The tree-top disease is an altered behavioral state, displayed by baculovirus-infected lepidopteran larvae, and characterized by climbing to an elevated position before death. The detailed molecular mechanism underlying this phenomenal behavior change has not been reported yet. Our study focused on the transcriptomic changes in the host larvae due to baculovirus infection from pre-symptomatic to tree-top disease stage. Enrichment map visualization of the gene sets grouped based on the functional annotation similarity revealed 34 enriched pathways in signaling mechanism cluster during LdMNPV induced tree-top disease in third instar Lymantria dispar asiatica larvae. Directed light bioassay demonstrated the positively phototactic larvae during tree-top disease and the gene expression analysis showed altered rhythmicity of the host’s core circadian genes (per and tim) during the course of infection emphasizing the role of Circadian entrainment and Phototransduction pathways in the process, which also showed maximum interactions (>50% shared genes with 24 and 23 pathways respectively) among other signaling pathways in the enrichment map. Our study provided valuable insights into different pathways and genes, their coordinated response and molecular regulation during baculovirus infection and also improved our understanding regarding signaling mechanisms in LdMNPV induced tree-top disease.

Baculoviruses are effective biological control agents for many insect pests. They not only efficiently challenge the host immune system but also make them hyperactive for better virus dispersal. Some investigations have focused on the viral mechanisms for induction of such altered response from the host. However, there are no current studies monitoring changes in gene expression during this altered phenotype in infected larvae. The L. dispar multiple nucleopolyhedrovirus (LdMNPV) induces hyperactivity in third instar L. dispar larvae at 3-days post infection (dpi), to continued till 6 dpi. The transcriptome profiles of the infected and uninfected larvae at these time points were analyzed to provide new clues on the response of the larvae towards infection during hyperactivity. Gene ontology enrichment analysis revealed, most of the differentially expressed genes (DEGs) were involved in proteolysis, extracellular region, and serine-type endopeptidase activity. Similarly, Kyoto Encyclopedia of Genes and Genome enrichment analysis showed maximum enrichment of 487 genes of the signal transduction category and neuroactive ligand-receptor interaction sub-category with 85 annotated genes. In addition, enrichment map visualization of gene set enrichment analysis showed the coordinated response of neuroactive ligand-receptor interaction genes with other functional gene sets, as an important signal transduction mechanism during the hyperactive stage. Interestingly all the DEGs in neuroactive ligand-receptor interactions were serine proteases, their differential expression during the hyperactive stage correlated with their conceivable involvement in disease progression and the resulting altered phenotype during this period. The outcome provides a basic understanding of L. dispar larval responses to LdMNPV infection during the hyperactive stage and helps to determine the important host factors involved in this process.

Entomopathogenic fungi are considered to be a safe microbiological pesticide alternative to chemical control. Efforts are underway to understand precisely their taxonomy and natural distribution through mycological and biodiversity studies based on molecular markers. Here, we present descriptions of the diversity of the entomopathogenic fungi in the genera Metarhizium and Beauveria found along the elevational gradients of the Qinling subtropical and temperate forests of Shaanxi province in China, using morphological aspects and molecular markers. Molecular characterization using the Mz_IGS3 intergenic region revealed that Metarhizium isolates phylogenetically clustered in the PARB clade with four different distinguishable species, but the 5'-TEF gene allowed only ambiguous delimitation of Metarhizium species. Beauveria isolates were characterized by sequence analyses of the translation elongation factor 1-α and the Bloc region. The richness of Metarhizium species decreased with increasing elevation, with Metarhizium robertsii s.l. being the most abundant species along the elevational gradient. Our bioassay suggests that certain species of Metarhizium are significantly pathogenic to the insect model Tenebrio molitor at both the adult and larvae stages and could potentially serve as a control of insect pests of forests.

Exploring prospective mates can be risky. Males of the spider mite Tetranychus urticae approach and guard immobile (quiescent) female nymphs to increase their chances of fathering offspring, this being a first-male sperm priority species. We investigated the behaviour of male T. urticae towards females killed by the mite pathogenic fungus Neozygites floridana, letting them choose between a fungal killed and a healthy quiescent female. The dead female (called cadaver) was in one of three stages: (1) non-sporulating; (2) sporulating with primary conidia (non-infective); (3) surrounded/partly covered by secondary capilliconidia (infective). When the cadaver was in stage 1 or 2, males were significantly more often observed near the cadaver than near the healthy female. When the cadaver was in stage 3 (infective capilliconidia), males preferred the vicinity of healthy females. The frequency of two male behaviours, touching and guarding, was also recorded. Touching the cadaver tended to decrease as cadaver developed, whereas touching the healthy females increased. Guarding of non-sporulating cadavers and healthy females was equally common, while guarding of sporulating cadavers was only observed once (stage 2) or not at all (stage 3). To differentiate between the effect of fungal infection and sex, we also let males choose between a non-sporulating cadaver of each sex. Males then preferred to approach the female cadaver. Touching behaviour followed the same pattern, and guarding of male cadavers was not observed. Our results indicate that T. urticae males are more attracted to non-infective female cadavers than to healthy females, only detecting their mistake when very close. Moreover, males approach and explore cadavers surrounded by infective conidia. Whether the results of host manipulation by the pathogen or just sensory constraints in the host, this inability to detect unsuitable and indeed infective mates promotes transmission of the pathogen.