Crystal structure and molecular characterization of NADP+-farnesol dehydrogenase from cotton bollworm, Helicoverpa armigera

Nano-delivery systems have been applied to deliver various synthetic/botanical pesticides to increase the efficiency of pesticide use and reduce the volumes of pesticides applied. Previous studies have supported the hypothesis that the nanocarriers can help expand the insecticidal target of pesticides to include non-target pests. However, the potential mechanism underlying this interesting phenomenon remains unclear. Herein, a widely applied star polycation (SPc) nanocarrier was synthesized to construct a thiamethoxam (TMX) nano-delivery system. The SPc-based delivery system could promote the translocation of exogenous substances across the membrane of Sf9 cells, increase the cytotoxicity of TMX against Sf9 cells by nearly 20%, and expand the insecticidal target of TMX to include Spodoptera frugiperda (the fall armyworm), with a 27.5% mortality increase at a concentration of 0.25 mg/mL. Moreover, the RNA-seq analysis demonstrated that the SPc could upregulate various transport-related genes, such as Rab, SORT1, CYTH, and PIKfyve, for the enhanced cellular uptake of TMX. Furthermore, enhanced cell death in larvae treated with the TMX-SPc complex was observed through changes in the expression levels of death-related genes, such as Casp7, BIRC5, MSK1, and PGAM5. The SPc-based nano-delivery system improved the cellular uptake of TMX and expanded its insecticidal target by adjusting the expression levels of death-related genes. The current study mainly identified the transport and cell death genes related to nanocarrier-based insecticidal target expansion, which is beneficial for understanding the bioactivity enhancement of the nano-delivery system.

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“…, 2009; Kumar et al. , 2022). Juvenile hormone diol kinase (JHDK) is an enzyme involved in JH metabolism (Zeng et al.…”

Section: Discussionmentioning

confidence: 99%

A nano‐delivery system expands the insecticidal target of thiamethoxam to include a devastating pest, the fall armyworm

Jiang

Liu

et al. 2022

Insect Science

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“…Before RNA extraction, larvae were starved overnight, as detailed in our previous article (Kumar et al, 2022). RNA was extracted using PureLink™ RNA Mini Kit (Invitrogen™, Thermo Fisher Scientific Corporation) as per the manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

“…The primer sequences used for quantitative real‐time PCR (qRT‐PCR) experiments were the same as were used in the aforementioned study (Table 1). The qPCR conditions were followed as described in our previous article (Das et al, 2022; Kumar et al, 2022). Briefly, the qRT‐PCR program was set as: 95°C for 5 min, 37 cycles of 95°C for 30 s, 60°C for 30 s and 72°C for 40 s, final extension at 72°C for 7 min.…”

Section: Methodsmentioning

confidence: 99%

“…The expression level of ago1 was subjected to analysis of variance (ANOVA) presented as mean relative gene expressions along with standard errors (Kumar et al, 2019). Actin was used as normalizing gene (Kumar et al, 2022; Raghavendra et al, 2021). The qRT‐PCR data were analysed using the ΔΔ C t method (Livak & Schmittgen, 2001).…”

Section: Methodsmentioning

confidence: 99%

“…Before RNA extraction, larvae were starved overnight, as detailed in our previous article (Kumar et al, 2022). RNA was extracted using…”

Section: Rna Extraction and Cdna Synthesismentioning

confidence: 99%

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Simple cost‐effective larval injection method for dsRNA delivery to induce RNAi response in Helicoverpa armigera (Hübner)

Das

Kumar

Shah

et al. 2023

J Applied Entomology

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Microinjection is considered as an effective method for dsRNA delivery in insects. It also facilitates the delivery of a precise quantity of dsRNA in the host insect, inducing an efficient RNAi response. However, the microinjection method needs prior optimization of several parameters like concentration of dsRNA, site of injection, developmental stage of insect etc. for inducing effective RNAi response. Moreover, sophisticated microinjection devices are largely expensive with high maintenance cost. The Old‐World bollworm, Helicoverpa armigera (Hübner) is known to be a detrimental polyphagous pest with widespread infestations across the globe. In the present study, we demonstrate a low‐cost and effective dsRNA delivery method for inducing RNAi response in H. armigera with the aid of basic insulin injection syringe and fabricated micropipette tip. In order to validate the RNAi response following dsRNA injection, we have selected three key genes from the chitin biosynthesis pathway of the insect. Besides these, argonaute 1 (ago1) was also used as an indicator gene for dsRNA‐mediated RNAi induction. Delivery of dsRNA using injection with insulin syringe caused significant upregulation of the ago1 gene in the insect irrespective of any of the three target genes concerned viz. HaNAGK (3.9 fold; p < 0.001), HaAGM (6.3 fold; p < 0.001) and HaUAP (5.9 fold; p < 0.01) respectively, as compared to control injected with nuclease‐free water. The dsRNA‐injected insects showed aberrant developmental symptoms typical of impeded chitin synthesis, eventually leading to anomalous ecdysis with substantial mortality (up to 69.04%), as compared to controls. The described protocol reduces insect injury, enabling easy restraining of larva and quick execution of dsRNA injection with efficient RNAi response.

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Farnesol dehydrogenase from Helicoverpa armigera (Hübner) as a promising target for pest management: molecular docking, in vitro and insect bioassay studies using geranylgeraniol as potential inhibitor

et al. 2023

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Crystal structure and molecular characterization of NADP+-farnesol dehydrogenase from cotton bollworm, Helicoverpa armigera

Cited by 9 publications

References 49 publications

A nano‐delivery system expands the insecticidal target of thiamethoxam to include a devastating pest, the fall armyworm

A nano‐delivery system expands the insecticidal target of thiamethoxam to include a devastating pest, the fall armyworm

Simple cost‐effective larval injection method for dsRNA delivery to induce RNAi response in Helicoverpa armigera (Hübner)

Farnesol dehydrogenase from Helicoverpa armigera (Hübner) as a promising target for pest management: molecular docking, in vitro and insect bioassay studies using geranylgeraniol as potential inhibitor

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