Showing papers on "Bacillus thuringiensis published in 2018"
TL;DR: This study is the first documented confirmation and characterization of S. frugiperda Cry1F field-evolved resistance in Argentina, which is autosomal and incompletely recessive similar to the resistance reported in Brazil.
Abstract: BACKGROUND
Transgenic maize (Zea mays L.) event TC1507 (Herculex® I insect protection), expressing Cry1F δ-endotoxin derived from Bacillus thuringiensis var. aizawai, was commercialized in 2003 in the Americas. Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) susceptibility to Cry1F was monitored annually across several regions in Argentina using diagnostic concentration bioassays. Reduced performance of TC1507 maize against S. frugiperda was reported in 2013. A resistant population was established in the laboratory and the dominance of Cry1F resistance was characterized. RESULTS: During 2012–2015, high-survivorship of several populations was observed in the resistance monitoring program. Reciprocal crosses of a Cry1F-resistant population with a Cry1F-susceptible population were evaluated to calculate effective dominance (DML) based on mortality levels observed at 100 µg/ml Cry1F. Two additional dominance levels (DLC and DEC) were calculated using lethal (LC50) or effective concentration (EC50) derived from concentration-response bioassays. Estimates indicated that Cry1F resistance in S. frugiperda in Argentina was either highly-recessive (DML=0.005) or incompletely recessive (DLC < 0.26 and DEC<0.19).
CONCLUSION
This study is the first documented confirmation and characterization of S. frugiperda Cry1F field-evolved resistance in Argentina. The resistance to Cry1F in S. frugiperda populations collected in Argentina, is autosomal and incompletely recessive similar to the resistance reported in Brazil.
103 citations
TL;DR: Proactive identification and tracking of the tetraspanin mutation demonstrate the potential for genomic analysis, gene editing, and molecular monitoring to improve management of resistance and enhance sustainability of Bt cotton for millions of smallholder farmers in China.
Abstract: Extensive planting of crops genetically engineered to produce insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) has suppressed some major pests, reduced insecticide sprays, enhanced pest control by natural enemies, and increased grower profits. However, rapid evolution of resistance in pests is reducing these benefits. Better understanding of the genetic basis of resistance to Bt crops is urgently needed to monitor, delay, and counter pest resistance. We discovered that a point mutation in a previously unknown tetraspanin gene in the cotton bollworm (Helicoverpa armigera), a devastating global pest, confers dominant resistance to Cry1Ac, the sole Bt protein produced by transgenic cotton planted in China. We found the mutation using a genome-wide association study, followed by fine-scale genetic mapping and DNA sequence comparisons between resistant and susceptible strains. CRISPR/Cas9 knockout of the tetraspanin gene restored susceptibility to a resistant strain, whereas inserting the mutation conferred 125-fold resistance in a susceptible strain. DNA screening of moths captured from 23 field sites in six provinces of northern China revealed a 100-fold increase in the frequency of this mutation, from 0.001 in 2006 to 0.10 in 2016. The correspondence between the observed trajectory of the mutation and the trajectory predicted from simulation modeling shows that the dominance of the mutation accelerated adaptation. Proactive identification and tracking of the tetraspanin mutation demonstrate the potential for genomic analysis, gene editing, and molecular monitoring to improve management of resistance.
97 citations
TL;DR: How endophytic Bacillus species are a valuable multifunctional toolbox to be integrated with crop management practices for achieving higher crop yield is discussed.
Abstract: One of the major challenges of agriculture currently is to obtain higher crop yield. Environmental conditions, cultivar quality, and plant diseases greatly affect plant productivity. On the other hand, several endophytic Bacillus species have emerged as a complementary, efficient, and safe alternative to current crop management practices. The ability of Bacillus species to form spores, which resist adverse conditions, is an advantage of the genus for use in formulations. Endophytic Bacillus species provide plants with a wide range of benefits, including protection against phytopathogenic microorganisms, insects, and nematodes, eliciting resistance, and promoting plant growth, without causing damage to the environment. Bacillus thuringiensis, B. subtilis, B. amyloliquefaciens, B. velezensis, B. cereus, B. pumilus, and B. licheniformis are the most studied Bacillus species for application in agriculture, although other species within the genus have also shown great potential. Due to the increasing number of whole-genome sequenced endophytic Bacillus spp. strains, various bioactive compounds have been predicted. These data reveal endophytic Bacillus species as an underexploited source of novel molecules of biotechnological interest. In this review, we discuss how endophytic Bacillus species are a valuable multifunctional toolbox to be integrated with crop management practices for achieving higher crop yield.
