scholarly journals Binary Toxins from Bacillus thuringiensis Active against the Western Corn Rootworm, Diabrotica virgifera virgifera LeConte

2004 ◽  
Vol 70 (8) ◽  
pp. 4889-4898 ◽  
Author(s):  
James A. Baum ◽  
Chi-Rei Chu ◽  
Mark Rupar ◽  
Gregory R. Brown ◽  
William P. Donovan ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Insect Pest ◽  
Diabrotica Virgifera Virgifera ◽  
The United States ◽  
Western Corn Rootworm ◽  
Binary Toxin ◽  
Corn Rootworm ◽  
Diabrotica Virgifera ◽  
Crystal Proteins ◽  
Genes Encoding

ABSTRACT The western corn rootworm, Diabrotica virgifera virgifera LeConte, is a significant pest of corn in the United States. The development of transgenic corn hybrids resistant to rootworm feeding damage depends on the identification of genes encoding insecticidal proteins toxic to rootworm larvae. In this study, a bioassay screen was used to identify several isolates of the bacterium Bacillus thuringiensis active against rootworm. These bacterial isolates each produce distinct crystal proteins with approximate molecular masses of 13 to 15 kDa and 44 kDa. Insect bioassays demonstrated that both protein classes are required for insecticidal activity against this rootworm species. The genes encoding these proteins are organized in apparent operons and are associated with other genes encoding crystal proteins of unknown function. The antirootworm proteins produced by B. thuringiensis strains EG5899 and EG9444 closely resemble previously described crystal proteins of the Cry34A and Cry35A classes. The antirootworm proteins produced by strain EG4851, designated Cry34Ba1 and Cry35Ba1, represent a new binary toxin. Genes encoding these proteins could become an important component of a sustainable resistance management strategy against this insect pest.

scholarly journals Functional validation of DvABCB1 as a receptor of Cry3 toxins in western corn rootworm, Diabrotica virgifera virgifera

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Xiping Niu ◽  
Adane Kassa ◽  
James Hasler ◽  
Samantha Griffin ◽  
Claudia Perez-Ortega ◽  
...  
Keyword(s):  
Resistant Strain ◽  
Insect Pest ◽  
Amino Acid Level ◽  
Diabrotica Virgifera Virgifera ◽  
Western Corn Rootworm ◽  
Hek293 Cells ◽  
Corn Rootworm ◽  
Diabrotica Virgifera ◽  
Functional Validation ◽  
Bt Toxins

Abstract Western corn rootworm (WCR), Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae), is a serious insect pest in the major corn growing areas of North America and in parts of Europe. WCR populations with resistance to Bacillus thuringiensis (Bt) toxins utilized in commercial transgenic traits have been reported, raising concerns over their continued efficacy in WCR management. Understanding the modes of action of Bt toxins is important for WCR control and resistance management. Although different classes of proteins have been identified as Bt receptors for lepidopteran insects, identification of receptors in WCR has been limited with no reports of functional validation. Our results demonstrate that heterologous expression of DvABCB1 in Sf9 and HEK293 cells conferred sensitivity to the cytotoxic effects of Cry3A toxins. The result was further validated using knockdown of DvABCB1 by RNAi which rendered WCR larvae insensitive to a Cry3A toxin. However, silencing of DvABCB2 which is highly homologous to DvABCB1 at the amino acid level, did not reduce the sensitivity of WCR larvae to a Cry3A toxin. Furthermore, our functional studies corroborate different mode-of-actions for other insecticidal proteins including Cry34Ab1/35Ab1, Cry6Aa1, and IPD072Aa against WCR. Finally, reduced expression and alternatively spliced transcripts of DvABCB1 were identified in a mCry3A-resistant strain of WCR. Our results provide the first clear demonstration of a functional receptor in the molecular mechanism of Cry3A toxicity in WCR and confirmed its role in the mechanism of resistance in a mCry3A resistant strain of WCR.

