Calcineurin-Modulated Antimicrobial Peptide Expression Is Required for the Development of Helicoverpa armigera

Wei, Jizhen; Li, Linhong; Yao, Shuangyan; Yang, Shuo; Zhou, Shuai; Liu, Xiaoguang; Du, Mingliang; An, Shiheng

doi:10.3389/fphys.2019.01312

Cited by 8 publications

(32 citation statements)

References 46 publications

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“…The reliability and reproducibility of RT-PCR results were ensured by employing three biological replicates of each sample. Melting curve analysis and the calculation method for expression level of the target genes was done according to the description in Liu et al (2015) and Wei et al (2019).…”

Section: Quantitative Real-time Pcrmentioning

confidence: 99%

Supplemental Sugar Is Required for Sex Pheromone Biosynthesis in Mythimna separata

Zhang

Yao

et al. 2020

Front. Physiol.

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Supplemental nutrients of adult moths maximize moth fitness and contribute to the pollination of many plants. Previous reports have revealed that sugar feeding promotes to sex pheromone biosynthesis by increasing the haemolymph trehalose concentration in mating moths. Here, Mythimna separata adults were employed as a model to investigate the effect of sugar feeding on sex pheromone biosynthesis. Results showed that in virgin females, sugar feeding markedly increased the concentrations of trehalose, pyruvic acid, and acyl-CoA in pheromone glands (PGs), which in turn led to an increase in sex pheromone titer, female ability to attract males and successfully mating frequency in sugar-fed females. Consistently, sugar-fed females laid more eggs than water-fed females. Furthermore, the refeeding of starved females also caused significantly increase in the concentrations of trehalose, pyruvic acid, and acyl-CoA in PGs, thus facilitating a significant increase in sex pheromone production. Most importantly, RNAi-mediated knockdown of trehalase (leading to PG starvation) resulted in an increase in trehalose content, and decrease in the concentrations of pyruvic acid, and acyl-CoA in PGs, which in turn led to a decrease of sex pheromone titer, female ability to attract males and successful mating efficacy. Altogether, results revealed a mechanism by which sugar feeding contributed to trehalose utilization in PGs, promoted to significantly increased sex pheromone precursor by increasing the concentrations of pyruvic acid and acyl-CoA, and facilitated to sex pheromone biosynthesis and successful mating.

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Section: Quantitative Real-time Pcrmentioning

confidence: 99%

Supplemental Sugar Is Required for Sex Pheromone Biosynthesis in Mythimna separata

Zhang

Yao

et al. 2020

Front. Physiol.

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“…After the Sfi I digestion of the PCR products, Cry1Ac was cloned into the bait vector pBT3-N (DUALsystems BioTech). HaCAN was cloned into expression vector pPR3-N ( Figure 4B ), the construction of HaCAN was prepared in our previous study, and it had been shown that pPR3-N-CAN can express in yeast cells and work well ( Wei et al, 2019a ).…”

Section: Methodsmentioning

confidence: 99%

“…Four treatments were introduced into competent cells of the Yeast strain NMY51 (DUAL systems BioTech) with lithium acetate (LiAc)/PEG-mediated transformation, including 100 ng pTSU2-APP and 100 ng pNubG-Fe65, 100 ng pBT3-N-Cry1Ac and 100 ng pOst1-NubI, 100 ng pBT3-N-Cry1Ac and 100 ng pPR3-N, and 100 ng pBT3-N-Cry1Ac and 100 ng pPR3-N-HaCAN. The detail of co-transformation method followed the previous description ( Wei et al, 2019a ). Then, the interaction pairs were screened on four SD (Synthetic Defined Drop-out Medium) plates as the previous description ( Wei et al, 2019a ).…”

Section: Methodsmentioning

confidence: 99%

“…In fact, feeding with Bt toxins themselves activates a series of immune signal pathways ( Chai et al, 2008 ; Xia et al, 2016 ; Portugal et al, 2017 ). Meanwhile, the genes that are involved in the immune pathway can significantly affect insect development ( Luo and Dearolf, 2010 ; Wei et al, 2019a ). Thus, immune genes have attracted the attention of researchers, and are considered as candidate genes for the RNAi + Bt for insect control strategy.…”

