Death-Associated LIM-Only Protein Reduces Cry1Ac Toxicity by Sequestration of Cry1Ac Protoxin and Activated Toxin in Helicoverpa armigera.

The extensive use of Bacillus thuringiensis (Bt) in pest management has driven the evolution of pest resistance to Bt toxins, particularly Cry1Ac. Effective management of Bt resistance necessitates a good understanding of which pest proteins interact with Bt toxins. In this study, we screened a Helicoverpa armigera larval midgut cDNA library and captured 208 potential Cry1Ac-interacting proteins. Among these, we further examined the interaction between Cry1Ac and a previously unknown Cry1Ac-interacting protein, HaDALP (H. armigera death-associated LIM-only protein), as well as its role in toxicology. The results revealed that HaDALP specifically binds to both the Cry1Ac protoxin and activated toxin, significantly enhancing cell and larval tolerance to Cry1Ac. Additionally, HaDALP was overexpressed in a Cry1Ac-resistant H. armigera strain. These findings reveal a greater number of Cry1Ac-interacting proteins than previously known and demonstrate, for the first time, that HaDALP reduces Cry1Ac toxicity by sequestering both the protoxin and activated toxin.

ATP Synthase Subunit α from Helicoverpa armigera Acts as a Receptor of Bacillus thuringiensis Cry1Ac and Synergizes Cry1Ac Toxicity.

Insect resistance to Bacillus thuringiensis (Bt) toxins has led to an urgent need to explore the insecticidal mechanisms of Bt. Previous studies indicated that Helicoverpa armigera ATP synthase subunit α (HaATPs-α) is involved in Cry1Ac resistance. In this study, a real-time quantitative polymerase chain reaction (RT-PCR) confirmed that HaATPs-α expression was significantly reduced in the Cry1Ac-resistant strain (BtR). Cry1Ac feeding induced the downregulated expression of HaATPs-α in the susceptible strain, but not in the BtR strain. Furthermore, the interaction between HaATPs-α and Cry1Ac was verified by ligand blotting and homologous competition experiments. The in vitro gain and loss of function analyses showed HaATPs-α involved in Cry1Ac toxicity by expressing endogenous HaATPs-α and HaATPs-α double-stranded RNAs in Sf9 and midgut cells, respectively. Importantly, purified HaATPs-α synergized Cry1Ac toxicity to H. armigera larvae. These findings provide the first evidence that HaATPs-α is a potential receptor of Cry1Ac, it shows downregulated participation in Cry1Ac resistance, and it exhibits higher enhancement of Cry1Ac toxicity to H. armigera larvae.

ABCC2 is a functional receptor of Bacillus thuringiensis Cry1Ca in Spodoptera litura.

Spodoptera litura is a serious polyphagous pest in the whole world, which has developed resistance to most conventional insecticides and even some Bacillus thuringiensis (Bt) toxins. Cry1Ca has excellent insecticide activity against S. litura with potential application to control S. litura and delay the development of insect resistance. However, the mode of action of Cry1Ca in S. litura is poorly understood. Here, Cry1Ca-binding proteins were identified from S. litura by using pull down assays and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The results indicated that aminopeptidase-N (APN), ATP binding cassette subfamily C member 2 (ABCC2), polycalin, actin and V-type proton ATPase subunit A may bind with Cry1Ca. Further study confirmed that ABCC2 fragment expressed in vitro can bind to Cry1Ca as demonstrated by Ligand blot and homologous competition experiments. The over-expression of endogenous SlABCC2 in Sf9 cells increased Cry1Ca cytotoxicity. Correspondingly, the vivo loss of function analyses by SlABCC2 small interfering RNAs (siRNAs) in S. litura larvae decreased the toxicity of Cry1Ca to larvae. Altogether, these results show that ABCC2 of S. litura is a functional receptor that is involved in the action mode of Cry1Ca.

Efficient Mechanochemical Synthesis of Polyoxometalate⊂ZIF Complexes as Reusable Catalysts for Highly Selective Oxidation.

One-pot mechanochemical synthesis was demonstrated to be an efficient strategy to synthesize host-guest POM⊂rho-ZIF complexes (POM = polyoxometalate; rho-ZIF = zeolitic imidazolate framework with rho topology) with high crystallinity. In this work, the metastable rho-ZIF with large interior cavities and windows was used as host matrix for encapsulating and immobilizing bulky guest molecules with high loading efficiency and chemical stability. As novel catalysts, POM⊂rho-ZIF complexes were found effective for the selective oxidation of a series of sulfides to sulfoxides. Moreover, the heterogeneity of these composite catalysts was confirmed by leaching tests, and they can be recycled at least four times without significant loss of activity.

The solution chemistry and reactivity of lacunary Keggin silicotungstates monitored in real-time by a combination of mass spectrometry and electrochemistry.

The solution chemistry of a series of mono-, di- and trilacunary Keggin silicotungstates was investigated by electrospray ionization mass spectrometry (ESI-MS) and general electrospray features especially for lacunary POMs are summarized. The reactions of vanadium incorporation into the lacunary structures were successfully monitored in real-time by a combination of ESI-MS and differential pulse voltammetry (DPV). It was found that all the reactions took place instantaneously and that a subtle speciation change occurred at prolonged reaction times for the pair of reactants, monovacant silicotungstate and sodium metavanadate, suggesting a conversion of mono- to divanadium substituted derivatives. This was shown to result from a solution process, not an ESI-induced reaction, by DPV measurements. The relative stabilities of the V-substituted products were assessed in both solution and gas phases.

Application of the standard addition method for the determination of acrylamide in heat-processed starchy foods by gas chromatography with electron capture detector.

A gas chromatography electron capture detector (GC-ECD) using the standard addition method was developed for the determination of acrylamide in heat-processed foods. The method entails extraction of acrylamide with water, filtration, defatting with n-hexane, derivatization with hydrobromic acid and saturated bromine-water, and liquid-liquid extraction with ethyl acetate. The sample pretreatment required no SPE clean-up and concentration steps prior to injection. The final extract was analyzed by GC-ECD. The chromatographic analysis was performed on polar columns, e.g. Supelcowax-10 capillary column, and good retention and peak response of the analyte were achieved under the optimal conditions. The qualification of the analyte was by identifying the peak with same retention time as standard compound 2,3-DBPA and confirmed by GC-MS. GC-MS analysis confirmed that 2,3-DBPA was converted to 2-BPA nearly completely on the polar capillary column, whether or not treated with triethylamine. A four-point standard addition protocol was used to quantify acrylamide in food samples. The limit of detection (LOD) was estimated to be 0.6µg/kg on the basis of ECD technique. Validation and quantification results demonstrated that the method should be regarded as a low-cost, convenient, and reliable alternative for conventional investigation of acrylamide.