Archives of Insect Biochemistry and Physiology最新文献

Histone acetylation levels maintained by histone acetyltransferases (HATs) and histone deacetylases play important roles in maintaining local chromatin accessibility and expression of genes that regulate many biological processes, including development and reproduction. N-terminal acetylation of proteins catalyzed by N-terminal acetyltransferases (NATs) also regulates gene expression. We identified 25 HATs/NATs genes in the yellow fever mosquito, Aedes aegypti, and investigated their function in female reproduction using RNA interference (RNAi). Among the HATs/NATs studied, the knockdown of AANAT1 (Arylamine N-acetyltransferase), NAA40 (N-alpha-acetyltransferase 40), NAA80 (N-alpha-acetyltransferase 80), KAT7 (Histone lysine acetyltransferase 7), ACNAT (Acyl-CoA N-acyltransferase), and MCM3AP (Minichromosome maintenance complex component 3 associated protein) significantly reduced egg laying and caused severe problems in oocyte development compared to that in control insects injected with dsGFP. Gene expression analysis using RT-qPCR revealed that vitellogenin and its receptor genes are downregulated in mosquitoes injected with dsAANAT1, dsNAA40, dsNAA80, dsKAT7, dsACNAT, and dsMCM3AP compared to that in control animals. Also, the knockdown of HATs/NATs genes ATAT1 (Alpha-tubulin N-acetyltransferase 1), AANAT1, TAFIID (Transcription initiation factor TFIID subunit 1), HATB (Histone acetyltransferase type B) and NAT9 (N-acetyltransferase 9) decreased more than 50% egg hatch by blocking embryonic development. These results suggest that the acetylation of proteins, especially histones mediated by NATs and HATs, plays an important role in regulating female reproduction and embryonic development of Ae. aegypti.

The gut microbiota plays a vital role in nutrient and energy utilization, as well as in the host's ability to adapt its immune system to environmental changes. As a biological control agent for the invasive Parthenium weed, the Parthenium beetle Zygogramma bicolorata (Z. bicolorata) Pallister is often exposed to fluctuating temperatures, which may induce stress in its natural habitat. This study utilized 16S amplicon sequencing to explore the impact of temperature stress on the gut microbiome of Z. bicolorata under cold (15°C), control (27°C), and hot (35°C) conditions. A total of 11 bacterial phyla and 149 genera were identified, with Firmicutes, Proteobacteria, and Cyanobacteria being the most abundant. Temperature treatments significantly influenced the diversity of the gut microbiota, as evidenced by alpha diversity measures. Principal coordinate analysis further revealed substantial variations in microbiome composition across the different temperature conditions. Additionally, PICRUSt2 analysis suggested that the gut microbiota is linked to metagenomic functions related to amino acid and carbohydrate transport, inorganic ion metabolism, and cellular processes. Our findings suggest that thermal stress alters the gut microbiome of Parthenium beetles, offering new insights into how these beetles may have ecologically adapted to temperature fluctuations, while also highlighting the potential role of gut microbes in maintaining beetle health under environmental stress.

The low excretory rates of secreted digestive enzymes, such as trypsins, in insect species with peritrophic membranes led to the hypothesis of ectoperitrophic countercurrent water fluxes causing enzyme recycling. The midgut water flux model of Tenebrio molitor (T. molitor) is revisited and supported by in vivo experiments. Sequences from proteins putatively involved in water transport were retrieved from the T. molitor transcriptome by Blast and analyzed using bioinformatics tools. Gene expression of selected proteins was determined in three midgut sections (anterior, AM; middle, MM; posterior, PM) by RNA-seq, and transporter proteins were verified in microvillar-membrane-enriched midgut samples by proteomics. Genes encoding three cation chloride cotransporters (CCC) and four aquaporins were expressed in the midgut. TmNaCCC2, TmPrip, and TmEglp1 showed higher expression in the front half, while TmKCC, TmNKCC1, TmDrip, and TmEglp2 were more highly expressed in the back half. However, only TmNaCCC2 was found by proteomics. Midgut water fluxes were quantified by feeding T. molitor larvae with nonabsorbable dye and measuring its concentration along the midgut. The results suggest water absorption in AM and secretion in MM and PM, potentially caused by TmNaCCC2 and TmPrip in AM, and TmKCC and TmDrip in PM, whereas MM serves as a transition region. Larvae fed on furosemide, an NKCC and KCC inhibitor, showed altered midgut water fluxes, resulting in higher trypsin excretion into the hindgut, thus reinforcing the hypothesis of a countercurrent water flux generated by CCCs powering enzyme recycling in insect midguts.