Strategies for controlling western corn rootworm (Diabrotica virgifera virgifera)

2005 ◽  
Vol 53 (1) ◽  
pp. 71-79 ◽  
Author(s):  
E. Széll ◽  
I. Zsellér ◽  
G. Ripka ◽  
J. Kiss ◽  
G. Princzinger
Keyword(s):  
Life Cycle ◽  
Seed Treatment ◽  
Large Scale ◽  
Insect Pest ◽  
Diabrotica Virgifera Virgifera ◽  
Western Corn Rootworm ◽  
Corn Rootworm ◽  
Diabrotica Virgifera ◽  
Non Profit ◽  
Small Plot

Western corn rootworm (WCR) first appeared in Europe near Belgrade airport in 1992. The first adult of this species was found near Mórahalom, in the southern part of Hungary, on 30th June 1995. Small-plot trials were started in Szeged (Cereal Research Non-Profit Co.) and large-scale trials in Mezohegyes (Ménesbirtok Ltd.) in 1996 to elaborate strategies to control this insect pest. To make the work more efficient, eight experts working at different institutions formed a team. At the same time, monitoring was begun on when and where the insect appeared and in what numbers. The experiments were adapted to the life cycle of the species. WCR has one generation per year, laying eggs that overwinter. Larvae begin to hatch in late May to mid-June. Adults emerge by mid-July and survive till early October. They lay eggs in abundance from mid-July to late August.  Trials were conducted to control WCR both with and without chemicals. For adult control pesticides were sprayed from an aircraft or using a field sprayer. Larval control involved seed treatment with insecticide or the use of soil insecticide at planting or at cultivation.

Oviposition site selected by the western corn rootworm (Diabrotica virgifera virgifera Leconte) in southern Ontario strip plots

2005 ◽  
Vol 85 (4) ◽  
pp. 949-954 ◽  
Author(s):  
C. C. Knight ◽  
K. P. Pauls ◽  
M. K. Sears ◽  
A. W. Schaafsma
Keyword(s):  
Insect Pest ◽  
Adult Emergence ◽  
Diabrotica Virgifera Virgifera ◽  
Conclusive Evidence ◽  
Western Corn Rootworm ◽  
Egg Laying ◽  
Larval Feeding ◽  
Corn Rootworm ◽  
Diabrotica Virgifera ◽  
Southern Ontario

Western corn rootworm (WCR), Diabrotica virgifera virgifera L., is the most serious insect pest of corn, Zea mays L., in North America. Crop rotation had successfully controlled this pest until the late 1980s when a new biotype that lays its eggs in non-corn crops was discovered in Illinois. This biotype has spread to parts of Indiana, Michigan, and Ohio, and threatens Ontario. By using emergence cages, Pherocon Am® sticky traps and corn root ratings, egg-laying behaviour of WCR in Ontario was assessed in strip crop settings that represent typical cropping systems of southern Ontario. WCR adults were active in both soybean and winter wheat fields in July and August. Larval feeding and adult emergence in first-year corn fields following soybean and wheat were similar, confirming that oviposition did occur in both crops. These data do not, however, provide conclusive evidence for the presence of the rotation resistant WCR in Ontario because previous studies have demonstrated that WCR are capable of utilizing other crops for oviposition. Key words: Western corn rootworm, Diabrotica virgifera virgifera, strip plots, corn, soybean, wheat (winter)

scholarly journals Engineering of Bacillus thuringiensis Cry Proteins to Enhance the Activity against Western Corn Rootworm

Toxins ◽  
2019 ◽  
Vol 11 (3) ◽  
pp. 162 ◽  
Author(s):  
Jingtong Hou ◽  
Ruth Cong ◽  
Michi Izumi-Willcoxon ◽  
Hana Ali ◽  
Yi Zheng ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
3D Structure ◽  
Diabrotica Virgifera Virgifera ◽  
Western Corn Rootworm ◽  
Corn Rootworm ◽  
Diabrotica Virgifera ◽  
Cry Protein ◽  
Neutral Ph ◽  
Highly Active ◽  
Cry Toxin

