scholarly journals Independent and Synergistic Effects of Knocking out Two ABC Transporter Genes on Resistance to Bacillus thuringiensis Toxins Cry1Ac and Cry1Fa in Diamondback Moth

Toxins ◽  
2020 ◽  
Vol 13 (1) ◽  
pp. 9
Author(s):  
Shan Zhao ◽  
Dong Jiang ◽  
Falong Wang ◽  
Yihua Yang ◽  
Bruce E. Tabashnik ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Abc Transporter ◽  
Insect Pests ◽  
Synergistic Effects ◽  
Diamondback Moth ◽  
Cross Resistance ◽  
Vegetable Crops ◽  
Double Knockout ◽  
Bt Toxins ◽  
Abc Transporter Genes

Insecticidal proteins from Bacillus thuringiensis (Bt) are used widely in sprays and transgenic crops to control insect pests. However, evolution of resistance by pests can reduce the efficacy of Bt toxins. Here we analyzed resistance to Bt toxins Cry1Ac and Cry1Fa in the diamondback moth (Plutella xylostella), one of the world’s most destructive pests of vegetable crops. We used CRISPR/Cas9 gene editing to create strains with knockouts of the ATP-binding cassette (ABC) transporter genes PxABCC2, PxABCC3, or both. Bioassay results show that knocking out either gene alone caused at most 2.9-fold resistance but knocking out both caused >10,320-fold resistance to Cry1Ac and 380-fold resistance to Cry1Fa. Cry1Ac resistance in the double knockout strain was recessive and genetically linked with the PxABCC2/PxABCC3 loci. The results provide insight into the mechanism of cross-resistance to Cry1Fa in diamondback moth. They also confirm previous work with this pest showing that mutations disrupting both genes cause higher resistance to Cry1Ac than mutations affecting either PxABCC2 or PxABCC3 alone. Together with previous work, the results here highlight the value of using single and multiple gene knockouts to better understand the independent and synergistic effects of putative Bt toxin receptors on resistance to Bt toxins.

scholarly journals Insect resistance to Bacillus thuringiensis: uniform or diverse?

1998 ◽  
Vol 353 (1376) ◽  
pp. 1751-1756 ◽  
Author(s):  
B. E. Tabashnik ◽  
Y. Liu ◽  
T. Malvar ◽  
D. G. Heckel ◽  
L. Masson ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Insect Resistance ◽  
Diamondback Moth ◽  
Genetically Engineered ◽  
Cross Resistance ◽  
Recessive Inheritance ◽  
Insecticidal Proteins ◽  
Bt Toxins ◽  
Evolutionary Pathways ◽  
Genetically Engineered Crops

Resistance to the insecticidal proteins produced by the soil bacterium Bacillus thuringiensis (Bt) has been documented in more than a dozen species of insect. Nearly all of these cases have been produced primarily by selection in the laboratory, but one pest, the diamondback moth ( Plutella xylostella ), has evolved resistance in open–field populations. Insect resistance to Bt has immediate and widespread significance because of increasing reliance on Bt toxins in genetically engineered crops and conventional sprays. Furthermore, intense interest in Bt provides an opportunity to examine the extent to which evolutionary pathways to resistance vary among and within species of insect. One mode of resistance to Bt is characterized by more than 500–fold resistance to at least one Cry1A toxin, recessive inheritance, little or no cross–resistance to Cry1C, and reduced binding of at least one Cry1A toxin. Analysis of resistance to Bt in the diamondback moth and two other species of moth suggests that although this particular mode of resistance may be the most common, it is not the only means by which insects can attain resistance to Bt.

scholarly journals Bacillus thuringiensis Vip3Aa Toxin Resistance in Heliothis virescens (Lepidoptera: Noctuidae)

2017 ◽  
Vol 83 (9) ◽  
Author(s):  
Brian R. Pickett ◽  
Asim Gulzar ◽  
Juan Ferré ◽  
Denis J. Wright
Keyword(s):  
Bacillus Thuringiensis ◽  
Heliothis Virescens ◽  
Insect Pests ◽  
Management Strategies ◽  
Resistance Management ◽  
Cross Resistance ◽  
Inheritance Of Resistance ◽  
Resistant Parent ◽  
Bt Crops ◽  
Content Type

