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1.
Sci Rep ; 11(1): 17944, 2021 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-34504232

RESUMO

The northern corn rootworm, Diabrotica barberi Smith & Lawrence, has a univoltine life cycle that typically produces one generation a year. When rearing the northern corn rootworm in the laboratory, in order to break diapause, it is necessary to expose eggs to a five month cold period before raising the temperature. By selective breeding of the small fraction of eggs that hatched without cold within 19-32 days post oviposition, we were able to develop a non-diapausing colony of the northern corn rootworm within five generations of selection. Through selection, the percentages of adult emergence from egg hatch without exposure to cold treatment significantly increased from 0.52% ± 0.07 at generation zero to 29.0% ± 2.47 at generation eight. During this process, we developed an improved method for laboratory rearing of both the newly developed non-diapausing strain as well as the diapausing strain. The development of the non-diapausing colony along with the improvements to the rearing system will allow researchers to produce up to six generations of the northern corn rootworm per year, which would facilitate research and advance our knowledge of this pest at an accelerated rate.

2.
BMC Genomics ; 22(1): 639, 2021 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-34479486

RESUMO

BACKGROUND: Resistance of pest insect species to insecticides, including B. thuringiensis (Bt) pesticidal proteins expressed by transgenic plants, is a threat to global food security. Despite the western corn rootworm, Diabrotica virgifera virgifera, being a major pest of maize and having populations showing increasing levels of resistance to hybrids expressing Bt pesticidal proteins, the cell mechanisms leading to mortality are not fully understood. RESULTS: Twenty unique RNA-seq libraries from the Bt susceptible D. v. virgifera inbred line Ped12, representing all growth stages and a range of different adult and larval exposures, were assembled into a reference transcriptome. Ten-day exposures of Ped12 larvae to transgenic Bt Cry3Bb1 and Gpp34/Tpp35Ab1 maize roots showed significant differential expression of 1055 and 1374 transcripts, respectively, compared to cohorts on non-Bt maize. Among these, 696 were differentially expressed in both Cry3Bb1 and Gpp34/Tpp35Ab1 maize exposures. Differentially-expressed transcripts encoded protein domains putatively involved in detoxification, metabolism, binding, and transport, were, in part, shared among transcripts that changed significantly following exposures to the entomopathogens Heterorhabditis bacteriophora and Metarhizium anisopliae. Differentially expressed transcripts in common between Bt and entomopathogen treatments encode proteins in general stress response pathways, including putative Bt binding receptors from the ATP binding cassette transporter superfamily. Putative caspases, pro- and anti-apoptotic factors, as well as endoplasmic reticulum (ER) stress-response factors were identified among transcripts uniquely up-regulated following exposure to either Bt protein. CONCLUSIONS: Our study suggests that the up-regulation of genes involved in ER stress management and apoptotic progression may be important in determining cell fate following exposure of susceptible D. v. virgifera larvae to Bt maize roots. This study provides novel insights into insect response to Bt intoxication, and a possible framework for future investigations of resistance mechanisms.


Assuntos
Bacillus thuringiensis , Besouros , Praguicidas , Animais , Bacillus thuringiensis/genética , Sobrevivência Celular , Besouros/genética , Endotoxinas/toxicidade , Resistência a Inseticidas , Larva/genética , Controle Biológico de Vetores , Plantas Geneticamente Modificadas/genética , Regulação para Cima , Zea mays/genética
3.
J Econ Entomol ; 114(5): 2220-2228, 2021 Oct 13.
Artigo em Inglês | MEDLINE | ID: mdl-34453170

