Dual mode of action of Bt proteins: protoxin efficacy against resistant insects
TL;DR: Evidence is reported from seven resistant strains of three major crop pests showing that Cry1Ab and Cry1Ac protoxins were generally more potent than the corresponding activated toxins.
Abstract: Transgenic crops that produce Bacillus thuringiensis (Bt) proteins for pest control are grown extensively, but insect adaptation can reduce their effectiveness. Established mode of action models assert that Bt proteins Cry1Ab and Cry1Ac are produced as inactive protoxins that require conversion to a smaller activated form to exert toxicity. However, contrary to this widely accepted paradigm, we report evidence from seven resistant strains of three major crop pests showing that Cry1Ab and Cry1Ac protoxins were generally more potent than the corresponding activated toxins. Moreover, resistance was higher to activated toxins than protoxins in eight of nine cases evaluated in this study. These data and previously reported results support a new model in which protoxins and activated toxins kill insects via different pathways. Recognizing that protoxins can be more potent than activated toxins against resistant insects may help to enhance and sustain the efficacy of transgenic Bt crops.
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TL;DR: This work reviews the available data on specificity determinants of crystal (Cry) insecticidal proteins as the Bt toxins most well characterized and used in transgenic crops and defines seven levels that could influence specificity.
122 citations
TL;DR: Understanding how Bt toxins kill insects will help to understand the nature of both problems and the recent realization that ABC transporters play a central role in the killing mechanism will play an important role in devising solutions.
Abstract: Three-domain Cry toxins from the bacterium Bacillus thuringiensis (Bt) are increasingly used in agriculture to replace chemical insecticides in pest control. Most chemical insecticides kill pest insects swiftly, but are also toxic to beneficial insects and other species in the agroecosystem. Cry toxins enjoy the advantages of high selectivity and the possibility of the application by sprays or transgenic plants. However, these benefits are offset by the limited host range and the evolution of resistance to Bt toxins by insect pests. Understanding how Bt toxins kill insects will help to understand the nature of both problems. The recent realization that ABC transporters play a central role in the killing mechanism will play an important role in devising solutions.
64 citations
TL;DR: It is confirmed for the first time that Sf-SR-C acts as a receptor for Vip3Aa protoxin and provides an insight into the mode of action of Vip 3Aa that will significantly facilitate the study of its insecticidal mechanism and application.
Abstract: The vegetative insecticidal proteins (Vip), secreted by many Bacillus thuringiensis strains during their vegetative growth stage, are genetically distinct from known insecticidal crystal proteins (ICPs) and represent the second-generation insecticidal toxins. Compared with ICPs, the insecticidal mechanisms of Vip toxins are poorly understood. In particular, there has been no report of a definite receptor of Vip toxins to date. In the present study, we identified the scavenger receptor class C like protein (Sf-SR-C) from the Spodoptera frugiperda (Sf9) cells membrane proteins that bind to the biotin labeled Vip3Aa, via the affinity magnetic bead method coupled with HPLC-MS/MS. We then certified Vip3Aa protoxin could interact with Sf-SR-C in vitro and ex vivo. In addition, downregulation of SR-C expression in Sf9 cells and Spodoptera exigua larvae midgut reduced the toxicity of Vip3Aa to them. Coincidently, heterologous expression of Sf-SR-C in transgenic Drosophila midgut significantly enhanced the virulence of Vip3Aa to the Drosophila larvae. Moreover, the complement control protein domain and MAM domain of Sf-SR-C are involved in the interaction with Vip3Aa protoxin. Furthermore, endocytosis of Vip3Aa mediated by Sf-SR-C correlates with its insecticidal activity. Our results confirmed for the first time that Sf-SR-C acts as a receptor for Vip3Aa protoxin and provides an insight into the mode of action of Vip3Aa that will significantly facilitate the study of its insecticidal mechanism and application.
45 citations
TL;DR: It is shown that the C-terminal region of Cry1Ab protoxin provides additional binding sites for alkaline phosphatase (ALP) and aminopeptidase N (APN) insect receptors, suggesting an active role of the C.terminal protoxin fragment in the mode of action of Cry toxins.
30 citations
Cites background from "Dual mode of action of Bt proteins:..."
...Conversely, they are consistent with the proposition that the C-terminal regions of protoxins contribute to the binding and toxicity of Cry proteins (18, 21)....
