Nutritional symbionts confer structural defence against predation and fungal infection in a grain pest beetle

Many insects benefit from bacterial symbionts that provide essential nutrients and thereby extend the hosts’ adaptive potential and their ability to cope with challenging environments. However, the implications of nutritional symbioses for the hosts’ defence against natural enemies remain largely unstudied. Here, we investigated if the cuticle-enhancing nutritional symbiosis of the saw-toothed grain beetle Oryzaephilus surinamensis confers protection against predation and fungal infection. We exposed age-defined symbiotic and symbiont-depleted (aposymbiotic) beetles to two antagonists that must actively penetrate the cuticle for a successful attack: wolf spiders (Lycosidae) and the fungal entomopathogen Beauveria bassiana. While young beetles suffered from high predation and fungal infection rates regardless of symbiont presence, symbiotic beetles were able to escape this period of vulnerability and reach high survival probabilities significantly faster than aposymbiotic beetles. To understand the mechanistic basis underlying these differences, we conducted a time-series analysis of cuticle development in symbiotic and aposymbiotic beetles by measuring cuticular melanisation and thickness. The results reveal that the symbionts accelerate their host's cuticle formation and thereby enable it to quickly reach a cuticle quality threshold that confers structural protection against predation and fungal infection. Considering the widespread occurrence of cuticle enhancement via symbiont-mediated tyrosine supplementation in beetles and other insects, our findings demonstrate how nutritional symbioses can have important ecological implications reaching beyond the immediate nutrient provisioning benefits.

Transcriptome Analyses Provide Insights into the Aggressive Behavior toward Conspecific and Heterospecific in Thitarodes xiaojinensis (Lepidoptera: Hepialidae)

Aggressive behavior in animals is important for survival and reproduction. It is well studied in adult insects, such as flies, ants, honey bees, and crickets. However, the larvae of Lepidopteran insects are also aggressive, studies of which are still lacking. Here, RNA-seq was used to generate a high-quality database for the aggressive behavior of Thitarodes xiaojinensis toward conspecifics and heterospecifics. Although there was similar aggressive behavior between the conspecific group and heterospecific group, significant differences were identified at the transcriptional level. When there was aggressive behavior toward conspecifics, T. xiaojinensis trended toward higher expression at the respiratory chain, while cuticle development and metabolism may have interfered. On the other hand, when there was aggressive behavior toward H. armigera, genes related to neuron and cuticle development, cellular processes, and its regulated signaling pathways were significantly upregulated, while the genes associated with oxidation-reduction and metabolism were downregulated. Weighted gene co-expression networks analysis (WGCNA) was performed, and two modules with properties correlating to the aggressive behavior of T. xiaojinensis were identified. Several hub genes were predicted and confirmed by qRT-PCR, such as CLTC, MYH, IGF2BP1, and EMC. This study provides a global view and potential key genes for the aggressive behavior of T. xiaojinensis toward conspecifics and heterospecifics. Further investigation of the hub genes would help us to better understand the aggressive behavior of insects.

Allomones confer resistance to Musa cultivar Pisanglilin against infestation by Odoiporus longicollis [Oliver] and characterization of Allomones

In bioassay guided extraction of pseudostem powder of Pisanglilin by organic solvents we found the larvicidal activity in acetone extract, whose column chromatography by methanol-chloroform mixture separated the extract into 9-fractions, of which the 8th fraction showed larvicidal activity. Subfractionation of the active fraction by column chromatography resulted in the isolation of two larvicidal molecules [Stigmasterol-3-O-glucoside (SOG) and Sulfoquinovosyl diacylglycerol (SQDG)]. Yield of SOG was 0.002 % and SQDG was 0.005 % and both were highly toxic to O. longicollis larvae with LD50 of 0.40 and 0.378 ppm, respectively. Larvae fed these compounds stopped feeding on third day and died within one week. SOG inhibited the amylase and protease activity of gut and induced histolysis in the mid gut. While SQDG inhibited the leucine amino peptidase and trypsin like serine protease activities, which decreased the content of total free amino acids. Imbalance in the activities of aspartate amino transferase and alanine amino transferase disrupted the aminoacid metabolism and the compound inhibited the activity of tyrosinase (an enzyme involved in cuticle development). SQDG toxicity caused accumulation of 20-hydroxyecdysone, the active moulting hormone in the hemolymph. Simultaneous action of two allomones present in Pisanglilin effectively resisted the attack of endophytic larvae in the pseudostem and thereby conferred resistance against infestation by O.longicollis. Preliminary study by intrapseudostem injection of Pisanglilin extract in susceptible M.paradisiaca cultivar Kappa, gave complete protection to it from attack by this pest, under field condition.