RESUMO

The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), poses a serious threat to maize (Zea mays L.) growers in the U.S. Corn Belt. Transgenic corn expressing Bacillus thuringiensis (Bt) Berliner is the major management tactic along with crop rotation. Bt crops targeting WCR populations have been widely planted throughout the Corn Belt. Rootworms have developed resistance to nearly all management strategies including Bt corn. Therefore, there is a need for new products that are not cross-resistant with the current Bt proteins. In this study, we evaluated the susceptibility of WCR strains resistant and susceptible to Cry3Bb1 to the biological insecticide Spear-T (GS-omega/kappa-Hexatoxin-Hv1a) alone and combined with Cry3Bb1 protein. The activity of Hv1a alone was similar between Cry3Bb1-resistant and susceptible strains (LC50s = 0.95 mg/cm2 and 1.50 mg/cm2, respectively), suggesting that there is no cross-resistance with Cry3Bb1 protein. Effective concentration (EC50), molt inhibition concentration (MIC50), and inhibition concentration (IC50) values of Hv1a alone were also similar between both strains, based on non-overlapping confidence intervals. Increased mortality (64%) was observed on resistant larvae exposed to Hv1a (0.6 mg/cm2) + Cry3Bb1 protein (170.8 µg/cm2) compared to 0% mortality when exposed to Cry3Bb1 alone and 34% mortality to Hv1a alone (0.3 mg/cm2). The time of larval death was not significantly different between Hv1a alone (3.79 mg/cm2) and Hv1a (0.6 mg/cm2) + Cry3Bb1 (170.8 µg/cm2). New control strategies that are not cross-resistant with current insecticides and Bt proteins are needed to better manage the WCR, and Hv1a together with Cry3Bb1 may fit this role.

4.
Mol Ecol ; 2021 Mar 08.
Artigo em Inglês | MEDLINE | ID: mdl-33683750

RESUMO

Evolution of resistance to transgenic crops producing toxins from Bacillus thuringiensis (Bt) threatens the sustainability of the technology. Examination of resistance mechanisms has largely focused on characterization of mutations in proteins serving as Bt toxin binding sites. However, insect microbial communities have the potential to provide host resistance to pesticides in a myriad of ways. Previous findings suggest the killing mechanism of Bt relies on enteric bacteria becoming pathogenic in the disrupted gut environment of the insect following Bt intoxication. Thus, here we hypothesized that resistance to Bt would alter the microbiome composition of the insect. Previous studies have manipulated the microbiome of susceptible insects and monitored their response to Bt. In our study, we characterized the associated bacterial communities of Bt-resistant and -susceptible western corn rootworms, a widespread pest of maize in the United States. We found resistant insects harbor a bacterial community that is less rich and distinct from susceptible insects. After feeding on Bt-expressing maize, susceptible insects exhibited dysbiosis of the associated bacterial community, whereas the community within resistant insects remained relatively unchanged. These results suggest resistance to Bt produces alterations in the microbiome of the western corn rootworm that may contribute to resistance. We further demonstrated that by itself, feeding on Bt toxin-expressing seedlings caused a shift in the microbiota. This work provides a broader picture of the effect stressors have on microbiome composition, and the potential heritable changes induced as a result of intense selection.

5.
Insects ; 12(2)2021 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-33671118

RESUMO

The western corn rootworm, Diabrotica virgifera virgifera LeConte, is resistant to four separate classes of traditional insecticides, all Bacillius thuringiensis (Bt) toxins currently registered for commercial use, crop rotation, innate plant resistance factors, and even double-stranded RNA (dsRNA) targeting essential genes via environmental RNA interference (RNAi), which has not been sold commercially to date. Clearly, additional tools are needed as management options. In this review, we discuss the state-of-the-art knowledge about biotic factors influencing herbivore success, including host location and recognition, plant defensive traits, plant-microbe interactions, and herbivore-pathogens/predator interactions. We then translate this knowledge into potential new management tools and improved biological control.

