Blue light stimulates light stress and phototactic behavior when received in the brain of Diaphorina citri.

Blue light with a wavelength of 400-470 nm is the composition of the visible light. However, in recent years, blue light contributed the most significance to light pollution due to the artificial light at night. Previously, we have demonstrated that the Asian citrus psyllid (ACP), Diaphorina citri, an important pest in citrus production, has significant positive phototaxis with a light-emitting diode light of 400 nm. In this study, ACP with positive phototactic behavior to 400 nm light (PH) and non-phototactic behavior to 400 nm light (NP) were collected, individually. Transcriptome dynamics of head tissues of PH and NP groups were captured by using RNA-sequencing technology, respectively. Forty-three to 46 million clean reads with high-quality values were obtained, and 1773 differential expressed genes (DEGs) were detected. Compared with the NP group, there were 841 up-regulated DEGs and 932 down-regulated DEGs in the PH group. Eight pathways were significantly enriched in the PH group in the KEGG database, while 43 up-regulated pathways and 25 down-regulated pathways were significantly enriched in the PH group in the GO database. The DGE approach was reliable validated by real time quantitative PCR. Results indicated that the blue light acted as an abiotic stress causing physiological and biochemical responses such as oxidative stress, protein denaturation, inflammation and tumor development in ACPs. Additionally, the light was absorbed by photoreceptors of ACPs, and converted into electrical signal to regulate neuromodulation. This study provides basic information for understanding the molecular mechanisms of ACP in response to blue light and provides a reference for further studies to elucidate phototactic behavior.

Potential role of lysine succinylation in the response of moths to artificial light at night stress.

Artificial light at night (ALAN) is a widespread environmental pollutant and stressor. Many nocturnal insects have been shown to experience ALAN stress. However, few studies have been conducted to uncover the mechanism by which nocturnal insects respond to ALAN stress. Previous studies suggest that lysine succinylation (Ksuc) is a potential mechanism that coordinates energy metabolism and antioxidant activity under stressful conditions. Mythimna separata (Walker) (M. separata) is a nocturnal insect that has been stressed by ALAN. In this study, we quantified the relative proteomic Ksuc levels in ALAN-stressed M. separata. Of the 466 identified Ksuc-modified proteins, 103 were hypersuccinylated/desuccinylated in ALAN-stressed moths. The hypersuccinylated/desuccinylated proteins were shown to be involved in various biological processes. In particular, they were enriched in metabolic processes, reactive oxygen species (ROS) homeostasis and the neuromuscular system. Furthermore, we demonstrated that Ksuc might affect moth locomotion by intervening with and coordinating these systems under ALAN stress. These findings suggest that Ksuc plays a vital role in the moth response to ALAN stress and moth locomotion behavior and provide a new perspective on the impact of ALAN on nocturnal insect populations and species communities.

Molecular characterization and juvenile hormone-regulated transcription of the vitellogenin receptor in the cabbage beetle Colaphellus bowringi.

The vitellogenin receptor (VgR) is highly expressed in the ovaries where it is responsible for vitellogenin (Vg) deposition during oogenesis in insects. Therefore, identifying the VgR of insect pests, and understanding the mechanism regulating its expression, could lead to the development of pest management strategies based on disrupting reproduction. We cloned and identified VgR in the cabbage beetle, Colaphellus bowringi, which is a serious pest of cruciferous vegetables in Asia. The regulation of VgR transcription by juvenile hormone (JH) was also investigated. The results show that C. bowringi VgR cDNA contains an open reading frame of 5310 bp encoding 1769 amino acid residues. Protein domain prediction indicates that C. bowringi VgR belongs to the LDLR gene superfamily, having the same group of structural domains that has been well characterized in other insects. VgR mRNA was highly expressed in the ovaries of reproductive female cabbage beetles. Knockdown of VgR reduced yolk deposition in the ovaries, increased the accumulation of Vg proteins in the hemolymph and decreased the transcription of Vg1 and Vg2 in the fat body. RNA interference and hormone challenge experiments showed that JH induced VgR transcription via the JH intracellular receptor methoprene-tolerant (Met) and the JH-responsive transcription factor Krüppel homolog 1 (Kr-h1). Our results suggest that there is a feedback loop between VgR transcription in the ovaries and Vg transcription in the fat body. JH acting through Met-Kr-h1 pathway induces the transcription of the VgR that is essential for Vg uptake and reproductive development. These findings not only reveal the potential JH signaling mechanism regulating VgR transcription, but may also contribute to the development of pest control strategies based on disrupting endocrine-regulated reproduction.

