- Background: Vespa velutina, one of the most aggressive and fearful wasps in China, can cause grievous allergies and toxic reactions, leading to organ failure and even death. - Therefore, we aimed to provide an insight into the transcripts expressed in the venom gland of wasps.. - In addition, c-type, p1 type, p2 type and p3 type were the most commonly found simple sequence repeat (SSR) types in the four species of wasp. - There were differences in the distribution of SSRs and single nucleotide polymorphisms (SNPs) among the four wasp species.. - The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. - If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. - 2 CDC of Western Theater Command, PLA, Chengdu 610021, China Full list of author information is available at the end of the article. - velutina, one of the most aggressive and fearful wasps in China, can cause grievous allergies and toxic reactions, leading to organ failure and even death [14]. - Many bioactive peptides and macromolecular proteins, including enzymes, allergens, and toxins, are abundant in the venom of these wasps [20–22].. - velu- tina has been deciphered, and related genes in the venom gland, such as the putative toxin sequences, have been revealed [14]. - Protein and peptide com- pounds are regarded as the bioactive substances in the wasp venom, and 398 wasp venom-related proteins were annotated in the UniProt database including mastoparan-like peptide, tachykinin-like peptide, vespin, melittin, venom protein and peptide, phospholipase, polybine, dominulin, and sodium channel subunit (https://www.uniprot.org. - However, verification of the specific venom is significant for the clinical treatment of wasp injury.. - Furthermore, DNA barcoding has also been reported for the identification and taxo- nomic classification of the wasp subfamily [35–37].. - In particular, transcriptome-wide studies have provided insights into caste systems, and the phenotypic plasticity of the genome has been studied in the facultatively eusocial bee, Megalopta genalis [38], Apis cerana cerana [39] and bumblebee, Bombus terres- tris [40] by using conventional and high-throughput se- quencing technologies. - Based on COI sequence identification, the NJ tree revealed the unique lineage of these individuals, and the clustering information clarified the differences and similarities in the molecular sequences (Fig. - analis fabricius 7 were the factors that con- tribute to the group sequence variation of the other six unanimous individuals, indicating the occurrence of probable mutation or evolution process in this species (V. - clean and high-quality reads were obtained by deleting redundant transcripts, and the filtering rates of the sequencing reads ranged from 87.75 to 91.70% (Additional file 2). - mandarinia smith. - The open reading frame (ORF) and coding domain se- quence (CDS) of the wasps were predicted using the se- quence information and reference structures obtained from ORFfinder. - mandarinia smith had and 332 unigenes annotated into all databases, respectively Table 1 The statistics of the sequencing data after quality trimming. - Annotation results of unigenes from the four wasp species of V. - In the nr database, the species of the annotated homologous sequences of V. - In the Swiss-Prot database, the species hits of the an- notated homologous sequences of V. - Moreover, in the Tox-Prot database, the species of the annotated homologous sequences of V. - These results indicated that the unigenes of the four different wasps (V. - mandarinia smith) were annotated in the nr, Swiss-. - Venom serine protease Bi-VSP, Acetylcholinesterase, Scoloptoxin SSD976, Probable phospholipase A1 magni- fin, and Alpha-latrocrustotoxin-Lt1a (Fragment) accounted for a high proportion in the V. - In the V. - the species and proportion of toxins contained in the four venom glands were different and may vary from species to species.. - a GO enrichment analysis of unigenes from V. - b GO enrichment analysis of unigenes from V. - c GO enrichment analysis of unigenes from V. - Only the top 10 GO-terms are displayed in the categories of biological process (GO-BP), cellular component (GO-CC) and molecular function (GO-MF). - We further carried out GO enrichment analysis on the shared and specific unigenes of the four species of wasp.. - GO enrichment analysis of unigenes specific to each of the four wasp species showed that only V. - 5 GO enrichment analysis of common and specific unigenes of the four wasp species. - mandarinia smith (VMS). - b GO enrichment analysis of unigenes shared by the four wasp species. - The results showed that and 196,691 SSRs were detected in the V. - Among them, c-type, p1 type, p2 type and p3 type were the most commonly found SSR types in the transcriptome of the four wasp species.. - The results showed that for transition and 71,400 for transversion for transition and 70,164 for transversion for transition and 75,420 for transver- sion) and for transition and 66,580 for transversion) SNPs were identified in the V. - There were differences in the distribution of SNPs among the four species of wasp, and V. - 6 Identification of molecular markers in the venom gland of the four wasp