66 citations
TL;DR: This study shows that many B. thuringiensis biopesticide strains exhibit mid-level cytotoxicity in a Vero cell assay and that some of these strains cannot be differentiated from isolates collected from foods or in association with outbreaks.
Abstract: While the relevance of Bacillus (B.) cereus as a major cause of gastroenteritis is undisputed, the role of the closely related B. thuringiensis in foodborne disease is unclear. B. thuringiensis strains frequently harbor enterotoxin genes. However, the organism has only very rarely been associated with foodborne outbreaks, possibly due to the fact that during outbreak investigations, B. cereus is routinely not differentiated from B. thuringiensis. A recent EFSA scientific opinion stresses the urgent need for further data allowing for improved risk assessment, in particular as B. thuringiensis is a commonly used biopesticide. Therefore, the aim of this study was to gain further insights into the hazardous potential of B. thuringiensis. To this end, 39 B. thuringiensis isolates obtained from commercially used biopesticides, various food sources, as well as from foodborne outbreaks were characterized by panC typing, panC-based SplitsTree analysis, toxin gene profiling, FTIR spectroscopic analysis, a cytotoxicity assay screening for enterotoxic activity, and a sphingomyelinase assay. The majority of the tested B. thuringiensis isolates exhibited low (23%, n = 9) or mid level enterotoxicity (74%, n = 29), and produced either no (59%, n = 23) or low levels (33%, n = 13) of sphingomyelinase, which is reported to act synergistically with enterotoxins Nhe and Hbl. One strain isolated from rosemary was however classified as highly enterotoxic surpassing the cytotoxic activity of the high-level reference strain by a factor of 1.5. This strain also produced vast amounts of sphingomyelinase. Combining all results obtained in this study into a fingerprint pattern, several enterotoxic biopesticide strains were indistinguishable from those of isolates from foods or collected in association with outbreaks. Our study shows that many B. thuringiensis biopesticide strains exhibit mid-level cytotoxicity in a Vero cell assay and that some of these strains cannot be differentiated from isolates collected from foods or in association with outbreaks. Thus, we demonstrate that the use of B. thuringiensis strains as biopesticides can represent a food safety risk, underpinning the importance of assessing the hazardous potential of each strain and formulation used.
65 citations
TL;DR: It is discovered that resistance to Cry2Ab is associated with mutations disrupting the same ATP-binding cassette transporter gene (PgABCA2) in a laboratory-selected strain from Arizona, USA, and in field-selected populations from India, implying that mutations in the same gene confer Bt resistance in laboratory- and field- selected strains.
Abstract: Evolution of pest resistance threatens the benefits of genetically engineered crops that produce Bacillus thuringiensis (Bt) insecticidal proteins. Strategies intended to delay pest resistance are most effective when implemented proactively. Accordingly, researchers have selected for and analyzed resistance to Bt toxins in many laboratory strains of pests before resistance evolves in the field, but the utility of this approach depends on the largely untested assumption that laboratory- and field-selected resistance to Bt toxins are similar. Here we compared the genetic basis of resistance to Bt toxin Cry2Ab, which is widely deployed in transgenic crops, between laboratory- and field-selected populations of the pink bollworm (Pectinophora gossypiella), a global pest of cotton. We discovered that resistance to Cry2Ab is associated with mutations disrupting the same ATP-binding cassette transporter gene (PgABCA2) in a laboratory-selected strain from Arizona, USA, and in field-selected populations from India. The most common mutation, loss of exon 6 caused by alternative splicing, occurred in resistant larvae from both locations. Together with previous data, the results imply that mutations in the same gene confer Bt resistance in laboratory- and field-selected strains and suggest that focusing on ABCA2 genes may help to accelerate progress in monitoring and managing resistance to Cry2Ab.
59 citations
TL;DR: There is a worldwide controversy about the safety of Bt crops to the environment and mammals, and some researchers support the cultivation of BT crops depending upon the results of their laboratory and field studies on the safety, while others, however, are against Bt crop as they may cause risk to human.
Abstract: Bacillus thuringiensis (Bt) crops are plants genetically engineered (modified) to contain the endospore (or crystal) toxins of the bacterium, Bt to be resistant to certain insect pests. In 1995, the Environmental Protection Agency (EPA) in USA approved the commercial production and distribution of the Bt crops: corn, cotton, potato, and tobacco. Currently, the most common Bt crops are corn and cotton. The crystal, referred to as Cry toxins, is proteins formed during sporulation of some Bt strains and aggregate to form crystals. Such Cry toxins are toxic to specific species of insects belongs to orders: Lepidoptera, Coleoptera, Hymenoptera, Diptera, and Nematoda. In 2016, the total world area cultivated with genetically modified crops (GM crops) reached about 185 million ha. This review shows that there is a worldwide controversy about the safety of Bt crops to the environment and mammals. Some researchers support the cultivation of Bt crops depending upon the results of their laboratory and field studies on the safety of such crops. Others, however, are against Bt crops as they may cause risk to human.