The Influence of Allogrooming Behavior on Individual Innate Immunity in the Subterranean Termite Reticulitermes chinensis (Isoptera: Rhinotermitidae).

Insect societies have evolved a series of disease defenses against pathogens, including social sanitary behavior and individual innate immunity. However, whether sanitary behavior can affect individual innate immunity remains unknown. Here, we exposed the termite Reticulitermes chinensis Snyder to the entomopathogenic fungus Metarhizium anisopliae (Metchnikoff) Sorokin(Ascomycota: Hypocreales), and then measured their allogrooming behavior, conidia load, infection mortality, antifungal activity and immune gene expressions . Our results showed that most of the fungal conidia were fast removed from the cuticles of the grouped termites by intensive allogrooming behavior, resulting in low mortality. The antifungal activity and immune gene expressions (termicin and transferrin) in grouped exposed termites were significantly lower than those in single exposed termite but not significantly different from those in unexposed treatments. These results suggest that allogrooming behavior can fast remove fungal conidia from termite cuticles and then decrease their physiological investment in individual innate immunity.

The gut bacteria across life stages in the synanthropic fly Chrysomya megacephala.

BACKGROUND: Gut bacteria are closely associated with host. Chrysomya megacephala, as a vector and resource insect, can transmit various pathogenic bacteria and consume manure to produce biofertilizer and larva biomass. However, the gut bacteria composition and abundance of C. megacephala remain unclear. RESULTS: Illumina MiSeq platform was used to compare composition of gut bacterial community in eggs, 1-day-old larvae, 5-day-old larvae, pupae, adult females and males by sequencing with variation in V4 region of 16S ribosomal DNA gene. In total, 928 operational taxonomic units (OTUs) were obtained. These OTUs were annotated into 19 phyla, 42 classes, 77 orders, 153 families and 289 genera. More than 0.5% abundance of 32 OTU core genera were found across all life stages. At class level, Alphaproteobacteria, Bacilli, Bacteroidia, Betaproteobacteria, Flavobacteriia and Gammaproteobacteria were the most abundant in C. megacephala. Eight species were identified to have significantly different abundance between 1-d-larvae and 5-day-larvae and took 28.95% of shared species between these two groups. Sex-specific bacterial species were identified that Faecalibacterium prausnitzii was merely present in females, while Rhodococcus fascians was merely present in males. CONCLUSION: Gut bacteria of C. megacephala varied across life stages. The composition and community structure of the bacterial community differed from young larvae to mature larvae, while that were similar in adult females and males. These data will provide an overall view of bacterial community across life stages in C. megacephala with attention on manure associated and pathogenic bacteria.

A transgenic strategy for controlling plant bugs (Adelphocoris suturalis) through expression of double-stranded RNA homologous to fatty acyl-coenzyme A reductase in cotton.

