The Asian citrus psyllid, D. citri Kuwayama is the primary vector for Candidatus Liberibacter asiaticus (CLas), which causes a destructive disease in citrus plants. Bacterial symbionts are important determinants of insect physiology, and they can be impacted by many external factors. Temperature is an important abiotic factor affecting insect physiology, and it is also known that differences in symbiont proportions may vary in different insect genders. To date, it is unclear how the symbionts of D. citri are affected by temperature and gender. This study used high-throughput sequencing of 16S ribosomal RNA amplicons to determine how temperature and gender affect the bacterial communities present in D. citri. We identified 27 amplicon sequence variants (ASVs) belonging to 10 orders, seven classes, and five phyla. The dominant phylum was Proteobacteria (99.93%). Other phyla, including Firmicutes, Bacteroidota, Deinococcota, Cyanobacteria, and Actinobacteriota, were less abundant (<0.1%). Profftella (71.77–81.59%) and Wolbachia (18.39–28.22%) were the predominant taxa in all samples. Under high-temperature treatment, Profftella was more common in females, while Wolbachia had a higher abundance in males. In males, Profftella was more abundant under low-temperature treatments than under high-temperature treatments. In contrast, Wolbachia showed a higher abundance under high-temperature treatments than under low-temperature treatments. An RT-qPCR (quantitative real-time PCR) approach confirmed the results obtained with high-throughput DNA sequencing. Our results provide a basis for understanding the co-adaptation of D. citri and its symbionts to environmental temperature stress.

The citrus leaf beetle, Clitea metallica, is a specialized citrus pest through feeding on fresh leaves by larva and adults, and causes nicks and holes into leaves, leaving only a waxy surface layer. Insect cuticle is a complex exoskeleton that is not only involved in development but also protects the insect from environmental contaminations. Due to these key roles of the cuticle, cuticle-related genes are currently investigated in understanding the insect physiology in adaptation. Therefore, in this study, we built two libraries, transcriptomic (43 million clean reads) and small RNA (17 million clean reads), of C. metallica to identify cuticle-related genes and possibly associated miRNAs, being as an example to explore these data sets. Our results showed that a total of 47 cuticular protein genes were identified and most of these genes harbored a conserved motif (the Rebers and Riddiford motif) and belonged to the CPR family. Unigenes encoding proteins involved in chitin synthesis and degradation were also identified, including chitin synthase (2 unigenes), chitinase (14 unigenes), chitinase-like protein (2 unigenes), and chitin deacetylase (5 unigenes). Based on the small RNA library, we identified 30 miRNAs conserved across insect species. Among these miRNAs, 14 were predicted to be target genes associated with cuticle synthesis and degradation. In summary, 70 cuticle-related genes and 14 cuticle-related miRNAs were identified based on the transcriptome and small RNA library of C. metallica. These data sets will promote the understanding of cuticle molecular regulation in C. metallica as well as provide new potential targets for pest control.