparition des barbes et l'émission du pollen; 2) lors de la présence de barbes et de l'émission de pollen; 3) après la pollinisation et quand les barbes ont bruni. Un quatrième régime a été fourni pendant toute l'expérience comprenant des barbes vertes, du pollen et des feuilles. Les pontes moyennes des femelles pendant les 12 semaines de l'étude on été 125, 235, 179 et 441 pour les régimes 1, 2, 3 et 4. Les dates de mort de la moitié des adultes a été 7,2, 7,2, 6,7 et 8,8 semaines pour respectivement les mêmes régimes. Les femelles du régime 1 pondent à un âge plus avancé que les femelles des régimes 2 et 3. L'influence des changements de la qualité alimentaire du maïs au fur et à mesure du vieillissement des plantes et deDiabrotica, a été déterminée par la survie au bout de 48 heures d'insectes éclos depuis peu et d'autres maintenus en cage sur des lots de maïs à différents stades poussant en serre depuis des temps plus ou moins longs. Plus les plantes sont âgées, plus la survie des 2 groupes deDiabrotica diminue, mais plus vite chez les lots d'insectes âgés.

     

Evaluating multiple approaches for managing western corn rootworm larvae with seed blends

Corn rootworm, Diabrotica spp., larvae represent a significant and widespread economic threat to corn, Zea mays (L.), production in the United States, where control costs and yield losses associated with these insect pests exceed $1 billion annually. Preventing root injury and associated yield loss caused by corn rootworm larvae may be accomplished by the independent use of planting time soil insecticides or transgenic Bt hybrids. However, recent reports of both confirmed and suspected Bt resistance in corn rootworm populations throughout the Corn Belt have led to significant interest in the use of these two management tactics simultaneously. Although this approach has been investigated to some extent previously, information is lacking on how the use of a soil insecticide in tandem with a Bt seed blend—Bt and refuge (non‐Bt) seed mixed into a single product—may affect root protection and yield. We describe an experiment including six trial sites conducted over a three‐year period where various seed blends and soil insecticide/seed blend combinations were evaluated. The predominant species contributing to root injury across all sites was the western corn rootworm (Diabrotica virgifera virgifera LeConte). A weighted technique is presented for evaluating root injury for seed blends that offers a reliable estimate of product performance. The addition of a soil insecticide to the seed blend treatments never resulted in significantly improved root protection and failed to provide a consistent yield benefit. Our results suggest that a soil insecticide/seed blend combination approach is not warranted. Additionally, a subanalysis performed on individual refuge and nearby Bt root systems for seed blend treatments provides insight into the spatial characteristics of root injury in seed blend scenarios.     

Microbial Symbionts in Insects Influence Down-Regulation of Defense Genes in Maize

Diabrotica virgifera virgifera larvae are root-feeding insects and significant pests to maize in North America and Europe. Little is known regarding how plants respond to insect attack of roots, thus complicating the selection for plant defense targets. Diabrotica virgifera virgifera is the most successful species in its genus and is the only Diabrotica beetle harboring an almost species-wide Wolbachia infection. Diabrotica virgifera virgifera are infected with Wolbachia and the typical gut flora found in soil-living, phytophagous insects. Diabrotica virgifera virgifera larvae cannot be reared aseptically and thus, it is not possible to observe the response of maize to effects of insect gut flora or other transient microbes. Because Wolbachia are heritable, it is possible to investigate whether Wolbachia infection affects the regulation of maize defenses. To answer if the success of Diabrotica virgifera virgifera is the result of microbial infection, Diabrotica virgifera virgifera were treated with antibiotics to eliminate Wolbachia and a microarray experiment was performed. Direct comparisons made between the response of maize root tissue to the feeding of antibiotic treated and untreated Diabrotica virgifera virgifera show down-regulation of plant defenses in the untreated insects compared to the antibiotic treated and control treatments. Results were confirmed via QRT-PCR. Biological and behavioral assays indicate that microbes have integrated into Diabrotica virgifera virgifera physiology without inducing negative effects and that antibiotic treatment did not affect the behavior or biology of the insect. The expression data and suggest that the pressure of microbes, which are most likely Wolbachia, mediate the down-regulation of many maize defenses via their insect hosts. This is the first report of a potential link between a microbial symbiont of an insect and a silencing effect in the insect host plant. This is also the first expression profile for a plant attacked by a root-feeding insect.     