6.
Pest Manag Sci ; 77(5): 2385-2394, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33415809

RESUMO

BACKGROUND: Management of the corn pest, western corn rootworm (WCR), Diabrotica virgifera virgifera (LeConte) (Coleoptera: Chrysomelidae), relies heavily on the planting of transgenic corn expressing toxins produced by the bacterium Bacillus thuringiensis (Bt). This has resulted in the evolution of resistance to all of the four commercially available Bt toxins targeting coleopteran insects. In this study, we evaluated the susceptibility of a Cry34/35Ab1-resistant WCR colony in seedling and diet toxicity assays after removal from selection for six and nine generations. In addition, female fecundity, egg fertility, adult lifespan, larval development, and adult emergence were evaluated in two Cry34/35Ab1-resistant and two susceptible WCR colonies to assess fitness costs. RESULTS: Susceptibility to Cry34/35Ab1 was restored in a colony removed from selection after six and nine generations based on diet toxicity assays and comparisons of relative survival, head capsule width, and dry weight in plant assays. Thus, pronounced fitness costs associated with resistance to Cry34/35Ab1 were documented by susceptibility being restored within six generations. In separate studies evaluating specific fitness costs, larval fitness when reared on isoline corn did not differ between resistant and susceptible colonies. However, beetles from susceptible colonies lived longer than resistant beetles which resulted in females from susceptible colonies producing significantly more eggs than resistant colonies, with no differences in egg fertility. CONCLUSIONS: The presence of a fitness cost that may contribute to the restoration of susceptibility to Bt has not been documented in other Cry3-resistant WCR populations and could have significant impact on the deployment of resistance management practices. Published 2021. This article is a U.S. Government work and is in the public domain in the USA.


Assuntos
Bacillus thuringiensis , Besouros , Animais , Bacillus thuringiensis/genética , Proteínas de Bactérias/genética , Besouros/genética , Endotoxinas/genética , Endotoxinas/farmacologia , Feminino , Resistência a Inseticidas/genética , Larva/genética , Controle Biológico de Vetores , Plantas Geneticamente Modificadas/genética , Zea mays/genética
7.
Artigo em Inglês | MEDLINE | ID: mdl-33310708

RESUMO

This study describes three closely related proteins, cloned from Brevibacillus laterosporus strains, that are lethal upon feeding to Diabrotica virgifera virgifera LeConte, the western corn rootworm (WCR). Mpp75Aa1, Mpp75Aa2 and Mpp75Aa3 were toxic to WCR larvae when fed purified protein. Transgenic plants expressing each mMpp75Aa protein were protected from feeding damage and showed significant reduction in adult emergence from infested plants by both susceptible and Cry3Bb1 and Cry34Ab1/Cry35Ab1-resistant WCR. These results demonstrate that proteins from B. laterosporus are as efficacious as the well-known Bacillus thuringiensis (Bt) insecticidal proteins in controlling major insect pests such as WCR. The deployment of transgenic maize expressing mMpp75Aa along with other active molecules lacking cross-resistance have the potential to be a useful tool for control of WCR populations resistant to current Bt traits.IMPORTANCE Insects feeding on roots of crops can damage the plant roots resulting in yield loss due to poor water and nutrient uptake and plant lodging. In maize the western corn rootworm (WCR) can cause severe damage to the roots resulting in significant economic loss for farmers. Genetically modified (GM) expressing Bacillus thuringiensis (Bt) insect control proteins, has provided a solution for control of these pests. In recent years populations of WCR resistant to the Bt proteins in commercial GM maize have emerged. There is a need to develop new insecticidal traits for the control of WCR populations resistant to current commercial traits. New proteins with commercial level efficacy on WCR from sources other than Bt are becoming more critical. The Mpp75Aa proteins, from B. laterosporus, when expressed in maize, are efficacious against the resistant populations of WCR and have the potential to provide solutions for control of resistant WCR.

8.
PLoS One ; 15(11): e0242791, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33253273

RESUMO

The Western corn rootworm (WCR) Diabrotica virgifera virgifera LeConte is one of the most economically important insect pests in North America. Since 2003, transgenic maize expressing WCR-active proteins from Bacillus thuringiensis (Bt) have been widely adopted as the main approach to controlling WCR in the U.S. However, the emergence of field resistance to the Bt proteins in current commercial products has been documented in recent years, highlighting the need to develop additional tools for controlling this devasting pest. Here we report the discovery of Vpb4Da2 (initially assigned as Vip4Da2), a new insecticidal protein highly selective against WCR, through high-throughput genome sequencing of a Bt strain sourced from grain dust samples collected in the eastern and central regions of the US. Vpb4Da2 contains a sequence and domain signature distinct from families of other WCR-active proteins. Under field conditions, transgenic maize expressing Vpb4Da2 demonstrates commercial-level (at or below NIS 0.25) root protection against WCR, and reduces WCR beetle emergence by ≥ 97%. Our studies also conclude that Vpb4Da2 controls WCR populations that are resistant to WCR-active transgenic maize expressing Cry3Bb1, Cry34Ab1/Cry35Ab1 (reassigned as Gpp34Ab1/Tpp35Ab1), or DvSnf7 RNA. Based on these findings, Vpb4Da2 represents a valuable new tool for protecting maize against WCR.