Section: Introductionmentioning

confidence: 99%

“…In fact, although many immune-related genes in Helicoverpa armigera have been reported ( Wang et al, 2010 ), their functions in Bt toxicity are rarely reported. Our previous study screened one important Ca 2+ -dependent phosphatase, calcineurin (CAN), which serves as an immune activator by binding with relish, a key transcription factor of the immune deficiency (IMD) pathway, to affect the development of cotton bollworm by regulating antimicrobial peptide (AMP) expression ( Wei et al, 2019a ). CAN has been identified as a dephosphatase of protein phosphatase 2B family and participates in various biological pathways ( Furman and Norris, 2014 ), including immunity pathways ( Bueno et al, 2002 ; Kang et al, 2007 ; Furman and Norris, 2014 ; Ballesteros-Martinez et al, 2017 ).…”

Section: Introductionmentioning

confidence: 99%

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Suppression of Calcineurin Enhances the Toxicity of Cry1Ac to Helicoverpa armigera

et al. 2021

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Insect resistance to Bacillus thuringiensis (Bt) insecticidal proteins has rapidly evolved with the expansion of the planting area of transgenic Bt crops. Pyramiding RNA interference (RNAi) and Bt in crops is urgently needed to counter the rapid increase in pest resistance. The ideal “pyramid” strategy simultaneously targets different action pathways that exert synergetic effects on each other. Here, we identified a dephosphatase, namely, Helicoverpa armigera calcineurin (HaCAN), which might enhance the insecticidal activity of Cry1Ac against Helicoverpa armigera by regulating immune gene expression via dephosphatase activity, but not by acting as a receptor. Notably, blocking enzyme activity or knocking down endogenous HaCAN significantly promoted the enhancement in Cry1Ac toxicity to insect larvae and cells. Correspondingly, the increase in HaCAN activity reduced the cytotoxicity of Cry1Ac as shown by the heterologous expression of HaCAN. Our results provide a probable that HaCAN is an important candidate gene for pyramiding RNAi and Cry1Ac crops to control cotton bollworm.

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Suppressing calcineurin activity increases the toxicity of Cry2Ab to Helicoverpa armigera

Wei

Yang

Zhou

et al. 2021

Pest Management Science

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BACKGROUND Extensive planting of transgenetic Bacillus thuringiensis crops has driven the evolution of pest resistance to Cry1Ac. Adjustment of cropping structures has promoted further outbreak of Helicoverpa armigera in China. To control this pest, a combination of pyramiding RNA interference (RNAi) and Cry2Ab is considered a promising strategy for countering cross‐resistance and enhancing the toxicity of Cry2Ab to cotton bollworm. We explored the possibility of using calcineurin (CAN) as a target RNAi gene, because it is involved in cotton bollworm responses to the toxicity of Cry2Ab. RESULTS Cry2Ab treatment led to a significant increase in HaCAN mRNA level and HaCAN activity. Suppression of HaCAN activity due to RNAi‐mediated knockdown of HaCAN increased the susceptibility of midgut cells to Cry2Ab. The increase in HaCAN activity shown by heterologous expression of HaCAN reduced the cytotoxicity of Cry2Ab to Sf9 cells. Moreover, ingestion of HaCAN‐specific inhibitor FK506 increased the toxicity of Cry2Ab in larvae. Interestingly, HaCAN does not function as a Cry2Ab direct binding protein that participates in Cry2Ab toxicity. CONCLUSIONS The results in this study provide evidence that suppression of HaCAN not only affected the development of the cotton bollworm, but also enhanced the toxicity of Cry2Ab to the pest. HaCAN is therefore an important candidate gene in cotton bollworm that can be targeted for pest control when the pest infests RNAi+Cry2Ab crops. Meanwhile, the mechanism of action of HaCAN in Cry2Ab toxicity suggested that protein dephosphorylation was involved. © 2020 Society of Chemical Industry

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Calcineurin-Modulated Antimicrobial Peptide Expression Is Required for the Development of Helicoverpa armigera

Cited by 8 publications

References 46 publications

Supplemental Sugar Is Required for Sex Pheromone Biosynthesis in Mythimna separata

Supplemental Sugar Is Required for Sex Pheromone Biosynthesis in Mythimna separata

Suppression of Calcineurin Enhances the Toxicity of Cry1Ac to Helicoverpa armigera

Suppressing calcineurin activity increases the toxicity of Cry2Ab to Helicoverpa armigera

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