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Tachycines meditationis (Orthoptera: Rhaphidophoridae: Tachycines) is a widely distributed insect in eastern Asia. This species is common in urban environments, and its unique omnivorous diet may contribute to its success in various habitats. However, molecular studies on the species are scarce. Here, we obtained the first transcriptome sequence of T. meditationis and performed preliminary analyses to test whether the evolution of coding sequences fits the expectations based on the species’ ecology. We retrieved 476,495 effective transcripts and annotated 46,593 coding sequences (CDS). We analysed the codon usage and found that directional mutation pressure was the leading cause of codon usage bias in this species. This genome-wide relaxed codon usage pattern in T. meditationis is surprising, given the potentially large population size of this species. Moreover, despite the omnivorous diet, the chemosensory genes of this species do not exhibit codon usage deviating significantly from the genome-level pattern. They also do not seem to experience more gene family expansion than other cave cricket species do. A thorough search for rapidly evolved genes using the dN/dS value showed that genes associated with substance synthesis and metabolic pathways, such as retinol metabolism, aminoacyl-tRNA biosynthesis, and fatty acid metabolism, underwent species-specific positive selection. While some results seem to contradict the species ecology, our transcriptome assembly provides a valuable molecular resource for future studies on camel cricket evolution and molecular genetics for feeding ecology in insects, in general.

Gene arrangement (relative location of genes) is another evolutionary marker of the mitogenome that can provide extensive information on the evolutionary mechanism. To explore the evolution of gene arrangements in the mitogenome of diversified Ensifera, we sequenced the mitogenome of the unique dune cricket species found in China and used it for phylogenetic analysis, in combination with 84 known Ensiferan mitogenomes. The mitogenome of Schizodactylus jimo is a 16,428-bp circular molecule that contains 37 genes. We identified eight types of gene arrangement in the 85 ensiferan mitogenomes. The gene location changes (i.e., gene translocation and duplication) were in three gene blocks: I-Q-M-ND2, rrnl-rns-V, and ND3-A-R-N-S-E-F. From the phylogenetic tree, we found that Schizodactylus jimo and most other species share a typical and ancient gene arrangement type (Type I), while Grylloidea has two types (Types II and III), and the other five types are rare and scattered in the phylogenetic tree. We deduced that the tandem replication–random loss model is the evolutionary mechanism of gene arrangements in Ensifera. Selection pressure analysis revealed that purifying selection dominated the evolution of the ensiferan mitochondrial genome. This study suggests that most gene rearrangements in the ensiferan mitogenome are rare accidental events.

Human-induced ecological and climatic changes have led to the decline and even local extinction of many formerly widely distributed temperate and cold-adapted species. Determining the exact causes of this decline remains difficult. Bryodemella tuberculata was a widely distributed orthopteran species before the mid-19th century. Since then, many European populations have suffered drastic declines and are now considered extinct or critically endangered. We used ecological niche modelling based on a large dataset of extant and extinct occurrence data to investigate whether poor climatic suitability in the periphery of its global range was a possible cause of the local extinction of the European populations of B. tuberculata. We also used population genetics based on the COI marker to estimate and compare the genetic diversity of extant populations. We found that Europe still provides highly suitable habitats close to the climatic optimum, contradicting the assumption of climate change as major driver of this decline. Instead, changes in land-cover and other anthropogenic modifications of the habitats at the local scale seem to be the major reasons for local extinctions. Genetic analysis suggests Central Asia as center of diversity with a stable population size, whereas the effective sizes of the remaining European populations are decreasing. We found European genetic lineages nested within Central Asian lineages, suggesting a Central Asian source distribution area. Our results suggest that the declining European populations represent relics of a formerly wider distribution, which was fragmented by changes in land-use. These relics are now threatened by limited connectivity and small effective population sizes. Specific conservation actions, such as the restoration of former or potential new habitats, and translocation of individuals from extant populations to these restored sites may help slow, stall, or even revert the extinction process.

Background: While most adult butterflies are nectarivores, some brush-footed species evolved saprophagy. High-quality genomes will provide a backbone for untangling this unique feeding strategy's genetic mechanism and evolutionary history. Results: We assembled a chromosome-level genome for a typical saprophagous butterfly species, Hestina assimilis (Lepidoptera: Nymphalidae). The assembled genome was 423.87 Mb long with the scaffold N50 of 15.84 Mb, and 96.2% scaffolds were anchored onto 30 chromosomes. The BUSCO (version 5.2.2, lineage insecta) completeness was 99.3%. We predicted 16,815 coding genes and found that 46.37% of the genome were repetitive sequences. The populations of H. assimilis increased dramatically after the last interglacial period of the Pleistocene. Comparative genomics analysis showed that H. assimilis clustered with another saprophagous species, Vanessa tameamea, and they diverged from a nectarivorous butterfly species, Heliconius erato, approximately 78.4 million years ago, suggesting that the transition to saprophagy was an ancient evolutionary event. The unique and significantly expanded gene families were relevant to feeding strategies and food intake functions, such as digestion, detoxification, chemoreception, immunity, nerves and development. The expansion events were also involved in processes of acid chemical response and alcohol biosynthesis. Furthermore, we detected 23 odorant-binding protein (OBP) genes-- two key antennal OBP genes, PBP-A and PBP-B, were absent. Conclusions: Our results provided insights into the genetic mechanism of feeding habits as well as provides an important resource for future butterfly genome sequencing projects.