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...Furthermore, both forms promoted different post-binding events in the toxic pathway; two different pathways of oligomerization and pore formation have been described that are based on the interaction of protoxin or the activated toxin with the CAD receptor (18, 21)....
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TL;DR: How Bt crop development has moved past single-gene events, toward multigene stacked varieties that often contain herbicide resistance genes in addition to multiple Bt genes, and how their use (in conjunction with co-technology such as glyphosate/Roundup) may impact and interact with aquatic ecosystems is discussed.
Abstract: The term Bt crops collectively refers to crops that have been genetically modified to include a gene (or genes) sourced from Bacillus thuringiensis (Bt) bacteria. These genes confer the ability to produce proteins toxic to certain insect pests. The interaction between Bt crops and adjacent aquatic ecosystems has received limited attention in research and risk assessment, despite the fact that some Bt crops have been in commercial use for 20 yr. Reports of effects on aquatic organisms such as Daphnia magna, Elliptio complanata, and Chironomus dilutus suggest that some aquatic species may be negatively affected, whereas other reports suggest that the decreased use of insecticides precipitated by Bt crops may benefit aquatic communities. The present study reviews the literature regarding entry routes and exposure pathways by which aquatic organisms may be exposed to Bt crop material, as well as feeding trials and field surveys that have investigated the effects of Bt-expressing plant material on such organisms. The present review also discusses how Bt crop development has moved past single-gene events, toward multigene stacked varieties that often contain herbicide resistance genes in addition to multiple Bt genes, and how their use (in conjunction with co-technology such as glyphosate/Roundup) may impact and interact with aquatic ecosystems. Lastly, suggestions for further research in this field are provided. Environ Toxicol Chem 2016;35:2891-2902. © 2016 SETAC.
28 citations
Cites background from "Dual mode of action of Bt proteins:..."
...It is theorized that bacterial production of protoxins is a strategy to impede resistance development [51]....
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...In fact, in some cases, the protoxin was more effective than the activated toxin [51,53]....
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References
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TL;DR: This assay is very reproducible and rapid with the dye binding process virtually complete in approximately 2 min with good color stability for 1 hr with little or no interference from cations such as sodium or potassium nor from carbohydrates such as sucrose.
225,085 citations
TL;DR: The understanding of the biochemical and genetic basis of resistance to Bt can help design appropriate management tactics to delay or reduce the evolution of resistance in insect populations.
Abstract: Bacillus thuringiensis (Bt) is a valuable source of insecticidal proteins for use in conventional sprayable formulations and in transgenic crops, and it is the most promising alternative to synthetic insecticides. However, evolution of resistance in insect populations is a serious threat to this technology. So far, only one insect species has evolved significant levels of resistance in the field, but laboratory selection experiments have shown the high potential of other species to evolve resistance against Bt. We have reviewed the current knowledge on the biochemical mechanisms and genetics of resistance to Bt products and insecticidal crystal proteins. The understanding of the biochemical and genetic basis of resistance to Bt can help design appropriate management tactics to delay or reduce the evolution of resistance in insect populations.
907 citations
TL;DR: Field outcomes support theoretical predictions that factors delaying resistance include recessive inheritance of resistance, low initial frequency of resistance alleles, abundant refuges of non-Bt host plants and two-toxin Bt crops deployed separately from one-t toxin BT crops.
Abstract: Evolution of resistance in pests can reduce the effectiveness of insecticidal proteins from Bacillus thuringiensis (Bt) produced by transgenic crops We analyzed results of 77 studies from five continents reporting field monitoring data for resistance to Bt crops, empirical evaluation of factors affecting resistance or both Although most pest populations remained susceptible, reduced efficacy of Bt crops caused by field-evolved resistance has been reported now for some populations of 5 of 13 major pest species examined, compared with resistant populations of only one pest species in 2005 Field outcomes support theoretical predictions that factors delaying resistance include recessive inheritance of resistance, low initial frequency of resistance alleles, abundant refuges of non-Bt host plants and two-toxin Bt crops deployed separately from one-toxin Bt crops The results imply that proactive evaluation of the inheritance and initial frequency of resistance are useful for predicting the risk of resistance and improving strategies to sustain the effectiveness of Bt crops
838 citations