Diabrotica Research Articles

Cross-resistance and synergism bioassays suggest multiple mechanisms of pyrethroid resistance in western corn rootworm populations.

Recently, resistance to the pyrethroid bifenthrin was detected and confirmed in field populations of western corn rootworm, Diabrotica virgifera virgifera LeConte from southwestern areas of Nebraska and Kansas. As a first step to understand potential mechanisms of resistance, the objectives of this study were i) to assess adult mortality at diagnostic concentration-LC99 to the pyrethroids bifenthrin and tefluthrin as well as DDT, ii) estimate adult and larval susceptibility to the same compounds as well as the organophosphate methyl-parathion, and iii) perform synergism experiments with piperonyl butoxide (PBO) (P450 inhibitor) and S,S,S-tributyl-phosphorotrithioate (DEF) (esterase inhibitor) in field populations. Most of the adult field populations exhibiting some level of bifenthrin resistance exhibited significantly lower mortality to both pyrethroids and DDT than susceptible control populations at the estimated LC99 of susceptible populations. Results of adult dose-mortality bioassays also revealed elevated LC50 values for bifenthrin resistant populations compared to the susceptible control population with resistance ratios ranging from 2.5 to 5.5-fold for bifenthrin, 28 to 54.8-fold for tefluthrin, and 16.3 to 33.0 for DDT. These bioassay results collectively suggest some level of cross-resistance between the pyrethroids and DDT. In addition, both PBO and DEF reduced the resistance ratios for resistant populations although there was a higher reduction in susceptibility of adults exposed to PBO versus DEF. Susceptibility in larvae varied among insecticides and did not correlate with adult susceptibility to tefluthrin and DDT, as most resistance ratios were < 5-fold when compared to the susceptible population. These results suggest that both detoxifying enzymes and target site insensitivity might be involved as resistance mechanisms.

An Alcaligenes strain emulates Bacillus thuringiensis producing a binary protein that kills corn rootworm through a mechanism similar to Cry34Ab1/Cry35Ab1

Crops expressing Bacillus thuringiensis (Bt)-derived insecticidal protein genes have been commercially available for over 15 years and are providing significant value to growers. However, there remains the need for alternative insecticidal actives due to emerging insect resistance to certain Bt proteins. A screen of bacterial strains led to the discovery of a two-component insecticidal protein named AfIP-1A/1B from an Alcaligenes faecalis strain. This protein shows selectivity against coleopteran insects including western corn rootworm (WCR). Transgenic maize plants expressing AfIP-1A/1B demonstrate strong protection from rootworm injury. Surprisingly, although little sequence similarity exists to known insecticidal proteins, efficacy tests using WCR populations resistant to two different Cry proteins show that AfIP-1A/1B and mCry3A differ in their mode of action while AfIP-1A/1B and the binary Cry34Ab1/Cry35Ab1 protein share a similar mode. These findings are supported by results of competitive binding assays and the similarity of the x-ray structure of AfIP-1A to Cry34Ab1. Our work indicates that insecticidal proteins obtained from a non-Bt bacterial source can be useful for developing genetically modified crops and can function similarly to familiar proteins from Bt.

Genome Sequence of a Novel Positive-Sense, Single-Stranded RNA Virus Isolated from Western Corn Rootworm, Diabrotica virgiferavirgifera LeConte.

ABSTRACTThe genome sequence of a novel small RNA virus was assembled from the transcriptome of the western corn rootworm, Diabrotica virgifera virgifera. The assembled genome has 13,182 nucleotides with a 3′ polyadenylated tail. Two open reading frames are predicted to encode polyproteins of 2,838 and 1,073 amino acids.

Genome Sequence of Diabrotica virgifera virgifera virus2, a Novel Small RNA Virus of the Western Corn Rootworm, Diabrotica virgifera virgifera LeConte.

