By employing a culture-dependent and -independent 16S rRNA and ITS gene high-throughput sequencing analyses, comprehensive information was obtained on the gut bacterial and fungal communities in the ghost moth larvae of three different geographic locations from high-altitude on Tibet plateau and from low-altitude laboratory. Twenty-six culturable bacterial species belonging to 21 genera and 14 fungal species belonging to 12 genera were identified from six populations by culture-dependent method. Carnobacterium maltaromaticum was the most abundant bacterial species from both the wild and laboratory-reared larvae. The most abundant OTUs in the wild ghost moth populations were Carnobacteriaceae, Enterobacteriaceae for bacteria, and Ascomycota and Basidiomycota for fungi. Larval microbial communities of the wild ghost moth from different geographic locations were not significantly different from each other but significant difference in larval microbial community was detected between the wild and laboratory-reared ghost moth. The larval gut of the wild ghost moth was dominated by the culturable Carnobacterium. However, that of the laboratory-reared ghost moth exhibited significantly abundant Wolbachia, Rhizobium, Serratia, Pseudomonas, and Flavobacterium. Furthermore, the larval gut of the wild ghost moth had a significantly higher abundance of Ophiocordyceps but lower abundance of Candida and Aspergillus than that of the laboratory-reared ghost moth.

Thitarodes armoricanus is a medicinal and economically important Lepidopteran insect species. The larvae infected by Paecilomyces hepiali survive no more than four days, while those infected by Ophiocordyceps sinensis can survive for several months before mummification. This provides a valuable comparative system to study interactions between an insect host and different pathogenic fungi. By using the T. armoricanus genome, a time-course transcriptome analysis of the whole larvae without guts was performed to explore the larvae response to P. hepiali and O. sinensis infection. A total of 3106 differentially expressed genes in five clusters were identified. The genes involved in coagulation and multiple metabolisms were both suppressed after P. hepiali or O. sinensis infection, whereas those related to environmental information responses, cell processes, biotic stimulus, and immunity (such as cecropin (CEC)) were elevated. The rapid death of T. armoricanus after P. hepiali infection might be caused by osmotic imbalance, immunocompromise (such as DEFs and GLVs), and nervous system dysfunction (glutamatergic synapse). Up-regulation of the genes related to cuticle structure, nervous system (such as neurotrophin signal pathway and dopaminergic synapse) and immune effectors (such as attacin (ATT) and proline-rich antimicrobial peptide 1 (PRAMP1)) in T. armoricanus, may contribute to the co-existence of T. armoricanus and O. sinensis. This study provides a global view and potential key genes of the interaction between T. armoricanus and two fungal entomopathogens.

Pharmacogenomic analysis based on drug transcriptome characteristics is widely used to identify mechanisms of action. The purpose of this study was to elucidate the molecular mechanism of protective effect against adriamycin (ADM)-induced mpc5 cell injury of Chinese cordyceps aqueous extracts (WCCs) by a systematic transcriptomic analysis. The phytochemicals of WCCs were analyzed via the “phenol–sulfuric acid method”, high-performance liquid chromatography (HPLC), and HPLC–mass spectrometry (MS). We analyzed the drug-reaction transcriptome profiles of mpc5 cell after treating them with WCCs. RNA-seq analysis revealed that WCCs alleviated ADM-induced mpc5 cell injury via restoring the expression of certain genes to normal level mainly in the one-carbon pool by the folate pathway, followed by the relaxin, apelin, PI3K-Akt, and nucleotide-binding, oligomerization domain (NOD)-like receptor signaling pathway, enhancing DNA synthesis and repair, cell proliferation, fibrosis reduction, and immune regulation. Otherwise, WCCs also modulated the proliferation and survival of the mpc5 cell by regulating metabolic pathways, and partially restores the expression of genes related to human disease pathways. These findings provide an innovative understanding of the molecular mechanism of the protective effect of WCCs on ADM-induced mpc5 cell injury at the molecular transcription level, and Mthfd2, Dhfr, Atf4, Creb5, Apln, and Serpine1, etc., may be potential novel targets for treating nephrotic syndrome.