Plant bugs (Miridae species), which are sap-sucking insects, have emerged as major pests of cotton in China. Most Miridae species are not sensitive to commercial Bacillus thuringiensis (Bt) cotton, resulting in significant economic losses and an increased application of insecticide, which eventually may compromise the future of Bt cotton. We demonstrate that FATTY ACYL-COA REDUCTASE (AsFAR) plays an essential role in the reproduction of the bug Adelphocoris suturalis. Down-regulation of AsFAR expression by injection of double-stranded RNA suppresses ovarian development and female fertility, resulting in females producing few viable offspring. To determine the viability of an RNA interference approach to limit FAR expression and reproductive ability in A. suturalis, a dsRNA targeting the AsFAR gene (dsAsFAR) of A. suturalis was expressed in transgenic cotton plants. AsFAR transcription levels were significantly downregulated in A. suturalis feeding on the transgenic plants. In contained field trials, the transgenic cotton lines significantly suppressed the development of A. suturalis populations and were resistant to damage caused by plant bug infestation. These results suggest a new strategy for the management of plant bug pests of cotton.

RNA interference knockdown of aminopeptidase N genes decrease the susceptibility of Chilo suppressalis larvae to Cry1Ab/Cry1Ac and Cry1Ca-expressing transgenic rice.

Transgenic rice expressing Bacillus thuringiensis (Bt) Cry toxins are resistant to lepidopteran pests, such as Chilo suppressalis, a major insect pest of rice in Asia. Understanding how these toxins interact with their hosts is crucial to understanding their insecticidal action. In this study, knockdown of two aminopeptidase N genes (APN1 and APN2) by RNA interference resulted in decreased susceptibility of C. suppressalis larvae to the Bt rice varieties TT51 (Cry1Ab and Cry1Ac fusion genes) and T1C-19 (Cry1Ca), but not T2A-1 (Cry2Aa). This suggests that APN1 and APN2 are receptors for Cry1A and Cry1C toxins in C. suppressalis.

The developmental transcriptome of the synanthropic fly Chrysomya megacephala and insights into olfactory proteins.

BACKGROUND: Chrysomya megacephala (Fabricius) is a prevalent and synanthropic blowfly which has two sides, for being a pathogenic vector, an efficient pollinator, a promising resource of proteins, lipids, chitosan, biofuel et al., and an important forensic indicator. Moreover olfactory proteins are crucial component to function in related processes. However, the genomic platform of C. megacephala remains relatively unavailable. Developmental transcriptomes of eggs, larvae from 1st instar to before pupa stage and adults from emergence to egg laying period were built by RNA-sequencing to establish sequence background of C. megacephala with special lights on olfactory proteins. RESULTS: Clean reads in eggs, larvae and adults were annotated into 59486 transcripts. Transcripts were assembled into 22286, 17180, 18934 and 35900 unigenes in eggs, larvae, adults and the combined datasets, respectively. Unigenes were annotated using Nr (NCBI non-redundant protein sequences), Nt (NCBI non-redundant nucleotide sequences), GO (Gene Ontology), PFAM (Protein family), KOG/COG (Clusters of Orthologous Groups of proteins), Swiss-Prot (A manually annotated and reviewed protein sequence database), and KO (KEGG Orthology). Totally 12196 unigenes were annotated into 51 sub-categories belonging to three main GO categories; 8462 unigenes were classified functionally into 26 categories to KOG classifications; 5160 unigenes were functionally classified into 5 KEGG categories. Moreover, according to RSEM, the number of differentially expressed genes between larvae and eggs, adults and eggs, adults and larvae, and the common differentially expressed genes were 2637, 1804, 2628 and 258, respectively. Among them, 17 odorant-binding proteins (OBPs), 7 chemosensory proteins (CSPs) and 8 ionotropic receptors (IRs) were differently expressed in adults and larvae. Ten were confirmed as significant differentially expressed genes. Furthermore, OBP Cmeg32081-c4 was highly expressed in the female head and Cmeg33593_c0 were up-regulated with the increase of larval age. CONCLUSIONS: A comprehensive sequence resource with desirable quality was built by comparative transcriptome of eggs, larvae and adults, enriching the genomic platform of C. megacephala. The identified differentially expressed genes would facilitate the understanding of metamorphosis, development and the fitness to environmental change of C. megacephala. OBP Cmeg32081-c4 and Cmeg33593_c0 might play a crucial role in the interactions between olfactory system and biological processes.