ABSTRACTThe genome of a novel small RNA virus, tentatively named Diabrotica virgifera virgifera virus 2 (DvvV2), was identified in the western corn rootworm, Diabrotica virgifera virgifera LeConte, through transcriptome sequencing and confirmed by reverse transcription-PCR. Here, we report the near-complete nucleotide sequence and the genome organization of DvvV2.

Mycotoxin contamination in maize triggered by arthropod pests and the related protection possibilities

Mycotoxin contamination in harvested maize has increased in the last decades, which can be unequivocally back to the plant health troubles caused by global warming. The increasing of wounds in maize crops was occurred by climate change both on direct (hailstorm) and indirect(newly appeared pests) ways. In additional, the settling phytopathogenic microfungi on these plant wounds inflict serious human and animal health problems.The changing of Hungarian arthropod pests assemblages stand in the background of this dangerous nuisance complex. The spreading of European corn borer (Ostrinia nubilalis Hbn.) bivoltine ecotype as well as the newly appeared adventive species [cotton bollworm Helicoverpa armigera Hbn.), western corn rootworm (Diabrotica v. virgifera LeConte), fourspotted-sapbeetle (Glischrochilus quadrisignatus Say)] in Hungary can be responsible for this situation. In total, all technological elements, which obstruct the damage of these chewing mouthparts pests, as well as moderate the mechanical damage of maize, can be contribute to the reduction of both these phytopathogens injuries and mycotoxin contaminations.

Assessment of Inheritance and Fitness Costs Associated with Field-Evolved Resistance to Cry3Bb1 Maize by Western Corn Rootworm.

The western corn rootworm, Diabrotica virgifera virgifera LeConte, is among the most serious insect pests of maize in North America. One strategy used to manage this pest is transgenic maize that produces one or more crystalline (Cry) toxins derived from the bacterium Bacillus thuringiensis (Bt). To delay Bt resistance by insect pests, refuges of non-Bt maize are grown in conjunction with Bt maize. Two factors influencing the success of the refuge strategy to delay resistance are the inheritance of resistance and fitness costs, with greater delays in resistance expected when inheritance of resistance is recessive and fitness costs are present. We measured inheritance and fitness costs of resistance for two strains of western corn rootworm with field-evolved resistance to Cry3Bb1 maize. Plant-based and diet-based bioassays revealed that the inheritance of resistance was non-recessive. In a greenhouse experiment, in which larvae were reared on whole maize plants in field soil, no fitness costs of resistance were detected. In a laboratory experiment, in which larvae experienced intraspecific and interspecific competition for food, a fitness cost of delayed larval development was identified, however, no other fitness costs were found. These findings of non-recessive inheritance of resistance and minimal fitness costs, highlight the potential for the rapid evolution of resistance to Cry3Bb1 maize by western corn rootworm, and may help to improve resistance management strategies for this pest.

Histopathological Effects of Bt and TcdA Insecticidal Proteins on the Midgut Epithelium of Western Corn Rootworm Larvae (Diabrotica virgifera virgifera).

Western corn rootworm (WCR, Diabrotica virgifera virgifera LeConte) is a major corn pest in the United States, causing annual losses of over $1 billion. One approach to protect against crop loss by this insect is the use of transgenic corn hybrids expressing one or more crystal (Cry) proteins derived from Bacillus thuringiensis. Cry34Ab1 and Cry35Ab1 together comprise a binary insecticidal toxin with specific activity against WCR. These proteins have been developed as insect resistance traits in commercialized corn hybrids resistant to WCR feeding damage. Cry34/35Ab1 is a pore forming toxin, but the specific effects of Cry34/35Ab1 on WCR cells and tissues have not been well characterized microscopically, and the overall histopathology is poorly understood. Using high-resolution resin-based histopathology methods, the effects of Cry34/35Ab1 as well as Cry3Aa1, Cry6Aa1, and the Photorhabdus toxin complex protein TcdA have been directly visualized and documented. Clear symptoms of intoxication were observed for all insecticidal proteins tested, including swelling and sloughing of enterocytes, constriction of midgut circular muscles, stem cell activation, and obstruction of the midgut lumen. These data demonstrate the effects of these insecticidal proteins on WCR midgut cells, and the collective response of the midgut to intoxication. Taken together, these results advance our understanding of the insect cell biology and pathology of these insecticidal proteins, which should further the field of insect resistance traits and corn rootworm management.