Relationship of Cuticle Development with Water Loss and Texture of Pepper Fruit

Postharvest water loss in pepper fruit (Capsicum annuum L.) reduces its shelf life. Fruit texture is one of the most important components of fruit quality for consumers. In this study, the anatomical traits of pepper fruit related to postharvest water loss and texture were assessed. There was a strong positive relationship between postharvest water loss and the thickness of the cuticular membrane, cuticular weight, total cutin weight, and polysaccharide-cutan weight. An amorphous fibrous structure that forms a path for diffusion and increases water loss was observed in the thick cuticle of the pericarp. In addition, positive correlations between the hardness of the exocarp and the weight of cuticular membrane, cutin content, and polysaccharide-cutan content were found. These results indicate that the thickness of the cuticular membrane wedged between subepidermal cells may influence water loss through the pericarp of pepper fruit and fruit with a high cutin and polysaccharide content have a hard tough texture.

Gut bacteria are essential for normal cuticle development in herbivorous turtle ants

Across the evolutionary history of insects, the shift from nitrogen-rich carnivore/omnivore diets to nitrogen-poor herbivorous diets was made possible through symbiosis with microbes. The herbivorous turtle ants Cephalotes possess a conserved gut microbiome which enriches the nutrient composition by recycling nitrogen-rich metabolic waste to increase the production of amino acids. This enrichment is assumed to benefit the host, but we do not know to what extent. To gain insights into nitrogen assimilation in the ant cuticle we use gut bacterial manipulation, 15N isotopic enrichment, isotope-ratio mass spectrometry, and 15N nuclear magnetic resonance spectroscopy to demonstrate that gut bacteria contribute to the formation of proteins, catecholamine cross-linkers, and chitin in the cuticle. This study identifies the cuticular components which are nitrogen-enriched by gut bacteria, highlighting the role of symbionts in insect evolution, and provides a framework for understanding the nitrogen flow from nutrients through bacteria into the insect cuticle.

Transcriptome analysis of Caenorhabditis elegans lacking heme peroxidase SKPO-1 reveals an altered response to Enterococcus faecalis

The nematode Caenorhabditis elegans is commonly used as a model organism in studies of the host immune response. The worm encodes twelve peroxidase-cyclooxygenase superfamily members, making it an attractive model in which to study the functions of heme peroxidases. In previous work, loss of one of these peroxidases, SKPO-1 (ShkT-containing peroxidase), rendered C. elegans more sensitive to the human, Gram-positive pathogen Enterococcus faecalis. SKPO-1 was localized to the hypodermis of the animals where it also affected cuticle development as indicated by a morphological phenotype called “dumpy.” In this work, a better understanding of how loss of skpo-1 impacts both sensitivity to pathogen as well as cuticle development was sought by subjecting a deletion mutant of skpo-1 to transcriptome analysis using RNA sequencing following exposure to control (Escherichia coli) and pathogenic (E. faecalis) feeding conditions. Loss of skpo-1 caused a general upregulation of genes encoding collagens and other proteins related to cuticle development. On E. faecalis, these animals also failed to upregulate guanylyl cyclases that are often involved in environmental sensing. Hoechst straining revealed increased permeability of the cuticle and atomic force microscopy exposed the misalignment of the cuticular annuli and furrows. These findings provide a basis for better understanding of the morphological as well as the pathogen sensitivity phenotypes associated with loss of SKPO-1 function.

Class I TCP transcription factors regulate trichome branching and cuticle development in Arabidopsis

Trichomes and the cuticle are two specialized structures of the aerial epidermis that are important for plant organ development and interaction with the environment. In this study, we report that Arabidopsis thaliana plants affected in the function of the class I TEOSINTE BRANCHED 1, CYCLOIDEA, PCF (TCP) transcription factors TCP14 and TCP15 show overbranched trichomes in leaves and stems and increased cuticle permeability. We found that TCP15 regulates the expression of MYB106, a MIXTA-like transcription factor involved in epidermal cell and cuticle development, and overexpression of MYB106 in a tcp14 tcp15 mutant reduces trichome branch number. TCP14 and TCP15 are also required for the expression of the cuticle biosynthesis genes CYP86A4, GPAT6, and CUS2, and of SHN1 and SHN2, two AP2/EREBP transcription factors required for cutin and wax biosynthesis. SHN1 and CUS2 are also targets of TCP15, indicating that class I TCPs influence cuticle formation acting at different levels, through the regulation of MIXTA-like and SHN transcription factors and of cuticle biosynthesis genes. Our study indicates that class I TCPs are coordinators of the regulatory network involved in trichome and cuticle development.