ABSTRACT Laboratory selection with Vip3Aa of a field-derived population of Heliothis virescens produced >2,040-fold resistance in 12 generations of selection. The Vip3Aa-selected (Vip-Sel)-resistant population showed little cross-resistance to Cry1Ab and no cross-resistance to Cry1Ac. Resistance was unstable after 15 generations without exposure to the toxin. F1 reciprocal crosses between Vip3Aa-unselected (Vip-Unsel) and Vip-Sel insects indicated a strong paternal influence on the inheritance of resistance. Resistance ranged from almost completely recessive (mean degree of dominance [h] = 0.04 if the resistant parent was female) to incompletely dominant (mean h = 0.53 if the resistant parent was male). Results from bioassays on the offspring from backcrosses of the F1 progeny with Vip-Sel insects indicated that resistance was due to more than one locus. The results described in this article provide useful information for the insecticide resistance management strategies designed to overcome the evolution of resistance to Vip3Aa in insect pests. IMPORTANCE Heliothis virescens is an important pest that has the ability to feed on many plant species. The extensive use of Bacillus thuringiensis (Bt) crops or spray has already led to the evolution of insect resistance in the field for some species of Lepidoptera and Coleoptera. The development of resistance in insect pests is the main threat to Bt crops. The effective resistance management strategies are very important to prolong the life of Bt plants. Lab selection is the key step to test the assumption and predictions of management strategies prior to field evaluation. Resistant insects offer useful information to determine the inheritance of resistance and the frequency of resistance alleles and to study the mechanism of resistance to insecticides.

scholarly journals Insecticidal Effects of Organotin(IV) Compounds on Plutella Xylostella (L.) Larvae. II. Inhibitory Potencies Against Acetylcholinesterase and Evidence for Synergism in Tests With Bacillus Thuringiensis(BER.) and Malathion

1994 ◽  
Vol 1 (1) ◽  
pp. 1-17 ◽  
Author(s):  
Nazni W. Ahmad ◽  
Tay Siew Huang ◽  
S. Balabaskaran ◽  
K. M. Lo ◽  
V. G. Kumar Das
Keyword(s):  
Bacillus Thuringiensis ◽  
Plutella Xylostella ◽  
Resistant Strain ◽  
Synergistic Effects ◽  
Diamondback Moth ◽  
Ache Inhibition ◽  
Microbial Insecticide ◽  
In Vitro Inhibition ◽  
Sublethal Concentrations

Features of pesticide synergism and acetylcholinesterase (AChE) inhibition (in vitro) were studied using a selected range of organotin compounds against the early 4th instar larvae of a highly resistant strain of the diamondback moth (DBM), Plutella xylostella, a major universal pest of cruciferous vegetables.Fourteen triorganotin compounds were evaluated for their ability to enhance the toxicity of the microbial insecticide, Bacillus thuringiensis (BT) and of the commercial insecticide, Malathion to Plutella xylostella larvae. Supplemental synergism was observed with triphenyl- and tricyclopentyltin hydroxides in combinations with Bacillus thuringiensis. Increased synergism was observed with an increase in the number of cyclopentyl groups on tin in the mixed series, CypnPh3-n SnX, where X = OH, and 1-(1,2,4-triazolyl). The combination of (p-chlorophenyl)diphenyltin N,N-dimethyldithiocarbamate at LD10 and LD25 concentrations with sublethal concentrations of Malathion as well as of tricyclohexyltin methanesulphonate at the 0.01% (w/v) concentration with Malathion exerted strong synergistic effects (supplemental synergism) with toxicity index (T.I) values of 7.2, 19.8 and 10.1, respectively.Studies on the in vitro inhibition of acetylcholinesterase prepared from the DBM larvae showed that while most of the triorganotin Compounds tested were without effect on the enzyme, compounds containing the thiocarbamylacetate or the dithiocarbamylacetate moieties demonstrated appreciable levels of inhibition, being comparable in efficacy to commercial grades of Malathion and Methomyl.

Different Cross-Resistance Patterns in the Diamondback Moth (Lepidoptera: Plutellidae) Resistant to Bacillus thuringiensis Toxin Cry1C

2001 ◽  
Vol 94 (6) ◽  
pp. 1547-1552 ◽  
Author(s):  
Jian-Zhou Zhao ◽  
Ya-Xin LI ◽  
Hilda L. Collins ◽  
Jun Cao ◽  
Elizabeth D. Earle ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Diamondback Moth ◽  
Cross Resistance ◽  
Resistance Patterns

scholarly journals Fishing for New Bt Receptors in Diamondback Moth

2017 ◽  
Author(s):  
Yazhou Chen ◽  
Yuping Huang ◽  
Qun Liu ◽  
Jun Xu ◽  
Saskia Hogenhout ◽  
...  
Keyword(s):  
Brush Border ◽  
Molecular Mechanisms ◽  
Brush Border Membrane ◽  
Insect Pests ◽  
Diamondback Moth ◽  
Genetically Modified Crops ◽  
Ongoing Research ◽  
Bt Toxins ◽  
Insect Gut ◽  
Immunoblotting Technique