Gut bacteria exert effects on the health and fitness of their insect hosts. Grasshoppers are an important part of the grassland ecosystem and provide important ecosystem services. As the most valuable feature in grassland ecosystem, the compositions and potential influences of gut bacterial in herbivorous grasshoppers in the same ecological environment are essential but undetermined. To facilitate such studies, we collected nine species of grasshoppers (n = 110) from a rebuild grassland on the Loess Plateau in northern Shaanxi, China, which is a representative area of ecosystem restoration model. We characterized the composition and function of the gut bacteria. We found that 326 OTUs were exhibited in all grasshoppers in which Enterobacter, Pantoea, Bacillus, and Spiroplsma are dominant. Among them, 18 OTUs were shared across all nine species of grasshoppers. The predicted function showed that the majority function of those OTUs were involved in survival dependent processes including membrane transport, carbohydrate metabolism, amino acid metabolism, and DNA replication and repair. The composition of gut bacteria is specific to each grasshopper species, and the bacteria community is most various in Trilophidia annulata. These results highlight the gut bacterial community diversity in different grasshopper species. Our findings are necessary for better understanding the relationships between this important herbivorous insect and their microbiomes and have the potential contribution of evaluating the revegetation and ecosystem management in this area.

Most butterflies feed on nectar, while some saprophagous butterflies forage on various non-nectar foods. To date, little is known about the genomic and molecular shifts associated with the evolution of the saprophagous feeding strategy. Here, we assembled the high-quality chromosome-level genome of Hestina assimilis to explore its saprophagous molecular and genetic mechanisms. This chromosome-level genome of H. assimilis is 412.82 Mb, with a scaffold N50 of 15.70 Mb. In total, 98.11% of contigs were anchored to 30 chromosomes. Compared with H. assimilis and other Nymphalidae butterflies, the genes of metabolism and detoxification experienced expansions. We annotated 80 cytochrome P450 (CYP) genes in the H. assimilis genome, among which genes belonging to the CYP4 subfamily were significantly expanded (p < 0.01). These P450 genes were unevenly distributed and mainly concentrated on chromosomes 6–9. We identified 33 olfactory receptor (OR), 20 odorant-binding protein (OBP), and six gustatory receptor (GR) genes in the H. assimilis genome, which were fewer than in the nectarivorous Danaus plexippus. A decreased number of OBP, OR, and GR genes implied that H. assimilis should resort less to olfaction and gustation than their nectarivorous counterparts, which need highly specialized olfactory and gustatory functions. Moreover, we found one site under positive selection occurred in residue 996 (phenylalanine) of GR genes exclusive to H. assimilis, which is conservative in most lineages. Our study provides support for the adaptive evolution of feeding habits in butterflies.

The mole cricket Gryllotalpa orientalis is an evolutionarily, medicinal, and agriculturally significant insect that inhabits underground environments and is distributed globally. This study measured genome size by flow cytometry and k-mer based on low-coverage sequencing, and nuclear repetitive elements were also identified. The haploid genome size estimate is 3.14 Gb by flow cytometry, 3.17 Gb, and 3.77 Gb-based two k-mer methods, respectively, which is well within the range previously reported for other species of the suborder Ensifera. 56% of repetitive elements were found in G. orientalis, similar to 56.83% in Locusta migratoria. However, the great size of repetitive sequences could not be annotated to specific repeat element families. For the repetitive elements that were annotated, Class I-LINE retrotransposon elements were the most common families and more abundant than satellite and Class I-LTR. These results based on the newly developed genome survey could be used in the taxonomic study and whole genome sequencing to improve the understanding of the biology of G. orientalis.

Grasshoppers have been identified as excellent monitors of landscape use. Despite their importance, their composition and distribution in the highly disturbed Littoral Cameroon is still unknown. The aim of this study was to determine the effect of human activities on diversity, abundance and distribution of grasshopper species in the Littoral region of Cameron. We investigated three types of vegetation differing remarkably on the level of anthropogenic impact (farmlands, fallows and forests), using sweep netting. The eight non-parametric estimators for specific richness, abundance, α and β diversity indices and species abundance distribution models, were used to compare the structure of communities among vegetation. Overall, 38 species belonging to three families and ten subfamilies were recorded. The Acrididae was the most diverse family. The species richness, abundance and diversity were higher in farmlands than in fallows and in forests. Five species occurred exclusively in farmlands, one in fallows and four in forests. Eyprepocnemis plorans, Coryphosima stenoptera, Serpusia opacula were overall the most abundant species respectively in cultivated farms, fallows and forests. Species abundance distribution fitted the Motomura model in all sites. Serpusia opacula is considered as a useful indicator since its presence and abundance significantly depend on the rate of forest naturalness. The farmlands were characterized by short vegetation while the fallows and forests were dominated by tall grasses and tall trees respectively. Anthropogenic disturbances promote the species richness, diversity and abundance of open meadow species, while it is detrimental to forest species which are sensitive, specialized and have limited dispersal abilities. Key words: Grasshopper, diversity, abundance, distribution, bioindicator

