Additional file 6: Figure S2. of Transcriptional cellular responses in midgut tissue of Aedes aegypti larvae following intoxication with Cry11Aa toxin from Bacillus thuringiensis

Characterization of JNK expression by Western Blot analysis using α-JNK antibody. Presence of JNK was determined on protein extracts of midguts of A. aegypti larvae exposed to Cry11Aa for 9 or 12 h (lanes 4 and 5, respectively). Proteins were separated by SDS-PAGE and transferred to PVDF membranes for Western Blotting with human αJNK2 antibody. Non-toxin exposed larvae dissected at corresponding times were used as controls (lanes 1 to 3). (TIF 1042 kb

Additional file 5: Figure S1. of Transcriptional cellular responses in midgut tissue of Aedes aegypti larvae following intoxication with Cry11Aa toxin from Bacillus thuringiensis

RNAseq and log2 transformed RT-qPCR fold changes. Data is presented for RNAseq and RT-qPCR at 3, 6, 9, and 12 h of an LC50 Cry11Aa treatment. Also shown are RT-qPCR values for 9 and 12 h of unexposed larvae or Cry11Aa non-toxic mutants treatments. All log2 fold changes are referred to control larvae at the start of respective treatment. Panel A and B show genes with high correlation between RNAseq data and RT-qPCR of Cry11Aa. Panel C shows the three genes with low or no positive correlation between these data. (ZIP 779 kb

Toxicity of 5 Bt toxins against the Asian corn borer strains ACB-BtS and ACB-FR.

<p>Toxicity of 5 Bt toxins against the Asian corn borer strains ACB-BtS and ACB-FR.</p

Response of F₁ progenies of Asian corn borer to Cry1F, Cry1Ab and Cry1Ac.

<p>Response of F₁ progenies of Asian corn borer to Cry1F, Cry1Ab and Cry1Ac.</p

Development of an online genome sequence comparison resource for bacillus cereus sensu lato strains using the efficient composition vector method

An automated method was developed for differentiating closely related B. cereus sensu lato (s.l.) species, especially biopesticide Bacillus thuringiensis, from other human pathogens, B. anthracis and B. cereus sensu stricto (s.s.). In the current research, four typing methods were initially compared, including multi-locus sequence typing (MLST), single-copy core genes phylogenetic analysis (SCCGPA), dispensable genes content pattern analysis (DGCPA) and composition vector tree (CVTree), to analyze the genomic variability of 23 B. thuringiensis strains from aizawai, kurstaki, israelensis, thuringiensis and morrisoni serovars. The CVTree method was the best option to be used for typing B. thuringiensis strains since it proved to be the fastest method, whilst giving high-resolution data about the strains. In addition, CVTree agrees well with ANI-based method, revealing the relationship between B. thuringiensis and other B. cereus s.l. species. Based on these data, an online genome sequence comparison resource was built for Bacillus strains called the Bacillus Typing Bioinformatics Database to facilitate strain identification and characterization

Effective of dominance (<i>h</i>) of resistance to Cry1F in Cry1F-selected Asian corn borer.

<p>Effective of dominance (<i>h</i>) of resistance to Cry1F in Cry1F-selected Asian corn borer.</p

Resistance to <i>Bacillus thuringiensis</i> Mediated by an ABC Transporter Mutation Increases Susceptibility to Toxins from Other Bacteria in an Invasive Insect

<div><p>Evolution of pest resistance reduces the efficacy of insecticidal proteins from the gram-positive bacterium <i>Bacillus thuringiensis</i> (Bt) used widely in sprays and transgenic crops. Recent efforts to delay pest adaptation to Bt crops focus primarily on combinations of two or more Bt toxins that kill the same pest, but this approach is often compromised because resistance to one Bt toxin causes cross-resistance to others. Thus, integration of Bt toxins with alternative controls that do not exhibit such cross-resistance is urgently needed. The ideal scenario of negative cross-resistance, where selection for resistance to a Bt toxin increases susceptibility to alternative controls, has been elusive. Here we discovered that selection of the global crop pest, <i>Helicoverpa armigera</i>, for >1000-fold resistance to Bt toxin Cry1Ac increased susceptibility to abamectin and spineotram, insecticides derived from the soil bacteria <i>Streptomyces avermitilis</i> and <i>Saccharopolyspora spinosa</i>, respectively. Resistance to Cry1Ac did not affect susceptibility to the cyclodiene, organophospate, or pyrethroid insecticides tested. Whereas previous work demonstrated that the resistance to Cry1Ac in the strain analyzed here is conferred by a mutation disrupting an ATP-binding cassette protein named ABCC2, the new results show that increased susceptibility to abamectin is genetically linked with the same mutation. Moreover, RNAi silencing of <i>HaABCC2</i> not only decreased susceptibility to Cry1Ac, it also increased susceptibility to abamectin. The mutation disrupting ABCC2 reduced removal of abamectin in live larvae and in transfected Hi5 cells. The results imply that negative cross-resistance occurs because the wild type ABCC2 protein plays a key role in conferring susceptibility to Cry1Ac and in decreasing susceptibility to abamectin. The negative cross-resistance between a Bt toxin and other bacterial insecticides reported here may facilitate more sustainable pest control.</p></div

Efficacy of Bt toxins Cry1AbMod, Cry1Ac and Cry2Ab singly and in combinations against a susceptible strain of pink bollworm (APHIS-S) (see Methods for details).

a<p>All mortality values are adjusted for control mortality.</p>b<p>Observed mortality</p>c<p>Expected mortality for combinations of two or three toxins</p>d<p>Observed mortality - expected mortality; synergism causes positive values and antagonism causes negative values</p>e<p>Probability that the difference between observed and expected mortality occurred by chance based on Fisher’s exact test </p

Responses to spineotram, endosulfan, phoxim, and cyhalothrin in the Cry1Ac-resistant strain (LF60) of <i>H</i>. <i>armigera</i> and its Cry1Ac-susceptible parent strain (LF).

<p>Responses to spineotram, endosulfan, phoxim, and cyhalothrin in the Cry1Ac-resistant strain (LF60) of <i>H</i>. <i>armigera</i> and its Cry1Ac-susceptible parent strain (LF).</p

Suppression of <i>HaABCC2</i> transcription by RNAi in the Cry1Ac-susceptible LF strain of <i>H</i>. <i>armigera</i>.

<p>Early third instar larvae were fed individually with water (control), dsRNA from <i>GFP</i> (control) or dsRNA from <i>HaABCC2</i>. <i>HaABCC2</i> transcription was monitored using qRT-pCR at 1, 3 and 5 days after treatment. The bars show mean transcript levels relative to reference genes (actin and GAPDH) and standard errors from three biological replicates (n = 5 larvae per replicate). For 1, 3 or 5 days after treatment, different letters indicate significantly different means (P < 0.05 by Duncan’s multiple range tests).</p