The 70 kDa heat shock proteins play important roles in protecting organisms against environmental stresses, which are divided into stress-inducible forms (HSP70s) and heat shock cognates (HSC70s). In this study, heat shock protein 70 family was identified in the whole genome of the silkworm. Based on the known nomenclature and phylogenetic analysis, four HSP70s and five HSC70s were classified. Relatively, heat shock cognates were more conservative and were constitutively expressed in various tissues of the silkworm larvae. Under thermal (37 °C and 42 °C) and cold (2 °C) stresses, the expressions of HSP70–1, HSP70–2, and HSP70–3 were up-regulated, and the highest induction reached 4147.3, 607.1, and 1987.3 times, respectively. Interestingly, HSC70–1, HSC70–4, and HSC70–5 also showed slight induced expressions in the fat body and/or midgut under thermal stresses. In addition, the expression of HSP70–1 was induced by dichlorvos and phoxim insecticides, while most HSC70 genes were inhibited. The results suggested that stress-inducible forms play more important roles in adaptation to various stresses than HSC70s.

Embryonic diapause is a reversible arrest of embryonic development that enables insects to survive adverse environmental conditions. In the silkworm (Bombyx mori), diapause is established during early egg development and can be efficiently terminated by corona discharge; however, the transcriptional responses associated with this physical stimulus remain insufficiently characterized. Here, we present a strand-specific RNA-seq dataset generated from diapause-destined silkworm eggs subjected to early or late corona discharge treatment. The dataset comprises 36 libraries from three experimental groups across eight developmental time points, with two biological replicates per condition. In total, 283.9 Gb of high-quality clean data were obtained, with average Q20 and Q30 values of 97.60% and 94.20%, respectively, and an average genome-mapping rate of 82.62%. The dataset includes raw reads, gene-expression matrices, lists of differentially expressed genes, novel transcript annotations, alternative splicing events, and SNP/InDel calls. This resource provides a time-resolved transcriptomic framework for investigating physical-stress-induced diapause termination and supports comparative studies of diapause regulation in silkworms and other insects.

The silkworm (Bombyx mori) is rich in germplasm resources, including thermotolerant strains that live in tropical/subtropical humid climates. In this study, two thermotolerant strains and one sensitive strain were used as materials, with the former exhibiting higher critical thermal maximum (CTmax) values. Under different temperature and humidity stresses, physiological and transcriptomic responses of the fifth instar larvae were compared. It was confirmed that high humidity exacerbates harmful effects only under high temperature conditions. Based on transcriptome and co-expression network analysis, 88 evolved thermoplastic genes (Evo_TPGs) and 1338 evolved non-plastic genes (Evo_non-PGs) were identified, which exhibited specific responses or expressions in the two thermotolerant strains. Eighteen of the Evo_TPGs encode cuticular proteins, 17 of which were specifically downregulated in the two thermotolerant strains after short-term exposure to 35 °C. This may promote cuticular transpiration to dissipate internal heat, thus compensating for the suppression of tracheal ventilation in hot and humid climates. For the Evo_non-PGs, most of the metabolic genes showed lower expression at background levels in the thermotolerant strains, while oxidative stress genes showed the opposite trend, suggesting that silkworms can enhance heat tolerance by suppressing metabolic rates and allocating more resources to overcome heat-induced oxidative damage. Furthermore, the heat resistance-related genes showed higher single nucleotide polymorphisms (SNPs) between resistant and sensitive strains compared to randomly selected genes, suggesting that they may have been subjected to natural selection. Through long-term adaptive evolution, thermotolerant silkworms may reduce their internal temperature by dynamically regulating cuticle respiration in response to high temperature and humidity, while allocating more energy to cope with and repair heat-induced damage. Overall, these findings provide insights into the evolution of heat-resistant adaptations to climate change in insects.

Embryonic diapause, a state of developmental arrest in silkworm (Bombyx mori) eggs, poses a challenge for year-round sericulture. While physical stimuli like corona discharge can effectively terminate diapause, the underlying molecular mechanisms, particularly the initial events, remain poorly understood. This study employed an integrated transcriptomic and proteomic approach to analyze silkworm eggs within 48 h after corona treatment. Our time-series analysis revealed that the Hippo and Wnt signaling pathways were specifically activated as early as 1 h post-treatment, preceding the previously reported FoxO pathway response. We identified two temporally distinct gene clusters within the Hippo pathway, including immediate–early genes (e.g., Dachs_17/25/29, Ft_10) and late-phase effector genes, coordinating the exit from cell cycle arrest. Concurrently, the Wnt pathway was rapidly initiated, marked by the sustained upregulation of key regulators Notum and Pontin52, suggesting its role in unlocking the cell cycle. We propose a synergistic model wherein corona discharge triggers the concurrent, early activation of Hippo and Wnt signaling, which collectively reprogram the cell cycle and reinstate the developmental trajectory by promoting proliferation and suppressing apoptosis. These findings provide crucial insights into the initial molecular events of diapause termination, identifying Hippo and Wnt pathways as master regulators in transducing the physical corona stimulus into a developmental signal.