Glyphodes Pyloalis Research Articles

Influence of Biotic and Abiotic Factor on Incidence of Glyphodes pyloalis and its Natural Enemy’s Population in Jorhat District, Assam, India

Influence of Biotic and Abiotic Factor on Incidence of Glyphodes pyloalis and its Natural Enemy’s Population in Jorhat District, Assam, India

Binding characteristics and structural dynamics of two general odorant-binding proteins with plant volatiles in the olfactory recognition of Glyphodes pyloalis

Glyphodes pyloalis Walker (Lepidoptera: Pyralidae) is the most destructive pest, causing severe damage to mulberry production in China's sericulture industry. The insecticide application in mulberry orchards poses a significant risk of poisoning to Bombyx mori. Shifting from insecticides to odor attractants is a beneficial alternative, but not much data is available on the olfactory system of G. pyloalis. We identified 114 chemosensory genes from the antennal transcriptome database of G. pyloalis, with 18 odorant-binding protein (OBP) and 17 chemosensory protein (CSP) genes significantly expressed in the antennae. Ligand-binding assays for two antennae-biased expressed general odorant-binding proteins (GOBPs) showed high binding affinities of GOBP1 to hexadecanal, β-ionone, and 2-ethylhexyl acrylate, while GOBP2 exhibited binding to 4-tert-octylphenol, benzyl benzoate, β-ionone, and farnesol. Computational simulations indicated that van der Waal forces predominantly contributed to the binding free energy in the binding processes of complexes. Among them, Phe12 of GOBP1 and Phe19 of GOBP2 were demonstrated to play crucial roles in their bindings to plant volatiles using site-directed mutagenesis experiments. Moreover, hexadecanal and β-ionone attracted G. pyloalis male moths in the behavioral assays, while none of the candidate plant volatiles significantly affected female moths. Our findings provide a comprehensive understanding of the molecular mechanisms underlying olfactory recognition in G. pyloalis, setting the groundwork for novel mulberry pests control strategies based on insect olfaction.

Insecticidal efficacy of some apiaceae plant metabolites against glyphodes pyloalis walker (lepidoptera: pyralidae)

Botanical based-insecticides are presently recognized to interfere behavioral and physiological performances of herbivorous insects. In the current study, the insecticidal properties of Pimpinella anisum, Foeniculum vulgare and Heracleum persicum essential oils (EOs), and their main component, trans-anethole, were studied on a destructive pest of mulberry leaves, Glyphodes pyloalis Walker. The effect of essential oils (EOs) on the larval feeding was evaluated by the leaf-dipping method and demonstrated the highest insecticidal and antifeedant activities of F. vulgare and H. persicum. Apart from negative effects on feeding indices, feeding treatment with the EOs significantly inhibited the activities of digestive enzymes and acetylcholinesterase (AChE). At nearly, all the tested compounds increased the activities of detoxifying and antioxidant enzymes. Thus, these EOs demonstrate potential as new control products to combat G. pyloalis, and they may function as an insecticide, antifeedant, or inhibitor of physiological performance.

Cadmium Exposure Impairs Development, Detoxification Mechanisms and Gene Expression of Glyphodes pyloalis Walker (Lepidoptera: Pyralidae)

Cadmium (Cd) is the most serious heavy metal pollutant in the agricultural soil of China and can transfer and accumulate through the food chain and affect the growth, development and physiological processes of phytophagous insect. Glyphodes pyloalis Walker (G. pyloalis) is one of the most important pests of mulberry, and there are few studies on the adverse effects of heavy metals on insects, especially mulberry pests. To understand the toxicology of Cd exposure on G. pyloalis, we investigated the effects of three different concentrations of Cd (0, 3.89 and 51.69 mg/kg, labeled as control check (CK), low dose (LD) and high dose (HD)) on the development and detoxification mechanism of G. pyloalis and explored the molecular mechanism of Cd on G. pyloalis larvae using RNA-seq technology. Transcriptome analysis showed that compared with the CK, a total of 63 differentially expressed genes (DEGs) were identified in LD exposure, including 24 upregulated and 39 downregulated candidates. In CK versus HD groups, 395 upregulated DEGs and 436 downregulated DEGs were identified and the expression patterns of 12 genes related to detoxification and metabolism were verified using qPCR. These DEGs were relevant to multiple specific peroxisome and drug metabolism-cytochrome P450 pathways. Gene annotation and quantitative real-time PCR revealed that a high concentration of Cd significantly stimulated the expression of metabolic detoxification enzyme genes. The results revealed that Cd exposure changed the pupal weight, adult emergence rate and the activities of AKP, ACP and GST in G. pyloalis and induced notable adverse effects at the molecular level on detoxification and metabolism such as concentration and time differences. In addition, we silenced CYP12A2 by RNA interference. Bioassays showed that after silencing CYP12A2, the survival rate of G. pyloalis under Cd exposure was lower than that of the control group.