55 citations
TL;DR: The proteins Cry1F, Cry2Aa, and the herbicide glyphosate were highly toxic to the stingless bee M. quadrifasciata, causing lethal or sublethal effects which can severely impair colony growth and viability, and reduce pollination ability.
52 citations
TL;DR: Data support an important role for SeP‐gp in susceptibility of S. exigua to insecticides and imply that overexpression of SeP-gp may contribute to abamectin and EB resistance in S.Exigua.
Abstract: P-glycoprotein [P-gp or the ATP-binding cassette transporter B1 (ABCB1)] is an important participant in multidrug resistance of cancer cells, yet the precise function of this arthropod transporter is unknown. The aim of this study was to determine the importance of P-gp for susceptibility to insecticides in the beet armyworm (Spodoptera exigua) using clustered regularly interspaced short palindromic repeats/CRISPR-associated 9 (CRISPR/Cas9) gene-editing technology. We cloned an open reading frame (ORF) encoding the S. exigua P-gp protein (SeP-gp) predicted to display structural characteristics common to P-gp and other insect ABCB1 transporters. A knockout line with a frame shift deletion of four nucleotides in the SeP-gp ORF was established using the CRISPR/Cas9 gene-editing system to test its potential role in determining susceptibility to chemical insecticides or insecticidal proteins from the bacterium Bacillus thuringiensis (Bt). Results from comparative bioassays demonstrate that knockout of SeP-gp significantly increases susceptibility of S. exigua by around threefold to abamectin and emamectin benzoate (EB), but not to spinosad, chlorfenapyr, beta-cypermethrin, carbosulfan indoxacarb, chlorpyrifos, phoxim, diafenthiuron, chlorfluazuron, chlorantraniliprole or two Bt toxins (Cry1Ca and Cry1Fa). Our data support an important role for SeP-gp in susceptibility of S. exigua to abamectin and EB and imply that overexpression of SeP-gp may contribute to abamectin and EB resistance in S. exigua.
49 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: Comparison between clades suggests that co‐evolutionary interactions may drive association between plasmids and chromosomes and limit wider transfer of key virulence traits in the B. cereus group.
Abstract: Bacterial plasmids can vary from small selfish genetic elements to large autonomous replicons that constitute a significant proportion of total cellular DNA. By conferring novel function to the cell, plasmids may facilitate evolution but their mobility may be opposed by co-evolutionary relationships with chromosomes or encouraged via the infectious sharing of genes encoding public goods. Here, we explore these hypotheses through large-scale examination of the association between plasmids and chromosomal DNA in the phenotypically diverse Bacillus cereus group. This complex group is rich in plasmids, many of which encode essential virulence factors (Cry toxins) that are known public goods. We characterized population genomic structure, gene content and plasmid distribution to investigate the role of mobile elements in diversification. We analysed coding sequence within the core and accessory genome of 190 B. cereus group isolates, including 23 novel sequences and genes from 410 reference plasmid genomes. While cry genes were widely distributed, those with invertebrate toxicity were predominantly associated with one sequence cluster (clade 2) and phenotypically defined Bacillus thuringiensis. Cry toxin plasmids in clade 2 showed evidence of recent horizontal transfer and variable gene content, a pattern of plasmid segregation consistent with transfer during infectious cooperation. Nevertheless, comparison between clades suggests that co-evolutionary interactions may drive association between plasmids and chromosomes and limit wider transfer of key virulence traits. Proliferation of successful plasmid and chromosome combinations is a feature of specialized pathogens with characteristic niches (Bacillus anthracis, B. thuringiensis) and has occurred multiple times in the B. cereus group.
43 citations
TL;DR: Results from assays of responses of the T. ni strains to different Bt toxins revealed that the cadherin and ABCC2 play independent roles in the mode of action of Cry1A toxins and that BT toxins exert insecticidal activity through multiple redundant pathways of toxicity in insects.
TL;DR: Expression of Cry2Aa in transgenic pigeon pea and its effectiveness towards H. armigera is reported by employing Agrobacterium-mediated in planta transformation approach and can be used in pigeonpea improvement programmes for pod borer resistance.
