Characterization of bacterial diversity and screening of cellulose-degrading bacteria in the gut system of Glenea cantor (Fabricius) larvae.

The intestinal bacteria of longhorn beetles would be ideal targets for pest control and lignocellulosic resources by destroying or exploiting their cellulose-degrading function. This article aims to investigate the diversity and community structure of intestinal bacteria the oligophagous longhorn beetle Glenea cantor. Additionally, it seeks to identify the presence of lignocellulose-degrading bacteria in the gut, and explore their role in consuming host kapok trees Bombax malabaricum. In this study, the bacterial community from G. cantor was examined by Illumina sequencing of 16S ribosomal RNA (rRNA) targeting the V3 and V4 regions. A total of 563,201 valid sequences and 814 OTUs were obtained. The dominant phyla were Proteobacteria, and the dominant genera were Acinetobacter and Lactococcus. The analysis of microbial diversity revealed a high bacterial diversity in the samples, with the gut bacteria playing a crucial role in the physiological activities of the host, particularly, 9 genera of intestinal bacteria with cellulose degradation function were found, highlighting their vital role in cellulose degradation. Five strains of cellulose-degrading bacteria, belonging to the genus Pseudomonas, were obtained from the intestinal tract of G. cantor larvae using traditional isolation and culture techniques as well as 16S rDNA sequencing. Among these strains, A4 exhibited a cellulase activity of 94.42 ± 0.42 U/mL, while A5 displayed the highest filter paper enzyme activity of 127.46 ± 3.54 U/mL. These results offered valuable insights into potential targets for pest control through internal attack digestion and cellulose-degrading bacteria in longhorn beetles.

Observations on the ultrastructure of the oral appendages of Glenea cantor Fabricius (Coleoptera: Lamiinae).

Using scanning electron microscopy, we examined the gross and ultrastructure morphology of the mandibles, labial palpi, and mandibular palpi of adult male and female Glenea cantor beetles. The morphology of these parts, both in their gross and ultrastructure, varied significantly between males and females. The lengths of the mandible, labial palpi, and mandibular palpi were clearly noticeably longer in females than in men, which is mostly related to the oviposition mechanism. In terms of the ultrastructural morphology of the mandibles, labial palpi, and mandibular palpi, seven types of sensilla were found on these parts: sensilla twig basiconica (STB I, II, III, and IV), sensilla chaetica (Scheme IV and V), Böhm's bristles (Bb), sensilla placodea (SP), sensilla trichodea (ST II), sensilla plate (SP), sensilla coeloconica (SC), and sensilla campaniformia (SCa). Females have significantly more ST I and Scheme III on their mandibles than males. The mechanical sensors SCh and Bb, olfactory sensor ST II, taste sensor STB IV, and carbon dioxide sensor and temperature and humidity sensors SC, SP, and SCa were much more developed on the labial and maxillary palpi of females compared to those of males, which further clarified the significance of the oral appendages in oviposition behavior. These findings will advance information-based technique design and the creation of information-based pest control strategies by assisting in our understanding of the host preference and oviposition behavior of adult G. cantor. RESEARCH HIGHLIGHTS: The external morphology and distribution of the oral appendages of Glenea cantor between sexes has been researched through scanning electron microscopy (SEM) for the first time. Several significant differences between males and females have been found by analyzing the oral appendages of G. cantor with SEM. The differences in the structure of oral appendages of G. cantor between sexes reflect functional differences in reproductive behaviors.

Antennal Transcriptome Analysis and Identification of Olfactory Genes in Glenea cantor Fabricius (Cerambycidae: Lamiinae).

Glenea cantor Fabricius (Cerambycidae: Lamiinae) is a pest that devastates urban landscapes and causes ecological loss in southern China and Southeast Asian countries where its main host kapok trees are planted. The olfactory system plays a vital role in mating, foraging, and spawning in G. cantor as an ideal target for pest control. However, the olfactory mechanism of G. cantor is poorly understood at the molecular level. In this study, we first established the antennal transcriptome of G. cantor and identified 76 olfactory-related genes, including 29 odorant binding proteins (OBPs), 14 chemosensory proteins (CSPs), 13 odorant receptors (ORs), 18 ionotropic receptors (IRs) and 2 sensory neuron membrane proteins (SNMPs). Furthermore, the phylogenetic trees of olfactory genes were constructed to study the homology with other species of insects. We also verified the reliability of transcriptome differential genes by qRT-PCR, which indicated the reliability of the transcriptome. Based on the relative expression of 30 d adults, GcanOBP22 and GcanOBP25 were highly expressed not only in the antennae, but also in the wings and legs. In addition, GcanCSP4 was the highest expression on the female antennae at 12 d. These findings laid the foundation for further research on the mechanism of G. cantor olfactory mechanism at the molecular level.

