The pederin-producing bacteria density dynamics in Paederus fuscipes at different developmental stages.

Pederin, a defensive toxin in Paederus fuscipes, is produced by an uncultured Gram-negative symbiont, which establishes a stable symbiotic relationship with a female host before completion of metamorphosis. However, the transmission process of pederin-producing bacteria (PPB) in P. fuscipes at different life stages remains unknown. Herein, the PPB population dynamics and transcriptome atlas for P. fuscipes development (egg, first-instar larva, second-instar larva, pupa, and newly emerged female and male) were characterised. We found that a microbial layer containing PPB covered the eggshell, which could be sterilised by smearing the eggshell with streptomycin. Maternal secretions over the eggshell are likely the main PPB acquisition route for P. fuscipes offspring. The PPB density in eggs was significantly higher than that in other life stages (p < 0.05), which demonstrated that the beetle mothers gave more PPB than the larvae acquired. Physiological changes (hatching and eclosion) led to a decreased PPB density in P. fuscipes. Pattern recognition receptors related to Gram-negative bacteria recognition were identified from P. fuscipes transcriptomes across various life stages, which might be used to screen genes involved in PPB regulation. These results will help advance future efforts to determine the molecular mechanisms of PPB colonisation of P. fuscipes.

Field investigation- and dietary metabarcoding-based screening of arthropods that prey on primary tea pests.

Predatory natural enemies play key functional roles in biological control. Abundant predatory arthropod species have been recorded in tea plantation ecosystems. However, few studies have comprehensively evaluated the control effect of predatory arthropods on tea pests in the field. We performed a 1-year field investigation and collected predatory arthropods and pests in the tea canopy. A total of 7931 predatory arthropod individuals were collected, and Coleosoma blandum (Araneae, Theridiidae) was the most abundant species in the studied tea plantation. The population dynamics between C. blandum and four main tea pest species (Aleurocanthus spiniferus, Empoasca onukii, Ectropis grisescens, and Scopula subpunctaria) were established using the individual number of predators and pests in each month. The results showed that C. blandum appeared to co-occur in the tea canopy with A. spiniferus, Em. onukii, and Ec. grisescens in a longer period. The prey spectrum of C. blandum was further analyzed using DNA metabarcoding. Among prey species, A. spiniferus, Em. onukii, and Ec. grisescens were included, and the relative abundance and positive rates of target DNA fragments of A. spiniferus were greater than that of other two pests. Combined with the high dominance index of C. blandum, co-occurrence between C. blandum and A. spiniferus in time and space and high positive rate and relative abundance of target DNA fragments of A. spiniferus, C. blandum was identified to prey on A. spiniferus, and C. blandum may be an important predator of A. spiniferus. Thus, C. blandum has potential as a biological control agent of A. spiniferus in an integrated pest management strategy.

Differences in microbiome composition and transcriptome profiles between male and female Paederus fuscipes harbouring pederin-producing bacteria.

Pederin, a group of antitumor compounds, is produced by an endosymbiotic bacterium of Paederus fuscipes. Pederin content differed between male and female P. fuscipes, but the reason why these differences are maintained remains unexplored. Here, the pederin-producing bacteria (PPB) infection rate in P. fuscipes was investigated. Furthermore, we assessed the microbiota structure differences in male and female P. fuscipes harbouring PPB and sequenced the transcriptome of both sexes to shed light on genes of interest. Of the 625 analysed beetles (275 females, 350 males), 96.36% of females and 31.14% of males were positive for PPB infection. PPB accounted for 54.36%-82.70% of the bacterial population in females but showed a much lower abundance in males (0.92%-3.87%). Reproductive organs possessed the highest PPB abundance compared with other parts of females, but no such relationships existed in males. Moreover, we provide the first transcriptome analysis of male and female P. fuscipes harbouring PPB and identified 8893 differentially expressed unigenes. Our results indicated that the pederin content difference between males and females might be caused by the PPB density difference in hosts. The biosequence data would be helpful for illustrating the mechanism that regulates PPB density in P. fuscipes.

A comparative analysis of spider prey spectra analyzed through the next-generation sequencing of individual and mixed DNA samples.