Xanthium sibiricum has been used widely in Chinese medicine for many years. Although of medicinal value, some species in the Xanthium are reported to be toxic and thus correct identification is essential. Whereas previous identification methods based on molecular characteristics are insufficient for in-depth authentication, chloroplast genome sequences provide more information that can assist in genome-wide evolutionary research. In this study, we sequenced complete chloroplast (cp) genome of X. sibiricum using next-generation sequencing technology and the complete sequence was deposited in Genbank with the accession number MH473582. This is the first reported sequence for the complete cp genome of Xanthium. The complete X. sibiricum cp genome is 151,897 bp in length, comprising one pair of inverted repeats (IRs) of 24,905 bp, separated by one small single copy region (SSC; 17,900 bp) and one large single copy region (LSC; 84,187 bp). The genome contains 115 unique genes, which code for 81 proteins, four rRNAs and 30 tRNAs. All four rRNA genes and seven tRNA genes, ycf15, ycf2, rpl2, rps12, rpl23, ndhB and rps7, are duplicated in the IR regions. The X. sibiricum cp genome has 40 repeat sequences and 59 simple sequence repeat (SSR) loci. Both Maximum parsimony (MP) and maximum likelihood (ML) trees provide very strong bootstrap support (100%) for the sister relationship of X. sibiricum with Ambrosia in the subtribe Ambrosiinae. Our study provides molecular information for further studies with which to identify X. sibiricum among closely related species and other angiosperms.

The short-video platform industry is a relatively new global industry with a considerable number of users attracted by electronic word-of-mouth (eWOM). User-generated content, which is a form of customer co-creation value (CCCV), is considered as the essence of short-video platforms. This study investigated the effects of CCCV on psychological ownership, customer-based brand equity, and eWOM in relation to short-video platforms. Examination of 337 users on Chinese short-video platforms revealed the underlying mechanism of the shift from CCCV to eWOM in the short-video platform industry by identifying the direct, indirect, and total effects of four research constructs. This study demonstrated that a well-designed co-creation value strategy is a potent reflection of a platform’s customers that enhances CCCV for short-video platforms and transforms customers into intangible assets for eWOM.

The medicinal utility of leeches has been demonstrated through decades of use in modern hospital settings, mainly as relievers of venous congestion following flap or digit replantation surgery. In the present study, we sequence and annotate (through BLAST- and Gene Ontology-based approaches) the salivary transcriptome of the nonblood feeding hirudinid Whitmania pigra and assess the differential gene expression of anticoagulation factors (through both quantitative real-time PCR [qRT-PCR] and in silico-based methods) during feeding and fasting conditions. This was done in order to evince the diversity of putative anticoagulation factors, as well as estimate the levels of upregulation of genes immediately after feeding. In total, we found sequences with demonstrated orthology (via both phylogenetic analyses and BLAST-based approaches) to seven different proteins that have previously been linked to anticoagulatory capabilities-eglin C, bdellin, granulin, guamerin, hyaluronidase, destabilase I, and lipocalin. All of these were recovered from leeches both in the fasting and in the feeding conditions, but all show signs of upregulation in the feeding leeches. Interestingly, our RNA-seq effort, coupled with a hypergeometric test, indicated that the differentially expressed genes were disproportionately involved in three main immunological pathways (endocytosis, peroxisome regulation, and lysosome regulation). The results and implications of the finding of anticoagulants in this nonblood feeding leech and the putative upregulation of anticoagulation factors after feeding are briefly discussed in an evolutionary context.

Sphingonotus Fieber, 1852 (Orthoptera: Acrididae) is a species-rich grasshopper genus with ~146 species. All species of this genus prefer dry environments, such as: desert, steppe, sand, and stony benchland. This genomic study aimed to understand the evolution and ecology of these grasshopper species. Here, the genome size of Sphingonotus tsinlingensis was estimated using flow cytometry and the first high-quality full-length transcriptome of this species is presented, which may serve as a reference genetic resource for the drought-adapted grasshopper species of Sphingonotus Fieber. The genome size of Sphingonotus tsinlingensis was ~12.8 Gb. Based on the 146.98 Gb Pacbio isoform sequencing data, 221.47 Mb full-length transcripts were assembled. Among these transcripts, 88,693 non-redundant isoforms were identified with an average length of 2,497 bp and an N50 value of 2,726 bp, which was much longer than the formal grasshopper transcriptome assemblies. A total of 48,502 protein coding sequences were determined, and 37,569 were annotated in public gene function databases. A total of 36,488 simple tandem repeats, 12,765 long non-coding RNAs, and 414 transcription factors were also identified. According to gene functions, 70 heat shock proteins and 61 P450 genes that may correspond to drought adaptation of S. tsinlingensis were identified. The genome of Sphingonotus tsinlingensis is an ultra-large and complex genome. Full-length transcriptome sequencing is an ideal strategy for genomic research. This is the first full-length transcriptome of the genus Sphingonotus. The assembly parameters were better than all known grasshopper transcriptomes. This full-length transcriptome may be used to understand its genetic background and the evolution and ecology of grasshoppers.