Binding characteristics of pheromone-binding protein 1 in Glyphodes pyloalis to organophosphorus insecticides: Insights from computational and experimental approaches.

Glyphodes pyloalis (Lepidoptera: Pyralidae) is one of the major pests in mulberry production in China, which has developed resistance to various insecticides. Chemoreception is one of the most crucial physiological tactics in insects, playing a pivotal role in recognizing chemical stimuli in the environment, including noxious stimuli such as insecticides. Herein, we obtained recombinant pheromone-binding protein 1 (GpylPBP1) that exhibited antennae-biased expression in G. pyloalis. Ligand-binding assays indicated that GpylPBP1 had the binding affinities to two organophosphorus insecticides, with a higher binding affinity to chlorpyrifos than to phoxim. Computational simulations showed that a mass of nonpolar amino acid residues formed the binding pocket of GpylPBP1 and contributed to the hydrophobic interactions in the bindings of GpylPBP1 to both insecticides. Furthermore, the binding affinities of three GpylPBP1 mutants (F12A, I52A, and F118A) to both insecticides were all significantly reduced compared to those of the GpylPBP1-wild type, suggesting that Phe12, Ile52, and Phe118 residues were crucial binding sites and played crucial roles in the bindings of GpylPBP1 to both insecticides. Our findings can be instrumental in elucidating the effects of insecticides on olfactory recognition in moths and facilitating the development of novel pest management strategies using PBPs as targets based on insect olfaction.

Impact of Glyphodes pyloalis walker infestation on macronutrient and biochemical contents of mulberry (Morus spp.)

Impact of Glyphodes pyloalis walker infestation on macronutrient and biochemical contents of mulberry (Morus spp.)

Identification and functional study of detoxification-related genes in response to tolfenpyrad stress in Glyphodes pyloalis Walker (Lepidoptera: Pyralidae)

Glyphodes pyloalis Walker (G. pyloalis) is a common destructive mulberry pest. Due to the long-term and frequent use of insecticides, it has developed tolerance to commonly used insecticides. Tolfenpyrad (TFP) is a novel pyrazole heterocyclic insecticide. In order to understand the TFP detoxification mechanism of G. pyloalis larvae, we first estimated the LC30 dose of TFP for 3rd instar G. pyloalis larvae. Next, we identified genes that were differentially expressed in 3rd instar G. pyloalis larvae treated with TFP compared to the control group by transcriptome sequencing. In total, 86,949,569 and 67,442,028 clean reads were obtained from TFP-treated and control G. pyloalis larvae, respectively. A total of 5588 differentially expressed genes (DEGs) were identified in TFP-treated and control G. pyloalis larvae, of which 3084 genes were upregulated and 2504 genes were downregulated. We analyzed the expression of 43 candidate detoxification enzyme genes associated with insecticide tolerance using qPCR. According to the spatiotemporal expression pattern of DEGs, we found that CYP6ABE1, CYP333A36 and GST-epsilon8 were highly expressed in the midgut, while CarEs14 was strongly expressed in haemolymph. Furthermore, we successfully knocked down these genes by RNA interference. After silencing CYP6ABE1 and CYP333A36, bioassay showed that the mortality rate of TFP-treated G. pyloalis larvae was significantly higher compared to the control group. This study provides a theoretical foundation for understanding the sensitivity of G. pyloalis to TFP and establish the basis for the effective and green management of this pest.