As one of the most abundant predators of insects in terrestrial ecosystems, spiders have long received much attention from agricultural scientists and ecologists. Do spiders have a certain controlling effect on the main insect pests of concern in farmland ecosystems? Answering this question requires us to fully understand the prey spectrum of spiders. Next-generation sequencing (NGS) has been successfully employed to analyze spider prey spectra. However, the high sequencing costs make it difficult to analyze the prey spectrum of various spider species with large samples in a given farmland ecosystem. We performed a comparative analysis of the prey spectra of Ovia alboannulata (Araneae, Lycosidae) using NGS with individual and mixed DNA samples to demonstrate which treatment was better for determining the spider prey spectra in the field. We collected spider individuals from tea plantations, and two treatments were then carried out: (1) The DNA was extracted from the spiders individually and then sequenced separately (DESISS) and (2) the DNA was extracted from the spiders individually and then mixed and sequenced (DESIMS). The results showed that the number of prey families obtained by the DESISS treatment was approximately twice that obtained by the DESIMS treatment. Therefore, the DESIMS treatment greatly underestimated the prey composition of the spiders, although its sequencing costs were obviously lower. However, the relative abundance of prey sequences detected in the two treatments was slightly different only at the family level. Therefore, we concluded that if our purpose were to obtain the most accurate prey spectrum of the spiders, the DESISS treatment would be the best choice. However, if our purpose were to obtain only the relative abundance of prey sequences of the spiders, the DESIMS treatment would also be an option. The present study provides an important reference for choosing applicable methods to analyze the prey spectra and food web compositions of animal in ecosystems.

Characterization of the complete mitochondrial genome of Ectropis grisescens (Lepidoptera, Geometridae).

Ectropis grisescens (Lepidoptera, Geometridae) is one of the main leaf-eating pests in tea plantations in China. In this study, the complete mitochondrial genome of this species was sequenced and assembled. The total length of the mitochondrial genome of E. grisescens was 15,794 bp (GenBank accession No. MW337302). The base composition was 41.26% for A, 39.49% for T, 7.92% for G, and 11.33% for C. The circular mitogenome contained 13 protein-coding genes, 2 ribosomal RNA genes, and 22 transfer RNA genes. Phylogenetic analysis performed using 13 protein-coding genes of 15 species of Geometridae and an out-group Pieris melete (Lepidoptera, Pieridae) showed that E. grisescens is closely related to species of E. obliqua, and this is consistent with the morphological identification.

Comparison of the Microsatellite Distribution Patterns in the Genomes of Euarchontoglires at the Taxonomic Level.

Microsatellite or simple sequence repeat (SSR) instability within genes can induce genetic variation. The SSR signatures remain largely unknown in different clades within Euarchontoglires, one of the most successful mammalian radiations. Here, we conducted a genome-wide characterization of microsatellite distribution patterns at different taxonomic levels in 153 Euarchontoglires genomes. Our results showed that the abundance and density of the SSRs were significantly positively correlated with primate genome size, but no significant relationship with the genome size of rodents was found. Furthermore, a higher level of complexity for perfect SSR (P-SSR) attributes was observed in rodents than in primates. The most frequent type of P-SSR was the mononucleotide P-SSR in the genomes of primates, tree shrews, and colugos, while mononucleotide or dinucleotide motif types were dominant in the genomes of rodents and lagomorphs. Furthermore, (A)n was the most abundant motif in primate genomes, but (A)n, (AC)n, or (AG)n was the most abundant motif in rodent genomes which even varied within the same genus. The GC content and the repeat copy numbers of P-SSRs varied in different species when compared at different taxonomic levels, reflecting underlying differences in SSR mutation processes. Notably, the CDSs containing P-SSRs were categorized by functions and pathways using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes annotations, highlighting their roles in transcription regulation. Generally, this work will aid future studies of the functional roles of the taxonomic features of microsatellites during the evolution of mammals in Euarchontoglires.

A checklist of spiders in tea plantations of China.

BACKGROUND: Spiders are the most dominant predatory natural enemies of insect pests in the tea plantation ecosystem. There has been a large amount of literature published about the investigation of spider species in Chinese tea plantations from 1982 to 2020. Here, the spider species in Chinese tea plantations has been summarised and the dominant spider species in each regional tea plantation recorded. To date, there were 535 spider species from 40 families reported in Chinese tea plantations. NEW INFORMATION: There are 245 spider species from 13 families now being added to the checklist. A total of 89 spider species from 19 families were the dominant species, amongst them, Agelena labyrinthica, Allagelena difficilis, Neoscona theisi, Clubiona deletrix, Clubiona japonicola, Hylyphantes graminicola, Pardosa laura, Oxyopes sertatus, Evarcha albaria, Plexippus paykulli, Coleosoma octomaculatum, Ebrechtella tricuspidata and Xysticus ephippiatus were recorded in many tea plantations. The checklist will provide important data for the biodiversity and distribution of spiders in tea plantations of China.

