Genome-Wide Identification and Transcriptome Analysis of the Hsp70 Gene Family in Monodonta labio Reveals Its Role in Response to Nanoplastics Stress.

For marine invertebrates, the disruption of organismal physiology and behavior by nanoplastics (NPs) has been extensively reported. Heat shock proteins (Hsps) are important for redundant protein breakdown, environmental changes, and intracellular protein transport. An exhaustive identification of Hsp70 genes and an experiment where different concentrations of NPs were stressed were performed to study how Hsp70 genes respond to NPs stress in Monodonta labio. Our results identified 15 members of Hsp70 within the genome of M. labio and provided insights into their responses to different concentrations of acute NP stress. Phylogenetic analyses revealed extensive amplification of the Hsp70 genes from the Hsc70 subfamily, with gene duplication events. As a result of NP stress, five of fifteen genes showed significant upregulation or downregulation. Three Hsp70 genes were highly expressed at an NP concentration of 0.1 mg/L, and no genes were downregulated. At 10 mg/L, they showed significant upregulation of two genes and significant downregulation of two genes. At 1 mg/L treatment, three genes were significantly downregulated, and no genes were significantly upregulated. Moreover, a purifying selection was revealed using a selection test conducted on duplicate gene pairs, indicating functional redundancy. This work is the first thorough examination of the Hsp70s in Archaeogastropoda. The findings improve knowledge of Hsp70s in molluscan adaptation to NP stress and intertidal living and offer essential data for the biological study of M. labio.

Chromosomal-scale genome assembly and annotation of the land slug (Meghimatium bilineatum).

Meghimatium bilineatum is a notorious pest land slug used as a medicinal resource to treat ailments in China. Although this no-model species is unique in terms of their ecological security and medicinal value, the genome resource of this slug is lacking to date. Here, we used the Illumina, PacBio, and Hi-C sequencing techniques to construct a chromosomal-level genome of M. bilineatum. With the Hi-C correction, the sequencing data from PacBio system generated a 1.61 Gb assembly with a scaffold N50 of 68.08 Mb, and anchored to 25 chromosomes. The estimated assembly completeness at 91.70% was obtained using BUSCO methods. The repeat sequence content in the assembled genome was 72.51%, which mainly comprises 34.08% long interspersed elements. We further identified 18631 protein-coding genes in the assembled genome. A total of 15569 protein-coding genes were successfully annotated. This genome assembly becomes an important resource for studying the ecological adaptation and potential medicinal molecular basis of M. bilineatum.

Fitness of Nutrition Regulation in a Caterpillar Pest Mythimna separata (Walker): Insights from the Geometric Framework.

In nature, plants can contain variable nutrients depending upon the species, tissue, and developmental stage. Insect herbivores may regulate their nutrient intake behaviorally and physio- logically when encountering different foods. This study examined the nutritional regulation of the oriental armyworm, Mythimna separata, for the first time. In one experiment, we allowed the cater-pillars to choose between two nutritionally balanced but complementary diets. The caterpillars did not randomly consume the paired foods, but instead chose between the nutritionally balanced but complementary diets. This intake behavior was found to change with their developmental stages. Furthermore, the nutrient concentrations in food significantly impacted the insect's performance. In the other experiment, caterpillars were given one of eleven diets that reflected the different nutrient conditions in the field. The results showed that proteins were significantly associated with developmental time and fecundity. For example, by consuming protein-biased food, the caterpillars developed faster and produced more eggs. In contrast, carbohydrates were more strongly linked to lipid accumulation, and caterpillars accumulated more lipids when consuming the carbohydrate-biased food. Moreover, the caterpillars were also found to actively regulate their intake of proteins and carbohydrates based on food quality and to physiologically prepare for subsequent life stages. These findings enhance our understanding of how M. separata feeds and responds to different nutritional environments in the field, which could have implications for managing insect herbivores in agricultural settings.