species. - a Identification of SSRs in the venom gland of V. - b Identification of SNPs in the venom gland of V. - mandarinia smith.. - The transcriptome of the venom glands of the four wasp species were analyzed using the Illumina Hiseq4000 RNA-seq platform. - mandarinia smith). - Thus, our study might dir- ectly or indirectly promote or contribute to the comple- tion of the sequencing of wasp venom.. - The unigenes of the four wasp species were searched for the Tox-Prot database by blastx to anno- tate the venom protein. - Studies have shown that factor V activator and thrombin-like enzyme are involved in the blood coagulation cascade, which has been reported in the V. - Acetylcholinesterase has been found in the venom of V. - velutina, which plays key roles in neurotransmission through inactivating the acetylcholine, ultimately resulting in paralysis of the prey [14, 48]. - Due to the influence of factors such as individual differences, sex differences and geographical envir- onment differences, it is easy to confuse similar spe- cies in the traditional identification of wasps. - Similar barcoding procedures have been used in snakes [31], spiders [51], and parasitoid wasps [52], showing the feasibility and efficiency of this method in distin- guishing specific DNA sequences in the venom. - They were homologous and all clustered in the same big branch, indicating that they were from the same subspecies, the V.. - It also revealed the unique lineage of these individuals, and the clustering infor- mation clarified the differences and similarities in the molecular sequences.. - In this study, 8 types of SSR were detected in the four wasp species, of which c, p1, p2 and p3 were the most common SSR types in the four species of wasp. - It can fully reflect the genetic and variational levels of the population by studying its distribution in the biological genome. - These data are valuable for further molecular researches of the wasps. - In addition, most of the genes could not be annotated in the databases. - These observations indicated that there is a large number of genes in the venom gland of wasp species that need to be identi- fied. - This will play a significant role in the applications of wasp species to pest control and medical purposes.. - Until then, the annotated genes discovered in the present study will be useful for current studies but the validation of these genes by additional molecular biology techniques and functional analysis by experi- mentation will be performed in our future studies.. - The “Wildlife Protection Law of the People’s Republic of China” stipulates that:. - After investigation, we found that wasp species are not included in the “National List of Key Wildlife Protection”, and in fact, there is a large number of wasps species, which are ubiquitous and are not en- dangered species, so they are not considered as “wild animals” protected by the “Wild Animal Protection Law”. - BLAST search was performed by inputting the FASTA file in the nucleotide database in GenBank (https://blast.. - All of the insertions and deletions were tested by the bootstrap method with 1000 replicates, and the nu- cleotide sequences with homologies of 100% were isolated as representative to construct the phylogen- etic tree.. - Illumina TruSeq adapters were li- gated to the 5 ′ and 3 ′ ends of the cDNA of both samples. - For Illumina sequencing, the cDNA samples were fractionated on agarose gels ranging in the size from 300 to 500 bp. - The operating parameters of Trinity program were: k-mer = 25, the minimum k- mer coverage = 2, the maximum length expected be- tween the pair of fragments = 500, the minimum over- lap with the reading of the transcript PE = 75 and the maximum number of readings anchored in a single figure . - Within the significant category, the relative enrichment was given as follows: Re = (n f /n)/(N f /N), where n f is the num- ber of unigenes in a particular category, n is the total number of genes in the same category, N f is the number of unigenes in the entire array, and N is the total num- ber of genes in the array.. - The density and distribution of different SSR types were determined in the whole genome. - The statistics of the sequencing data of 12 wasp individuals.. - (A) The intersection of the significant hits for V. - (B) The intersection of the significant hits for V. - (C) The inter- section of the significant hits for V. - tropica ducalis (D) The intersection of the significant hits for V. - The GO enrichment of unigenes shared by the venom gland of the four wasp species.. - JT participated in the design of the study, sample collection, experimental work, data analysis, interpretation of the results and manuscript editing. - WW, YL and FW carried out the experimental work and contributing to the writing of the manuscript. - All authors read and approved the final version of the manuscript.. - The “ Wildlife Protection Law of the People ’ s Republic of China ” stipulates that:. - After investigation, we found that wasp species are not included in the. - Taxonomy, distribution and nesting biology of species of the genus Dolichovespula (Hymenoptera, Vespidae). - 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