Abstract: Pigeon pea is an important legume infested by a plethora of insect pests amongst which gram pod borer Helicoverpa armigera is very prominent. Imparting resistance to this insect herbivore is of global importance in attaining food security. Expression of insecticidal crystal proteins (ICP) in diverse crops has led to increased resistance to several pests. We report in this paper, expression of Cry2Aa in transgenic pigeon pea and its effectiveness towards H. armigera by employing Agrobacterium-mediated in planta transformation approach. Approximately 0.8% of T1 generation plants were identified as putative transformants based on screening in the presence of 70 ppm kanamycin as the selection agent. Promising events were further recognized in advanced generations based on integration, expression and bioefficacy of the transgenes. Seven T3 lines (11.8% of the selected T1 events) were categorized as superior as these events demonstrated 80–100% mortality of the challenged larvae and improved ability to prevent damage caused by the larvae. The selected transgenic plants accumulated Cry2Aa in the range of 25–80 µg/g FW. The transgenic events developed in the study can be used in pigeon pea improvement programmes for pod borer resistance.
TL;DR: Denaturing gradi ent gel electrophoresis (DGGE) fingerprint patterns showed that Bt rice had little effect on the dominant rhizo sphere bacterial, fungal and actinobacterial communities.
Abstract: In contrast to other transgenic Bacillus thuringiensis (Bt) crops (e.g. Bt maize and cotton), risk assessments of Bt rice on soil ecosystem are few. To assess the influence of Bt rice on rhizosphere soil ecosystems, soil samples from Bt, non-Bt and controls were taken at seedling, tillering, booting, heading and maturing stages. The activities of dehydrogenases, invertase, phenol oxidases, acid phosphatases, ureases and proteases showed no significant differ ences between Bt and non-Bt rice. A Biolog system was used to evaluate the effect of Bt rice on functional diversity of microbial communities. Although there were differences in carbon substrate utilization between Bt and non-Bt rice at seedling, tillering and heading stages, these differences were transient and not persistent. Denaturing gradi ent gel electrophoresis (DGGE) fingerprint patterns showed that Bt rice had little effect on the dominant rhizo sphere bacterial, fungal and actinobacterial communities. The richness and consistency of microbial communities according to carbon substrate utilizations and DGGE band patterns did not differ significantly between Bt and nonBt rice, and were close to that of control soil. There was no evidence to indicate apparent effects of Bt rice on soil enzyme activities, microbial community composition and functional diversity in this study.
TL;DR: Qatari soil ecology provides a good collection and diversity of Bt isolates, the majority of which are different and very promising for the search of novel insecticidal endotoxins.
Abstract: Bacillus thuringiensis (Bt) based insecticidal formulations have been recognized as one of the most successful, environmentally safe and sustainable method of controlling insect pests. Research teams worldwide are in search of Bt diversity giving more choices of bio-insecticides and alternatives to address insect resistance. In fact, there are many unexplored ecologies that could harbor novel Bt strains. This study is the first initiative to explore Bt strain diversity in Qatar. A collection of 700 Bt isolates was constructed. Scanning electron microscopy of Bt crystals showed different crystal forms, with a high abundance of spherical crystals compared to the bipyramidal ones. Among the spherical crystals, four different morphologies were observed. The δ-endotoxin content of parasporal crystals from each Bt isolate revealed that there are 16 different protein profiles among the isolates of the collection. On the other hand, plasmid pattern analysis showed seven different plasmid profiles. Their insecticidal activity was predicted by exploring the δ-endotoxin coding genes and conducting qualitative insecticidal bioassays. 19 smooth spherical crystal producing isolates have been identified that could be possible candidates for endotoxin production targeting Dipteran insects. Another group of 259 isolates producing bipyramidal and cuboidal crystals could target Lepidopteran and Coleopteran insects. The remaining 422 isolates have novel profiles. In conclusion, Qatari soil ecology provides a good collection and diversity of Bt isolates. In addition to strains harboring genes encoding common endotoxins, the majority are different and very promising for the search of novel insecticidal endotoxins.
TL;DR: The first case of a cadherin transmembrane mutation associated with insect resistance to Bt is reported, discovered in a strain of the devastating global cotton pest, the pink bollworm, derived from a field population in the Yangtze River Valley of China.
TL;DR: In this paper, the authors reviewed published and unpublished data from field trials of Bt cotton in the eastern and central Cotton Belt of the USA through 2015 to evaluate the effectiveness of bt cotton (Bollgard, Bollgard II, Widestrike, WideStrike 3, and TwinLink).