The specimens studied were collected with sweep net and pitfall in the forests, agro-forests, herbaceous fallows, and crop fields of 14 localities in the southern part of Cameroon, from August 2015 to February 2018. The results show that Heteracris hannai sp. nov. and Heteracris lecoqi sp. nov. differ from previously known species of the same genus by the pattern of coloration and the details or shape of the phallic complex. H. lecoqi sp. nov. is characterized by light brownish body; male cercus with flattened, downcurved and obtuse apex; lophus strongly curved; interlophal space with V shape; apodeme of cingulum bow, convergent, with V-shape; valve of cingulum in lateral view longer than apical valve of penis; ramus in ventral view joined. H. hannai sp. nov. is distinct to other species by brown to grey body; male cercus with apex rounded, curved inside; lophus curved; interlophal space with U-shape; apodeme of cingulum thick, slightly parallel, with U shape; valve of cingulum in lateral view hardly longer than apical valve of penis; ramus in ventral view opened. The characteristics of H. guineensis are closer to H. hannai sp. nov. than to H. lecoqi sp. nov. H. lecoqi sp. nov. was collected only in the fallows while H. hannai sp. nov. was collected in the forests, agro-forests, fallows, and crop fields. Compared to H. guineensis, both new species are scarce in the natural vegetation and their distribution area is limited to two and four localities respectively for H. lecoqi sp. nov. and H. hannai sp. nov. All these three grasshopper’s species were recorded as accidental species in all types of vegetation.

With unique climate, topography, and vegetation, the Qinghai-Tibet plateau is a special biogeographic region with richness of endemic species. However, the taxonomy and distribution of many insect groups on this plateau are still poorly known. Here, we synthesized a species checklist of subfamily Gomphomastacinae (Orthoptera: Acridoidea: Eumastacidae) for this region, while describing a new species: Myrmeleomastax wideis Qiu, sp. nov.. Images of the new species and the distribution map of all the known species are provided. Type specimens for the new species are deposited in the Zoological and Botanical Museum, Shaanxi Normal University, Xi’an, China (SNNU).

Background: The Asian buffalo leech Hirudinaria manillensis is a valuable animal widely used in traditional Chinese medicine to cure blood-clotting. However, although the content of the anticoagulant genes in H. manillensis were identified, their actual expression profile remains undetermined. Herein, in this study we conducted transcriptome analysis on this species to address this subject.Results: A high qualified accurate gene expression profile of H. manillensis transcripts was obtained. By identifying genes upregulated during feeding, anticoagulant-related genes and signaling pathways were identified. The assembled Unigenes were compared in various mainstream databases, and a total of 16,682 Unigenes received annotation information. In addition, the Unigenes were evaluated in terms of length distribution, GC content, and expression level. The data showed good sequencing quality and high reliability. A total of 155 anticoagulant-related genes were found in this study, including those involved in different types of degradation of the extracellular matrix and inhibition of platelet aggregation.Conclusions：Substantial transcriptome information was obtained by transcriptome sequencing of H. manillensis. This information should help to provide a further molecular theoretical basis for functional gene analysis, genomics, genetic diversity analysis, and molecular marker development of H. manillensis and for the anticoagulation-related genes of this species and its medicinal value as an anticoagulant.

Whitmania pigra is a unique, fluid-sucking ectoparasite and an anticoagulant medical leech. The codon usage bias (CUB) is the nonuniform usage of synonymous codons in which some codons are more preferred than others. Here, we performed a comprehensive analysis of CUB of genes in W. pigra, analyzing 140 780 transcripts, 59 553 unigenes, and 20 304 qualified coding sequences (CDSs) from the transcriptomic data of W. pigra. The effective number of codons values suggested that the CUB was low in these genes. We recognized profoundly favored codons in W. pigra that have a G/C-ending. Parity rule two-bias plots suggested that both mutation pressure and natural selection might have influenced the CUB. However, neutrality plots revealed that natural selection might have played a major role while mutation pressure might have played a minor role in shaping the CUB. We applied principal component analysis to relative synonymous codon usage values for divided CDSs based on GC content and codon-ending bases. Codon usage in W. pigra had a general inclination toward C-ending codons and natural selection rather than mutation pressure is the dominant force in the genetic evolution of W. pigra. To our knowledge, this is the first study to describe a complete codon usage analysis of W. pigra; this will increase the understanding of CUB and evolution in W. pigra. The analysis of codon usage patterns in W. pigra aids in understanding its evolution and genetic architecture.

