Background: Insects harbor a myriad of microorganisms, many of which can affect the sex ratio and manipulate the reproduction of the host. Leptocybe invasa is an invasive pest that causes serious damage to eucalyptus plantations, and the thelytokous parthenogenesis, low temperature resistance, protection in galls, generation overlap and small body of L. invasa contribute to its rapid invasion and population growth. However, the endosymbiotic bacterial composition, abundance and sex differences of L. invasa remain unclear. Therefore, this research aimed to identify the bacterial communities in L. invasa adults and compare them between the sexes of L. invasa lineage B. Results: The Illumina MiSeq platform was used to compare bacterial community composition between females and males of L. invasa by sequencing the V3–V4 region of the 16S ribosomal RNA gene. A total of 1,320 operational taxonomic units (OTUs) were obtained. These OTUs were subdivided into 24 phyla, 71 classes, 130 orders, 245 families and 501 genera. At the genus level, the dominant bacteria in females and males were Rickettsia and Rhizobium, respectively. Conclusion: The endosymbiotic bacteria of L. invasa females and males were highly diverse. There were differences in the bacterial community of L. invasa between sexes, and the bacterial diversity in male specimens was greater than that in female specimens. This study presents a comprehensive comparison of bacterial communities in L. invasa and these data will provide an overall view of the bacterial community in both sexes of L. invasa with special attention on sex-related bacteria.

Leptocybe invasa (Hymenoptera: Eulophidae) is a globally intrusive pest. Despite extensive research into the physiological responses of this pest, our understanding of the molecular mechanisms still needs to be improved. We want to accurately investigate the expression of L. invasa’s target genes, so it is imperative to select fitting reference genes. In this study, eight housekeeping genes’ stability (RPS30, ACTR, 18S rRNA, ACT, RPL18, GAPDH, 28S rRNA, and TUB) was tested under five different experimental conditions, including male or female adults, somites (head, thorax, and abdomen), temperatures (0 °C, 25 °C, and 40 °C), diets (starvation, clear water, 10% honey water, Eucalyptus sap), and pesticides (acetone was used as a control, imidacloprid, monosultap). Gene stability was calculated using RefFinder, which integrates four algorithms (the ∆Ct method, geNorm, NormFinder, and BestKeeper). The findings implied that ACT and ACTR were the most accurate when comparing sexes. For analyzing different somites, 28S rRNA and RPL18 were ideal; the 28S rRNA and RRS30 were perfect for analyzing at different temperatures. The combination of ACT and GAPDH helped to analyze gene expression in different diets, and GAPDH and 28S rRNA were suitable for various pesticide conditions. Overall, this research offers a complete list of reference genes from L. invasa for precise analysis of target gene expression, which can improve the trustworthiness of RT-qPCR and lay the foundation for further investigations into the gene function of this pest.

Karst rocky desertification (KRD) is a typical fragile ecological environment with its key and difficult management point being vegetation restoration. Therefore, it is crucial to determine the adaptation mechanisms of suitable plants for ecological restoration in KRD areas. D. odorifera is a tall leguminous, woody plant with high medicinal and wood value. This study aimed to explore the adaptation strategy of the D. odorifera root system to the shallow karst fissure-soil (SKF-S) habitats. The growth, biomass, spatial root distribution, morphological characteristics, and physiological responses of D. odorifera seedlings under different treatments were studied in pots simulating SKF-S habitats. Through the experiments conducted, the following conclusions were obtained: (I) D. odorifera enhanced its ability to acquire limited resources through an allocation adjustment strategy (adjusting the biomass allocation strategy, increasing the root shoot ratio, prioritizing organ leaves and 3-level roots), which effectively offset some of the adverse effects; (II) with an increase in the stress severity, D. odorifera improved its resource acquisition adaptive strategy by reducing the root diameter and increasing the contact area with soil; (III) the spatial development characteristics of its root system were mainly manifested in the ability to grow vertically, deeper, compared to a horizontal extension; (IV) D. odorifera did not passively endure rocky desertification stress but actively improved its metabolism through root metabolic activity and SOD enzyme activity.