Transcriptomic analysis of the fat body of resistant and susceptible silkworm strains, Bombyx mori (Lepidoptera), after oral treatment with fenpropathrin

The widespread use of pyrethroid pesticides has brought serious economic losses in sericulture, but there is still no viable solution. The key to solving the problem is to improve silkworm resistance to pesticides, which depends on understanding the resistance mechanism of silkworms to pesticides. This study aimed to use transcriptomes to understand the underlying mechanism of silkworm resistance to fenpropathrin, which will provide a theoretical molecular reference for breeding pesticide-resistant silkworm varieties. In this study, the fat bodies of two strains with differential resistance after 12 h of fenpropathrin feeding were analyzed using RNA-Seq. After feeding fenpropathrin, 760 differentially expressed genes (DEGs) were obtained in the p50(r) strain and 671 DEGs in the 8y strain. The DEGs involved in resistance to fenpropathrin were further identified by comparing the two strains, including 207 upregulated DEGs in p50(r) and 175 downregulated DEGs in 8y. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that these fenpropathrin-related DEGs are mainly enriched in the metabolism and transporter pathways. Moreover, 28 DEGs involved in the metabolic pathway and 18 in the transporter pathway were identified. Furthermore, organic cation transporter protein 6 (BmOCT6), a transporter pathway member, was crucial in enhancing the tolerance of BmN cells to fenpropathrin. Finally, the knockdown of the expression of the homologs of BmOCT6 in Glyphodes pyloalis (G. pyloalis) significantly decreased the resistant level of larvae to fenpropathrin. The findings showed that the metabolism and transporter pathways are associated with resistance to fenpropathrin in silkworm, and OCT6 is an effective and potential target not only for silkworm breeding but also for pest biocontrol.

Integrated Pest and Disease Management (IPDM) Approach in Mulberry Sericulture of Jammu & Kashmir

Jammu and Kashmir is one of the traditional sericultural states of India, produces 735 MT of cocoons which is converted into 98 MT of raw silk. Spring is the major silkworm rearing season of J&K when fresh mulberry leaf is available for rearing the silkworm, Bombyx mori L. The food plant of silkworm, mulberry is affected by various insect pests viz., Glyphodes pyloalis, Spilosoma oblique, Amsacta lactinea, Hemerophilla atrilineata etc., and diseases like leaf spot and powdery mildew. In integrated pest and disease management of mulberry various factors such as cultural, mechanical, biological and bio-pesticides management practices have been adopted to control the pests as well as the diseases of mulberry. The major diseases are powdery mildew and leaf spot which affect the foliage of the mulberry plant thereby reducing the quality and quantity of the silkworm feed. The present study was conducted to identify the suitable IPDM approach to manage pest and diseases in mulberry sericulture of Jammu & Kashmir. Results showed that the impact of IPDM the incidence of insect pest to an extent of 43.66% and 48.82% by leaf Webber and semilooper respectively. Leaf spot and powdery mildew disease of mulberry was also reduced up to 39.56% and 49.92% respectively. By adopting 100% IPDM module cocoon yield was increased up to 4 kg 100 disease free laying (dfl).

Effect of thyme essential oil and its two components on toxicity and some physiological parameters in mulberry pyralid Glyphodes pyloalis Walker

Extensive usage of synthetic pesticides has proved to be destructive to all living being and the resurgence of pest resistance. Compounds derived from certain plants are usually safer compared to chemical control of pest. The present study thus intended to use Thymus vulgaris essential oil (EO) and two of its derivatives including thymol and carvacrol in order to see their deleterious effects on Glyphodes pyloalis (Walker). We also studied the oil components. This pest has recently become a serious concern for the silk industry. Our results showed that the thyme EO contain several components including thymol (26.9%), ρ-Cymene (14.54%), linalool (13.39%) and carvacrol (5.7%). Our toxicity tests revealed an estimated LD50 values for thyme EO, thymol and carvacrol 2.82, 32.18 and 56.54 μg/larva, respectively. However, the thyme EO was more toxic than its two tested compounds. The activity of certain detoxifying enzymes such as α- and β-esterase, glutathione S-transferase and cytochrome P450 were significantly inhibited by thymol-treated larvae compared to the control group. Similarly, the activity of alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase and alkaline phosphatases enzymes in thymol-treated larvae decreased while the activity of acid phosphatases increased. Our results suggest that thyme EO and its components have potential for the control of G. pyloalis larvae in mulberry orchards, where no synthetic chemicals are allowed.

