Genetically engineered crops simultaneously produce defensive allelochemicals and Bacillus thuringiensis (Bt) toxin proteins to kill some of the world's most devastating insect pests. How the two types of toxins, when ingested sequentially or simultaneously, interact at both lethal and sublethal doses in these pests remains underexplored. Here, we examined the toxicological interactions between the Bt toxin Cry1Ac and the flavonoid allelochemical flavone in Helicoverpa armigera. Simultaneous exposure of H. armigera neonates to lethal doses (LC25) of Cry1Ac and flavone caused a mortality significantly higher than that of either toxin alone and their expected additive mortality. Preexposure for 24 h to a sublethal dose (LC10) of Cry1Ac followed by 6‐d simultaneous exposure to the same dose of Cry1Ac plus a lethal dose (1.6 mg/g diets, LC50) of flavone resulted in a mortality significantly higher than that of the LC50 dose of flavone alone and the expected additive mortality of the LC50 dose of flavone plus the LC10 dose of Cry1Ac. One‐day preexposure to the sublethal dose (LC10) of flavone followed by 6‐d simultaneous exposure to the LC50 dose (6 ng/cm2) of Cry1Ac plus the LC10 dose of flavone yielded a mortality significantly higher than that of the LC50 dose of Cry1Ac but similar to the expected additive mortality of the LC50 dose of Cry1Ac plus the LC10 dose of flavone. The results suggest that Cry1Ac induces and synergizes the toxicity of flavone against H. armigera larvae.
Summary An adaptive sliding mode observer (SMO)–based fault‐tolerant control method taking into consideration of actuator saturation is proposed for a hypersonic scramjet vehicle (HSV) under a class of time‐varying actuator faults. The SMO is designed to robustly estimate the HSV states and reconstruct the fault signals. The adaptive technique is integrated into the SMO to approximate the unknown bounds of system uncertainties, actuator faults, and estimation errors. The robust SMO synthesis condition, which can be formulated as a set of linear matrix inequalities, is improved by relaxing structure constraints to the Lyapunov matrix. An anti‐windup feedback control law, which utilizes the estimated HSV states and the fault signals, is designed to counteract the negative effects of actuator saturation induced by actuator faults. Simulation results demonstrate that the proposed approach can guarantee stability and maintain L2 performance of the closed‐loop system in the presence of HSV actuator faults and saturation.
The high fecundity of the most destructive pest Helicoverpa armigera and its great resistance risk to insecticides and Bt crops make the reproductive-destruction-based control of this pest extremely appealing. To find suitable targets for disruption of its reproduction, we observed the testis and ovary development of H. armigera and conducted deep sequencing of the ovary and testis small RNAs of H. armigera and quantitative RT-PCR (RT-qPCR) validation to identify reproduction-related micro RNAs (miRNAs). A total of 7,592,150 and 8,815,237 clean reads were obtained from the testis and ovary tissue, respectively. After further analysis, we obtained 173 novel and 74 known miRNAs from the two libraries. Among the 74 known miRNAs, 60 miRNAs existed in the ovary and 72 existed in the testis. Further RT-qPCR validation of 5 miRNAs from the ovary and 6 miRNAs from the testis confirmed 8 of them were indeed ovary- (miR-989a, miR-263-5p, miR-34) or testis-biased (miR-2763, miR-998, miR-2c, miR-2765, miR-252a-5p). The 8 ovary- or testis-biased miRNAs had a total of 30,172 putative non-redundant target transcripts, as predicted by miRanda and RNAhybrid. Many of these target transcripts are assigned to reproduction-related GO terms (e.g., oocyte maturation, vitellogenesis, spermatogenesis) and are members of multiple reproduction-related KEGG pathways, such as the JAK-STAT signaling pathway, oocyte meiosis, the insulin signaling pathway, and insect hormone biosynthesis. These results suggest that the 8 gonad-biased miRNAs play important roles in reproduction and may be used as the targets for the development of reproductive-destruction-based control of H. armigera and, possibly, other lepidopteran pests.
Circular RNAs (circRNAs) are a group of covalently closed RNAs, and their biological function is largely unknown. In this study, we focused on circRNAs that are generated from exon back-splicing (exonic circRNAs). The linear RNA counterparts encode functional proteins, that we can compare and investigate the relationship between circular and linear RNAs. We compared circRNA expression profiles between untreated and Pseudomonas syringae-infected Arabidopsis, and identified and experimentally validated differentially expressed exonic circRNAs by multiple approaches. We found that exonic circRNAs are preferentially enriched in biological processes that associate with biotic and abiotic stress responses. We discovered that circR194 and circR4022 are involved in plant response against P. syringae infection, while circR11208 is involved in response against Botrytis cinerea infection. Intriguingly, our results indicate that these exonic circRNAs function synergistically with their corresponding linear RNAs. Furthermore, circR4022 and circR11208 also play substantial roles in Arabidopsis tolerance to salt stress. This study extends our understanding of the molecular functions of plant circRNAs.
MicroRNAs (miRNAs) are regulatory RNA molecules that bind to target messenger RNAs (mRNAs) and affect the stability or translational efficiency of the bound mRNAs. Single or dual‐luciferase reporter systems have been successfully used to identify miRNA target genes in mammalian cells. These reporter systems, however, are not sensitive enough to verify miRNA–target gene relationships in insect cell lines because the promoters of the target luciferase (usually Renilla) used in these reporter systems are too weak to drive sufficient expression of the target luciferase in insect cells. In this study, we replaced the SV40 promoter in the psiCHECK‐2 reporter vector, which is widely used with mammalian cell lines, with the HSV‐TK or AC5.1 promoter to yield two new dual‐luciferase reporter vectors, designated psiCHECK‐2‐TK and psiCHECK‐2‐AC5.1, respectively. Only psiCHECK‐2 and psiCHECK‐2‐AC5.1 had suitable target (Renilla)/reference (firefly) luciferase activity ratios in mammalian (HeLa and HEK293) and insect (Sf9, S2, Helicoverpa zea fat body and ovary) cell lines, while psiCHECK‐2‐TK had suitable Renilla/firefly luciferase activity ratios regardless of the cell line. Moreover, psiCHECK‐2‐TK successfully detected the interaction between Helicoverpa armigera miRNA9a and its target, the 3′‐untranslated region of heat shock protein 90, in both mammalian and H. zea cell lines, but psiCHECK‐2 failed to do so in H. zea cell lines. Furthermore, psiCHECK‐2‐TK with the target sequence, HzMasc (H. zea Masculinizer), accurately differentiated between H. zea cell lines with or without the negative regulation factor (miRNA or piRNA) of HzMasc. These data demonstrate that psiCHECK‐2‐TK can be used to functionally characterize small RNA target genes in both mammalian and insect cells.
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