Leptocybe invasa Fisher et LaSalle is a global invasive pest that seriously damages Eucalyptus plants. Studying the genetic diversity, genetic structure and introgression hybridization of L. invasa in China is of great significance for clarifying the breeding strategy, future invasion and diffusion trends of L. invasa in China and developing scientific prevention and control measures. Genetic diversity and phylogenetic analyses of 320 L. invasa female adults from 14 geographic populations in China were conducted using 10 polymorphic microsatellite loci (SSRs) and mitochondrial DNA cytochrome oxidase I gene sequences (COIs). (1) The Bayesian phylogenetic tree and haplotype network diagram showed that only haplotype Hap3 existed in L. invasa lineage B in China, while haplotypes Hap1 and Hap2 existed in lineage A, among which haplotype Hap2 was found for the first time. The nucleotide and haplotype diversities of lineage A were higher than those of lineage B. (2) The SSR genetic diversity of the Wuzhou Guangxi, Ganzhou Jiangxi and Panzhihua Sichuan populations was higher than that of the other 11 populations, and the SSR genetic diversity of lineage A was higher than that of lineage B. (3) The AMOVA analysis of mitochondrial COI data showed that 75.55% of the variation was among populations, and 99.86% of the variation was between lineages, while the AMOVA analysis of nuclear SSR data showed that 35.26% of the variation was among populations, and 47.04% of the variation was between lineages. There were obvious differences in the sources of variation between the COI and SSR data. (4) The optimal K value of COI and SSR data in structure analysis was 2, and PCoA analysis also divided the dataset into two obvious categories. The UPMGA phylogenetic tree based on SSR data clustered 14 geographic species into two groups. The results of genetic structure analysis supported the existence of two lineages, A and B, in China. (5) Structural analysis showed that there was obvious introgressive hybridization in Wuzhou Guangxi, Ganzhou Jiangxi, Panzhihua Sichuan and other populations. These results suggest that lineage introgressive hybridization has occurred in the L. invasa population in China. The introgressive hybridization degree and genetic diversity of lineage A are obviously higher than those of lineage B. Lineage introgressive hybridization may be the driving force for further L. invasa invasion and diffusion in China in the future.

Pheromone-binding proteins (PBPs) are specific odorant-binding proteins that can specifically recognize insect pheromones. Through transcriptional analysis of the antennae of adult Endoclita signifer, EsigPBP3 was discovered and identified, and EsigPBP3 was found to be highly expressed in the antennae of male moths. Based on the binding characteristics and ability of EsigPBP3, we can find the key ligands and binding site to consider as a target to control the key wood bore E. signifier. In this study, the fluorescence competitive binding assays (FCBA) showed that EsigPBP3 had a high binding affinity for seven key eucalyptus volatiles. Molecular docking analysis revealed that EsigPBP3 had the strongest binding affinity for the sexual pheromone component, (3E,7E)-4,7,11-trimethyl-1,3,7,10-dodecatetraene. Furthermore, same as the result of FCBA, the EsigPBP3 exhibited high binding affinities to key eucalyptus volatiles, eucalyptol, α-terpinene, (E)-beta-ocimene, (−)-β-pinene, and (−)-α-pinene, and PHE35, MET7, VAL10, PHE38, ILE52, and PHE118 are key sites. In summary, EsigPBP3 exhibits high binding affinity to male pheromones and key volatile compounds and the crucial binding sites PHE35, MET7, VAL10, PHE38, ILE52, and PHE118 can act as targets in the recognition of E. signifier pheromones.

[Objective] This study’s objective was to investigate the diversity of intestinal microorganisms in adult Vespa velutina (Asian hornet) killed by natural causes. This study investigates the composition of intestinal fungi and bacteria and predicts the pathogenic pathogen in adult Vespa velutina (Asian hornet). [Methods] We determined the ITS1 sequence of fungi and the V3–V4 variant region of 16S rRNA of bacteria using Illumina MiSeq technology. Operational taxonomic units (OTU) of gut symbiotic microorganisms were quantified, and the resulting data were subjected to analysis of species abundance, composition, and alpha diversity. OTU function was predicted using PICRUSt2/FUNGuild. In addition, cultured microorganisms from the gut microbiota of adult Vespa velutina were isolated and identified. A number of 3610 (fungi) and 8373 (bacteria) were identified via cluster analysis. A total of 13 strains, 51 classes, 126 orders, 285 families, and 586 genera were identified for fungi and 44 strains, 113 classes, 319 orders, 662 families, and 1394 genera were identified for bacteria. E. shigella, Herbaspirillum, and Aaaia were the most abundant classes of bacteria, and Fusarium, Mortierella, and Starmerella were the most abundant classes of fungi. In addition, 16 community genera of fungi and 11 of bacteria were outlined as core taxa. Species diversity and richness for the gut fungal and bacterial communities with VN were found to be higher than those with VA. Furthermore, bacterial species diversity and richness were found to be higher than those of fungi in VA and VN. Functional analysis revealed that Vespa velutina gut bacteria exhibited 20 functions, while fungi were classified into three types of nutrient modes. Cultivable bacteria were obtained from two phyla and two classes, but no fungi could be cultivated. [Conclusion] Variations in the species diversity and abundance of both fungi and bacteria in the gut were observed between the VA and the VN. The involvement of bacteria in the death of adult Vespa velutina was found to be significant. In addition, VA1 (the self-named strain) may be a pathogenic bacterium derived from the gut of the VA that exhibits virulence.