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Transboundary rural communities in West Africa play an important role in the exchange of goods, mainly food, among countries. The COVID-19 pandemic restricted these activities due to the closure of the borders. Semi-structured interviews were conducted in two regions of Senegal and Burkina Faso to examine the impacts of these restrictions on the pillars (availability, access, utilization, and stability) of food security in rural areas on men and women. The data set included 230 interviews, and they were analyzed through thematic content analysis. The results showed a decrease in agricultural production in all the communities due to mainly lack of labor force, and limited access to inputs, resulting in increased post-harvest losses. The disruption of trade and border and market closures affected rural families engaged in transboundary trade. Farmers experienced a sharp loss of household income leading to debts and decapitalization. Availability and diversity of and access to food was also heavily affected. Food security greatly varied among the communities and between countries. Perceptions also varied between men and women in terms of production, mobility, and food consumption. The restriction measures have triggered a spiral of effects and responses seriously impacting long-term food security in already highly vulnerable countries.

Summary: Understanding a population’s fitness heterogeneity and genetic basis of thermal adaptation is essential for predicting the responses to global warming. We examined the thermotolerance and genetic adaptation of Plutella xylostella to exposure to hot temperatures. The population fitness parameters of the hot-acclimated DBM strains varied in the thermal environments. Using genome scanning and transcription profiling, we find a number of genes potentially involved in thermal adaptation of DBM. Editing two ABCG transporter genes, PxWhite and PxABCG, confirmed their role in altering cuticle permeability and influencing thermal responses. Our results demonstrate that SNP mutations in genes and changes in gene expression can allow DBM to rapidly adapt to thermal environment. ABCG transporter genes play an important role in thermal adaptation of DBM. This work improves our understanding of genetic adaptation mechanisms of insects to thermal stress and our capacity to predict the effects of rising global temperatures on ectotherms.

Abstract Transdisciplinary sustainability scientists are called to conduct research with community actors to understand and improve relations between people and nature. Yet, research hierarchies and power relations continue to favour western academic researchers who remain the gatekeepers of knowledge production and validation. To counter this imbalance, in 2018 we structured a multi‐day workshop to co‐design a set of principles to guide our own transdisciplinary, international and intercultural community of practice for biocultural diversity and sustainability. This community includes community collaborators, partner organizations, and early career and established researchers from Argentina, Bolivia, Canada, Germany, Mexico and South Africa. In 2021, we undertook online critical reflection workshops to share our research experiences and deepen our intercultural understanding of the application of the principles. Through these exercises, we adopted seven principles for working together that include: honour self‐determination and nationhood; commit to reciprocal relationships; co‐create the research agenda; approach research in a good way: embed relational accountability; generate meaningful benefits for communities; build in equity, diversity and inclusion; and emphasize critical reflection and shared learning. We explain these principles and briefly highlight their application to our research practices. By sharing these principles and associated practices, we seek to facilitate debate and spur transformations in how we conduct international and intercultural sustainability research. Our efforts also illustrate a strategy for on‐going knowledge co‐production as we cultivate safe and ethical spaces for learning together. Lessons learned may be particularly useful to those who engage in intercultural, collaborative research to advance sustainability transformations. Read the free Plain Language Summary for this article on the Journal blog.

ABSTRACT The diamondback moth (DBM), Plutella xylostella, has successfully adapted to the potent chemical defenses of Brassicaceae plants that deter most other herbivores. Gut bacteria are increasingly recognized as key to the biology of many species but their role in DBM adaptation to plant defense compounds is not well known. In this study, the secondary metabolites of radish seedlings, rich in flavonoids, were identified by liquid chromatography-mass spectrometry. These secondary metabolites reduced the larval growth of DBM lacking gut bacteria. The effect was rapidly eclipsed by the re-introduction of gut microbiota, which was dominated by Enterobacter (Proteobacteria). Similarly, while treatment with the flavonoid kaempferol adversely affected growth and extended the development time, these were alleviated by the re-introduction of Enterobacter sp. EbPXG5 (EbPXG5) to the DBM gut. EbPXG5 not only degrades kaempferol both in vitro and DBM gut, but is also shown to colonize the gut epithelium, forming a protective biofilm. Genomic sequencing of EbPXG5 showed that metabolic genes were the most abundant, especially those involved in xenobiotic degradation, and the metabolism of terpenoids and polyketides, which could participate in the degradation of plant secondary metabolites such as kaempferol. Overall, our results showed that EbPXG5 is a bacterium common in the gut of DBM larvae and has the in vitro and in vivo capacity to detoxify a major secondary metabolite that is produced in brassica plants as a defense against herbivores. This insect-bacterial association may be an important contributor to the status of DBM as a major pest of brassica crops worldwide.IMPORTANCEIn this study, we identify an important role of gut bacteria in mediating the adaptation of diamondback moth (DBM) to plant secondary metabolites. We demonstrate that kaempferol’s presence in radish seedlings greatly reduces the fitness of DBM with depleted gut biota. Reinstatement of gut biota, particularly Enterobacter sp. EbPXG5, improved insect performance by degrading kaempferol. This bacterium was common in the larval gut of DBM, lining the epithelium as a protective film. Our work highlights the role of symbiotic bacteria in insect herbivore adaptation to plant defenses and provides a practical and mechanistic framework for developing a more comprehensive understanding of insect-gut microbe-host plant co-evolution.