Satyrinae is a 3000-species butterfly subfamily of Nymphalidae. The higher-level classification of this family is still controversial. In this research, we sequenced the complete mitogenomes of three satyrid butterfly species, Hipparchia autonoe, Paroeneis palaearctica, and Oeneis buddha, and studied the phylogeny of Satyrinae with all known complete mitogenomes. The results showed that the lengths of the three satyrid butterfly mitogenomes are 15,435 bp (H. Autonoe), 15,942 bp (P. palaearctica), and 15,259 bp (O. buddha). Gene content and arrangement of newly sequenced mitogenomes are highly conserved and are typical of Lepidoptera. These three mitogenomes were found to have a typical set of 37 genes and an A + T-rich region. The tRNA genes in these three mitogenomes showed a typical clover leaf structure, but the stem of tRNASer (AGN) was lacking dihydroacridine. In these three species, the lengths of the A + T-rich regions were different, which led to differences in mitochondrial genome sizes. The characterizations of the three mitogenomes enrich our knowledge on the Lepidopteran mitogenome and provide us genetic information to reconstruct the phylogenetic tree. Finally, the phylogenetic results confirmed the position of the genus Davidina in the subfamily Satyrini, had a closer phylogenetic relationship with Oeneis, and the phylogenetic analysis supported the formation of Oeneis buddha as an independent taxon in Oeneis.

The grasshopper Aiolopus thalassinus has long been used as an ecotoxicity indicator species of heavy metal pollution. The rapid molecular identification and genetic resource determination of this species would be of great helpful for the application in environmental monitoring. Mitochondrial DNA sequences are the widely applied molecular markers in insect taxonomic identification. In this study, we reported the complete mitochondrial genome of A. thalassinus. The whole genome was 15,753 bp in size and encoded 37 typical mitochondrial genes. The nucleotide composition was asymmetric, with an obvious bias towards A and T (75.3% combined). The typical ATN start codons were exhibited in the initiation of all protein-coding genes (PCGs). The typical TAA/TAG codons or the incomplete T(aa) codon were the terminal codons. The control region was determined by its remarkably high A+T content of 83.3%. This mitochondrial genome data for A. thalassinus will provide a useful resource for environmental monitoring.

The Asian Buffalo leech, Hirudinaria manillensis, is an aquatic sanguivorous species distributed widely in Southeast Asia. H. manillensis has long been used clinically for bloodletting and other medical purposes. Recent studies have focused on artificial culturing, strain optimization, and the identification and development new drugs based on the anticoagulant effects of H. manillensis bites; however, data regarding its genome remain unclear. This study aimed to determine the genome sequence of an adult Asian Buffalo leech. We generated a draft assembly of 151.8 Mb and a N50 scaffold of 2.28 Mb. Predictions indicated that the assembled genome contained 21,005 protein-coding genes. Up to 17,865 genes were annotated in multiple databases including Gene Ontology. Sixteen anticoagulant proteins with a Hirudin or Antistasin domain were identified. This study is the first to report the whole-genome sequence of the Asian Buffalo leech, an important sanguivorous leech of clinical significance. The quality of the assembly is comparable to those of other annelids. These data will help further the current understanding of the biological mechanisms and genetic characteristics of leeches and serve as a valuable resource for future studies.

The genus Lespedeza (tribe: Desmodieae) consists of about 40 species that have high medicinal and economic value. However, in this genus, using morphological characters, the species identification is quite complicated, which can be solved by the analysis of the complete chloroplast genomes. As primary organelle genomes, the complete genome sequences of chloroplasts (cp) provide unique molecular information to study the divergence of species, RNA editing, and phylogeny. Therefore, to the best of our knowledge, for the first time, we sequenced the complete cp genomes of two representative Lespedeza species: Lespedeza davurica and Lespedeza cuneata. The cp genomes of both the species were found to be 149,010 bp in length, exhibiting the typical angiosperm chloroplast structure containing four regions. The Lespedeza cp genomes showed similar conserved gene contents, order, and orientations with a total GC content of 35.0%. A total of 128 genes, including 83 protein-coding genes, 37 tRNAs, and eight rRNAs, were identified from each genome. Unique molecular features of the two Lespedeza cp genome sequences were obtained by performing the analysis of repeats, sequence divergence, codon usage, and predicting the RNA editing sites in addition to phylogenetic analysis with other key genera in tribe Desmodieae. Using the two datasets, the phylogenetic relationship of Lespedeza species among Deasmodieae was discovered, suggesting that whole cp genomes provided useful information for phylogenetic studies of these species.

Eragrostis of the tribe Eragrostideae is a taxonomically complex genus, because of its polyploid nature and the presence of similar morphological characters among its species. However, the relationship between these morphologically indistinguishable species at the genomic level has not yet been investigated. Here, we report the complete chloroplast genome of E. pilosa and compare its genome structures, gene contents, simple sequence repeats (SSRs), sequence divergence, codon usage bias, and Kimura 2-parameter (K2P) interspecific genetic distances with those of other Eragrostideae species. The E. pilosa chloroplast genome was 134,815 bp in length and contained 132 genes and four regions, including a large single-copy region (80,100 bp), a small single-copy region (12,661 bp), and a pair of inverted repeats (21,027 bp). The average nucleotide diversity between E. pilosa and E. tef was estimated to be 0.011, and 0.01689 among all species. The minimum and maximum K2P interspecific genetic distance values were identified in psaA (0.007) and matK (0.029), respectively. Of 45 SSRs, eight were shared with E. tef, all of which were in the LSC region. Phylogenetic analysis resolved the monophyly of the sampled Eragrostis species and confirmed the close relationship between E. pilosa and E. tef. This study provides useful chlorophyll genomic information for further species identification and phylogenetic reconstruction of Eragrostis species.