Abstract: The primary management tactic for lepidopteran pests of cotton in the United States of America (USA) is the use of transgenic cotton that produces Bacillus thuringiensis Berliner (Bt) toxins. The primary target pests of this technology are Helicoverpa zea (Boddie) and Heliothis virescens (F.) in the eastern and central Cotton Belt of the USA. Concerns over the evolution of resistance in H. zea to Bt toxins and scrutiny of the necessity of Bt crops has escalated. We reviewed published and unpublished data from field trials of Bt cotton in the eastern and central Cotton Belt of the USA through 2015 to evaluate the effectiveness of Bt cotton (Bollgard, Bollgard II, WideStrike, WideStrike 3, and TwinLink). Bt cotton reduced insecticide usage, reduced heliothine pest numbers and damage, and provided a yield benefit, but Bollgard II and WideStrike efficacy declined in the Midsouth over the period evaluated. In the Southeastern region, heliothine damage remained constant through 2015, but yield benefits declined from 2010 until 2015. Resistance of H. zea to several Bt toxins is the most plausible explanation for the observed changes in Bt cotton efficacy. The introduction of new Bt toxins such as found in Widestrike 3 and Twinlink may preserve the benefits of Bt crops. However, while both Widestrike 3 and Twinlink had less damage than Widestrike, damage levels of both were similar to Bollgard II.
TL;DR: In this review, the putative role of the mobile gene pool of B. thuringiensis serovar israelensis in its pathogenicity and dedicated lifestyle is reviewed, with specific emphasis on the nature, diversity, and potential mobility of its constituents.
Abstract: Bacillus thuringiensis is a well-known biopesticide that has been used for more than 80 years This spore-forming bacterium belongs to the group of Bacillus cereus that also includes, among others, emetic and diarrheic pathotypes of B cereus, the animal pathogen Bacillus anthracis and the psychrotolerant Bacillus weihenstephanensis Bacillus thuringiensis is rather unique since it has adapted its lifestyle as an efficient pathogen of specific insect larvae One of the peculiarities of B thuringiensis strains is the extent of their extrachromosomal pool, with strains harbouring more than 10 distinct plasmid molecules Among the numerous serovars of B thuringiensis, 'israelensis' is certainly emblematic since its host spectrum is apparently restricted to dipteran insects like mosquitoes and black flies, vectors of human and animal diseases such as malaria, yellow fever, or river blindness In this review, the putative role of the mobile gene pool of B thuringiensis serovar israelensis in its pathogenicity and dedicated lifestyle is reviewed, with specific emphasis on the nature, diversity, and potential mobility of its constituents Variations among the few related strains of B thuringiensis serovar israelensis will also be reported and discussed in the scope of this specialised insect pathogen, whose lifestyle in the environment remains largely unknown
TL;DR: By similarly and efficiently controlling lepidopteran strains susceptible and resistant to Bt toxins, the S. guianensis essential oil represents a promising management tool against key lepidpteran pests.
Abstract: Although the cultivation of transgenic plants expressing toxins of Bacillus thuringiensis (Bt) represents a successful pest management strategy, the rapid evolution of resistance to Bt plants in several lepidopteran pests has threatened the sustainability of this practice. By exhibiting a favorable safety profile and allowing integration with pest management initiatives, plant essential oils have become relevant pest control alternatives. Here, we assessed the potential of essential oils extracted from a Neotropical plant, Siparuna guianensis Aublet, for improving the control and resistance management of key lepidopteran pests (i.e., Spodoptera frugiperda and Anticarsia gemmatalis). The essential oil exhibited high toxicity against both lepidopteran pest species (including an S. frugiperda strain resistant to Cry1A.105 and Cry2Ab Bt toxins). This high insecticidal activity was associated with necrotic and apoptotic effects revealed by in vitro assays with lepidopteran (but not human) cell lines. Furthermore, deficits in reproduction (e.g., egg-laying deterrence and decreased egg viability), larval development (e.g., feeding inhibition) and locomotion (e.g., individual and grouped larvae walking activities) were recorded for lepidopterans sublethally exposed to the essential oil. Thus, by similarly and efficiently controlling lepidopteran strains susceptible and resistant to Bt toxins, the S. guianensis essential oil represents a promising management tool against key lepidopteran pests.
TL;DR: The data show that insect specificity of Cyt1Aa toxin can be modified by introduction of loop regions from another non-related toxin with different insect specificity.