Field Trial Performance of Herculex XTRA (Cry34Ab1/Cry35Ab1) and SmartStax (Cry34Ab1/Cry35Ab1 + Cry3Bb1) Hybrids and Soil Insecticides Against Western and Northern Corn Rootworms (Coleoptera: Chrysomelidae).

Western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), and northern corn rootworm, Diabrotica barberi Smith and Lawrence (Coleoptera: Chrysomelidae), are important insect pests in corn, Zea mays L. For more than a decade, growers have been using transgenic plants expressing proteins from the bacterium Bacillus thuringiensis (Bt) to protect corn roots from feeding. In 2011, western corn rootworm populations were reported to have developed resistance to Bt hybrids expressing Cry3Bb1 and later found to be cross-resistant to hybrids expressing mCry3A and eCry3.1Ab. The identification of resistance to Cry3 (Cry3Bb1, mCry3A, and eCry3.1Ab) hybrids led to concerns about durability and efficacy of products with single traits and of products containing a pyramid of a Cry3 protein and the binary Bt proteins Cry34Ab1 and Cry35Ab1. From 2012 to 2014, 43 field trials were conducted across the central United States to estimate root protection provided by plants expressing Cry34Ab1/Cry35Ab1 alone (Herculex RW) or pyramided with Cry3Bb1 (SmartStax). These technologies were evaluated with and without soil-applied insecticides to determine if additional management measures provided benefit where Cry3 performance was reduced. Trials were categorized for analysis based on rootworm damage levels on Cry3-expressing hybrids and rootworm feeding pressure within each trial. Across scenarios, Cry34Ab1/Cry35Ab1 hybrids provided excellent root protection. Pyramided traits provided greater root and yield protection than non-Bt plus a soil-applied insecticide, and only in trials where larval feeding pressure exceeded two nodes of damage did Cry34Ab1/Cry35Ab1 single-trait hybrids and pyramided hybrids show greater root protection from the addition of soil-applied insecticides.

Protandry of Western Corn Rootworm (Coleoptera: Chrysomelidae) Beetle Emergence Partially Due to Earlier Egg Hatch of Males

Abstract: The western corn rootworm, Diabrotica virgifera virgifera LeConte, exhibits protandry. The contribution of pre-hatch development to protandry in western corn rootworm was previously investigated with a small set of data from one population. To verify the contribution of pre-hatch development to protandry, more than 10,000 larvae from seven wild-type populations collected from across the Corn Belt were evaluated. Larvae were placed into containers on each day of egg hatch for each population and allowed to develop into adults. Duration of hatch for these populations ranged from 7 to 19 days with the percent of insects surviving to adulthood varying between 13 and 52%. For the first 25% of egg hatch, significantly more males emerge. Pre-hatch development does contribute to protandry, but overall the contribution is likely less than post-hatch development.