Mylabris aulica is a widely distributed blister beetle of the Meloidae family. It has the ability to synthesize a potent defensive secretion that includes cantharidin, a toxic compound used to treat many major illnesses. However, owing to the lack of genetic studies on cantharidin biosynthesis in M. aulica, the commercial use of this species is less extensive than that of other blister beetle species in China. This study reports a draft assembly and possible genes and pathways related to cantharidin biosynthesis for the M. aulica blister beetle using nanopore sequencing data. The draft genome assembly size was 288.5 Mb with a 467.8 Kb N50, and a repeat content of 50.62%. An integrated gene finding pipeline performed for assembly obtained 16,500 protein coding genes. Benchmarking universal single-copy orthologs assessment showed that this gene set included 94.4% complete Insecta universal single-copy orthologs. Over 99% of these genes were assigned functional annotations in the gene ontology, Kyoto Encyclopedia of Genes and Genomes, or Genbank non-redundant databases. Comparative genomic analysis showed that the completeness and continuity of our assembly was better than those of Hycleus cichorii and Hycleus phaleratus blister beetle genomes. The analysis of homologous orthologous genes and inference from evolutionary history imply that the Mylabris and Hycleus genera are genetically close, have a similar genetic background, and have differentiated within one million years. This M. aulica genome assembly provides a valuable resource for future blister beetle studies and will contribute to cantharidin biosynthesis.

The Asian Buffalo leech, Hirudinaria manillensis, is an aquatic sanguivorous species distributed widely in Southeast Asia. H. manillensis has long been used clinically for bloodletting and other medical purposes. Recent studies have focused on artificial culturing, strain optimization, and the identification and development new drugs based on the anticoagulant effects of H. manillensis bites; however, data regarding its genome remain unclear. This study aimed to determine the genome sequence of an adult Asian Buffalo leech. We generated a draft assembly of 151.8 Mb and a N50 scaffold of 2.28 Mb. Predictions indicated that the assembled genome contained 21,005 protein-coding genes. Up to 17,865 genes were annotated in multiple databases including Gene Ontology. Sixteen anticoagulant proteins with a Hirudin or Antistasin domain were identified. This study is the first to report the whole-genome sequence of the Asian Buffalo leech, an important sanguivorous leech of clinical significance. The quality of the assembly is comparable to those of other annelids. These data will help further the current understanding of the biological mechanisms and genetic characteristics of leeches and serve as a valuable resource for future studies.

International audience; The new species, Hilethera xinjiangensis sp. nov. (Orthoptera: Acrididae: Oedipodinae) is described based on specimens collected from Xinjiang, northern China. The new species is similar to H. brevipennis Zheng & Lu, 2002 and H. turanica Uvarov, 1925, but differs from: (1) dark brown in general coloration, (2) darker coloration in forewings, (3) forewings longer than H. brevipennis but shorter than H. turanica, (4) cubital area of forewings boarder than H. brevipennis and H. turanica, (5) hind tibiae dark brown with two light yellow pre-basal rings, while dark with one fade pre-basal ring in H. brevipennis and light yellow with three dark rings in H. turanica. In addition, the complete mitogenome of holotype was sequenced using next-generation sequencing technology. The total length of the assembled mitogenome is 16,145 bp, representing 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes and one noncoding region (D-loop region). Besides, the new mitogenome sequence is compared with published Oedipodinae mitogenomes and the phylogenetic relationships within the subfamily are reconstructed. The results infer that the gene cox1 could be a useful marker for higher phylogenetic level, while the genes nd5 and rrnL could be a potentially useful markers between closely related species.

Curculio davidi is a destructive weevil that threatens chestnut cultivation. Its rapid identification and genetic source determination, such as using mitochondrial sequences, may contribute to developing the effective control and the prevention of this species. In this study, the complete mitochondrial genome of C. davidi was assembled from high-coverage Illumina sequencing reads. The circular genome was 16,852 bp long and encoded 37 typical mitochondrial genes. The nucleotide composition was asymmetric with an obvious bias towards A and T (77.2% combined). All protein-coding genes (PCGs) were initiated with typical ATN start codons except for ND1, which was initiated with TTG. These PCGs were terminated by the typical TAA/TAG codons or the incomplete T(aa) codon. The control region was determined by its remarkably high A+T content of 83.6%, and it was located between 12S rRNA and tRNA-Ile.