The Effects of Diet on the Immune Responses of the Oriental Armyworm Mythimna separata.

Nutrients can greatly affect host immune defenses against infection. Possessing a simple immune system, insects have been widely used as models to address the relationships between nutrition and immunity. The effects of high versus low protein-to-carbohydrate ratio (P:C) diets on insect immune responses vary in different studies. To reveal the dietary manipulation of immune responses in the polyphagous agricultural pest oriental armyworm, we examined immune gene expression, phenoloxidase (PO) activity, and phagocytosis to investigate the immune traits of bacteria-challenged oriental armyworms, which were fed different P:C ratio diets. We found the oriental armyworms that were fed a 35:7 (P:C) diet showed higher phenoloxidase (PO) activity and stronger melanization, and those reared on a 28:14 (P:C) diet showed higher antimicrobial activity. However, different P:C diets had no apparent effect on the hemocyte number and phagocytosis. These results overall indicate that high P:C diets differently optimize humoral immune defense responses in oriental armyworms, i.e., PO-mediated melanization and antimicrobial peptide synthesis in response to bacteria challenge.

Neutral Processes Provide an Insight Into the Structure and Function of Gut Microbiota in the Cotton Bollworm.

Gut-associated microbes can influence insect health and fitness. Understanding the structure of bacterial communities provides valuable insights on how different species may be selected and their functional characteristics in their hosts. The neutral model is powerful in predicting the structure of microbial communities, but its application in insects remains rare. Here, we examined the contribution of neutral processes to the gut-associated bacterial communities in Helicoverpa armigera caterpillars collected from different maize varieties at four locations. The gut-associated bacteria can be assigned to 37 Phyla, 119 orders, and 515 genera, with each individual gut containing 17-75% of the OTUs and 19-79% of the genera in the pooled samples of each population. The distribution patterns of most (75.59-83.74%) bacterial taxa were in good agreement with the neutral expectations. Of the remaining OTUs, some were detected in more individual hosts than would be predicted by the neutral model (i.e., above-partition), and others were detected in fewer individual hosts than predicted by the neutral model (i.e., below-partition). The bacterial taxa in the above-partitions were potentially selected by the caterpillar hosts, while the bacteria in the below-partitions may be preferentially eliminated by the hosts. Moreover, the gut-associated microbiota seemed to vary between maize varieties and locations, so ecological parameters outside hosts can affect the bacterial communities. Therefore, the structure of gut microbiota in the H. armigera caterpillar was mainly determined by stochastic processes, and the bacteria in the above-partition warrant further investigation for their potential roles in the caterpillar host.

Understanding nutritive need in Harmonia axyridis larvae: Insights from nutritional geometry.

The multicolored Asian lady beetle, Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae), is an important natural enemy in agricultural ecosystems. In spite of being a carnivore consuming protein-rich preys, the lady beetles often consume carbohydrate-rich food like nectar or honeydew. However, most studies on nutrition regulation of carnivores mainly focus on protein and lipid, two major macronutrients in preys. In this study, nutrition regulation of protein and carbohydrate has been investigated in the 4th instar larvae of H. axyridis using Geometric Framework. We provided the insects two pairs of foods, one a protein-biased one and the second carbohydrate-biased, to determine the intake target. We then confined them to nutritionally imbalanced foods to examine how they regulated food intake to achieve maximal performance. The larvae performed well on the 2 foods that containing the closest P : C ratios to the intake target, but, surprisingly, the lipid content was much lower than that in the choice experiment. The lady beetles seemed to maintain the optimal lipid content by consuming carbohydrate-rich food. Moreover, consuming the carbohydrate-rich food was less metabolically expensive than the protein-rich food. Therefore, switching behavior between plant and animal foods actually reflects their nutritive needs. These findings extended our understanding of predator forage behavior and its influence on food web in ecosystems, and shed light on the role of agri-environment schemes in meeting the nutritional need of predators in field.

