Monochamus saltuarius (Coleoptera: Cerambycidae) is an important native pest in the pine forests of northeast China and a dispersing vector of an invasive species Bursaphelenchus xylophilus. To investigate the bacterial gut diversity of M. saltuarius larvae in different host species, and infer the role of symbiotic bacteria in host adaptation, we used 16S rRNA gene Illumina sequencing and liquid chromatography-mass spectrometry metabolomics processing to obtain and compare the composition of the bacterial community and metabolites in the midguts of larvae feeding on three host tree species: Pinus koraiensis, Pinus sylvestris var. mongolica, and Pinus tabuliformis. Metabolomics in xylem samples from the three aforementioned hosts were also performed. Proteobacteria and Firmicutes were the predominant bacterial phyla in the larval gut. At the genus level, Klebsiella, unclassified_f__Enterobacteriaceae, Lactococcus, and Burkholderia–Caballeronia–Paraburkholderia were most dominant in P. koraiensis and P. sylvestris var. mongolica feeders, while Burkholderia–Caballeronia–Paraburkholderia, Dyella, Pseudoxanthomonas, and Mycobacterium were most dominant in P. tabuliformis feeders. Bacterial communities were similar in diversity in P. koraiensis and P. sylvestris var. mongolica feeders, while communities were highly diverse in P. tabuliformis feeders. Compared with the other two tree species, P. tabuliformis xylems had more diverse and abundant secondary metabolites, while larvae feeding on these trees had a stronger metabolic capacity for secondary metabolites than the other two host feeders. Correlation analysis of the association of microorganisms with metabolic features showed that dominant bacterial genera in P. tabuliformis feeders were more negatively correlated with plant secondary metabolites than those of other host tree feeders.

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Background In many insect species, the larvae/nymphs are unable to disperse far from the oviposition site selected by adults. The Sakhalin pine sawyer Monochamus saltuarius (Gebler) is the newly discovered insect vector of the pine wood nematode (Bursaphelenchus xylophilus) in China. Adult M. saltuarius prefers to oviposit on the host plant Pinus koraiensis, rather than P. tabuliformis. However, the genetic basis of adaptation of the larvae of M. saltuarius with weaken dispersal ability to host environments selected by the adult is not well understood. Results In this study, the free amino and fatty acid composition and content of the host plants of M. saltuarius larvae, i.e., P. koraiensis and P. tabuliformis were investigated. Compared with P. koraiensis, P. tabuliformis had a substantially higher content of various free amino acids, while the opposite trend was detected for fatty acid content. The transcriptional profiles of larval populations feeding on P. koraiensis and P. tabuliformis were compared using PacBio Sequel II sequencing combined with Illumina sequencing. The results showed that genes relating to digestion, fatty acid synthesis, detoxification, oxidation-reduction, and stress response, as well as nutrients and energy sensing ability, were differentially expressed, possibly reflecting adaptive changes of M. saltuarius in response to different host diets. Additionally, genes coding for cuticle structure were differentially expressed, indicating that cuticle may be a potential target for plant defense. Differential regulation of genes related to the antibacterial and immune response were also observed, suggesting that larvae of M. saltuarius may have evolved adaptations to cope with bacterial challenges in their host environments. Conclusions The present study provides comprehensive transcriptome resource of M. saltuarius relating to host plant adaptation. Results from this study help to illustrate the fundamental relationship between transcriptional plasticity and adaptation mechanisms of insect herbivores to host plants.

BACKGROUND Anoplophora glabripennis (Coleoptera: Cerambycidae) is a major quarantine pest in forestry. It is widely distributed throughout many regions such as Asia, Europe, and North America, and has enormous destructive potential for forests. The larvae of A. glabripennis overwinter in a dormant state with strong cold tolerance, and whether the larvae survive winter determines the population density in the following year. However, the molecular mechanisms of this process are not clear. RESULTS RNA sequencing (RNA‐Seq) analysis of A. glabripennis larvae at five overwintering stages identified 6876 differentially expressed genes (DEGs). Among these, 46 functional genes that might respond to low temperature were identified. Weighted gene co‐expression network analysis revealed that the MEturquoise module was correlated with the overwintering process. The STPK, PP2A, DGAT, and HSF genes were identified as hub genes using visualization of gene network. In addition, four genes related to sugar transport, gluconeogenesis and glycosylation were screened, which may be involved in the metabolic regulation of overwintering larvae. The protein–protein interaction network indicated that ribosomal protein and ATP synthase may play an important role in connecting with other proteins. The expression levels of fifteen hub genes were further validated by quantitative RT‐PCR, and the results were consistent with RNA‐Seq. CONCLUSION This study demonstrates key genes that may reveal the molecular mechanism of overwintering in A. glabripennis larvae. The genes may be the potential targets to prevent larvae from surviving the cold winter by developing new biological agents using genetic engineering.

Pine wilt disease, caused by an invasive species native to North America, pinewood nematode Bursaphelenchus xylophilus (Steiner and Buhrer) (Nematoda: Aphelenchoididae), seriously harms pine trees and causes enormous economic and ecological losses, threatening the utilization of pine forests and the stability of forest ecosystems (Togashi & Jikumaru, 2007). With the increase in global trade, B. xylophilus was spread worldwide, leading to pine tree deaths over large areas in Japan (1905( ), China (1982