The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, is an important maize pest throughout most of the U.S. Corn Belt. Bacillus thuringiensis (Bt) insecticidal proteins including modified Cry3Aa and Cry34/35Ab1 have been expressed in transgenic maize to protect against WCR feeding damage. To date, there is limited information regarding the WCR midgut target sites for these proteins. In this study, we examined whether a cadherin-like gene from Diabrotica virgifera virgifera (DvvCad; GenBank accession # EF531715) associated with WCR larval midgut tissue is necessary for Cry3Aa or Cry34/35Ab1 toxicity. Experiments were designed to examine the sensitivity of WCR to trypsin activated Cry3Aa and Cry34/35Ab1 after oral feeding of the DvvCad dsRNA to knockdown gene expression. Quantitative real-time PCR confirmed that DvvCad mRNA transcript levels were reduced in larvae treated with cadherin dsRNA. Relative cadherin expression by immunoblot analysis and nano-liquid chromatography - mass spectrometry (nanoLC-MS) of WCR neonate brush border membrane vesicle (BBMV) preparations exposed to DvvCad dsRNA confirmed reduced cadherin expression when compared to BBMV from untreated larvae. However, the larval mortality and growth inhibition of WCR neonates exposed to cadherin dsRNA for two days followed by feeding exposure to either Cry3Aa or Cry34/35Ab1 for four days was not significantly different to that observed in insects exposed to either Cry3Aa or Cry34/35Ab1 alone. In combination, these results suggest that cadherin is unlikely to be involved in the toxicity of Cry3Aa or Cry34/35Ab1 to WCR.
With Next Generation Sequencing technologies, high-throughput RNA sequencing (RNAseq) was conducted to examine gene expression in neonates of Diabrotica virgifera virgifera (LeConte) (Western Corn Rootworm, WCR) challenged with individual proteins of the binary Bacillus thuringiensis insecticidal proteins, Cry34Ab1 and Cry35Ab1, and the combination of Cry34/Cry35Ab1, which together are active against rootworm larvae. Integrated results of three different statistical comparisons identified 114 and 1300 differentially expressed transcripts (DETs) in the Cry34Ab1 and Cry34/35Ab1 treatment, respectively, as compared to the control. No DETs were identified in the Cry35Ab1 treatment. Putative Bt binding receptors previously identified in other insect species were not identified in DETs in this study. The majority of DETs (75% with Cry34Ab1 and 68.3% with Cry34/35Ab1 treatments) had no significant hits in the NCBI nr database. In addition, 92 DETs were shared between Cry34Ab1 and Cry34/35Ab1 treatments. Further analysis revealed that the most abundant DETs in both Cry34Ab1 and Cry34/35Ab1 treatments were associated with binding and catalytic activity. Results from this study confirmed the nature of these binary toxins against WCR larvae and provide a fundamental profile of expression pattern of genes in response to challenge of the Cry34/35Ab1 toxin, which may provide insight into potential resistance mechanisms.
Conservation efforts leading to the recovery of the federally endangered American burying beetle (ABB), Nicrophorus americanus Olivier, have been challenging because of the unknown causes of its decline, difficulty in establishing habitat requirements, and unclear population distribution across the species' range. Extant populations of this widespread generalist species occur in broadly separated regions of North America with varying habitat characteristics. A habitat suitability model for ABB in the Nebraska Sandhills was developed over the course of 3 years resulting in a final crossvalidated spatial model. The succession of models from 2009 to 2011 indicated that most of the predictive variables stayed constant, but biased sampling and extrapolation areas affected classifier values differently. Variables associated with ABB occurrence were loamy sand, wetland and precipitation. Five variables, loam soil, agriculture, woodland, the average maximum temperature, and urban development, were associated with ABB absence. The 2011 cross-validated model produced an AUC value of 0.82 and provided areas designated as highly likely to support ABBs. By limiting the model extent to the Sandhills ecoregion and using threshold-dependent classifiers, the final habitat suitability model could be an important resource for wildlife managers engaged in the recovery of this habitat generalist.
The establishment and spread of non-native species often results in negative impacts on biodiversity and ecosystem function. Several species of saltcedar, Tamarix spp. L., have been recently naturalized in large portions of the United States where they have altered plant and animal communities. To test the prediction that saltcedar negatively affects invertebrates, we measured ant genera diversity and the activity density of the exotic isopod Armadillidium vulgare Latrielle (Isopoda: Oniscoidea) for 2 yr using pitfall traps located within 30 5-m2 plots with or without saltcedar at a south-central Nebraska reservoir. From 2005 to 2006, we collected 10,837 ants representing 17 genera and 4,953 A. vulgare. Per plot, the average number of ant genera was not different between saltcedar (x̅ = 3.9) and non-saltcedar areas ( x̅ = 3.9); however, saltcedar plots were compositionally different and more similar from plot to plot (i.e., they had lower beta diversity than control plots) in 2005, but not in 2006. Isopods were likewise temporally affected with higher activity density (+89%) in control plots in 2005, but higher activity density (+27%) in saltcedar plots in 2006. The observed temporal differences occurred as the drought that initially enabled the saltcedar invasion became less severe in 2006. Combined, our results suggest that invertebrate groups like ants, which are generally omnivorous, may be better equipped than more specialized taxa like detritivores to withstand habitat changes due to invasions by non-native species, especially during extreme weather events such as prolonged droughts.
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