The interaction between larval host plant quality and temperature can influence the short-term physiological rates and life-history traits of insect herbivores. These factors can vary locally, resulting in local adaptation in responses to diet and temperature, but the comparison of these interactions between populations is infrequently carried out. In this study, we examine how the macronutrient ratio of an artificial diet determines the larval growth, development, and survival of larval Pieris rapae (Lepidoptera: Pieridae) at different temperatures between two invasive North American populations from different climatic regions. We conducted a fully factorial experiment with three temperature treatments (18°C, 25°C, and 32°C) and three artificial diet treatments varying in terms of the ratio of protein to carbohydrate (low protein, balanced, and high protein). The effects of diet on life-history traits were greater at lower temperatures, but these differed between populations. Larvae from the subtropical population had reduced survival to pupation on the low-protein diet in the cold temperature treatment, whereas larval survival for the temperate population was equally high for all temperature and diet treatments. Overall, both populations performed more poorly (i.e., they showed slower rates of consumption, growth, and development, and had a smaller pupal mass) in the diet with the low protein ratio, but larvae from the temperate population were less sensitive to diet ratio changes at all temperatures. Our results confirm that the physiological and life-history consequences of imbalanced nutrition for insect herbivores may depend on developmental temperatures, and that different geographic populations of P. rapae within North America vary in their sensitivity to nutritional balance and temperature.

Knowledge of plant recognition of insects is largely limited to a few resistance (R) genes against sap-sucking insects. Hypersensitive response (HR) characterizes monogenic plant traits relying on R genes in several pathosystems. HR-like cell death can be triggered by eggs of cabbage white butterflies (Pieris spp.), pests of cabbage crops (Brassica spp.), reducing egg survival and representing an effective plant resistance trait before feeding damage occurs. Here, we performed genetic mapping of HR-like cell death induced by Pieris brassicae eggs in the black mustard Brassica nigra (B. nigra). We show that HR-like cell death segregates as a Mendelian trait and identified a single dominant locus on chromosome B3, named PEK (Pieris  egg- killing). Eleven genes are located in an approximately 50 kb region, including a cluster of genes encoding intracellular TIR-NBS-LRR (TNL) receptor proteins. The PEK locus is highly polymorphic between the parental accessions of our mapping populations and among B. nigra reference genomes. Our study is the first one to identify a single locus potentially involved in HR-like cell death induced by insect eggs in B. nigra. Further fine-mapping, comparative genomics and validation of the PEK locus will shed light on the role of these TNL receptors in egg-killing HR.

Research on insect microbiota has greatly expanded over the past decade, along with a growing appreciation of the microbial contributions to insect ecology and evolution. Many of these studies use DNA sequencing to characterize the diversity and composition of insect-associated microbial communities. The choice of strategies used for specimen collection, storage, and handling could introduce biases in molecular assessments of insect microbiota, but such potential influences have not been systematically evaluated. Likewise, although it is common practice to surface sterilize insects prior to DNA extraction, it is not known if this time-consuming step has any effect on microbial community analyses. To resolve these methodological unknowns, we conducted an experiment wherein replicate individual insects of four species were stored intact for two months using five different methods-freezing, ethanol, dimethyl sulfoxide (DMSO), cetrimonium bromide (CTAB), and room-temperature storage without preservative-and then subjected to whole-specimen 16S rRNA gene sequencing to assess whether the structure of the insect-associated bacterial communities was impacted by these different storage strategies. Overall, different insect species harbored markedly distinct bacterial communities, a pattern that was highly robust to the method used to store samples. Storage method had little to no effect on assessments of microbiota composition, and the magnitude of the effect differed among the insect species examined. No single method emerged as \"best,\" i.e., one consistently having the highest similarity in community structure to control specimens, which were not stored prior to homogenization and DNA sequencing. We also found that surface sterilization did not change bacterial community structure as compared to unsterilized insects, presumably due to the vastly greater microbial biomass inside the insect body relative to its surface. We therefore recommend that researchers can use any of the methods tested here, and base their choice according to practical considerations such as prior use, cost, and availability in the field, although we still advise that all samples within a study be handled in an identical manner when possible. We also suggest that, in large-scale molecular studies of hundreds of insect specimens, surface sterilization may not be worth the time and effort involved. This information should help researchers design sampling strategies and will facilitate cross-comparisons and meta-analyses of microbial community data obtained from insect specimens preserved in different ways.