ПРИМЕНЕНИЕ БИОТЕХНОЛОГИЧЕСКИХ ПОДХОДОВ И МЕТОДОВ В ШЕЛКОВОДСТВЕ, ТЕХНОЛОГИИ ШЕЛКА И РЕСУРСОСБЕРЕЖЕНИИ

This information presents research and development based on fundamental knowledge about the silkworm called Bombyx mori L. It highlights development of biotechnological approaches used in sericulture, cocoon processing and resource-saving. The research is devoted to a close development of artifi cial diets for silkworms, their use in various fields of practical application, including in outer-space experiments. The processes of respiration, digestion at B. mori L. and biotransformation of sericulture products and wastes at the enzymatic level have been explored; as well as methods for biocontrol of mulberry pyralid Glyphodes pyloalis Wlk. have been investigated and developed in the IPM system. Biotechnological approaches have been developed for rational use of raw materials and waste left from the silk-making processes, for production of fibroin and sericin, as well as composites and semi-finished products based on them - for pharmaceutical, biomedical, cosmetic and food industries and bionanotechnology. The structural organization of fibroin in fiber and composites has been researched. As the findings show, the research area in question proves to be quite fruitful and its further development will lead to a waste-less, diversified and profitable production of high-tech silk products and goods.

A Comparative Analysis of Bombyx mori (Lepidoptera: Bombycidae) β-fructofuranosidase Homologs Reveals Different Post-Translational Regulations in Glyphodes pyloalis Walker (Lepidoptera: Pyralidae).

Simple SummaryThe β-fructofuranosidase (β-FFase) encoding gene BmSuc1 regulates the glycometabolism of silkworm larvae, and it participates in the resistance of mulberry alkaloids. However, there is no molecular or biochemical information available about the mulberry pest Glyphodes pyloalis Walker β-FFase homologs. In this paper, we have obtained five β-FFase homologous genes in G. pyloalis and characterized the expression and the localization of GpSUC1a in the midgut. The β-FFase activity in the midgut of G. pyloalis larvae and GpSUC1a were both confirmed, while recombinant GpSUC1a displayed little activity as compared with the higher activity of BmSUC1. Some putative N-glycosylation sites were found in GpSUC1a but none in BmSUC1, while there was more methylation in BmSUC1 than in GpSUC1a. The results indicate that such post-translational modifications (PTMs) are differentially supporting that β-FFase are active in these two mulberry feeding caterpillars, and the activation of GpSUC1a may be controlled by a more complex post-translational regulatory system in G. pyloalis larvae. This is the first report on the characterization of β-FFase genes from G. pyloalis and the first comparison of expression regulation between two mulberry feeding insects B. mori and G. pyloalis. Moreover, this research may provide new ideas for the management of mulberry borers.The silk-spinning and Lepidopteran model insect Bombyx mori (Bombycidae) is a mulberry specialist. The BmSuc1 gene is the first β-fructofuranosidase (β-FFase) encoding gene identified in animals, and β-FFase acts as an essential sucrase for glycometabolism modulation in the silkworm larvae, involved in resistance to mulberry alkaloids. Glyphodes pyloalis Walker (Lepidoptera: Pyralidae) is an important mulberry pest leading to heavy economic loss of sericulture. However, no molecular or biochemical information is available about G. pyloalis β-FFase homologs. In this study, five β-FFase homologous genes in G. pyloalis were obtained. The genes GpSuc1a and GpSuc2c were expressed in the midgut; GpSuc2c encodes a truncated polypeptide. The expression and the localization of GpSUC1a in the midgut was characterized. Whereas recombinant GpSUC1a expressed in both Escherichia coli and BmN cells displayed little activity as compared with higher activity of BmSUC1, β-FFase activity in the larval midgut of G. pyloalis and GpSUC1a purified from the midgut were both confirmed. The data suggested that the activation of GpSUC1a is probably controlled by a more complicated post-translational regulation system in G. pyloalis larvae than that of BmSUC1 in B. mori. To study post-translational modifications (PTMs), GpSUC1a and BmSUC1 were purified from larval midguts using immunoprecipitation and subjected to LC-MS to perform PTMs analysis. Some putative N-glycosylated sites were found in GpSUC1a but none in BmSUC1, while there was more methylation in BmSUC1 than in GpSUC1a, indicating that such PTMs were supporting the differential β-FFases activities in these two mulberry feeding caterpillars.