English

Resistance in western corn rootworm to transgenic corn hybrids was first confirmed in 2011 in Midwestern USA, and threatens their continued use. Farmers are often the first line of resistance detection, so their understanding and attitudes toward this issue are critical for improving resistance management. We conducted telephone focus groups during 2013 with farmers who had experienced rootworm resistance. There were four stages in dealing with unexpected rootworm injury: Awareness of a problem, diagnosis, confirmation, and recommendations. Most farmers discovered the problem themselves, but this usually happened too late in the growing season to limit yield loss. Once aware of a problem, farmers first sought help diagnosing the problem from their seed dealer, chemical rep, and/or crop consultant. They considered the problem to be a significant one, both because of its severity and suddenness, and were concerned about their difficulty in obtaining a correct diagnosis. They eventually used extension entomology specialists to confirm the diagnosis. Farmers gathered recommendations from independent consultants, input suppliers, and extension and indicated that they would aggressively deal with the problem, because they were not sure of what would work to protect their crop. They recommended that public extension put more emphasis on increasing awareness of the problem, assessing the extent of the problem and being an unbiased source of information. However, farmers were unlikely to report rootworm injury if the perceived barriers to reporting outweighed the perceived incentives. These barriers were emotional ones, including being unsure who to trust, fear that reporting will be time-consuming, and shame that they did something wrong. The incentive was access to credible advice. They did not automatically acknowledge the broader social benefits of reporting. Thus, extension probably needs to be explicit about these broader benefits to obtain information about the extent of the problem. With the conflicting demands and multiple information sources, it will be a challenge for extension to involve farmers to improve resistance monitoring and management. Key words: Resistance management, focus group, qualitative analysis, Diabrotica virgifera, genetically modified organism, transgenic crop, extension.

Patterns of Gene Expression in Western Corn Rootworm (Diabrotica virgifera virgifera) Neonates, Challenged with Cry34Ab1, Cry35Ab1 and Cry34/35Ab1, Based on Next-Generation Sequencing.

With Next Generation Sequencing technologies, high-throughput RNA sequencing (RNAseq) was conducted to examine gene expression in neonates of Diabrotica virgifera virgifera (LeConte) (Western Corn Rootworm, WCR) challenged with individual proteins of the binary Bacillus thuringiensis insecticidal proteins, Cry34Ab1 and Cry35Ab1, and the combination of Cry34/Cry35Ab1, which together are active against rootworm larvae. Integrated results of three different statistical comparisons identified 114 and 1300 differentially expressed transcripts (DETs) in the Cry34Ab1 and Cry34/35Ab1 treatment, respectively, as compared to the control. No DETs were identified in the Cry35Ab1 treatment. Putative Bt binding receptors previously identified in other insect species were not identified in DETs in this study. The majority of DETs (75% with Cry34Ab1 and 68.3% with Cry34/35Ab1 treatments) had no significant hits in the NCBI nr database. In addition, 92 DETs were shared between Cry34Ab1 and Cry34/35Ab1 treatments. Further analysis revealed that the most abundant DETs in both Cry34Ab1 and Cry34/35Ab1 treatments were associated with binding and catalytic activity. Results from this study confirmed the nature of these binary toxins against WCR larvae and provide a fundamental profile of expression pattern of genes in response to challenge of the Cry34/35Ab1 toxin, which may provide insight into potential resistance mechanisms.

Influence of calcareous soil on Cry3Bb1 expression and efficacy in the field.

Greater than expected injury by western corn rootworm (WCR) (Diabrotica virgifera virgifera LeConte) to Cry3Bb1 expressing maize hybrids (Zea mays L.) has been reported in southwestern Nebraska. Affected areas of some fields are often associated with high pH calcareous soils where maize growth is poor and iron chlorosis is common. As part of a comprehensive study to understand potential causes of unexpected injury, experiments were conducted during 2013 and 2014 to ascertain whether the calcareous soil conditions and associated poor maize growth negatively affect the expression of Cry3Bb1. Quantitative determination of Cry3Bb1 protein expression levels in root tissues was carried out on plants at V5-V6 growth stage using the enzyme-linked immunosorbent assay. Cry3Bb1 and non-Bt near isoline maize hybrids were artificially infested with Cry3Bb1-susceptible WCR eggs to measure survival and efficacy of Cry3Bb1 maize in calcareous and non-calcareous soils. Results showed that there was not a significant difference in expression of Cry3Bb1 protein between plants from calcareous and non-calcareous soils (18.9-21.2µg/g fresh weight). Western corn rootworm survival was about sevenfold greater from the non-Bt isoline than Cry3Bb1 maize indicating that Cry3Bb1 performed as expected when infested with a Cry3Bb1 susceptible rootworm population. When survival from calcareous and non-calcareous soils was compared, no significant differences were observed in each soil. A significant positive correlation between soil pH and expression of Cry3Bb1 protein in roots was detected from samples collected in 2014 but not in 2013. No such correlation was found between soil pH and survival of WCR. Results suggest that Cry3Bb1 expression levels were sufficient to provide adequate root protection against WCR regardless of soil environment, indicating that lowered Cry3Bb1 expression is not a contributing factor to the greater than expected WCR injury observed in some southwestern Nebraska maize fields.