Assuntos
Toxinas de Bacillus thuringiensis/genética , Besouros/genética , Controle Biológico de Vetores , Zea mays/genética , Animais , Bacillus thuringiensis/genética , Besouros/patogenicidade , Proteínas Hemolisinas/genética , Humanos , Resistência a Inseticidas/genética , Inseticidas/efeitos adversos , Inseticidas/química , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/parasitologia , Zea mays/parasitologia
9.
J Econ Entomol ; 113(4): 1955-1962, 2020 08 13.
Artigo em Inglês | MEDLINE | ID: mdl-32789524

RESUMO

The northern corn rootworm (NCR), Diabrotica barberi Smith & Lawrence, is an economic pest of maize in the U.S. Corn Belt. The objective of this study was to determine the baseline susceptibility of a laboratory NCR strain to Bt proteins eCry3.1Ab, mCry3A, Cry3Bb1, and Cry34/35Ab1 using seedling, single plant, and diet-toxicity assays. Plant assays were performed in greenhouse using corn hybrids expressing one of the Bt proteins and each respective near-isoline. Diet-toxicity assays, consisting of Bt proteins overlaid onto artificial diet were also conducted. In both plant assays, significantly more larvae survived Cry34/35Ab1-expressing corn compared with all other Bt-expressing corn, and larvae that survived eCry3.1Ab-expressing corn had significantly smaller head capsule widths compared with larvae that survived Cry34/35Ab1-expressing corn. In seedling assays, larvae surviving eCry3.1Ab-expressing corn also had significantly smaller head capsule widths compared with larvae that survived mCry3A-expressing corn. Additionally, larvae that survived mCry3A-expressing corn weighed significantly more than larvae surviving eCry3.1Ab- and Cry34/35Ab1-expressing corn. In single plant assays, no significant differences in larval dry weight was observed between any of the Bt-expressing corn. In diet assays, LC50s ranged from 0.14 (eCry3.1Ab) to 10.6 µg/cm2 (Cry34/35Ab1), EC50s ranged from 0.12 (Cry34/35Ab1) to 1.57 µg/cm2 (mCry3A), IC50s ranged from 0.08 (eCry3.1Ab) to 2.41 µg/cm2 (Cry34/35Ab1), and MIC50s ranged from 2.52 (eCry3.1Ab) to 14.2 µg/cm2 (mCry3A). These results establish the toxicity of four Bt proteins to a laboratory diapausing NCR strain established prior to the introduction of Bt traits and are important for monitoring resistance evolution in NCR field populations.


Assuntos
Bacillus thuringiensis , Besouros , Animais , Bacillus thuringiensis/genética , Proteínas de Bactérias , Besouros/genética , Dieta , Endotoxinas , Proteínas Hemolisinas , Laboratórios , Larva , Controle Biológico de Vetores , Plantas Geneticamente Modificadas/genética , Plântula , Zea mays/genética
10.
Sci Rep ; 10(1): 746, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31937872

