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The European Commission requested the EFSA Panel on Plant Health to conduct a pest categorisation of Ceroplastes rubens Maskell (Hemiptera: Coccidae), following the commodity risk assessments of Acer palmatum plants grafted on A. davidii and Pinus parviflora bonsai plants grafted on P. thunbergii from China, in which C. rubens was identified as a pest of possible concern to the European Union (EU). The pest, which is commonly known as the pink, red or ruby wax scale, originates in Africa and is highly polyphagous attacking plants from more than 193 genera in 84 families. It has been present in Germany since 2010 in a single tropical glasshouse. It is known to attack primarily tropical and subtropical plants, but also other host plants commonly found in the EU, such as Malus sylvestris, Prunus spp., Pyrus spp. and ornamentals. It is considered an important pest of Citrus spp. The pink wax scale reproduces mainly parthenogenetically, and it has one or two generations per year. Fecundity ranges from 5 to 1178 eggs. Crawlers settle usually on young twigs and later stages are sessile. All life stages of C. rubens egest honeydew on which sooty mould grows. Host availability and climate suitability suggest that parts of the EU would be suitable for establishment. Plants for planting and cut branches provide the main pathways for entry. Crawlers could spread over short distances naturally through wind, animals, humans or machinery. C. rubens could be dispersed more rapidly and over long distances via infested plants for planting for trade. The introduction of C. rubens into the EU could lead to outbreaks causing damage to orchards, amenity ornamental trees and shrubs. Phytosanitary measures are available to inhibit the entry and spread of this species. C. rubens satisfies the criteria that are within the remit of EFSA to assess for it to be regarded as a potential Union quarantine pest.

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Insects encounter infection of microorganisms, and they also harbor endosymbiosis to participate in nutrition providing and act as a defender against pathogens. We previously found the Chinese white wax scale insect, Ericerus pela, was infected and killed by Cladosporium sp. (pathogen). We also found it harbored Cladosporium sp. (endogensis). In this study, we cultured these two Cladosporium fungi and sequenced their genome. The results showed Cladosporium sp. (endogensis) has a larger genome size and more genes than Cladosporium sp. (pathogen). Pan-genome analysis showed Cladosporium sp. (endogensis)-specific genes enriched in pathways related to nutrition production, such as amino acid metabolism, carbohydrate metabolism, and energy metabolism. These pathways were absent in that of Cladosporium sp. (pathogen). Gene Ontology analysis showed Cladosporium sp. (pathogen)-specific genes enriched in the biosynthesis of asperfuranone, emericellamide, and fumagillin. These terms were not found in that of Cladosporium sp. (endogensis). Pathogen Host Interactions analysis found Cladosporium sp. (endogensis) had more genes related to loss of pathogenicity and reduced virulence than Cladosporium sp. (pathogen). Cytotoxicity assay indicated Cladosporium sp. (pathogen) had cytotoxicity, while Cladosporium sp. (endogensis) had no cytotoxicity. These characters reflect the adaptation of endosymbiosis to host-restricted lifestyle and the invader of the entomopathogen to the host.

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The Chinese white wax scale, Ericerus pela, is an insect native to China. It harbors a variety of microbes. The Paraconiothyrium fungus was isolated from E. pela and genome sequenced in this study. A fungal cytotoxicity assay was performed on the Aedes albopictus cell line C6/36. The assembled Paraconiothyrium sp. genome was 39.55 Mb and consisted of 14,174 genes. The coding sequences accounted for 50.75% of the entire genome. Functional pathway analyses showed that Paraconiothyrium sp. possesses complete pathways for the biosynthesis of 20 amino acids, 10 of which E. pela lacks. It also had complementary genes in the vitamin B groups synthesis pathways. Secondary metabolism prediction showed many gene clusters that produce polyketide. Additionally, a large number of genes associated with 'reduced virulence' in the genome were annotated with the Pathogen-Host Interaction database. A total of 651 genes encoding carbohydrate-active enzymes were predicted to be mostly involved in plant polysaccharide degradation. Pan-specific genomic analyses showed that genes unique to Paraconiothyrium sp. were enriched in the pathways related to amino acid metabolism and secondary metabolism. GO annotation analysis yielded similar results. The top COG categories were 'carbohydrate transport and metabolism', 'lipid transport and metabolism', and 'secondary metabolite biosynthesis, transport and catabolism'. Phylogenetic analyses based on gene family and pan genes showed that Paraconiothyrium sp is clustered together with species from the Didymosphaeriaceae family. A multi-locus sequence analysis showed that it converged with the same branch as P. brasiliense and they formed one group with fungi from the Paraconiothyrium genus. To validate the in vitro toxicity of Paraconiothyrium sp., a cytotoxicity assay was performed. The results showed that medium-cultured Paraconiothyrium sp. had no harmful effect on cell viability. No toxins were secreted by the fungus during growth. Our results imply that Paraconiothyrium sp. may establish a symbiotic relationship with the host to supply complementary nutrition to E. pela.