Nitrogen Rate Effects on Cry3Bb1 and Cry3Bb1 + Cry34/35Ab1 Expression in Transgenic Corn Roots, Resulting Root Injury, and Corn Rootworm Beetle Emergence.

Nitrogen (N) application rates have been recommended historically for maximum economic yield of corn (Zea mays L.), but not for optimal expression or impacts of Bt (Bacillus thuringiensis Berliner) Cry protein(s) on target insects. This study explored the need to adjust N rates to optimize expression of corn rootworm-active Bt (Bt-RW) protein(s) in a single and a pyramided trait hybrid and resulting impacts on beetle emergence and root injury, under field conditions. The experiment featured a factorial treatment arrangement in a split-plot randomized complete block design with six N rates as the main plots and three hybrids (MON88017 expressing Cry3Bb1, MON88017 x DAS-59122 expressing Cry3Bb1 + Cry34/35Ab1, and a non-Bt-RW hybrid) as the subplots. Corn roots were sampled at the beginning of, and after, peak larval feeding to determine Bt-expression levels using an enzyme-linked immunosorbent assay. Beetles were collected every 2-3 d during emergence using cut-plant emergence cages. Cry3Bb1 expression was significantly reduced when little or no N was applied. Cry34Ab1 and Cry35Ab1 expression was highly variable and unaffected by N rate. Beetle emergence increased with N rate in the non-Bt-RW hybrid while root injury declined. Provided Bt-RW hybrids had sufficient applied N, root injury was relatively low. Results indicate that N management could affect Bt-RW expression and success of insect resistance management plans provided N is applied at rates that enhance production of susceptible beetles emerging from the non-Bt-RW (refuge) hybrid, and achieve optimal expression and efficacy of Bt traits.

Evaluation of SmartStax and SmartStax PRO maize against western corn rootworm and northern corn rootworm: efficacy and resistance management.

Cases of western corn rootworm (WCR) field-evolved resistance to Cry3Bb1 and other corn rootworm (CRW) control traits have been reported. Pyramid products expressing multiple CRW traits can delay resistance compared to single trait products. We used field studies to assess the pyramid CRW corn products, SmartStax (expressing Cry3Bb1 and Cry34Ab1/Cry35Ab1) and SmartStax PRO (expressing Cry3Bb1, Cry34Ab1/Cry35Ab1 and DvSnf7), at locations with high WCR densities and possible Cry3Bb1 resistance, and to assess the reduction in adult emergence attributable to DvSnf7 and other traits. Insect resistance models were used to assess durability of SmartStax and SmartStax PRO to WCR resistance. SmartStax significantly reduced root injury compared to non-CRW-trait controls at all but one location with measurable WCR pressure, while SmartStax PRO significantly reduced root injury at all locations, despite evidence of Cry3Bb1 resistance at some locations. The advantage of SmartStax PRO over SmartStax in reducing root damage was positively correlated with root damage on non-CRW-trait controls. DvSnf7 was estimated to reduce WCR emergence by approximately 80-95%, which modeling indicated will improve durability of Cry3Bb1 and Cry34Ab1/Cry35Ab1 compared to SmartStax. The addition of DvSnf7 in SmartStax PRO can reduce root damage under high WCR densities and prolong Cry3Bb1 and Cry34Ab1/Cry35Ab1 durability. © 2017 Society of Chemical Industry.