RESUMO

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

11.
Sci Rep ; 9(1): 16009, 2019 11 05.
Artigo em Inglês | MEDLINE | ID: mdl-31690801

RESUMO

The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, is an important pest of maize (Zea mays L.). Published WCR diets contain corn root powder, which is not available for purchase, thereby limiting the practical use of diets containing this ingredient. We applied response surface modeling combined with mixture designs to formulate a WCR diet that does not require corn root powder. We developed the new formulation by systematically exploring eight protein ingredients from animal, plant, and yeast sources based on simultaneous evaluation of three life history parameters (weight, molting, and survival). This formulation (WCRMO-2) without corn root powder supported approximately 97% of larval survival and successful molting. Larval weight gain after 10 days of feeding on WCRMO-2 was 4-fold greater than that of larvae feeding on the current best published WCR diet. Additionally, there was no significant difference in these larval performance traits when larvae were reared on WCRMO-2 and the best proprietary WCR diet. A commercial version of WCRMO-2 was tested and found to perform comparably for these traits. These improvements met our goal of a diet comprised of available ingredients that supports performance of WCR larvae equal to or better than publicly available formulations and proprietary formulations.


Assuntos
Besouros/crescimento & desenvolvimento , Dieta , Animais , Besouros/metabolismo , Dieta/veterinária , Proteínas do Ovo/química , Larva/crescimento & desenvolvimento , Larva/fisiologia , Modelos Biológicos , Raízes de Plantas/química , Raízes de Plantas/metabolismo , Proteínas do Soro do Leite/química , Zea mays/química , Zea mays/metabolismo
12.
Proc Natl Acad Sci U S A ; 116(46): 23174-23181, 2019 11 12.
Artigo em Inglês | MEDLINE | ID: mdl-31659056

RESUMO

Plants defend themselves against herbivores through the production of toxic and deterrent metabolites. Adapted herbivores can tolerate and sometimes sequester these metabolites, allowing them to feed on defended plants and become toxic to their own enemies. Can herbivore natural enemies overcome sequestered plant defense metabolites to prey on adapted herbivores? To address this question, we studied how entomopathogenic nematodes cope with benzoxazinoid defense metabolites that are produced by grasses and sequestered by a specialist maize herbivore, the western corn rootworm. We find that nematodes from US maize fields in regions in which the western corn rootworm was present over the last 50 y are behaviorally and metabolically resistant to sequestered benzoxazinoids and more infective toward the western corn rootworm than nematodes from other parts of the world. Exposure of a benzoxazinoid-susceptible nematode strain to the western corn rootworm for 5 generations results in higher behavioral and metabolic resistance and benzoxazinoid-dependent infectivity toward the western corn rootworm. Thus, herbivores that are exposed to a plant defense sequestering herbivore can evolve both behavioral and metabolic resistance to plant defense metabolites, and these traits are associated with higher infectivity toward a defense sequestering herbivore. We conclude that plant defense metabolites that are transferred through adapted herbivores may result in the evolution of resistance in herbivore natural enemies. Our study also identifies plant defense resistance as a potential target for the improvement of biological control agents.


Assuntos
Benzoxazinas/metabolismo , Besouros/parasitologia , Herbivoria , Interações Hospedeiro-Parasita , Rabditídios/fisiologia , Animais , Besouros/metabolismo , Cadeia Alimentar , Zea mays
13.
Sci Rep ; 9(1): 15332, 2019 10 25.
Artigo em Inglês | MEDLINE | ID: mdl-31653954

RESUMO

Western corn rootworm (Diabrotica virgifera virgifera LeConte) is a serious pest of maize (Zea mays L.) in North America and parts of Europe. With most of its life cycle spent in the soil feeding on maize root tissues, this insect is likely to encounter and interact with a wide range of soil and rhizosphere microbes. Our knowledge of the role of microbes in pest management and plant health remains woefully incomplete, yet that knowledge could play an important role in effective pest management strategies. For this study, insects were reared on maize in soils from different locations. Insects from two different laboratory colonies (a diapausing and a non-diapausing colony) were sampled at each life stage to determine the possible core bacteriome. Additionally, soil was sampled at each life stage and resulting bacteria were identified to determine the possible contribution of soil to the rootworm bacteriome, if any. We analyzed the V4 hypervariable region of bacterial 16S rRNA genes with Illumina MiSeq to survey the different species of bacteria associated with the insects and the soils. The bacterial community associated with insects was significantly different from that in the soil. Some differences appear to exist between insects from non-diapausing and diapausing colonies while no significant differences in community composition existed between the insects reared on different soils. Despite differences in the bacteria present in immature stages and in male and female adults, there is a possible core bacteriome of approximately 16 operational taxonomic units (i.e., present across all life stages). This research may provide insights into Bt resistance development, improved nutrition in artificial rearing systems, and new management strategies.