Abstract: The Cyt and Cry toxins are different pore-forming proteins produced by Bacillus thuringiensis bacteria, and used in insect-pests control. Cry-toxins have a complex mechanism involving interaction with several proteins in the insect gut such as aminopeptidase N (APN), alkaline phosphatase (ALP) and cadherin (CAD). It was shown that the loop regions of domain II of Cry toxins participate in receptor binding. Cyt-toxins are dipteran specific and interact with membrane lipids. We show that Cry1Ab domain II loop3 is involved in binding to APN, ALP and CAD receptors since point mutation Cry1Ab-G439D affected binding to these proteins. We hypothesized that construction of Cyt1A-hybrid proteins providing a binding site that recognizes gut proteins in lepidopteran larvae could result in improved Cyt1Aa toxin toward lepidopteran larvae. We constructed hybrid Cyt1Aa-loop3 proteins with increased binding interaction to Manduca sexta receptors and increased toxicity against two Lepidopteran pests, M. sexta and Plutella xylostella. The hybrid Cyt1Aa-loop3 proteins were severely affected in mosquitocidal activity and showed partial hemolytic activity but retained their capacity to synergize Cry11Aa toxicity against mosquitos. Our data show that insect specificity of Cyt1Aa toxin can be modified by introduction of loop regions from another non-related toxin with different insect specificity.
TL;DR: The results indicate that PIP‐47Aa is a novel insecticidal protein for controlling the corn rootworm pests.
Abstract: The coleopteran insect western corn rootworm (WCR, Diabrotica virgifera virgifera) is an economically important pest in North America and Europe. Transgenic corn plants producing Bacillus thuringiensis (Bt) insecticidal proteins have been useful against this devastating pest, but evolution of resistance has reduced their efficacy. Here, we report the discovery of a novel insecticidal protein, PIP-47Aa, from an isolate of Pseudomonas mosselii. PIP-47Aa sequence shows no shared motifs, domains or signatures with other known proteins. Recombinant PIP-47Aa kills WCR, two other corn rootworm pests (Diabrotica barberi and Diabrotica undecimpunctata howardi) and two other beetle species (Diabrotica speciosa and Phyllotreta cruciferae), but it was not toxic to the spotted lady beetle (Coleomegilla maculata) or seven species of Lepidoptera and Hemiptera. Transgenic corn plants expressing PIP-47Aa show significant protection from root damage by WCR. PIP-47Aa kills a WCR strain resistant to mCry3A and does not share rootworm midgut binding sites with mCry3A or AfIP-1A/1B from Alcaligenes that acts like Cry34Ab1/Cry35Ab1. Our results indicate that PIP-47Aa is a novel insecticidal protein for controlling the corn rootworm pests.
TL;DR: The results indicate that the Cry toxin–binding affinities of ABCC transporters are largely linked to the level of Cry susceptibility ofABCC-expressing cells and that the ABCC ECL structures determine the specificities to Cry toxins.
TL;DR: A novel strategy for creating abiotic Bacillus thuringiensis protein affinity ligands by biomimicry of the recognition process that takes place between Bt Cry1Ab/Ac proteins and insect receptor cadherin-like Bt-R1 proteins is reported.
Abstract: We report a novel strategy for creating abiotic Bacillus thuringiensis (Bt) protein affinity ligands by biomimicry of the recognition process that takes place between Bt Cry1Ab/Ac proteins and insect receptor cadherin-like Bt-R1 proteins. Guided by this strategy, a library of synthetic polymer nanoparticles (NPs) was prepared and screened for binding to three epitopes 280FRGSAQGIEGS290, 368RRPFNIGINNQQ379 and 436FRSGFSNSSVSIIR449 located in loop α8, loop 2 and loop 3 of domain II of Bt Cry1Ab/Ac proteins. A negatively charged and hydrophilic nanoparticle (NP12) was found to have high affinity to one of the epitopes, 368RRPFNIGINNQQ379. This same NP also had specific binding ability to both Bt Cry1Ab and Bt Cry1Ac, proteins that share the same epitope, but very low affinity to Bt Cry2A, Bt Cry1C and Bt Cry1F closely related proteins that lack epitope homology. To locate possible NP-Bt Cry1Ab/Ac interaction sites, NP12 was used as a competitive inhibitor to block the binding of 865NITIHITDTNNK876, a specifi...
TL;DR: A single amino acid residue located in loop 1 of ABCC2 transporter as responsible for the different levels of susceptibility to Cry1Ac among various lepidopteran species is identified.
TL;DR: Diet-based bioassays indicate that the corn-strains and hybrid populations are more tolerant to the Bt toxins, especially to Cry1F, than the rice-strain population.