Cadherin is involved in the action of Bacillus thuringiensis toxins Cry1Ac and Cry2Aa in the beet armyworm, Spodoptera exigua.

Bacillus thuringiensis (Bt) insecticidal crystal (Cry) proteins are effective against some insect pests in sprays and transgenic crops, although the evolution of resistance could threaten the long-term efficacy of such Bt use. One strategy to delay resistance to Bt crops is to "pyramid" two or more Bt proteins that bind to distinct receptor proteins within the insect midgut. The most common Bt pyramid in cotton (Gossypium hirsutum L.) employs Cry1Ac with Cry2Ab to target several key lepidopteran pests, including the beet armyworm, Spodoptera exigua (Hübner), which is a serious migratory pest of many vegetable crops and is increasingly important in cotton in China. While cadherin and aminopeptidase-N are key receptors of Cry1 toxins in many lepidopterans including S. exigua, the receptor for Cry2A toxins remains poorly characterized. Here, we show that a heterologous expressed peptide corresponding to cadherin repeat 7 to the membrane proximal extracellular domain (CR7-MPED) in the S. exigua cadherin 1b (SeCad1b) binds Cry1Ac and Cry2Aa. Moreover, SeCad1b transcription was suppressed in S. exigua larvae by oral RNA interference and susceptibility to Cry1Ac and Cry2Aa was significantly reduced. These results indicate that SeCad1b plays important functional roles of both Cry1Ac and Cry2Aa, having major implications for resistance management for S. exigua in Bt crops.

Morphology and chemical analysis of the metathoracic scent glands system in Adelphocoris suturalis (Hemiptera: Miridae).

The morphological structure of the metathoracic scent glands (MTGs) in Adelphocoris suturalis was observed by utilizing scanning electron microscope (SEM). Also, the secretions of MTGs in male and female were analyzed by using gas chromatography-mass spectrometry (GC-MS). The result showed that the MTGs comprised a reservoir and paired lateral glands, which are connected to a reservoir by duct. The MTGs belong to the diastomian type. A usually depressed channel extends from opening downward the middle of metathorax, a tongue-like structure was covered by bristles and mushroom-shaped cuticular structures, known as evaporative area. In GC-MS investigation, differences were found in quantitative or qualitative compositions of the substances between the two sexes. In our study, hexyl butyrate was the most abundant compound in the MTGs of A. suturalis, comprising ∼85% of total secretions in both female and male, respectively. 4-oxo-(E)-2-hexenal (5.22%) was the second most abundant compound in female MTGs secretions, whereas octacosane (2.42%) followed hexyl butyrate in male MTGs secretions.

Antiviral, immunomodulatory, and free radical scavenging activities of a protein-enriched fraction from the larvae of the housefly, Musca domestica.

In our previous study, protein-enriched fraction (PEF) that was isolated from the larvae of the housefly, Musca domestica L. (Diptera: Muscidae), showed excellent hepatoprotective activity as well as the potential for clinical application in therapy for liver diseases. In this study, antiviral, immunomodulatory, and free radical scavenging activities of PEF were evaluated. The antiviral results demonstrated that PEF inhibited the infection of avian influenza virus H9N2 and had a virucidal effect against the multicapsid nucleopolyhedrovirus of the alfalfa looper, Autographa californica Speyer (Lepidoptera: Noctuidae) in vitro. The mortality of silkworm larve in a PEF treatment group decreased significantly compared with a negative control. PEF showed excellent scavenging activity for 1,1-diphenyl-2-picrylhydrazyl and superoxide anion radicals, which were similar to those of ascorbic acid. The imunomodulatory results suggested that PEF could effectively improve immune function in experimental mice. Our results indicated that PEF could possibly be used for the prophylaxis and treatment of diseases caused by avian influenza virus infection. In addition, PEF with virucidal activity against insect viruses might provide useful for the development of antimicrobial breeding technology for economically important insects. As a natural product from insects, PEF could be a potential source for the discovery of potent antioxidant and immunomodulatory agents.