A novel Bacillus thuringiensis Cry protein, Cry8Hb, active against Diabrotica virgifera virgifera (Western corn rootworm, WCRW) was discovered. Unexpectedly, the anti-rootworm activity of the Cry8Hb toxin was enhanced significantly by fusing Escherichia coli maltose binding protein (MBP) to this Cry toxin. While the exact mechanism of the activity enhancement remains indefinite, it is probable that the enhancement is a result of increased solubility of the MBP-Cry8Hb fusion in the rootworm midgut. This hypothesis was examined using a synthetic Cry3 protein called IP3-1, which was not soluble at a neutral pH like Cry8Hb and marginally active to WCRW. When IP3-1 was fused to MBP, its anti-WCRW activity was enhanced 13-fold. To further test the hypothesis, DNA shuffling was performed on IP3-1 to increase the solubility without MBP. Screening of shuffled libraries found six new IP3 variants showing very high anti-WCRW activity without MBP. Sequence and 3D structure analysis of those highly active, shuffled IP3 variants revealed several charge-altering mutations such as Lys to Glu on the putative MBP-attaching side of the IP3 molecule. It is likely that those mutations make the protein acidic to substitute the functions of MBP including enhancing the solubility of IP3 at a neutral pH.

scholarly journals Cry75Aa (Mpp75Aa) Insecticidal Proteins for Controlling the Western Corn Rootworm, Diabrotica virgifera virgifera, (Coleoptera: Chrysomelidae), Isolated from the Insect Pathogenic Bacteria Brevibacillus laterosporus.

2020 ◽  
Author(s):  
David Bowen ◽  
Yong Yin ◽  
Stanislaw Flasinski ◽  
Catherine Chay ◽  
Gregory Bean ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Pathogenic Bacteria ◽  
Insect Pests ◽  
Diabrotica Virgifera Virgifera ◽  
Western Corn Rootworm ◽  
Cross Resistance ◽  
Corn Rootworm ◽  
Diabrotica Virgifera ◽  
Insecticidal Proteins ◽  
Brevibacillus Laterosporus

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.

EFFECTS OF AZADIRACHTIN ON THE WESTERN CORN ROOTWORM, DIABROTICA VIRGIFERA VIRGIFERA (LECONTE) (COLEOPTERA: CHRYSOMELIDAE)

1991 ◽  
Vol 123 (3) ◽  
pp. 707-710 ◽  
Author(s):  
Y.S. Xie ◽  
D. Gagnon ◽  
J.T. Arnason ◽  
B.J.R. Philogène ◽  
J.D.H. Lambert ◽  
...  
Keyword(s):  
United States ◽  
Diabrotica Virgifera Virgifera ◽  
The United States ◽  
Western Corn Rootworm ◽  
Crop Damage ◽  
Corn Rootworm ◽  
Diabrotica Virgifera ◽  
Corn Production ◽  
Main Method ◽  
Diabrotica Virgifera Virgifera Leconte

Corn rootworm (Diabrotica spp., Coleoptera: Chrysomelidae) is a serious pest insect of corn production. It is estimated that farmers in the United States have losses of over $1 billion each year as a result of crop damage and treatment costs for this pest (Metcalf 1986). Chemical control is the main method of suppressing corn rootworm populations and the amount of insecticide used against Diabrotica spp. is greater than for any other pests of corn in the United States (Suguiyama and Carlson 1985). The development of nontoxic and biodegradable alternatives to chemical insecticides is highly desirable.

scholarly journals Parameters for Successful Parental RNAi as An Insect Pest Management Tool in Western Corn Rootworm, Diabrotica virgifera virgifera

Genes ◽  
2016 ◽  
Vol 8 (1) ◽  
pp. 7 ◽  
Author(s):  
Ana Vélez ◽  
Elane Fishilevich ◽  
Natalie Matz ◽  
Nicholas Storer ◽  
Kenneth Narva ◽  
...  
Keyword(s):  
Pest Management ◽  
Insect Pest ◽  
Diabrotica Virgifera Virgifera ◽  
Western Corn Rootworm ◽  
Management Tool ◽  
Corn Rootworm ◽  
Insect Pest Management ◽  
Diabrotica Virgifera

scholarly journals The Use of Insecticides to Manage the Western Corn Rootworm, Diabrotica virgifera virgifera, LeConte: History, Field-Evolved Resistance, and Associated Mechanisms