Abstract: The fall armyworm, Spodoptera frugiperda (Smith; Lepidoptera: Noctuidae), is a highly polyphagous, multivoltine pest of commercial crops including corn (Zea mays L.), cotton (Gossypium spp. L.), rice (Oryza sativa L.), and pasture grasses. Fall armyworm has become a growing concern in agricultural communities across the Americas as field populations in many locales have evolved resistance to several Cry1 toxins derived from the bacterium Bacillus thuringiensis Berliner (Bt). An often overlooked aspect of fall armyworm biology is the existence of two host strains, the 'rice' and 'corn' strains. There has been little research devoted to the characterization of fall armyworm host strains, although there is evidence that the rice and corn-strains may differ in their tolerances to Bt toxins expressed by transgenic plants. In this study, diet-based bioassays were conducted to compare the susceptibilities of one rice-strain, two corn-strains, and one rice-corn hybrid population to Cry1Ab, Cry1Ac, and Cry1F protein. Results indicate that the corn-strains and hybrid populations are more tolerant to the Bt toxins, especially to Cry1F, than the rice-strain population. Results from this study, when combined with existing techniques for host strain identification, may aid in the development of regional insect resistance management programs for fall armyworm.
TL;DR: Increasing level of intracellular c-di-GMP exhibits similar activities as those in Gram-negative bacteria, including altered activities in cell motility, biofilm formation, and cell-cell aggregation, and a novel function exhibited by the increasing level of c- DiGMP to promote the insecticidal activity of this bacterium against Helicoverpa armigera is found.
Abstract: C-di-GMP has been well investigated to play significant roles in the physiology of many Gram-negative bacteria. However, its effect on Gram-positive bacteria is less known. In order to more understand the c-di-GMP functions in Gram-positive bacteria, we have carried out a detailed study on the c-di-GMP-metabolizing enzymes and their physiological functions in Bacillus thuringiensis, a Gram-positive entomopathogenic bacterium that has been applied as an insecticide successfully. We performed a systematic study on the ten putative c-di-GMP-synthesizing enzyme diguanylate cyclases (DGCs) and c-di-GMP-degrading enzyme phosphodiesterases (PDEs) in B. thuringiensis BMB171, and artificially elevated the intracellular c-di-GMP level in BMB171 by deleting one or more pde genes. We found increasing level of intracellular c-di-GMP exhibits similar activities as those in Gram-negative bacteria, including altered activities in cell motility, biofilm formation, and cell-cell aggregation. Unexpectedly, we additionally found a novel function exhibited by the increasing level of c-di-GMP to promote the insecticidal activity of this bacterium against Helicoverpa armigera. Through whole-genome transcriptome profile analyses, we found that 4.3% of the B. thuringiensis genes were differentially transcribed when c-di-GMP level was increased, and 77.3% of such gene products are involved in some regulatory pathways not reported in other bacteria to date. In summary, our study represents the first comprehensive report on the c-di-GMP-metabolizing enzymes, their effects on phenotypes, and the transcriptome mediated by c-di-GMP in an important Gram-positive bacterium.
TL;DR: It is indicated that the fibroblast growth factor receptor-like protein (Sf-FGFR) from the membrane of Sf9 cells could bind to Vip3Aa and is a novel receptor for Vip 3Aa.
Abstract: Vegetative insecticidal proteins (Vips), which are secreted by some Bacillus thuringiensis strains during vegetative growth, exhibit high virulence to many pests. Vip3A proteins have been used commercially both in some bio-insecticides and in transgenic crops; however, compared with insecticidal crystal proteins, the mechanism of action of Vip3A is still unclear. In this work, we indicated that the fibroblast growth factor receptor-like protein (Sf-FGFR) from the membrane of Sf9 cells could bind to Vip3Aa. The interaction between Vip3Aa and Sf-FGFR was confirmed by pull-down assays and dot blotting experiment in vitro. The binding affinity between Vip3Aa and extracellular regions of Sf-FGFR (GST-FGFR-N) was determined by microscale thermophoresis assay (MST). Moreover, Vip3Aa-Flag could be co-immunoprecipitated with Sf-FGFR-V5 ex vivo. Furthermore, knockdown of Sf-FGFR gene in Sf9 cells resulted in reducing the mortality of those cells to Vip3Aa. In summary, our data indicated that Sf-FGFR is a novel receptor for Vip3Aa.
TL;DR: Results of this study suggest that M. anisopliae ICIPE 30 was the most potent candidate and could be used in an autodissemination approach for management of adult S. recurvalis.