Fatty acid synthases and desaturases are essential for the biosynthesis of α-linolenic acid and metamorphosis in a major mulberry pest, Glyphodes pyloalis walker (Lepidoptera: Pyralidae).

α-linolenic acid is an essential unsaturated fatty acid in organisms. However, there is a large gap between α-linolenic acid accumulation and its synthesis mechanism in insects. Fatty acid synthases (FASs) and desaturases (Desats) are vital enzymes required for the synthesis of unsaturated fatty acids. The pupae of Glyphodes pyloalis (Lepidoptera: Pyralidae), which is a destructive pest of mulberry trees, contain the highest level of α-linolenic acid compared to other life-history stages. To further explore the synthesis mechanism of α-linolenic acid in G. pyloalis pupae, we constructed a pupal transcriptome dataset and identified 106 genes related to fatty acid metabolism from it. Among these, two fatty acid synthases (GpylFAS) and five desaturases (GpylDesat) were identified. A qRT-PCR validation revealed that GpylFAS1 and GpylDesat1, 2, 3, 5 were expressed highest at pupal stages. Furthermore, the content of α-linolenic acid decreased significantly after silencing GpylFAS1 and GpylDesat5, respectively. Besides, knocking down GpylFAS1 or GpylDesat5 resulted in more malformed pupae and adults, as well as lower emergence rates. Meanwhile, silencing GpylFAS1 or GpylDesat5 affected the expressions of the other GpylFASs and GpylDesats. The present results illustrate the pivotal function of FASs and Desats in α-linolenic acid biosynthesis and metamorphosis in insects. Our research also broadens the sources of unsaturated fatty acids, especially for α-linolenic acid from insects, and provides novel insights for the management of mulberry insect pests from the perspective of utilization rather than control. © 2022 Society of Chemical Industry.

Hemocytic cell line from the moth Glyphodes pyloalis (Lepidoptera: Crambidae) response to essential oils from Artemisia annua (Asterales: Asteraceae).

Extensive usage of synthetic chemical pesticides has collateral effects in harming human and animal health and the environment and promoting the development of resistance in pests. The potential of plant compounds as bio insecticides has been described as a promising field of agricultural development. The present study involved the use of Artemisia annua essential oils to evaluate their cytotoxic activities against an established cell line of lesser mulberry pyralid. Five types of hemocytes were recognized (prohaemocytes, plasmatocytes, granulocytes, oenocytoids, and spherulocytes) in the primary cultures maintained in Ex-Cell media with 10% fetal bovine serum (FBS). Artemisia annua essential oils produced noticeable cytotoxicity against the insect cell lines. Applied at a concentration 500ppm, oils extracted from the vegetative or flowering stages of A. annua produced 71% and 80% cell death, respectively. Nanoemulsions of EOs from the vegetative or flowering stages of A. annua killed 67 and 60% of the cells, respectively. This study has clearly shown significant bioactivities of A. annua secondary metabolites to insect cell lines.

The role of Glutathione-S-transferases in phoxim and chlorfenapyr tolerance in a major mulberry pest, Glyphodes pyloalis walker (Lepidoptera: Pyralidae)

Glyphodes pyloalis Walker is a destructive pest on mulberry trees and poses a significant threat to the sericultural industry in China. Phoxim and chlorfenapyr are two commonly used insecticides in mulberry fields. Glutathione-S-transferases (GSTs) comprise a multifunctional protein superfamily that plays important roles in the detoxification of insecticides and xenobiotic compounds in insects. However, whether GSTs participate in the tolerance of phoxim and chlorfenapyr in G. pyloalis is still unknown. To better understand the mechanism of insecticide tolerance in G. pyloalis, the enzymatic activity of GSTs was evaluated under phoxim and chlorfenapyr exposure, respectively. GST enzyme activity was significantly increased after 12, 36 and 48 h of phoxim treatment and 12, 24, 36 and 48 h of chlorfenapyr treatment. Subsequently, eighteen GST genes were identified from the larvae transcriptome of G. pyloalis. Among these, ten GpGSTs had GSH-binding sites and fifteen GpGSTs had variable hydrophobic substrate-binding sites. The expression levels of Delta-GpGST and Epsilon-GpGST genes were significantly influenced by phoxim and chlorfenapyr treatment, and by the time post insecticide application. Furthermore, after silencing GpGST-E4, the mortality rate of G. pyloalis larvae was increased when they were exposed to chlorfenapyr, but it did not significantly alter when the larvae were exposed to phoxim. Our results indicated the vital roles of GpGSTs in the tolerance of insecticides and this action depends on the categories of insecticides. The present study provides a theoretical basis for elucidating insecticide susceptibility and promotes functional research on GST genes in G. pyloalis.