Mirid plant bugs (Hemiptera: Miridae), including Lygus hesperus (western tarnished plant bug), are key pests of numerous agricultural crops. While management of this pest relies heavily on chemical insecticides, the evolution of resistance and environmental concerns underscore the need for new and more effective approaches. Genetic-based strategies that target male fertiliy are currently being evaluated for population suppression. However, a lack of candidate gene targets with appropriate function, specifically in non-model species like L. hesperus, has hindered progress in the development and application of such approaches. Given their conserved role in stabilization of the flagella axoneme and association with sperm motility in many organisms, members of the tektin gene family represent logical targets for genetic-based sterilization. Here, we identified four homologs of the non-vertebrate tektin family from L. hesperus and used RNA interference-mediated knockdown to assess their roles in male fertility. Although transcription of the four tektins was predominantly in the testis, knockdown had negligible effects on either sperm abundance or male fertility. Our results suggest that tektins do not play a critical role in sperm fertilization of eggs in L. hesperus and are thus likely poor targets for genetic-based sterilization approaches in this species.

Transgenerational immune priming (TGIP) is a phenomenon in which parental exposure to pathogen infection enhances resistance to pathogens in their offspring. TGIP has been reported in several insects, including Lepidoptera, but it has not yet been documented in silkworms. In this study, we demonstrated the existence of TGIP in silkworms by exposing the parental generation to a low dose of Bombyx mori nucleopolyhedrovirus (BmNPV). Notably, when maternal or paternal-primed moths were mated to produce the F1 generation, the F1 generation larvae from both primed groups were more resistant to the BmNPV challenge than silkworm larvae with only maternal or paternal priming. Importantly, both maternal and paternal contributions to offspring immunity were essential for TGIP. However, due to the characteristics of the BmNPV itself, no within-generation immune responses were detected following BmNPV priming. Further analysis revealed that immune-related genes might play a role in mediating specific TGIP in silkworms after BmNPV priming. These results broaden our understanding of TGIP and the antiviral memory of insects in their offspring.

As a key acetyltransferase, Bombyx mori cAMP response element-binding protein (CREB)-binding protein (BmCBP) plays an important role in post-translational modifications, affecting protein stability and immune response in silkworm; however, its mechanism of action remains unclear. In this study, we designed and synthesized Nano-dsRNA nanoparticles targeting BmCBP and validated their impact on BmApoLp-III protein expression. Using an E. coli expression system, we successfully synthesized nano-dsRNA targeting BmCBP, which significantly reduced BmCBP transcription level, thereby decreasing the acetylation and protein expression levels of BmApoLp-III. The knock-down of BmCBP by nano-dsRNA also increased the ubiquitination level of BmApoLp-III, suggesting a competitive relationship between acetylation and ubiquitination. These findings show that design and synthesis of efficient nano-dsRNA nanoparticles targeting BmCBP provide a novel tool and approach for studying gene expression and regulation in silkworm and other insects.

Technologies for controlled protein targeting allow the selective manipulations of proteins resulting in their degradation and/or loss of function. Over the past two decades, these technologies have overcome the limitations of genetic methods and have become powerful tools in biological research and the search for new therapeutic approaches to disease treatment. Various methods of protein degradation and inactivation have been successfully applied to a model organism such as Drosophila melanogaster. In this article, we overview the capabilities and prospects of the Drosophila in vivo model for testing and developing modern methods of controlled protein targeting, analyzing their efficacy at the organism level and solving fundamental biological problems.

Feeding bees carbohydrates, as a substitute for nectar, has become essential in modern beekeeping. We compared the effects of 65% sucrose (SS) and 65% invert sugar (IS) syrups on the survival and enzyme activity of caged honey bees. Specifically, we analyzed glycosidase activity in the head and midgut of the bees and compared the composition of sucrose-based (SH) and invert sugar-based (IH) “honey” stored by the bees and collected from the comb. Glycosidase activity was similar in head in contrast to midgut where it was higher in IS fed bees, which appeared to be residual yeast β-fructofuranosidase activity. Fructose to glucose ratio in SH were 60.84/39.16 and a presence of some other sugars were detected, while ratio in IH were 48.49/51.51, almost exactly the same as in start fed (IS) (48.57/51.43). It has been demonstrated that glycosidase activity in IH was residual yeast β-fructofuranosidase activity. Zymogram detected α-amylase band in SH, in contrast to IH, which suggest that honey bees do not add amylase into IS. In contrast to SH, no crystallization occurred in IH. SS and IS densities were 1.23 and 1.24 g/mL, respectively, increasing to 1.35 g/mL in SH and 1.28 g/mL in IH after processing. This suggests that higher humidity and restricted cleansing flight make it harder for bees to remove excess water from IH, leading to increased midgut and hindgut weight, which correlates with higher mortality in the third week for the IS-fed group and fourth week for the SS-fed group.