Assuntos
Bactérias/crescimento & desenvolvimento , Besouros/crescimento & desenvolvimento , Besouros/microbiologia , Estágios do Ciclo de Vida , Solo/parasitologia , Animais , Biodiversidade , Análise de Componente Principal , Microbiologia do Solo
14.
J Econ Entomol ; 112(6): 2737-2743, 2019 12 09.
Artigo em Inglês | MEDLINE | ID: mdl-31550358

RESUMO

The northern corn rootworm, Diabrotica barberi Smith & Lawrence (Coleoptera: Chrysomelidae), is one of the most important insect pests in the U.S. Corn Belt. Efforts to obtain eggs from wild northern corn rootworm populations using techniques developed for other rootworm species have been unsuccessful due to lack of oviposition. In 2016, we evaluated four oviposition media in choice tests within each of three female densities in 30.5 × 30.5 × 30.5 cm BugDorm cages. The number of eggs laid per female was significantly affected by female density and the interaction of female density × oviposition media, but oviposition was relatively poor in all oviposition media (1.2 eggs per female when averaging the three female densities and all oviposition media). Single females were also evaluated in nonchoice assays in 6 cm × 6 cm × 8 cm clear plastic boxes and averaged up to 108 eggs per female depending on the oviposition media. In 2017, the cumulative number of eggs laid per female in boxes with one female was not significantly different from the number of eggs laid per female in boxes with 3 females. In 2018, the cumulative number of eggs laid per female was not significantly different between female densities of 1, 3, 5, or 10 females per box. Total egg production per box therefore increased as female density increased. More than 27,000 wild northern corn rootworm eggs were collected from just 190 females when collected relatively early in the field season. We now have an efficient and robust system for obtaining eggs from wild northern corn rootworm females.


Assuntos
Besouros , Animais , Feminino , Oviposição , Plantas Geneticamente Modificadas , Zea mays
15.
J Insect Sci ; 19(2)2019 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-30953583

RESUMO

The western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is an important economic pest of maize (Zea mays L.) in North America and Europe. Previous efforts to formulate an artificial diet for western corn rootworm larvae highlighted an important role of corn root powder, which had a significant positive impact on several larval developmental traits. Unfortunately, this ingredient is not available for purchase. Toward the goal of developing an artificial diet for western corn rootworm larvae with all ingredients readily accessible, we conducted research to isolate essential growth factors for larval development from corn root powder to improve the performance of diet without corn root powder. For all experiments, multiple life history parameters (survival, weight, and molting) were recorded from 15-d diet bioassays. Corn roots may contain factors that assist in larval growth, but some of these factors were not fully extracted by methanol and remained in the extracted root. Methanolic extracts significantly increased molting to second instar, but did not significantly increase survival, dry weight, or molting to third instar, suggesting the primary corn root substituents affecting these factors cannot be extracted or other extraction methods may be required to extract the essential factors from corn roots. We showed that whole corn root powder was best when used in combination with all the other nutrient sources in the published western corn rootworm formulation. Corn root powder made from proprietary seed and Viking seed has similar value.


Assuntos
Ração Animal/análise , Besouros/crescimento & desenvolvimento , Zea mays/química , Animais , Besouros/fisiologia , Dieta , Larva/crescimento & desenvolvimento , Larva/fisiologia , Muda/fisiologia , Raízes de Plantas/química
16.
Sci Rep ; 9(1): 3127, 2019 02 28.
Artigo em Inglês | MEDLINE | ID: mdl-30816250