New artificial diet for continuous rearing of the bean pod borer, Maruca vitrata.

The bean pod borer, Maruca vitrata Fabricius (Lepidoptera: Crambidae), is a serious pantropical pest of grain legumes. A suitable artificial diet is desirable for producing uniform insects for commercial purposes or research. Three previously described artificial diets, 1 newly-developed artificial diet, and cowpea (Vigna unguiculata (L.) Walp. (Fabales: Fabaceae)), the natural hostplant of M. vitrata, were used for rearing M. vitrata, and the life parameters were examined. The results indicated that insects completed a full life cycle only when the larvae were fed cowpea or the diet reported by Onyango and Ochieng'-Odero ( 1993 ), called the "D-OO diet." However, the rearing efficiency (i.e., larval and pupal survival, longevity of adults, and fecundity) on the D-OO diet was inferior to the rearing efficiency on cowpea. Subsequently, a new artificial diet was formulated based on soybean powder, Glycine max (L.) Merr. (Fabales: Fabaceae), and wheat germ, Triticum aestivum L. (Poales: Poaceae). The egg production, egg hatching, larval developmental duration, and pupal survival of the M. vitrata reared on the new artificial diet were found to be significantly improved relative to the D-OO diet, but were not significantly better than on the host-plant cowpea. The optimum rearing density was 15-25 larvae per box. There were no significant changes in reproductive potential after 8 successive generations of rearing on the new diet. These results indicated that the newly developed diet could serve as a viable alternative to cowpea plant for continuous rearing of M. vitrata.

In vitro anti-influenza activity of a protein-enriched fraction from larvae of the housefly (Musca domestica).

CONTEXT: Insects are a large, unexplored and unexploited source of potentially useful compounds for modern medicine. The larvae of the housefly (Musca domestica) have been used to study immune-induced molecules because they can survive in pathogenic environments. OBJECTIVE: The antiviral activity of a protein-enriched fraction (PEF) from the larvae of the housefly was evaluated in vitro and the possible antiviral mechanism was studied. MATERIALS AND METHODS: PEF was isolated from the larvae of the housefly. The cytotoxicity of PEF was detected by the MTT assay. The in vitro antiviral activity of PEF against influenza virus was investigated. PEF was incubated with the virus and its target cells under various conditions, and its antiviral effects were examined by reduction in virus yield in cell cultures. Experiments with ribavirin were performed in parallel under the same conditions. RESULTS: The results indicated that PEF had minimal cytotoxicity against MDCK cells and the CC50 value was calculated to be 284.45 µg/ml. The antiviral results showed the loss of infectious capacity was more than two log (2) units in cell cultures compared with virus control. The effect of PEF was direct virucidal activity and the interference on the adsorption of cell and virus. The antiviral mechanism of PEF is different from ribavirin. CONCLUSION: The results indicate that PEF showed strong antiviral activity against influenza virus at a very early stage of the interaction with virus particles or their entry into the cells. PEF has a great potential as a resource of healthy products.

The complete mitochondrial genome of the flea Hystrichopsylla weida qinlingensis (Siphonaptera: Hystrichopsylla).

The complete mitogenome sequence of the flea, Hystrichopsylla weida qinlingensis (Siphonaptera: Hystrichopsylla) was sequenced. The 17,173 bp long genome has the standard metazoan complement of 37 genes. These genes contain 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and one control region. The nucleotide composition of the H. weida qinlingensis mitogenome was A: 39.10%, T: 41.49%, G:7.56%, and C: 11.85%. The A + T content is 80.59%, showing strong AT bias. Phylogenetic analysis indicates that Hystrichopsylla has a close affinity with a branch of Dorcadia.

Tick (Acari: Ixodoidea) fauna and zoogeographic division of Jiangxi Province, China.