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The Chinese white wax scale insect, Ericerus pela, is an important resource insect in China. The rapid response of E. pela to decreasing temperatures plays key roles in the population distribution. In this study, we analyzed the gene expression of E. pela treated with low temperature using transcriptome analyses and weighted gene coexpression network analysis (WGCNA). The results showed that the cold resistance of E. pela involved changes in the expression of many genes. The genes were mainly involved in alcohol formation activity, lipid metabolism, membrane and structure, and oxidoreductase activity. According to the WGCNA results, some pathways related to cold resistance were found in the genes in the modules, such as cytoskeleton proteins, cytoskeleton protein pathway, biosynthesis of unsaturated fatty acids, glycerophospholipid metabolism, ether lipid metabolism, and thermogenesis. Some of the hub genes were nonspecific lipid-transfer proteins, DnaJ homolog subfamily C member 13, paramyosin, tropomodulin, and tubulin beta chain. In particular, the hub genes of the tan module included the heat shock protein (hsp) 10, hsp 60, hsp 70, and hsp 90 genes. Thirty-five antifreeze protein (afp) genes were identified according to the annotation results. Three afp genes were further identified among the hub genes. Six of these genes were selected for heterogeneous protein expression. One of them was expressed successfully. The thermal hysteresis activity (THA) analyses showed that the THA was 1.73°C. These results showed that the cytoskeleton, lipid metabolism, thermogenesis, HSPs and AFPs may play important roles in the cold resistance of E. pela.

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BACKGROUND: The Chinese white wax scale insect, Ericerus pela, is best known for producing wax, which has been widely used in candle production, casting, Chinese medicine, and wax printing products for thousands of years. The secretion of wax, and other unusual features of scale insects, is thought to be an adaptation to their change from an ancestral ground-dwelling lifestyle to a sedentary lifestyle on the higher parts of plants. As well as helping to improve its economic value, studies of E. pela might also help to explain the adaptation of scale insects. However, no genomic data are currently available for E. pela. FINDINGS: To assemble the E. pela genome, 303.92 Gb of data were generated using Illumina and Pacific Biosciences sequencing, producing 277.22 Gb of clean data for assembly. The assembled genome size was 0.66 Gb, with 1,979 scaffolds and a scaffold N50 of 735 kb. The guanine + cytosine content was 33.80%. A total of 12,022 protein-coding genes were predicted, with a mean coding sequence length of 1,370 bp. Twenty-six fatty acyl-CoA reductase genes and 35 acyltransferase genes were identified. Evolutionary analysis revealed that E. pela and aphids formed a sister group and split ∼241.1 million years ago. There were 214 expanded gene families and 2,219 contracted gene families in E. pela. CONCLUSION: We present the first genome sequence from the Coccidae family. These results will help to increase our understanding of the evolution of unique features in scale insects, and provide important genetic information for further research.

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Parasitoids of Ceroplastes Gray were surveyed in the State of São Paulo, Brazil. Among the parasitoids reared, seven species of Metaphycus Mercet (Hymenoptera, Encyrtidae) were obtained. Metaphycusanaluciaesp. nov., M. floridensissp. nov., and M. grandissp. nov., are described as new species, and M. ceros Noyes, M. comes Noyes, M. dardanus Noyes, M. opis Noyes are recorded for the first time from Brazil. Ten new host-parasitoid associations are reported: M. analuciaesp. nov. parasitizing C. formicarius Hempel, C. glomeratus Peronti and C. iheringi Cockerel; M. ceros parasitizing C. cirripediformis Comstock; M. comes parasitizing C. formicarius; M. dardanus parasitizing C. glomeratus; M. floridensissp. nov. parasitizing C. floridensis; M. grandissp. nov. parasitizing C. grandis Hempel; and M. opis parasitizing C. glomeratus and C. janeirensis (Gray). A checklist of Metaphycus species associated with Ceroplastes, their distribution and host ranges are summarized and discussed.

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The scale insect, Ericerus pela Chavannes, shows a typical sexual dimorphism. Males and females are different not only in morphology, but also in their ability to secrete wax and ecological adaptability. Here we report the morphological and structural characteristics of wax glands on E. pela females and males. The differences in wax glands and wax secretion between females and males reflect their different needs for living habitats and different ecological strategies. Sciophilous male nymphs are with five types of wax glands, and the wax glands on the dorsum secrete a layer of wax filaments plausibly for protection against direct light irradiation. On the other hand, five types of wax glands were found on the abdomen of females. Heliophilous female nymphs hardly secrete any wax, but the wax glands located along the spiracle on the abdomen may help this insect to breathe. Female adults secrete wax filaments on eggs to protect them from predators and prevent themselves from sticking to each other. In summary, males appear to secreted wax for creating a shaded niche that fits their sciophilous life style, whereas females are likely to adopt an ecological strategy with thickened epidermis for heliophilous acclimatization and overwintering.