Characterization, and Functional Analysis of Hsp70 and Hsp90 Gene Families in Glyphodes pyloalis Walker (Lepidoptera: Pyralidae).

Glyphodes pyloalis Walker (Lepidoptera: Pyralidae) is a destructive mulberry pest, causing great damage to mulberry in China. Heat shock proteins (Hsps) are involved in various signal pathways and regulate lots of physiological processes in insects. The function of Hsps in G. pyloalis, however, has still received less attention. Here, we identified five Hsp genes from G. pyloalis transcriptome dataset including two Hsp70 family genes (GpHsp71.3 and GpHsp74.9) and three Hsp90 family genes (GpHsp82.4, GpHsp89, and GpHsp93.4). Quantitative Real-time PCR validation revealed that all Hsps of G. pyloalis have significant expression in pupal and diapause stage, at which the larvae arrest the development. Expressions of GpHsp71.3 and GpHsp82.4 were increased significantly after thermal treatment at 40°C, and this upregulation depended on heat treatment duration. Furthermore, silencing GpHsp82.4 by RNA interference led to a significant increase in mortality of G. pyloalis larvae under the heat stress compared to the control group. After starvation stress, the expression levels of GpHsp82.4 and GpHsp93.4 were significantly increased. At last, after being parasitized by the parasitoid wasp Aulacocentrum confusum, Hsp70 and Hsp90 genes of G. pyloalis were decreased significantly in the early stage of parasitization and this moderation was affected by time post-parasitization. This study highlights the function of G. pyloalis Hsps in response to environmental stress and provides a perspective for the control of this pest.

A role of peptidoglycan recognition protein in mediating insecticide detoxification in Glyphodes pyloalis.

Glyphodes pyloalis Walker has become one of the most significant mulberry pests, and it has caused serious economic losses in major mulberry growing regions in China. Peptidoglycan recognition proteins (PGRPs) are responsible for initiating and regulating immune signalling pathways in insects. However, their roles responding to chemical pesticides is still less known. This study aimed to investigate the possible detoxication function of GpPGRP-S2 and GpPGRP-S3 in G. pyloalis in response to chlorfenapyr and phoxim. The chlorfenapyr and phoxim treatment significantly induced the expression level of GpPGRP-S3 at 48 h. In addition, the expression levels of GpPGRP-S2 and GpPGRP-S3 in the chlorfenapyr/phoxim treatment group were significantly higher in midgut than those in the control group at 48 h. The results of the survival experiment showed that silencing either GpPGRP-S2 or GpPGRP-S3 would not influence the survival rate of G. pyloalis which treated with phoxim, however, silencing GpPGRP-S2 or GpPGRP-S3 would cause G. pyloalis to be more easily killed by chlorfenapyr. The expression of carboxylesterase GpCXE1 was significantly induced by chlorfenapyr/phoxim treatment, while it was suppressed once silenced GpPGRP-S2 followed with chlorfenapyr treatment or silenced GpPGRP-S3 followed with phoxim treatment. These results might suggest that under the chlorfenapyr/phoxim treatment condition, the connection between GpPGRPs and detoxification genes in insect was induced to maintain physiological homeostasis; and these results may further enrich the mechanisms of insects challenged by insecticides.