Transcriptome analyses to reveal the dynamic change mechanism of pigeon magnum during one egg-laying cycle.

We analyzed the transcriptome of pigeon magnum in three stages (C1: pre-ovulation, C2: post-ovulation, C3: 5-6 days after ovulation) to elucidate the molecular and cellular events associated with morphological changes during the laying cycle. We observed that C1 was highly developed, apoptosis rate was highest in C2, and C3 attained the smallest size. Through RNA-sequencing, we obtained 54,764,938 (97.2%) high-quality clean reads that aligned to 20,767 genes. Gene expression profile analysis showed the greatest difference between C1 and C3; 3966 differentially expressed genes (DEGs) were identified, of which 2250 genes were upregulated and 1716 genes were downregulated in C1. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses revealed that protein processing and transport activities were prominent in C1, and upregulated genes included those related to signal recognition particle (SRP), signal recognition particle receptor (SRPR), translocon, GRP78, RRBP1, TRAP, TRAM1, and OST. Egg white protein-related gene expression was highest, with OVALY being the most highly expressed. In C2, apoptosis-related gene expression was higher than in C1, and fatty acid metabolism was active, which may be correlated with magnum tissue regression. Collagen- and laminin-related gene expression was prominent in C1 and C3, indicating roles in egg white protein generation and magnum reconstruction. PR gene expression was highest and exhibited drastic change in the three groups, indicating that PR and its regulation may be involved in changes in magnum morphology and function. Through the identification and functional analysis of DEGs and other crucial genes, this may contribute to understand the egg white protein production, magnum tissue regression, and magnum regeneration mechanisms.

One new species of the genus Sinopoda from Hubei Province, with description of the male of Sinopoda angulata (Araneae, Sparassidae).

BACKGROUND: In the past year, Prof. Jian Chen conducted several spider collections in Hubei Province. Almost 1000 spiders were collected. After diagnosis, two of them were found to belong to the genus Sinopoda Jäger, 1999. NEW INFORMATION: Two Sinopoda Jäger, 1999 species, both from Hubei Province, including one new species, are treated in the current paper: S. angulata Jäger, Gao & Fei, 2002 and S. yichangensis sp. n. (♂). The male of S. angulata is described for the first time from Enshi Tujia and Miao Autonomous Prefecture, Hubei Province. New geographic records are provided as well as photos of the copulatory organs and habitus.

Compared with conventional PCR assay, qPCR assay greatly improves the detection efficiency of predation.

Studies of predation can contribute greatly to understanding predator-prey relationships and can also provide integral knowledge concerning food webs and multi-trophic level interactions. Both conventional polymerase chain reaction (cPCR) and quantitative PCR (qPCR) have been employed to detect predation in the field because of their sensitivity and reproducibility. However, to date, few studies have been used to comprehensively demonstrate which method is more sensitive and reproducible in studies of predation. We used a Drosophila melanogaster-specific DNA fragment (99 bp) to construct a tenfold gradient dilution of standards. Additionally, we obtained DNA samples from Pardosa pseudoannulata individuals that fed on D. melanogaster at various time since feeding. Finally, we compared the sensitivity and reproducibility between cPCR and qPCR assays for detecting DNA samples from feeding trials and standards. The results showed that the cPCR and qPCR assays could detect as few as 1.62 × 103 and 1.62 × 101 copies of the target DNA fragment, respectively. The cPCR assay could detect as few as 48 hr post-feeding of the target DNA fragment. But the qPCR assay showed that all spiders were positive after consuming prey at various time intervals (0, 24, 48, 72, and 96 hr). A smaller proportion of the technical replicates were positive using cPCR, and some bands on the agarose gel were absent or gray, while some were white and bright for the same DNA samples after amplification by cPCR. By contrast, a larger proportion of the technical replicates were positive using qPCR and the coefficients of variation of the Ct value for the three technical replicates of each DNA sample were less than 5%. These data showed that qPCR was more sensitive and highly reproducible in detecting such degraded DNA from predator's gut. The present study provides an example of the use of cPCR and qPCR to detect the target DNA fragment of prey remains in predator's gut.