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Atopic dermatitis (AD) is a common inflammatory skin disease with high rates of morbidity and is associated with erythema, pruritus, scaling of affected areas of skin. It is extremely important to introduce a therapeutic agent which has significant anti-inflammatory effect with less side-effect for treatment of AD. This study evaluated the effect of a natural compound from herbal extracts, the crude polysaccharide extracted from the white wax scale (CWPS), on AD-like mice. Repeated applications of 2,4-dinitrochlorobenzene (DNCB) were performed on ear and dorsal skin of BALB/c mice to induce AD-like symptoms and skin lesions. Oral administration of CWPS decreased serum IgE level and limited the infiltration of mast cells and eosinophils to the dermal tissues in the DNCB-induced AD mice. In addition, CWPS reduced Th1 and Th17 responses, leading to an attenuated cutaneous inflammatory response. Furthermore, in vitro study also demonstrated that CWPS limited T cell activation and cytokines (i.e. IFN-γ and IL-17) production induced by DNCB. We conclude that CWPS attenuates DNCB-induced AD-like skin lesion through modulating T cell-elicited immune responses and CD4+ T cell polarization, and could be exploited as a new therapeutic approach for AD.

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The wax scale Ceroplastes glomeratus Peronti shows seasonal behavioral and phenotypic differences. Individuals develop gregariously during autumn, whereas solitarily in spring. Besides such behavioral differences, spring ("isolated form") and autumn ("aggregated form") will also produce different morphotypes. We provide additional data on the morphological data of the species based on the microscopic and macroscopic characteristics of adult females and of first-instars of the two adult morphotypes of C. glomeratus.

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BACKGROUND: The Chinese white wax scale insect, Ericerus pela, is a well-known resource insect. The females and males are dramatically distinct at each developmental stage. We sought to identify suitable reference genes to use as internal controls in molecular research on E. plea. RESULTS: geNorm, RefFinder and Normfinder analyses showed that ßTub-2 was the best reference gene throughout different developmental stages; SdhA-1 was the most stable reference gene in different tissues, and ßTub-1 was the most reliable reference gene under treatment with different temperatures. The results also showed that the optimal number of reference genes for analyzing target gene expression levels in the three experimental conditions was two. CONCLUSIONS: The identified reference genes are suitable reference genes for normalization in RT-qPCR of E. pela samples.

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The Chinese white wax scale insect, Ericerus pela, can survive at extremely low temperatures, and some overwintering individuals exhibit supercooling at temperatures below -30°C. To investigate the deep supercooling ability of E. pela, transcriptomic and proteomic analyses were performed to delineate the major gene and protein families responsible for the deep supercooling ability of overwintering females. Gene Ontology (GO) classification and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated that genes involved in the mitogen-activated protein kinase, calcium, and PI3K-Akt signaling pathways and pathways associated with the biosynthesis of soluble sugars, sugar alcohols and free amino acids were dominant. Proteins responsible for low-temperature stress, such as cold acclimation proteins, glycerol biosynthesis-related enzymes and heat shock proteins (HSPs) were identified. However, no antifreeze proteins (AFPs) were identified through sequence similarity search methods. A random forest approach identified 388 putative AFPs in the proteome. The AFP gene ep-afp was expressed in Escherichia coli, and the expressed protein exhibited a thermal hysteresis activity of 0.97°C, suggesting its potential role in the deep supercooling ability of E. pela.

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The Chinese white wax scale insect (Ericerus pela) is sexually dimorphic with holometabolous males and hemimetabolous females. Holometabolous insects were assumed to originate from hemimetabolous ancestors. Therefore, the male pupal stage is a major innovation compared with hemimetabolous female insects. Here, the protein profiles of the male pupae were obtained by high-throughput proteomics and analyzed using bioinformatics methods. A total of 1,437 peptides were identified and assigned to 677 protein groups. Most of the proteins had molecular weights below 40 kDa and isoelectric points from 4 to 7. Gene Ontology terms were assigned to 331 proteins, including metabolic process, developmental process, and cellular process. Kyoto Encyclopedia of Genes and Genomes annotations identified 142 pathways and most proteins were assigned to metabolism events. Pathways involved in cell growth and death, signal transduction, folding, and sorting and degradation were also identified. Six proteins that had undergone positive selection were classified into four groups, protein biosynthesis, protein degeneration, signal transduction, and detoxification. Many of the high-abundance proteins were enzymes involved in carbohydrate, lipid, and amino acid metabolism; signal transduction; degradation; and immunization, which indicated that metabolism, disruption, and development occurred intensely at the pupal stage. These processes are closely related to the physiological status of pupae. The results also suggested that these related proteins may be fundamental factors in the formation of pupae. This study describes pupal characterization at the molecular level and provides a basis for further physiological studies.

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