Comparative 4D Label-Free Quantitative Proteomic Analysis of Bombus terrestris Provides Insights into Proteins and Processes Associated with Diapause.

Diapause, an adaptative strategy for survival under harsh conditions, is a dynamic multi-stage process. Bombus terrestris, an important agricultural pollinator, is declining in the wild, but artificial breeding is possible by imitating natural conditions. Mated queen bees enter reproductive diapause in winter and recover in spring, but the regulatory mechanisms remain unclear. Herein, we conducted a comparative 4D label-free proteomic analysis of queen bees during artificial breeding at seven timepoints, including pre-diapause, diapause, and post-diapause stages. Through bioinformatics analysis of proteomic and detection of substance content changes, our results found that, during pre-diapause stages, queen bees had active mitochondria with high levels of oxidative phosphorylation, high body weight, and glycogen and TAG content, all of which support energy consumption during subsequent diapause. During diapause stages, body weight and water content were decreased but glycerol increased, contributing to cold resistance. Dopamine content, immune defense, and protein phosphorylation were elevated, while fat metabolism, protein export, cell communication, signal transduction, and hydrolase activity decreased. Following diapause termination, JH titer, water, fatty acid, and pyruvate levels increased, catabolism, synaptic transmission, and insulin signaling were stimulated, ribosome and cell cycle proteins were upregulated, and cell proliferation was accelerated. Meanwhile, TAG and glycogen content decreased, and ovaries gradually developed. These findings illuminate changes occurring in queen bees at different diapause stages during commercial production.

Integrative Proteomic and Phosphoproteomic Analyses Revealed Complex Mechanisms Underlying Reproductive Diapause in Bombus terrestris Queens.

Reproductive diapause is an overwintering strategy for Bombus terrestris, which is an important pollinator for agricultural production. However, the precise mechanisms underlying reproductive diapause in bumblebees remain largely unclear. Here, a combination analysis of proteomics and phosphoproteomics was used to reveal the mechanisms that occur during and after diapause in three different phases: diapause (D), postdiapause (PD), and founder postdiapause (FPD). In total, 4655 proteins and 10,600 phosphorylation sites of 3339 proteins were identified. Diapause termination and reactivation from D to the PD stage were characterized by the upregulation of proteins associated with ribosome assembly and biogenesis, transcription, and translation regulation in combination with the upregulation of phosphoproteins related to neural signal transmission, hormone biosynthesis and secretion, and energy-related metabolism. Moreover, the reproductive program was fully activated from PD to the FPD stage, as indicated by the upregulation of proteins related to fat digestion and absorption, the biosynthesis of unsaturated fatty acids, fatty acid elongation, protein processing in the endoplasmic reticulum, and the upregulation of energy-related metabolism at the phosphoproteome level. We also predicted a kinase-substrate interaction network and constructed protein-protein networks of proteomic and phosphoproteomic data. These results will help to elucidate the mechanisms underlying the regulation of diapause in B. terrestris for year-round mass breeding.

Optimization via response surface methodology of the synthesis of a dust suppressant and its performance characterization for use in open cut coal mines.

To relieve dust pollution in open cut coal mines and reduce the hazards of coal dust pollution to the environment and workers we optimized the synthesis of a dust suppressant by graft copolymerization of environmentally friendly soy protein isolate with methyl methacrylate. This dust suppressant could effectively control dust pollution in open cut coal mines. The optimized conditions for graft copolymerization in this case were determined by a response surface experiment designed with Design-Expert 10 software. Characterization by scanning electron microscopy showed a significant morphology change of the dust suppressant and the generation of a rigid and dense layer on its surface after interacting with coal dust. The layer exhibited good bonding and dust suppression performance. The analysis with Fourier-transform infrared spectroscopy revealed the appearance of new absorption peaks near 1300, 1072, and 1631 cm-1, demonstrating effective graft copolymerization. The proposed dust suppressant exhibited excellent wind erosion resistance, with a resistance that exceeded 90% at a wind speed of 6.5 m/sec. The successful graft copolymerization and effective bonding and curing of the dust suppressant on coal dust were experimentally verified. This is of great significance to the control of coal dust pollution.