Insects ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 112 ◽  
Author(s):  
Lance J. Meinke ◽  
Dariane Souza ◽  
Blair D. Siegfried
Keyword(s):  
Diabrotica Virgifera Virgifera ◽  
The United States ◽  
Western Corn Rootworm ◽  
Cross Resistance ◽  
Corn Rootworm ◽  
Diabrotica Virgifera ◽  
Mechanisms Of Resistance ◽  
Pyrethroid Insecticides ◽  
Diabrotica Virgifera Virgifera Leconte ◽  
Over Time

The western corn rootworm, Diabrotica virgifera virgifera LeConte (Dvv) is a significant insect pest of maize in the United States (U.S.). This paper reviews the history of insecticide use in Dvv management programs, Dvv adaptation to insecticides, i.e., field-evolved resistance and associated mechanisms of resistance, plus the current role of insecticides in the transgenic era. In the western U.S. Corn Belt where continuous maize is commonly grown in large irrigated monocultures, broadcast-applied soil or foliar insecticides have been extensively used over time to manage annual densities of Dvv and other secondary insect pests. This has contributed to the sequential occurrence of Dvv resistance evolution to cyclodiene, organophosphate, carbamate, and pyrethroid insecticides since the 1950s. Mechanisms of resistance are complex, but both oxidative and hydrolytic metabolism contribute to organophosphate, carbamate, and pyrethroid resistance facilitating cross-resistance between insecticide classes. History shows that Dvv insecticide resistance can evolve quickly and may persist in field populations even in the absence of selection. This suggests minimal fitness costs associated with Dvv resistance. In the transgenic era, insecticides function primarily as complementary tools with other Dvv management tactics to manage annual Dvv densities/crop injury and resistance over time.

scholarly journals Structural and functional insights into the first Bacillus thuringiensis vegetative insecticidal protein of the Vpb4 fold, active against western corn rootworm

PLoS ONE ◽  
2021 ◽  
Vol 16 (12) ◽  
pp. e0260532
Author(s):  
Jean-Louis Kouadio ◽  
Meiying Zheng ◽  
Michael Aikins ◽  
David Duda ◽  
Stephen Duff ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Protective Antigen ◽  
Domain Architecture ◽  
Diabrotica Virgifera Virgifera ◽  
The United States ◽  
Western Corn Rootworm ◽  
Corn Rootworm ◽  
Insecticidal Proteins ◽  
Amino Terminal ◽  
Carbohydrate Binding Modules

The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, is a major maize pest in the United States causing significant economic loss. The emergence of field-evolved resistant WCR to Bacillus thuringiensis (Bt) traits has prompted the need to discover and deploy new insecticidal proteins in transgenic maize. In the current study we determined the crystal structure and mode of action (MOA) of the Vpb4Da2 protein (formerly known as Vip4Da2) from Bt, the first identified insecticidal Vpb4 protein with commercial level control against WCR. The Vpb4Da2 structure exhibits a six-domain architecture mainly comprised of antiparallel β-sheets organized into β-sandwich layers. The amino-terminal domains 1–3 of the protein share structural homology with the protective antigen (PA) PA14 domain and encompass a long β-pore forming loop as in the clostridial binary-toxB module. Domains 5 and 6 at the carboxyl-terminal half of Vpb4Da2 are unique as this extension is not observed in PA or any other structurally-related protein other than Vpb4 homologs. These unique Vpb4 domains adopt the topologies of carbohydrate-binding modules known to participate in receptor-recognition. Functional assessment of Vpb4Da2 suggests that domains 4–6 comprise the WCR receptor binding region and are key in conferring the observed insecticidal activity against WCR. The current structural analysis was complemented by in vitro and in vivo characterizations, including immuno-histochemistry, demonstrating that Vpb4Da2 follows a MOA that is consistent with well-characterized 3-domain Bt insecticidal proteins despite significant structural differences.

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