Assembly and annotation of full mitochondrial genomes for the corn rootworm species, Diabrotica virgifera virgifera and Diabroticabarberi (Insecta: Coleoptera: Chrysomelidae), using Next Generation Sequence data

Complete mitochondrial genomes for two corn rootworm species, Diabrotica virgifera virgifera (16,747 bp) and Diabrotica barberi (16,632; Insecta: Coleoptera: Chrysomelidae), were assembled from Illumina HiSeq2000 read data. Annotation indicated that the order and orientation of 13 protein coding genes (PCGs), and 22 tRNA and 2 rRNA sequences were in typical of insect mitochondrial genomes. Non-standard nad4 and cox3 stop codons were composed of single T nucleotides and likely completed by adenylation, and atypical TTT start codons was predicted for both D. v. virgifera and D. barberinad1 genes. The D. v. virgifera and D. barberi haplotypes showed 819 variable nucleotide positions within PCG regions (7.36% divergence), which suggest that speciation may have occurred ~ 3.68 million years ago assuming a linear rate of short-term substitution. Phylogenetic analyses of Coleopteran MtD genome show clustering based on family level, and may have the capacity to resolve the evolutionary history within this Order of insects.     

Survival and oviposition of a western corn rootworm variant feeding on soybean

The role of soybean, Glycine max (L.) Merril (Fabaceae), in the circumvention of crop rotation was evaluated by observing the effects of soybean herbivory on western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) survival and oviposition. Field collected D. virgifera adults resistant to annual rotation of corn, Zea mays (L.) (Poaceae), and soybean were given the opportunity to feed on a soybean pre‐treatment diet prior to diet combinations of corn, soybean, and/or water. The number of days that the adults survived after removal of the diet combinations was used as a measure of insect vigor and diet quality. Willingness to feed on a soybean foliage pre‐treatment did not indicate greater ability to benefit nutritionally from soybean herbivory. Adult D. virgifera previously feeding on corn, soybean, or water survived starvation for 8.3 ± 0.15 days, 5.1 ± 0.13 days, and 3.9 ± 0.14 days, respectively. Diabrotica virgifera recover from 2 days of starvation or poor diet if subsequently given access to corn tissues for 2 days. Eggs were laid within 1 day of access to poor diet or starvation. Other D. virgifera, captured as they flew from a cornfield into a soybean field, were maintained on soybean foliage or only water until they died. Of the beetles with access to soybean foliage, 24% fed within 24 h after capture and survived 1 day longer than insects given only water. Only 20% of these D. virgifera were able to lay eggs without consuming additional corn prior to death. Few D. virgifera enter soybean fields capable of immediate oviposition. Female reproductive status and diet quality influence the likelihood of oviposition following exposure to stress. The effects of soybean herbivory may contribute to the proximate mechanism of resistance to crop rotation.     

Generalist-feeding subterranean mites as potential biological control agents of immature corn rootworms

Predatory mites are important components of subterranean food webs and may help regulate densities of agricultural pests, including western corn rootworms (Chrysomelidae: Diabrotica virgifera virgifera). Implementing conservation and/or classical biocontrol tactics could enhance densities of specialist or generalist predatory mites and lead to pest suppression, but first relevant mite species must be identified and their predatory capabilities evaluated. We conducted lab assays to quantify consumption of immature rootworms and oviposition rates of various mite species. Our study indicates that rootworms are a sub-optimal food source for the mite taxa tested. However, all mite species fed upon rootworms to some degree, although consumption by nematophagous Eviphis ostrinus was extremely low. Predators consumed more rootworm larvae than eggs, and mite size was correlated with prey consumption, with larger predators eating more prey. Four mite taxa (Gaeolaelaps sp., S. miles, Gl. americana, and G. aculeifer) had detrimental effects on survival of rootworm larvae, and the latter two species also had negative impacts on densities of pest eggs. Although it is unlikely that any of these mite species by itself has a major impact on rootworm control, the community of generalist soil-dwelling mites may play an important role in regulating immature rootworm populations in the field.     

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.