Hormonal Regulation of Reproductive Diapause That Occurs in the Year-Round Mass Rearing of Bombus terrestris Queens.

Adult reproductive diapause is an adaptive strategy under adverse environments for insects and other arthropod species, including bumblebees, which enables queens to survive through a harsh winter and then build new colonies in the following spring. Little research has been done on the molecular regulatory mechanism of reproductive diapause in Bombus terrestris, which is an important pollinator of wild plants and crops. Our previous research identified the conditions that induced reproductive diapause during the year-round mass rearing of B. terrestris. Here, we performed combined transcriptomics and proteomics analyses of reproductive diapause in B. terrestris during and after diapause at three different ecophysiological phases, diapause, postdiapause, and founder postdiapause. The analyses showed that differentially expressed proteins/genes acted in the citrate cycle, insect hormone biosynthesis, insulin and mTOR signaling pathway. To further understand the mechanisms that regulated the reproductive diapause, genes involved in the regulation of JH synthesis, insulin/TOR signal pathway were determined. The BtRheb, BtTOR, BtVg, and BtJHAMT had lower expression levels in diapause queens. The JH III titer levels and the activities of the metabolic enzymes were significantly up-regulated in postdiapause queens. Also, after the microinjection of insulin-like peptides (ILPs) and JH analogue (JHA), hormones, cold-tolerance metabolites, metabolic enzymes, and reproduction showed significant changes. Together with results from other related research, a model of the regulation of reproductive diapause during the year-round mass rearing of B. terrestris was proposed. This study contributes to a comprehensive insight into the molecular regulatory mechanism of reproductive diapause in eusocial insects.

Preparation and Characterization of a Composite Dust Suppressant for Coal Mines.

In an effort to effectively control coal dust pollution and thereby reduce the harm of coal dust to human health, we prepared a highly efficient composite dust suppressant. First, dynamic contact angle and zeta potential measurements were used to select sodium dodecyl sulfonate (SDS) over sodium carboxymethyl cellulose and trisodium methyl silicon as the complementary additive to soy protein isolate for the dust suppressant. We employed viscosity and wind erosion resistance tests to compare the performance of the composite dust suppressant with three common, commercially available suppressants. As the concentration of the composite dust suppressant was increased, the viscosity increased, reaching a maximum value of 22.7 mPa·s at a concentration of 5 wt%. The 5 wt% concentration of the composite dust suppressant provided the lowest wind erosion rate (20.62%) at a wind speed of 12 m/s. The composite dust suppressant also had good bonding performance and wind erosion resistance. Scanning electron microscopy, X-ray diffraction, and thermogravimetric analysis were used to characterize the properties of the dust suppressants. The dust suppressant, which had a crystal-like structure, could easily capture coal dust and form an effective package. In addition, the density of the dust suppressant film increased as its crystallinity increased. The increased density was beneficial in that it enabled the dust suppressant to form a hard, solidified shell on the surface of coal dust, which improved dust suppression. The composite dust suppressant also had good thermal stability.

FoxO Transcription Factor Regulate Hormone Mediated Signaling on Nymphal Diapause.

Diapause is a complex physiological adaptation phenotype, and the transcription factor Forkhead-box O (FoxO) is a prime candidate for activating many of its diverse regulatory signaling pathways. Hormone signaling regulates nymphal diapause in Laodelphax striatellus. Here, the function of the FoxO gene isolated from L. striatellus was investigated. After knocking-down LsFoxO in diapausal nymphs using RNA interference, the titers of juvenile hormone III and some cold-tolerance substances decreased significantly, and the duration of the nymphal developmental period was severely shorted to 25.5 days at 20°C under short day-length (10 L:14 D). To determine how LsFoxO affects nymphal diapause, analyses of RNA-sequencing transcriptome data after treatment with LsFoxO-RNA interference was performed. The differentially expressed genes affected carbohydrate, amino acid and fatty acid metabolism, and phosphatidylinositol 3-kinase/protein kinase B signaling pathway. Thus, LsFoxO acts on L. striatellus nymphal diapause and is, therefore, a potential target gene for pest control. This study may lead to new information on the regulation of nymphal diapause in this important pest.