Comparison of RNAi Sequences in Insect-Resistant Plants to Expressed Sequences of a Beneficial Lady Beetle: A Closer Look at Off-Target Considerations.

Sequences obtained from transcriptomes of the lady beetle Coleomegilla maculata were compared to those designed for incorporation into crops. Searches of the transcriptomes identified sequences as the most likely to be closely similar to the sequences described in RNAi plant incorporated products. Some proposed prime RNAi pest management targets were also used to identify predicted orthologs from C. maculata. The lady beetle sequences were aligned with sequences from corn rootworms and Colorado potato beetles and, as appropriate in the case of targets, regions of similarity were compared with the genetic model organism for beetles, Tribolium castaneum. Some high levels of nucleotide identity were identified, particularly with an actin-derived sequence from Colorado potato beetle. This actin-derived sequence shared identical sequences with the lady beetle and a parasitic wasp.

Host Recognition Responses of Western (Family: Chrysomelidae) Corn Rootworm Larvae to RNA Interference and Bt Corn.

Western corn rootworm Diabrotica virgifera virgifera LeConte is an important pest of corn whose larvae exhibit particular quantifiable patterns of locomotion after exposure to, and removal from, host roots and nonhost roots. Using EthoVision software, the behavior and locomotion of the western corn rootworm larvae was analyzed to determine the level of host recognition to germinated roots of differing corn hybrids containing either rootworm targeted Bt genes, RNA interference (RNAi) technology, the stack of both Bt and RNAi, or the isoline of these. The behavior of the rootworm larvae indicated a significant host preference response to all corn hybrids (with or without insecticidal traits) compared to the filter paper and oat roots. A weaker host response to the RNAi corn roots was observed in the susceptible larvae when compared to the resistant larvae, but not for the Bt + RNAi vector stack. Additionally, the resistant larvae demonstrated a weaker host response to the isoline corn roots when compared to the susceptible larvae. Although weaker, these host responses were significantly different from those observed in the negative controls, indicating that all hybrids tested do contain the contact cues necessary to elicit a host preference response by both Cry3Bb1-resistant and Cry3Bb1-susceptible larvae that would work to hinder resistance development in refuge in a bag fields.

Dietary Risk Assessment of v-ATPase A dsRNAs on Monarch Butterfly Larvae.

By suppressing the expression of genes with essential biological functions, in planta RNAi can negatively affect the development and survival of target pests. As a part of a concerted effort to assess the risks of RNAi transgenic crops on non-target organisms, we developed an in vivo toxicity assay to examine the impacts of ingested dsRNAs incurred to the monarch butterfly, Danaus plexippus (L.), an iconic eco-indicator in North America. To create the worst case scenario, the full-length v-ATPase A cDNAs from the target pest, western corn rootworm, Diabrotica virgifera virgifera, and the non-target D. plexippus were respectively cloned. A 400 bp fragment with the highest sequence similarity between the two species was used as the template to synthesize dsRNAs for the subsequent dietary RNAi toxicity assay. Specifically, newly hatched neonates were provisioned with leaf disks surface-coated with v-ATPase A dsRNAs synthesized from D. v. virgifera and D. plexippus, respectively, a control dsRNA, β-glucoruronidase, from plants, and H2O. The endpoint measurements included gene expressions and life history traits. The 2283 bp D. plexippus v-ATPase A cDNA contains a 99 bp 5′-untranslated region, a 330 bp 3′-untranslated region, and an 1851 bp ORF encoding 617 amino acids. The temporal RNAi study did not detect any impact to D. plexippus v-ATPase A expression by the assay days and treatments. This was reflected in the phenotypic impacts of dietary RNAi, in which both survival rate and development time were not affected by the uptake of ingested dsRNAs. These combined results suggest that D. plexippus larvae are not susceptible to dietary RNAi, therefore, the impact of transgenic RNAi plants on this non-target organism is, likely, negligible.