A Semi-Synthetic Diet and the Potential Important Chemicals for Mythimna separata (Lepidoptera: Noctuidae).

Armyworm feeding in large, destructive groups is hugely difficult to control and the oriental armyworm, Mythimna separata (Walk), is one such pest. In this study, we reported a semisynthetic artificial diet for the oriental armyworm. This diet is based on Ritter's diet, a formula developed for Heliothis zea. The survival of M. separata was extremely low and only around 2% insects can reach the adult stage on Ritter's diet. But, it can reach up to 100% if corn leaf powder (CLP) was mixed, and insects grew faster and gained more mass. After testing a set of mixtures of Ritter's diet and CLP, we found that 14.3% was the optimal proportion of CLP for making the artificial diet. We then used chloroform to extract CLP. Insect performance was still much better on Ch-extracted CLP diets than that on Ritter's diet, but it was poorer than that on the diets containing unprocessed CLP, suggesting that the essential factor(s) was only partially extracted from corn leaf. We then used methanol and dichloromethane, two solvents differing in their polarity, to process the extractions and analyzed the extracted chemicals using gas chromatography-mass spectrometry (GC-MS). Insects had a better performance on dichloromethane-extracted CLP diet in comparison to methanol-extracted one, indicating that the important factor(s) is more prone to methanol extraction. The reported recipe here is useful for the research on M. separata and possibly other grain-crop eating armyworms. The functions of the chemicals extracted from corn leaf tissue can be investigated in the future studies.

Machine Learning Models Based on Molecular Fingerprints and an Extreme Gradient Boosting Method Lead to the Discovery of JAK2 Inhibitors.

Developing Janus kinase 2 (JAK2) inhibitors has become a significant focus for small-molecule drug discovery programs in recent years because the inhibition of JAK2 may be an effective approach for the treatment of myeloproliferative neoplasm. Here, based on three different types of fingerprints and Extreme Gradient Boosting (XGBoost) methods, we developed three groups of models in that each group contained a classification model and a regression model to accurately acquire highly potent JAK2 kinase inhibitors from the ZINC database. The three classification models resulted in Matthews correlation coefficients of 0.97, 0.94, and 0.97. Docking methods including Glide and AutoDock Vina were employed to evaluate the virtual screening effectiveness of our classification models. The R2 of three regression models were 0.80, 0.78, and 0.80. Finally, 13 compounds were biologically evaluated, and the results showed that the IC50 values of six compounds were identified to be less than 100 nM. Among them, compound 9 showed high activity and selectivity in that its IC50 value was less than 1 nM against JAK2 while 694 nM against JAK3. The strategy developed may be generally applicable in ligand-based virtual screening campaigns.

Phylogeny and evolution of the cholesterol transporter NPC1 in insects.

Sterols are essential nutrients for eukaryotes. Insects are obligate sterol auxotrophs and must acquire this key nutrient from their diets. The digestive tract is the organ for absorbing nutrients as well as sterols from food. In mice, the Niemann-Pick type C1 Like 1 (NPC1L1) gene is highly expressed in the intestine and is critical for cholesterol absorption. In contrast, the molecular mechanisms for the absorption of dietary sterols in insects have not been well studied. We annotated NPC1 genes in 39 insects from 10 orders using available genomic and transcriptomic information and inferred phylogenetic relationships. Insect NPC1 genes were grouped into two sister-clades, NPC1a and NPC1b, suggesting a likely duplication in the ancestor of insects. The former exhibited weaker gut-biased expression or a complete lack of tissue-biased expression, depending on the species, while the latter was highly enriched in the gut of three lepidopteran species. This result is similar to previous findings in Drosophila melanogaster. In insects, NPC1a accumulated non-synonymous substitutions at a lower rate than NPC1b. This pattern was consistent across orders, indicating that NPC1a evolved under stronger molecular constraint than NPC1b.