Curvipennis wixiensis is a rare species of terrestrial grasshopper with limited distribution in a few untraversed mesophorbium regions of the northwest Yunnan Province. This species has specific morphological characteristics, and the local complex ecological environment is considered to play an important role in the evolution of the species. Characterization of the sources of C. wixiensis is useful for local biodiversity conservation and ecological preservation. In this study, we determined the complete mitochondrial DNA genome of C. wixiensis. This circular genome (15,528 bp in length) contains 13 protein-coding genes (PCGs), 2 rRNA genes, 22 tRNA genes and one control region. All PCGs in C. wixiensis are distributed on the H-strand, except for the ND6 gene and eight tRNA genes, which are encoded on the L-strand. We expect that the complete mitochondrial DNA genome of C. wixiensis will provide valuable molecular data for species identification, conservation genetics research and evolutionary studies.

The Asian planthopper Ricania speculum is a major agricultural pest for a variety of crops in tropical and subtropical areas. Rapid species diagnosis and source determination of this pest would be essential to the formulation of effective management, control and prevention strategy, and mitochondrial DNA has proven powerful for such purposes. In this study, we assembled and characterized its complete mitochondrial genome from Illumina sequencing reads. The whole genome of R. speculum is 15,729 bp in length, and encodes the typical set of 37 mitochondrial genes (incl. 13 PCGs, 22tRNAs and 2 rRNAs) and a control region. All PCGs are initiated with the typical ATN codons, and are terminated with either the complete TAA/TAG codons or the incomplete T(aa) codon. The nucleotide composition is asymmetric (47.4 %A, 14.8 %C, 9.5 %G and 28.3 %T) with an obvious bias towards A and T. Phylogenetic analysis indicated that R. speculum is closely related to Ricania marginalis.

Damselflies and dragonflies, of the order Odonata, have distinct body plans and predatory abilities. Knowledge of their various evolutionary histories will allow for an understanding of the genetic and phenotypic evolution of insects. Mitogenomes are suitable materials to elucidate this, but the mitogenome of only a few odonates have been annotated. Herein, we report the complete mitogenome of nine odonates, including seven dragonflies and two damselflies, and a comprehensive analysis of the codon usage in 31 Odonata mitogenomes with the aim to estimate their evolutionary characteristics. Overall, a weak codon bias exists among odonate mitogenomes, although this favours AT-ending codons. Damselflies have a weaker codon usage bias than dragonflies, and 37 codons have significantly different usages. Both directional mutation and purifying selection shape damselfly and dragonfly mitogenomes. Although inevitable, directional mutation bias plays a minor role, whereas purifying selection pressure is the dominant evolutionary force. A higher selection pressure is observed in dragonflies than in damselflies, but it mainly acts on codon usage patterns rather than amino acid translation. Our findings suggest that dragonflies might have more efficient mitochondrial gene expression levels than damselflies, producing more proteins that support their locomotion and predatory abilities.

The medicinal utility of leeches has been demonstrated through decades of use in modern hospital settings, mainly as relievers of venous congestion following flap or digit replantation surgery. In the present study, we sequence and annotate (through BLAST‐ and Gene Ontology‐based approaches) the salivary transcriptome of the nonblood feeding hirudinid Whitmania pigra and assess the differential gene expression of anticoagulation factors (through both quantitative real‐time PCR [qRT‐PCR] and in silico‐based methods) during feeding and fasting conditions. This was done in order to evince the diversity of putative anticoagulation factors, as well as estimate the levels of upregulation of genes immediately after feeding. In total, we found sequences with demonstrated orthology (via both phylogenetic analyses and BLAST‐based approaches) to seven different proteins that have previously been linked to anticoagulatory capabilities—eglin C, bdellin, granulin, guamerin, hyaluronidase, destabilase I, and lipocalin. All of these were recovered from leeches both in the fasting and in the feeding conditions, but all show signs of upregulation in the feeding leeches. Interestingly, our RNA‐seq effort, coupled with a hypergeometric test, indicated that the differentially expressed genes were disproportionately involved in three main immunological pathways (endocytosis, peroxisome regulation, and lysosome regulation). The results and implications of the finding of anticoagulants in this nonblood feeding leech and the putative upregulation of anticoagulation factors after feeding are briefly discussed in an evolutionary context.

The grasshopper Uvaroviola multispinosa (Acrididae: Oedipodinae), is an agricultural pest to pasture and limitedly distributed in Qinghai-Tibet plateau of China. The complete mitochondrial genome of U. multispinosa is 15,620 bp long, which comprises of 13 protein-coding genes (PCGs), two ribosomal RNA genes, 22 transfer RNA (tRNA) genes and a putative non-coding control region (GenBank accession ID: MK829651). These genes are unequally distributed on different DNA chains, which 23 are located on the majority Chain. The nucleotide composition shows evidently bias (A, C, G, and T was 43.8, 14.8, 10.1, and 31.3%, respectively) with an overall AT content of 75.2%. All PCGs are initiated by ATN codons, among them ATG is the most preferred. Eleven PCGs use a common stop codon of TAA or TAG, whereas the remaining two were terminated with single T as incomplete stop codon. The phylogenetic relationships based on Bayes method showed that U. multispinosa is closely related to Compsorhipis davidiana, which is in accordance with its traditional morphological classification.