Tick fauna and zoogeographic distribution of Jiangxi Province remain largely unknown due to the lack of data on distribution, occurrence, and host associations of ticks. Considering this, we collected 1,817 individual samples from natural hosts, humans, and vegetation in 18 counties/districts throughout Jiangxi Province, China, from 2015 to 2021. These 1,817 individuals were found to 13 tick species, 4 genera, and 1 family. In addition, the tick sample data from 8 sampling localities (counties and districts) reported in previous studies were also included in our data. A total of 4,021 individuals, including our sample collection and the previously reported data, were assigned to at least 18 species, 6 genera, and 2 families. One newly recorded species Dermacentor sp. (near D. steini Schulze) was found; three misidentified species (Ixodes acuminatus, Haemaphysalis spinigera, and Haemaphysalis verticalis) reported previously were deleted; and one misidentified species Dermacentor auratus Supino was revised as Dermacentor steini Schulze. In addition, we divided the tick fauna in Jiangxi Province into 5 zoogeographic areas and assigned the 18 tick species collected from 26 localities to these 5 zoogeographic areas. To summarize, our findings provide valuable information on the distribution, tick-host associations, and zoogeographic division of ticks in Jiangxi Province, China. Their molecular characterizations, phylogenetic relationships, and tick-borne pathogens that they may transmit should be further explored.

Transcriptomics Reveals the Killing Mechanism by Which Entomopathogenic Fungi Manipulate the RNA Expression Profiles of Termites and Provides Inspiration for Green Pest Management.

As chemical pesticides have caused serious environmental pollution, fungus-based biological control has become a developing alternative to chemical control. Here, we aimed to determine the molecular mechanism underlying how Metarhizium anisopliae facilitated invasive infection. We found that the fungus increased its virulence by downregulating glutathione S-transferase (GST) and superoxide dismutase (SOD) throughout termite bodies. Among 13 fungus-induced microRNAs throughout termite bodies, miR-7885-5p and miR-252b upregulation significantly downregulated several mRNAs in response to toxic substances to increase the fungal virulence [e.g., phosphoenolpyruvate carboxykinase (GTP) and heat shock protein homologue SSE1]. In addition, nanodelivered small interfering RNA of GST and SOD and miR-7885-5p and miR-252b mimics increased the virulence of the fungus. These findings provide new insights into the killing mechanism of entomopathogens and their utilization of the host miRNA machinery to reduce host defenses, laying the groundwork to enhance virulence of biocontrol agents for green pest management.

Projecting overwintering regions of the beet armyworm, Spodoptera exigua in China using the CLIMEX model.

The beet armyworm, Spodoptera exigua Hübner (Lepidoptera: Noctuidae) is a serious agricultural pest worldwide. However, population sources of S. exigua in outbreak regions are still vague due to the lack of understanding the distribution of overwintering regions, especially in China. In the present study, the potential overwintering regions of S. exigua in China are projected using the method of Compare Location in the CLIMEX model in order to understand the population sources in outbreak regions and establish an accurate forecasting system. The results showed the southern and northern overwintering boundaries near the Tropic of Cancer (about 23.5 (°)N) and the Yangtze River valley (about 30 (°)N), respectively. Meanwhile, the projection was supported by the data of fieldwork in 14 countries/cities during winter from 2008-2010. In conclusion, results of this study indicated that the overwintering regions of S. exigua were accurately projected by the CLIMEX model.

The diversity and evolutionary relationships of ticks and tick-borne bacteria collected in China.