Comparison of microsatellite distribution patterns in twenty-nine beetle genomes.

Simple sequence repeats (SSRs) represent an important source of genetic variation that provides a basis for adaptation to different environments in organisms. In this study, we examined the distribution patterns of SSRs in twenty-nine beetle genomes and carried out Gene Ontology (GO) analysis of CDSs embedded with perfect SSRs (P-SSRs). The results demonstrated that imperfect SSRs (I-SSRs) represented the most abundant SSR category in beetle genomes and in different genomic regions (CDS, exon, and intron regions). The numbers of P-SSRs, I-SSRs, compound SSRs, and variable number tandem repeats were positively correlated with beetle genome size, whereas neither the frequency nor the density of the SSRs was correlated with genome size. Moreover, our results demonstrated that common genomic features of P-SSRs within the same suborder or family of Coleoptera were rare. Mono-, di-, tri-, or tetranucleotide SSRs were the most abundant P-SSR categories in beetle genomes. The preferred predominant repeat motif among the mononucleotide P-SSRs was (A)n, but the most frequent repeat motifs for other length classes varied differentially among these genomes. Furthermore, the P-SSR type with the highest GC content differed in the beetle genomes and in different genomic regions. CV (coefficient of variability) analysis demonstrated that the repeat copy numbers of P-SSRs presented relatively higher variation in introns than in CDSs and exons. The GO terms of CDSs containing P-SSRs for molecular functions were mainly enriched in "binding" and "transcription". Our findings will be useful for studying the functional roles of microsatellite heterogeneity in beetle adaptation.

Molecular identification of spiders preying on Empoasca vitis in a tea plantation.

Biological control using predators of key pest species is an attractive option in integrated pest management (IPM). Molecular gut analysis can provide an estimation of predator efficiency on a given prey. Here we use a combination of various experimental approaches, both in field and lab, to identify a potential biological control species of the common pest of commercially grown tea, Empoasca vitis (Göthe) (Hemiptera), in a Chinese plantation. We collected 2655 spiders from plantations and established relative abundances of spider species and their temporal overlap with the pest species in tea canopy. We analyzed DNA from 1363 individuals of the most common spider species using targeted RQ-PCR to quantify the potential efficiency of spiders as a predator on E. vitis. The results showed that, in the field, the jumping spider Evarcha albaria was the most abundant, had the closest temporal overlap with the pest, and frequently fed on it. Therefore, this spider may play a key role in pest suppression. The present study demonstrates the potential of our experimental approach to study predator-prey relationships in taxa that do not lend themselves to morphological identification of gut contents, such as spiders.

Main predators of insect pests: screening and evaluation through comprehensive indices.

BACKGROUND: Predatory natural enemies play key functional roles in integrated pest management. However, the screening and evaluation of the main predators of insect pests has seldom been reported in the field. Here, we employed comprehensive indices for evaluating the predation of a common pest (Ectropis obliqua) by nine common spider species in Chinese tea plantations. RESULTS: We established the relative dominance of the spider species and their phenological overlap with the pest species, and analyzed DNA from the nine spider species using targeted real-time quantitative polymerase chain reaction to identify the residual DNA of E. obliqua. The predation rates and predation numbers per predator were estimated by the positive rates of target fragments and the residual minimum number of E. obliqua in predators' guts, respectively. The results showed that only four spider species preyed on E. obliqua, and the order of potential of the spiders to control E. obliqua from greatest to smallest was Neoscona mellotteei, Xysticus ephippiatus, Evarcha albaria and Coleosoma octomaculatum by the Z-score method. CONCLUSION: The orb-weaving spider N. mellotteei has the maximum potential as a biological control agent of E. obliqua in an integrated pest management strategy. An approach of screening and evaluating main predators of insect pests through comprehensive indices was preliminarily established. © 2017 Society of Chemical Industry.

Four new species of Psechrus from Yunnan Province, China (Araneae, Psechridae).

Four new species of Psechrus are described from Yunnan Province: P. changminae sp. nov., P. conicus sp. nov., P. discoideus sp. nov. and P. spatulatus sp. nov. Herewith the number of species belonging to Psechrus in Yunnan Province extended from five to nine. Illustrations and colour photos are provided, with comparison of intraspecific variation of copulatory organs.