ABSTRACTBt toxins bind to receptors in the brush border membrane of the insect gut and create pores, leading to insect death. Bt-resistant insects demonstrate reduced binding of the Bt toxins to gut membranes. However, our understanding of the gut receptors involved in Bt toxin binding, and which receptors confer resistance to these toxins is incomplete, especially in diamondback moth (Plutella xylostella), a major agricultural pest. Identifying receptors has remained challenging because we lack sufficiently sensitive methods to detect Bt receptor interactions. Here, we report a modified far-immunoblotting technique, which revealed a broad spectrum of binding targets for the Bt toxins Cry1Ac, Cry1Ab, and Cry1Bd in diamondback moth. We confirm the role of the glucosinolate sulfatases GSS1 and GSS2 in Cry1Bd toxicity. GSS1 and GSS2 bind directly to Cry1Bd, and their expression is crucial for Cry1Bd toxicity. These results improve our understanding of the molecular mechanisms of Bt toxicity.AUTHOR SUMMARYThe Bt toxins, from the soil bacteriumBacillus thuringiensis, have wide applications in agriculture as insecticides applied to plants or expressed in genetically modified crops. Bt toxins bind to receptors in the brush border membrane of the insect gut and create pores leading to insect death. The success of the Bt toxins in controlling insect pests has been hindered by the emergence of resistant insects, which show reduced binding of Bt to their gut membranes. Although ongoing research has identified a few receptors, many remain unknown and the mechanisms by which these receptors cause resistance remain unclear. Here, we used a modified far-immunoblotting technique to identify proteins that bind to the toxins Cry1Ac, Cry1Ab, and Cry1Bd in the diamondback moth. This identified two glucosinolate sulfatases that bind directly to Cry1Bd; also, the toxicity of Cry1Bd requires expression of these glucosinolate sulfatases. Therefore, identification of these candidate receptors improves our understanding of Bt function and resistance.

scholarly journals Toxicity and Binding Studies of Bacillus thuringiensis Cry1Ac, Cry1F, Cry1C, and Cry2A Proteins in the Soybean Pests Anticarsia gemmatalis and Chrysodeixis (Pseudoplusia) includens

2017 ◽  
Vol 83 (11) ◽  
Author(s):  
Yolanda Bel ◽  
Joel J. Sheets ◽  
Sek Yee Tan ◽  
Kenneth E. Narva ◽  
Baltasar Escriche
Keyword(s):  
Bacillus Thuringiensis ◽  
Binding Sites ◽  
Specific Binding ◽  
Management Strategies ◽  
Anticarsia Gemmatalis ◽  
Cross Resistance ◽  
Binding Studies ◽  
Content Type ◽  
Bt Toxins ◽  
Competition Binding

ABSTRACT Anticarsia gemmatalis (velvetbean caterpillar) and Chrysodeixis includens (soybean looper, formerly named Pseudoplusia includens) are two important defoliating insects of soybeans. Both lepidopteran pests are controlled mainly with synthetic insecticides. Alternative control strategies, such as biopesticides based on the Bacillus thuringiensis (Bt) toxins or transgenic plants expressing Bt toxins, can be used and are increasingly being adopted. Studies on the insect susceptibilities and modes of action of the different Bt toxins are crucial to determine management strategies to control the pests and to delay outbreaks of insect resistance. In the present study, the susceptibilities of both soybean pests to the Bt toxins Cry1Ac, Cry1Fa, Cry1Ca, and Cry2Aa have been investigated. Bioassays performed in first-instar larvae showed that both insects are susceptible to all these toxins. Competition-binding studies carried out with Cry1Ac and Cry1Fa 125-iodine labeled proteins demonstrated the presence of specific binding sites for both of them on the midgut brush border membrane vesicles (BBMVs) of both A. gemmatalis and C. includens. Competition-binding experiments and specific-binding inhibition studies performed with selected sugars and lectins indicated that Cry1Ac and Cry1Fa share some, but not all, binding sites in the midguts of both insects. Also, the Cry1Ac- or Cry1Fa-binding sites were not shared with Cry1Ca or Cry2Aa in either soybean pest. This study contributes to the knowledge of Bt toxicity and midgut toxin binding sites in A. gemmatalis and C. includens and sheds light on the cross-resistance potential of Cry1Ac, Cry1Fa, Cry1Ca, and Cry2Aa Bt proteins as candidate proteins for Bt-pyramided crops. IMPORTANCE In the present study, the toxicity and the mode of action of the Bacillus thuringiensis (Bt) toxins Cry1Ac, Cry1Fa, Cry1Ca, and Cry2Aa in Anticarsia gemmatalis and Chrysodeixis includens (important defoliating pests of soybeans) have been investigated. These studies are crucial for determining management strategies for pest control. Bioassays showed that both insects were susceptible to the toxins. Competition-binding studies demonstrated the presence of Cry1Fa- and Cry1Ac-specific binding sites in the midguts of both pests. These results, together with the results from binding inhibition studies performed with sugars and lectins, indicated that Cry1Ac and Cry1Fa share some, but not all, binding sites, and that they were not shared with Cry1Ca or Cry2Aa in either soybean pest. This study contributes to the knowledge of Bt toxicity in A. gemmatalis and C. includens and sheds light on the cross-resistance potential of Cry1Ac, Cry1Fa, Cry1Ca, and Cry2Aa Bt proteins as candidate proteins for Bt-pyramided crops.