RESUMO

Diabrotica virgifera virgifera LeConte, the western corn rootworm (WCR), is the most destructive pest of maize in North America, and has recently spread across central Europe. Its subterranean larval stages are hard to reach with pesticides and it has evolved resistance to conventional management practices. The application of beneficial soil organisms is being considered as a sustainable and environmental friendly alternative. In a previous study, the combined application in wheat fields of arbuscular mycorrhizal fungi, entomopathogenic Pseudomonas bacteria, and entomopathogenic nematodes was found to promote growth and protection against a natural pest infestation, without negative cross effects. Because of the insect-killing capacity of the bacteria and nematodes, we hypothesized that the application of these organisms would have similar or even greater beneficial effects in WCR-infested maize fields. During three consecutive years (2015-2017), we conducted trials in Missouri (USA) in which we applied the three organisms, alone or in combinations, in plots that were artificially infested with WCR and in non-infested control plots. For two of the three trials, we found that in plots treated with entomopathogenic nematodes and/or entomopathogenic Pseudomonas bacteria, roots were less damaged than the roots of plants in control plots. During one year, WCR survival was significantly lower in plots treated with Pseudomonas than in control plots, and the surviving larvae that were recovered from these plots were lighter. The bacterial and nematodes treatments also enhanced yield, assessed as total grain weight, in one of the trials. The effects of the treatments varied considerable among the three years, but they were always positive for the plants.


Assuntos
Besouros/fisiologia , Controle Biológico de Vetores , Doenças das Plantas/parasitologia , Zea mays/parasitologia , Animais , Besouros/microbiologia , Besouros/parasitologia , Micorrizas/fisiologia , Nematoides/fisiologia , Controle Biológico de Vetores/métodos , Pseudomonas/fisiologia , Zea mays/fisiologia
17.
Sci Rep ; 9(1): 4896, 2019 03 20.
Artigo em Inglês | MEDLINE | ID: mdl-30894586

RESUMO

Diabrotica virgifera virgifera LeConte, the western corn rootworm (WCR) is one of the most destructive pests in the U.S. Corn Belt. Transgenic maize lines expressing various Cry toxins from Bacillus thuringiensis have been adopted as a management strategy. However, resistance to many Bt toxins has occurred. To investigate the mechanisms of Bt resistance we carried out RNA-seq using Illumina sequencing technology on resistant, eCry3.1Ab-selected and susceptible, unselected, whole WCR neonates which fed on seedling maize with and without eCry3.1Ab for 12 and 24 hours. In a parallel experiment RNA-seq experiments were conducted when only the midgut of neonate WCR was evaluated from the same treatments. After de novo transcriptome assembly we identified differentially expressed genes (DEGs). Results from the assemblies and annotation indicate that WCR neonates from the eCry3.1Ab-selected resistant colony expressed a small number of up and down-regulated genes following Bt intoxication. In contrast, unselected susceptible WCR neonates expressed a large number of up and down-regulated transcripts in response to intoxication. Annotation and pathway analysis of DEGs between susceptible and resistant whole WCR and their midgut tissue revealed genes associated with cell membrane, immune response, detoxification, and potential Bt receptors which are likely related to eCry3.1Ab resistance. This research provides a framework to study the toxicology of Bt toxins and mechanism of resistance in WCR, an economically important coleopteran pest species.


Assuntos
Proteínas de Bactérias/genética , Besouros/genética , Endotoxinas/genética , Proteínas Hemolisinas/genética , Resistência a Inseticidas/genética , Controle Biológico de Vetores , Zea mays/genética , Animais , Bacillus thuringiensis/genética , Toxinas de Bacillus thuringiensis , Larva/genética , Plantas Geneticamente Modificadas
18.
Sci Rep ; 9(1): 3709, 2019 03 06.
Artigo em Inglês | MEDLINE | ID: mdl-30842452

RESUMO

The northern corn rootworm (NCR), Diabrotica barberi Smith & Lawrence, is a major pest of maize (Zea mays L.). This pest has developed resistance to insecticides and adapted to crop rotation and may already be in the early stages of adaptation to toxins produced by Bacillus thuringiensis (Bt). Toxicity bioassays using artificial diet have proven to be valuable for monitoring resistance in many species, but no artificial diet has been developed specifically for NCR larvae. Toward this end, we first evaluated known Diabrotica diets to identify a starting media. We then developed a specialized diet for NCR using an iterative approach. Screening designs including 8 diet components were performed to identify the principal nutritional components contributing to multiple developmental parameters (survival, weight, and molting). We then applied mixture designs coupled with response surface modeling to optimize a blend of those components. Finally, we validated an improved NCR diet formulation that supports approximately 97% survival and molting, and a 150% increase in larval weight after 10 days of feeding compared with the best previously published artificial diet. This formulation appears suitable for use in diet bioassays as a tool for evaluating the resistance of NCR populations to insecticides.