Identification of chemosensory genes by antennal transcriptome analysis and expression profiles of odorant-binding proteins in parasitoid wasp Aulacocentrum confusum

The endoparasitoid wasp, Aulacocentrum confusum (Hymenoptera: Braconidae), is a preponderant natural enemy of the larvae of Glyphodes pyloalis Walker (Lepidoptera: Pyralidae), which is a destructive pest of mulberry trees. We first constructed the antennal transcriptome database of A. confusum. In total, we obtained 48,262,304 clean reads from the dataset and assembled 24,324 unigenes. A total of 12,690 (52.17%) unigenes indicated significant similarity (E-value < 10−5) compared to known protein sequences of other species from the NCBI non-redundant protein database. Gene ontology (GO) and cluster of orthologous groups (COG) analyses were used to determine the functional categories of these genes. A total of 84 putative chemosensory genes were identified from the antennal transcriptome of A. confusum, including 11 putative odorant-binding protein (OBP) genes, six chemosensory protein (CSP) genes, 44 olfactory receptor (OR) genes (including one olfactory co-receptor, Orco), 19 ionotropic receptor (IR) genes, and four sensory neuron membrane protein (SNMP) genes. Results of qPCR assays indicated that among of 11 AconOBPs, nine AconOBP genes were significantly expressed in the antennae of A. confusum adults. AconOBP8 was significantly expressed in the abdomen and AconOBP10 was highly expressed in the thorax. These findings can build a basis for further study on the processes of chemosensory perception in A. confusum at the molecular level.

Immunological and oxidative responses of the lesser mulberry pyralid, Glyphodes pyloalis by an aqueous extract of Artemisia annua L.

In this search for affordable and locally available biological substances both to farmers and environment, an aqueous extract of Artemisia annua L. was investigated for the first time against the lesser mulberry pyralid, Glyphodes pyloalis Walker a serious pest in mulberry orchards. The LC10, LC30 and LC50 values were estimated 12.82 %, 20.6 % and 27.35 % (W/V) respectively. The extract adversely affected oviposition, impaired immunity through reduced granulocytes and phenoloxidase activity. The increased activity of detoxifying enzymes including esterases and glutathione S-transferase (GST) were also observed. The enhanced antioxidant system including peroxidase (POX), catalase (CAT), glucose 6-phosphate dehydrogenase (GPDH) and superoxide dismutase (SOD) were also observed. The results of the present study may provide a very safe way to control this pest in mulberry orchard and deserve further studies.

Characterization and Functional Analysis of trehalase Related to Chitin Metabolism in Glyphodes pyloalis Walker (Lepidoptera: Pyralidae).

Glyphodes pyloalis Walker (G. pyloalis) is a serious pest on mulberry. Due to the increasing pesticide resistance, the development of new and effective environmental methods to control G. pyloalis is needed. Trehalase is an essential enzyme in trehalose hydrolysis and energy supply, and it has been considered a promising target for insect pest control. However, the specific function of trehalase in G. pyloalis has not been reported. In this study, two trehalase genes (GpTre1 and GpTre2) were identified from our previous transcriptome database. The functions of the trehalase in chitin metabolism were studied by injecting larvae with dsRNAs and trehalase inhibitor, Validamycin A. The open reading frames (ORFs) of GpTre1 and GpTre2 were 1,704 bp and 1,869 bp, which encoded 567 and 622 amino acid residues, respectively. Both of GpTre1 and GpTre2 were mainly expressed in the head and midgut. The highest expression levels of them were in 5th instar during different development stages. Moreover, knockdown both of GpTre1 and GpTre2 by the dsRNAs led to significantly decreased expression of chitin metabolism pathway-related genes, including GpCHSA, GpCDA1, GpCDA2, GpCHT3a, GpCHT7, GpCHSB, GpCHT-h, GpCHT3b, GpPAGM, and GpUAP, and abnormal phenotypes. Furthermore, the trehalase inhibitor, Validamycin A, treatment increased the expressions of GpTre1 and GpTre2, increased content of trehalose, and decreased the levels of glycogen and glucose. Additionally, the inhibitor caused a significantly increased cumulative mortality of G. pyloalis larvae on the 2nd (16%) to 6th (41.3%) day, and decreased the rate of cumulative pupation (72.3%) compared with the control group (95.6%). After the activities of trehalase were suppressed, the expressions of 6 integument chitin metabolism-related genes decreased significantly at 24 h and increased at 48 h. The expressions of GpCHSB and GpCHT-h, involved in chitin metabolism pathway of peritrophic membrane in the midgut, increased at 24 h and 48 h, and there were no changes to GpCHT3b and GpPAGM. These results reveal that GpTre1 and GpTre2 play an essential role in the growth of G. pyloalis by affecting chitin metabolism, and this provides useful information for insect pest control in the future.