Enhancement of Activity of Bacillus thuringiensis Berliner Against Four Lepidopterous Insect Pests by Nutrient-Based Phagostimulants1

1995 ◽  
Vol 30 (1) ◽  
pp. 29-42 ◽  
Author(s):  
Robert R. Farrar ◽  
Richard L. Ridgway
Keyword(s):  
Bacillus Thuringiensis ◽  
Ostrinia Nubilalis ◽  
Helicoverpa Zea ◽  
European Corn Borer ◽  
Insect Pests ◽  
Diamondback Moth ◽  
Lethal Dose ◽  
Corn Earworm ◽  
Corn Borer ◽  
Whole Plants

To help improve control of insect pests with microbial insecticides, we investigated the interactions of four commercial, nutrient-based phagostimulants (Pheast [AgriSense], Coax [CCT Corp.], Gusto [Atochem North America, Inc.], and Entice [Custom Chemicides] with Bacillus thuringiensis Berliner and four lepidopterous insect pests (gypsy moth, Lymantria dispar [L.] [Lymantriidae]; corn earworm, Helicoverpa zea [Boddie] [Noctuidae]; European corn borer, Ostrinia nubilalis [Hübner] [Pyralidae]; and diamondback moth, Plutella xylostella [L.] [Plutellidae]). Comparisons were made of treated foliage in Petri dishes in the laboratory and of sprayed whole plants in a greenhouse. In general, phagostimulants increased mortality of all species tested, but no consistent differences among phagostimulants were found for any species. Food consumption was generally lower on the treatments that contained phagostimulants causing the highest rates of mortality, possibly as a result of more rapid ingestion of a lethal dose on these treatments. Reduced rates of feeding by insects on treatments with B. thuringiensis alone were seen, probably due in part to intoxication and, possibly, to behavioral effects as well. Indications of potentially significant interactions between host plants and both B. thuringiensis and phagostimulants also were seen.

scholarly journals Insect resistance management in Bacillus thuringiensis cotton by MGPS (multiple genes pyramiding and silencing)

2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Muhammad Mubashar ZAFAR ◽  
Abdul RAZZAQ ◽  
Muhammad Awais FAROOQ ◽  
Abdul REHMAN ◽  
Hina FIRDOUS ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Insect Pests ◽  
Crop Protection ◽  
High Dose ◽  
Bt Cotton ◽  
Artificial Chromosomes ◽  
Cotton Pests ◽  
Multiple Genes ◽  
Bt Toxins ◽  
Evolution Of Resistance

AbstractThe introduction of Bacillus thuringiensis (Bt) cotton has reduced the burden of pests without harming the environment and human health. However, the efficacy of Bt cotton has decreased due to field-evolved resistance in insect pests over time. In this review, we have discussed various factors that facilitate the evolution of resistance in cotton pests. Currently, different strategies like pyramided cotton expressing two or more distinct Bt toxin genes, refuge strategy, releasing of sterile insects, and gene silencing by RNAi are being used to control insect pests. Pyramided cotton has shown resistance against different cotton pests. The multiple genes pyramiding and silencing (MGPS) approach has been proposed for the management of cotton pests. The genome information of cotton pests is necessary for the development of MGPS-based cotton. The expression cassettes against various essential genes involved in defense, detoxification, digestion, and development of cotton pests will successfully obtain favorable agronomic characters for crop protection and production. The MGPS involves the construction of transformable artificial chromosomes, that can express multiple distinct Bt toxins and RNAi to knockdown various essential target genes to control pests. The evolution of resistance in cotton pests will be delayed or blocked by the synergistic action of high dose of Bt toxins and RNAi as well as compliance of refuge requirement.

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.

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