Evaluation of Reference Genes in Glenea cantor (Fabricius) by Using qRT-PCR.

Kapok is the main host of Glenea cantor (Fabricius), which causes serious damage and is difficult to control. In severe cases, it often causes the kapok trees to die continuously, which seriously affects the results of urban landscaping. To provide reference for the functional research on related genes in G. cantor, we screened the stable expression of candidate reference genes at different developmental stages (i.e., eggs, larvae, pupae, and adults), in various adult tissues (i.e., head, thorax, abdomen, feet, antennae, and wings), and sexes (i.e., male pupae, female pupae, male adults, and female adults). In this study, 12 candidate reference genes (i.e., ACTINLIKE, ACTININ, TUB, RPL36, RPL32, RPS20, TBP, GAPDH, 18S rRNA, EF1A1, EF1A2, and UBQ) were evaluated using different adult tissues, developmental stages, and sexes. RefFinder, geNorm, NormFinder, and BestKeeper were used to evaluate and comprehensively analyze the stability of the expression of the candidate reference genes. The results show that RPL32 and EF1A1 were the most suitable reference genes in the different adult tissues, and RPL36 and EF1A1 were best at the different developmental stages. RPL36 and EF1A2 were the best fit for the qRT-PCR reference genes in the different sexes, while RPL36 and EF1A1 were the most appropriate qRT-PCR reference genes in all samples. Results from geNorm showed that the optimal number of reference genes was two. We also surveyed the expression of cellulase at the different developmental stages and in the different adult tissues. Results further verified the reliability of the reference genes, and confirmed the best reference genes under the different experimental conditions. This study provides a useful tool for molecular biological studies on G. cantor.

Observations on the Ultrastructure of Antennal Sensilla of Adult Glenea cantor (Cerambycidae: Lamiinae).

The external morphology and distribution of antennal sensilla of Glenea cantor Fabricius were studied with scanning electron microscopy. The antennae of G. cantor were observed to be filiform, consisting of scape, pedicel, and flagellum (nine flagellomeres). Four distinct types of sensory receptors were observed, including sensilla chaetica, sensilla trichodea, sensilla basiconica, and Böhm bristles. Three morphological subtypes of sensilla chaetica were found on the antennae, and sensilla trichodea were also categorized into three morphological subtypes. Sensilla basiconica was grouped into two morphological subtypes that were found on subsegments F2-F9 of the flagellum, and Böhm bristles were only found at the intersegmental joints between the scape and the head and between the scape and the pedicel. The antennae of male and female adults were similar in shape, length, and diameter. However, the length, diameter, distribution, and number of each of the four distinct types of sensilla on the males were significantly different from those on females. The types, lengths, diameters, numbers, and distributions of these sensilla were described, and their possible functions were also discussed. The results indicated that the base and end of an antennal segment have a similar sensillum density, but the middle section sensor density is significantly greater, especially for olfactory and gustatory sensilla, possibly because the joints are more involved in mechanical sensing. The density of sensors is closely related to its sensing function; so, future studies on the biology of olfaction and sexual communication in G. cantor will be facilitated by these observations.

The complete mitochondrial genome of an Asian longicorn beetle Glenea cantor (Coleoptera: Cerambycidae: Lamiinae).

The complete mitogenome sequence of an Asian longicorn beetle Glenea cantor, was sequenced. The 15,514 bp long genome has the standard metazoan complement of 38 genes. These genes contain 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, and 1 control region. The nucleotide composition of the G. cantor mitogenome was A: 39.4%, T: 37.7%, G: 8.9%, C: 14.0%. The A + T content is 77.1%, showing strong AT skew. Phylogenetic analysis indicated that G. cantor have sister relationship with Thyestilla gebleri.