Assuntos
Besouros/fisiologia , Dieta/métodos , Alimentos Formulados/parasitologia , Animais , Bacillus thuringiensis/genética , Proteínas de Bactérias , Besouros/metabolismo , Endotoxinas , Inseticidas , Larva/fisiologia , Raízes de Plantas , Plantas Geneticamente Modificadas , Zea mays/genética
19.
J Econ Entomol ; 112(2): 842-851, 2019 03 21.
Artigo em Inglês | MEDLINE | ID: mdl-30668732

RESUMO

The susceptibility of western corn rootworm, Diabrotica virgifera virgifera LeConte, larvae to nine insecticides from five different classes and to Bt proteins eCry3.1Ab and mCry3A in the presence or absence of feeding stimulants, was estimated in filter paper and diet toxicity assays, respectively. The use of a synthetic feeding stimulant blend of the sugars glucose, sucrose, and fructose plus linoleic acid at a ratio of 30:4:4:0.3 mg/ml of distilled water was evaluated to determine whether they increase the efficacy of insecticides and Bt proteins. The efficacy of thiamethoxam diluted in solutions with feeding stimulants was significantly increased when compared to thiamethoxam dilutions in water (>60-fold). Differences in the efficacy of the other insecticide classes when diluted in feeding stimulant solutions were no greater than fivefold when compared to the insecticides diluted in water. The presence of corn root juice as a natural feeding stimulant diminished toxicity of the insecticides, except for thiamethoxam, even though larval fresh weight was higher when fed on root juice compared to feeding stimulant or water. The use of feeding stimulants in diet toxicity assays did not enhance efficacy of eCry3.1Ab nor mCry3A proteins. Feeding stimulants can be recommended in combination with thiamethoxam to increase larval mortality. These results are discussed in terms of applicability of feeding stimulants to improve susceptibility of western corn rootworm larvae to pesticides in general.


Assuntos
Besouros , Inseticidas , Animais , Proteínas de Bactérias , Endotoxinas , Proteínas Hemolisinas , Recém-Nascido , Larva , Plantas Geneticamente Modificadas , Zea mays
20.
J Econ Entomol ; 112(2): 708-711, 2019 03 21.
Artigo em Inglês | MEDLINE | ID: mdl-30649522

RESUMO

Western corn rootworm, Diabrotica virgifera virgifera LeConte, became much easier to research with the development of a nondiapausing rootworm strain. In the event that the eggs cannot be used immediately researchers have been known to delay egg hatch by storing the eggs at low temperatures. It is not well known how this technique could affect egg hatch or larval development, which could alter the results of an experiment. To test for this nondiapausing eggs of the western corn rootworm were stored at low temperatures to test for potential negative effects on hatch and larval development. Eggs were stored in either soil or agar and placed in refrigerators set to 4 or 8.5°C. Nondiapausing eggs were exposed to the cold for 1, 2, or 4 wk and then placed in a chamber set to 25°C. Eggs were then tested for average hatch percentage in Petri dishes and average larval recovery from containers with seedling corn. Results showed a significant reduction in percent hatch for eggs stored at 4°C for 4 wk. Larval recovery was significantly reduced in eggs stored for 4 wk at both 4 and 8.5°C. Within the treatments tested, egg storage for less than 4 wk in soil at 8.5°C provided the best hatch and larval recovery. Researchers wishing to store eggs may use these results to improve their rearing or testing of western corn rootworm.


Assuntos
Besouros , Animais , Larva , Plantas Geneticamente Modificadas , Zea mays
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