Vietacheta, a cricket genus found across northern and southern Vietnam, is a rare taxon lacking any additional species discovered after the genus was established. This genus possesses features found across many other genera. Its general appearance is the same as Itaropsis; genitalia ectoparamere is very similar to Phonarellus; inner tympanum is absent, and particularly, the metanotum is armed with several hairs. Now, we found the first species of Vietacheta in China. The new species, Vietacheta harpophylla sp. nov., is close to Vietacheta aquila Gorochov, 1992, but they are different in details of anal plate, epiphallus, and ectoparamere. A description and illustration of this new species, a revision of other Vietacheta, and a key to the genus are provided.

Butterflies possess attributes that are sensitive to gradual environmental changes. Recently, the effects of environmental factors on the shapes of organisms, as well as the interactions of these elements, have been extensively examined, i.e., effects of seasonal changes on the colors of butterfly wings, and effects of landscape structure on butterfly distribution and morphology. However, few studies have dealt with variations in butterfly shapes in response to varying environmental conditions. Here we aimed to determine how body size and shape variations in butterflies are correlated to environmental heterogeneity. We used geometric morphometrics to quantify Pieris rapae wing shape variations. Results showed that forewing and hind wing sizes were significantly different among the 15 populations. P. rapae individuals with larger wing sizes were mainly distributed in mountainous areas, whereas those with smaller-sized wings were found on the plains. Canonical variate analysis was employed to examine the patterns of variation in wing shapes among and within the populations. Significant differences in shape were revealed in the forewings and the hind wings of P. rapae populations. All populations were divided into 2 groups on the first canonical variate axis (CV1), which followed the Qinling Mountains as an important boundary between the Palearctic and Oriental Realms in zoogeographical division of the world. The unweighted pair group method with arithmetic mean (UPGMA) clustered the 15 populations into 4 groups by forewing and hind wing shape in response to the 4 environment types in Qinling Mountains. We suggest that wing shapes of P.rapae are sensitive to environmental heterogeneity. The isolating effect of the Qinling Mountains on P. rapae population interactions was apparent.

A quantitative analysis of wing variation in grasshoppers can help us to understand how environmental heterogeneity affects the phenotypic patterns of insects. In this study, geometric morphometric methods were used to measure the differences in wing shape and size of Trilophidia annulata among 39 geographical populations in China, and a regression analysis was applied to identify the major environmental factors contributing to the observed morphological variations. The results showed that the size of the forewing and hindwing were significantly different among populations; the shape of the forewing among populations can be divided into geographical groups, however hindwing shape are geographical overlapped, and populations cannot be divided into geographical groups. Environmental PCA and thin-plate spline analysis suggested that smaller individuals with shorter and blunter-tip forewings were mainly distributed in the lower latitudes and mountainous areas, where they have higher temperatures and more precipitation. Correspondingly, the larger-bodied grasshoppers, those that have longer forewings with a longer radial sector, are distributed in contrary circumstances. We conclude that the size variations in body, forewing and hindwing of T. annulata apparently follow the Bergmann clines. The importance of climatic variables in influencing morphological variation among populations, forewing shape of T. annulata varies along an environmental gradient.

Toussaint, Emmanuel F. A. et al., The superfamily Dytiscoidea contains six families with an aquatic lifestyle, with most of its extant diversity in two families: the burrowing water beetles (Noteridae) and the diving beetles (Dytiscidae). The other families have few species (up to six) and generally highly disjunct extant distributions. Aspidytidae currently contains one genus with two species, one in China and one in South Africa. Here we provide the first molecular data for the Chinese species, allowing us to explore the phylogenetic relationships and position of both species of this small family for the first time. Based on a matrix of 11 genes we inferred a phylogenetic hypothesis for Dytiscoidea including all extant families. Unexpectedly, Aspidytidae were consistently recovered as paraphyletic relative to Amphizoidae, despite being well characterized by apparently synapomorphic adult features. A re-examination of larval characters in the two aspidytid species revealed that the larva of the Chinese species is strikingly similar to that of Amphizoidae. Both share a series of plesiomorphic features but also some potential synapomorphies, including a dense vestiture of short setae on the head capsule, anteriorly shifted posterior tentorial grooves and widely separated labial palps. Arguably these features may belong to the groundplan of the clade Aspidytidae + Amphizoidae, with far-reaching secondary modifications (including reversals) in the South African Aspidytes niobe. At present we retain the family Aspidytidae, however, due to the strong adult morphological synapomorphies of the two extant species, and the fact that the molecular paraphyly of the family may result from the highly divergent nature of the two extant species. This long evolutionary separation and strong divergence, in terms of gene sequences and larval features, is undeniable, substantial levels of saturation in third codon positions of protein-coding genes being present between the two taxa. We address this issue taxonomically by introducing the new genus Sinaspidytes gen. nov. for the Chinese Aspidytes wrasei. The continued contentious relationships amongst Dytiscidae, Hygrobiidae, Aspidytidae and Amphizoidae highlight the need for more data to address dytiscoid phylogenetics, possibly involving a genomic approach., This work was supported by the German Science Foundation (DFG) grants BA2152/6-1, 7-1 and 11-1.