Abstract: Spoladea recurvalis (Fabricius) is one of the most devastating pests of amaranths causing severe yield losses of 60%–100% to the crop. Unfortunately use of chemical pesticides is the most common control strategy that vegetable farmers rely on to control the pest. However, it is not effective and harmful to environmental and human health. Aiming to provide more environmentally friendly alternatives, this study evaluated the effects of various entomopathogenic fungal isolates and commercial based Bacillus thuringiensis Subsp. kurstaki product Halt®, on the pest. Twenty‐four entomopathogenic fungal (EPF) isolates from three genera (14 Metarhizium anisopliae, 9 Beauveria bassiana and 1 lsaria fumosorosea) were screened in the laboratory to assess their pathogenicity against second instar larvae of S. recurvalis. Only M. anisopliae ICIPE 30 reached a moderate threshold, causing 58.3% larval mortality. All the 11 isolates (8 M. anisopliae, 2 B. bassiana and 1 l. fumosorosea) tested against adult S. recurvalis were pathogenic, with M. anisopliae ICIPE 30 and B. bassiana ICIPE 725 causing the highest mortality of 92% and 83%, respectively. Metarhizium anisopliae ICIPE 30 had the shortest LT50 value of 4.8 days. Bacillus thuringiensis Subsp. kurstaki product Halt® caused <50% mortality on S. recurvalis larvae. A consecutive application of M. anisopliae ICIPE 30 and Bt did not cause a significant increase in larval mortality compared to separate applications of both products. Results of this study suggest that M. anisopliae ICIPE 30 was the most potent candidate and could be used in an autodissemination approach for management of adult S. recurvalis.
TL;DR: It is shown that the B. thuringiensis Cry9Aa and Vip3Aa toxins display potent synergy based on a specific interaction between them, which is highly relevant to the development of toxin combinations for effective insect control and for delaying development of insect resistance.
TL;DR: The reduction in pupal weight in the hybrids expressingCry1F and Cry1F + Cry1Ab significantly declined over time in South Carolina, possibly indicating developing resistance to these Bt toxins.
Abstract: The corn earworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), is only moderately susceptible to most toxins from the bacterium Bacillus thuringiensis (Bt) expressed in transgenic corn. To better understand the impact of Bt corn on the life cycle of H. zea, we collected pupae of H. zea during 2014-2016 in North Carolina, South Carolina, and Mississippi from corn hybrids expressing Cry1Ab, Cry1F, Cry1F + Cry1Ab, Cry1F + Cry1Ab + Vip3Aa20, Cry1A.105 + Cry2Ab2, and Cry1A.105 + Cry2Ab2 + Cry1F, as well as from non-Bt near-isolines. We investigated the effect of Bt corn on pupal weight, fecundity, and egg viability of H. zea. Pupal weights were significantly reduced for males and females from all Bt hybrids compared with non-Bt near-isolines. Female pupae from the hybrid expressing Cry1F + Cry1Ab were also significantly lighter relative to those from the near-isolines expressing only Cry1F. Reductions in pupal weight did not result in any detectable effects on fecundity or egg viability. The reduction in pupal weight in the hybrids expressing Cry1F and Cry1F + Cry1Ab significantly declined over time in South Carolina, possibly indicating developing resistance to these Bt toxins. These data can be incorporated into insect resistance management models used to improve risk management decisions regarding H. zea in Bt crops in the complex landscapes of the southern United States.
TL;DR: The results of the present study revealed the larvicidal potential of B. thuringiensis isolates found in the soils of the Cerrado region from Maranhao, although further research will be necessary to better elucidate and describe other genes associated with the production of insecticidal toxins in these isolates.
Abstract: This study investigated the potential of Bacillus thuringiensis isolates obtained in the Cerrado region of the Brazilian state of Maranhao for the biological control of Aedes aegypti larvae. The isolates were obtained from soil samples and the identification of the B. thuringiensis colonies was based on morphological characteristics. Bioassays were run to assess the pathogenicity and toxicity of the different strains of the B. thuringiensis against third-instar larvae of A. aegypti. Protein profiles were obtained by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Polymerase chain reaction assays were used to detect the toxin genes found in the bacterial isolates. Overall, 12 (4.0%) of the 300 isolates obtained from 45 soil samples were found to present larvicidal activity, with the BtMA-104, BtMA-401 and BtMA-560 isolates causing 100% of mortality. The BtMA-401 isolate was the most virulent, with the lowest median lethal concentration (LC50) (0.004 × 107 spores/mL), followed by the Bacillus thuringiensis var. israelensis standard (0.32 × 107 spores/mL). The protein profiles of BtMA-25 and BtMA-401 isolates indicated the presence of molecular mass consistent with the presence of the proteins Cry4Aa, Cry11Aa and Cyt1, similar to the profile of Bacillus thuringiensis var. israelensis IPS-82. Surprisingly, however, none of the cry and cyt genes analyzed were amplified in the isolate BtMA-401. The results of the present study revealed the larvicidal potential of B. thuringiensis isolates found in the soils of the Cerrado region from Maranhao, although further research will be necessary to better elucidate and describe other genes associated with the production of insecticidal toxins in these isolates.