BACKGROUND: Ticks (order Ixodida) are ectoparasites, vectors and reservoirs of many infectious agents affecting humans and domestic animals. However, the lack of information on tick genomic diversity leaves significant gaps in the understanding of the evolution of ticks and associated bacteria. RESULTS: We collected > 20,000 contemporary and historical (up to 60 years of preservation) tick samples representing a wide range of tick biodiversity across diverse geographic regions in China. Metagenomic sequencing was performed on individual ticks to obtain the complete or near-complete mitochondrial (mt) genome sequences from 46 tick species, among which mitochondrial genomes of 23 species were recovered for the first time. These new mt genomes data greatly expanded the diversity of many tick groups and revealed five cryptic species. Utilizing the same metagenomic sequence data we identified divergent and abundant bacteria in Haemaphysalis, Ixodes, Dermacentor and Carios ticks, including nine species of pathogenetic bacteria and potentially new species within the genus Borrelia. We also used these data to explore the evolutionary relationship between ticks and their associated bacteria, revealing a pattern of long-term co-divergence relationship between ticks and Rickettsia and Coxiella bacteria. CONCLUSIONS: In sum, our study provides important new information on the genetic diversity of ticks based on an analysis of mitochondrial DNA as well as on the prevalence of tick-borne pathogens in China. It also sheds new light on the long-term evolutionary and ecological relationships between ticks and their associated bacteria.

In silico promoter analysis and functional validation identify CmZFH, the co-regulator of hypoxia-responsive genes CmScylla and CmLPCAT.

Oxygen (O2) plays an essential role in aerobic organisms including terrestrial insects. Under hypoxic stress, the cowpea bruchid (Callosobruchus maculatus) ceases feeding and growth. However, larvae, particularly 4th instar larvae exhibit very high tolerance to hypoxia and can recover normal growth once brought to normoxia. To better understand the molecular mechanism that enables insects to cope with low O2 stress, we performed RNA-seq to distinguish hypoxia-responsive genes in midguts and subsequently identified potential common cis-elements in promoters of hypoxia-induced and -repressed genes, respectively. Selected elements were subjected to gel-shift and transient transfection assays to confirm their cis-regulatory function. Of these putative common cis-elements, AREB6 appeared to regulate the expression of CmLPCAT and CmScylla, two hypoxia-induced genes. CmZFH, the putative AREB6-binding protein, was hypoxia-inducible. Transient expression of CmZFH in Drosophila S2 cells activated CmLPCAT and CmScylla, and their induction was likely through interaction of CmZFH with AREB6. Binding to AREB6 was further confirmed by bacterially expressed CmZFH recombinant protein. Deletion analyses indicated that the N-terminal zinc-finger cluster of CmZFH was the key AREB6-binding domain. Through in silico and experimental exploration, we discovered novel transcriptional regulatory components associated with gene expression dynamics under hypoxia that facilitated insect survival.

Enhancement of supercooling capacity and survival by cold acclimation, rapid cold and heat hardening in Spodoptera exigua.

Insects can increase their resistance to cold stress by prior exposure to non-lethal cold temperatures. Here, we investigated the supercooling capacity and survival of eggs, 3rd and 5th instar larvae, and pupae of Spodoptera exigua (Lepidoptera: Noctuidae) during CA, and responses to various pre-treatment protocols, including constant temperatures, thermoperiods, and RCH, RHH, RCH+RHH and RHH+RCH combined with thermoperiods. Only acclimated eggs demonstrated a significant decrease in SCP, from -20.7±0.3 to -22.9±0.3°C, among all experimental groups compared to non-acclimated stages. Survival increased by 17.5% for eggs, 40.0% and 13.3% for 3rd and 5th instar larvae, and by 20.0% for pupae after CA. Compared to controls, survival of eggs under the conditions of thermoperiod (5:15°C), thermoperiod (5:15°C)+RHH, and thermoperiod (5:15, 10:20, and 15:25°C)+RCH significantly increased. In addition, survival of 3rd and 5th instar larvae and pupae increased under the conditions of thermoperiod (5:15°C) and thermoperiod (5:15°C)+RCH, possibly due to the induction of heat shock proteins or cryoprotectants. However, the pre-treatments of thermoperiod+RCH+RHH and thermoperiod+RHH+RCH did not significantly enhance survival of any developmental stage. These adaptive responses may allow S. exigua to enhance supercooling capacity and survival in response to seasonal or unexpected diurnal decreases in environmental temperatures.