Genome Sequence of the First Coleopteran Iflavirus Isolated from Western Corn Rootworm, Diabrotica virgiferavirgifera LeConte.

ABSTRACTThe genome sequence of a novel iflavirus was identified from the transcriptome of the western corn rootworm, Diabrotica virgifera virgifera. The RNA sequence consists of 9,823 nucleotides (nt) with a 3′ polyadenylated tail, containing a single open reading frame that encodes a 3,028-amino-acid polyprotein.

Minnesota field population of western corn rootworm (Coleoptera: Chrysomelidae) shows incomplete resistance to Cry34Ab1/Cry35Ab1 and Cry3Bb1

AbstractIn the United States of America, the western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is commonly managed with transgenic corn (Zea mays L.) expressing insecticidal proteins from the bacteria Bacillus thuringiensis Berliner (Bt). Colonies of this pest have been selected in the laboratory on each commercially available transformation event and several resistant field populations have also been identified; some field populations are also resistant. In this study, progeny of a western corn rootworm population collected from a Minnesota corn field planted to SmartStax® corn were evaluated for resistance to corn hybrids expressing Cry3Bb1 (event MON88017) or Cry34/35Ab1 (event DAS‐59122‐7) and to the individual constituent proteins in diet‐overlay bioassays. Results from these assays suggest that this population is resistant to Cry3Bb1 and is incompletely resistant to Cry34/35Ab1. In diet toxicity assays, larvae of the Minnesota (MN) population had resistance ratios of 4.71 and &gt;13.22 for Cry34/35Ab1 and Cry3Bb1 proteins, respectively, compared with the control colonies. In all on‐plant assays, the relative survival of the MN population on the DAS‐59122‐7 and MON88017 hybrids was significantly greater than the control colonies. Larvae of the MN population had inhibited development when reared on DAS‐59122‐7 compared with larvae reared on the non‐Bt hybrid, indicating resistance was incomplete. Overall, these results document resistance to Cry3Bb1 and an incomplete resistance to Cry34/35Ab1 in a population of WCR from a SmartStax® performance problem field.

Field efficacy of Heterorhabditis bacteriophora (Nematoda: Heterorhabditidae), Metarhizium brunneum (Hypocreales: Clavicipitaceae), and chemical insecticide combinations for Diabrotica virgifera virgifera larval management

A two-year field study using a blend of entomopathogens in conjunction with chemical insecticides was carried out to determine to which extent they affect western corn rootworm (WCR), Diabrotica virgifera virgifera, survival, maize root damages, and grain yield and to assess the potential for side-effects on natural arthropod enemies. The products tested were conducted on a maize crop and included maize seeds dressed with the neonicotinoid clothianidin (Poncho™), Belem™ (ai: cypermethrin), dianem™ (entomopathogenic nematode: Heterorhabditis bacteriophora), and a granular formulation of the insect pathogenic fungus Metarhizium brunneum. Trials were conducted at four (2014) and two (2015) sites heavily infested with WCR in the Southeast of Styria, Austria. The lowest number of WCR adults were recorded in 2015 at sites where the nematode and fungal biocontrol agents were used with clothianidin dressed maize seeds. The treatments did not significantly influence plant lodging and grain yield compared with the untreated (negative) control. The WCR population density rose during the investigation period and the average number of beetles in all treatments and in both study years far exceeded the stipulated economic threshold value for continuous maize of around one beetle per plant. Non-target evaluation using pitfall traps did not show any significant impact of the treatments on the abundance or diversity of indigenous predatory beetles and spiders. The lack of damage suggests that the WCR threshold is considerably higher than the stipulated economic threshold under favourable growing conditions, but a systematic crop rotation is recommended as an integral part of the WCR management both as a prophylaxis and as a background for the best treatment.