Tea green leafhopper (TGL), Empoasca onukii, is of biological and economic interest. Despite numerous studies, the mechanisms underlying its adaptation and evolution remain enigmatic. Here, we use previously untapped genome and population genetics approaches to examine how the pest adapted to different environmental variables and thus has expanded geographically. We complete a chromosome-level assembly and annotation of the E. onukii genome, showing notable expansions of gene families associated with adaptation to chemoreception and detoxification. Genomic signals indicating balancing selection highlight metabolic pathways involved in adaptation to a wide range of tea varieties grown across ecologically diverse regions. Patterns of genetic variations among 54 E. onukii samples unveil the population structure and evolutionary history across different tea-growing regions in China. Our results demonstrate that the genomic changes in key pathways, including those linked to metabolism, circadian rhythms, and immune system functions, may underlie the successful spread and adaptation of E. onukii. This work highlights the genetic and molecular basis underlying the evolutionary success of a species with broad economic impacts, and provides insights into insect adaptation to host plants, which will ultimately facilitate more sustainable pest management.

Vineyards are agroecosystems of great importance in the Niagara Region, Ontario (Canada). Due to its microclimate, this region is projected to be impacted by climate change with temperature increases, changes in precipitation patterns in all seasons, and greater frequency of extreme weather events. The aim of this review paper is to summarize which seasonal changes are expected to occur in the Niagara Region and assess how such changes are likely to affect the main components of the vineyard ecosystem (i.e., soil, vines, invertebrates, and pathogens). It is expected that by 2080 the region will experience an increase in temperature in all four seasons; an increase in precipitation during the fall, winter, and spring; and a decrease in precipitation during summer months. Impacts of the projected changes will likely lead to vine water stress, yield loss, increases in incidents of diseases, increases in the spread of new pests, and changes in grape quality ultimately resulting in lower wine quality and/or production. Current management practices will need to be better understood and adaptive strategies introduced to enhance grape growers’ ability to minimize these impacts.

Communities in coastal areas of Canada, including the Great Lakes, face a number of challenges, including increased water level variability and extreme weather events, causing flooding and localized erosion. To effectively respond to these coastal risks requires structured, deliberative approaches with the aim of fostering resilience and contributing to sustainability. A collaborative engagement process was used to explore community challenges. This included a project launch, key informant interviews, meetings, focus groups (agriculture, tourism, youth), and on-line methods (shoreline residents). Participatory social network analysis and theory of change were used for overall sense-making. As a result, community members identified six impact pathways moving forward with climate action: partnerships and collaboration; strategic engagement; water and watersheds; ecosystem-based adaptation; shoreline protection; and education. These themes are consistent with current theory on sustainability and theory of change development.

Choriogenesis is the last step of insect oogenesis, a process by which the chorion polypeptides are produced by the follicular cells and deposited on the surface of oocytes in order to provide a highly specialized protective barrier to the embryo. The essential features of chorion genes have yet to be clearly understood in the diamondback moth, Plutellaxylostella, a worldwide Lepidoptera pest attacking cruciferous crops and wild plants. In this study, complete sequences for 15 putative chorion genes were identified, and grouped into A and B classes. Phylogenetic analysis revealed that both classes were highly conserved and within each, branches are also species-specific. Chorion genes from each class were located in pairs on scaffolds of the P. xylostella genome, some of which shared the common promoter regulatory region. All chorion genes were highly specifically expressed in the P. xylostella adult females, mostly in the ovary with full yolk, which is a crucial period to build the shells of the eggs. RNAi-based knockdown of chorion-1, which is located on the Px_scaffold 6 alone, although had no effect on yolk deposition, resulted in smaller eggs and sharply reduced hatchability. Additionally, inhibition of PxCho-1 expression caused a less dense arrangement of the columnar layers, reduced exochorion roughness and shorter microvilli. Our study provides the foundation for exploring molecular mechanisms of female reproduction in P. xylostella, and for making use of chorion genes as the potential genetic-based molecular target to better control this economically important pest.

The diamondback moth is a cosmopolitan pest of significant economic importance. Here the authors analyse globally distributed genomic data to find evidence of climate-associated adaptive variation, and use an ecogenetic index to predict that it will maintain a global pest status under climate change.

Abstract Repeatability is the cornerstone of science, and it is particularly important for systematic reviews. However, little is known on how researchers’ choice of database, and search platform influence the repeatability of systematic reviews. Here, we aim to unveil how the computer environment and the location where the search was initiated from influence hit results. We present a comparative analysis of time‐synchronized searches at different institutional locations in the world and evaluate the consistency of hits obtained within each of the search terms using different search platforms. We revealed a large variation among search platforms and showed that PubMed and Scopus returned consistent results to identical search strings from different locations. Google Scholar and Web of Science's Core Collection varied substantially both in the number of returned hits and in the list of individual articles depending on the search location and computing environment. Inconsistency in Web of Science results has most likely emerged from the different licensing packages at different institutions. To maintain scientific integrity and consistency, especially in systematic reviews, action is needed from both the scientific community and scientific search platforms to increase search consistency. Researchers are encouraged to report the search location and the databases used for systematic reviews, and database providers should make search algorithms transparent and revise access rules to titles behind paywalls. Additional options for increasing the repeatability and transparency of systematic reviews are storing both search metadata and hit results in open repositories and using Application Programming Interfaces (APIs) to retrieve standardized, machine‐readable search metadata.

Genetic makeup of insect pest is informative for source-sink dynamics, spreading of insecticide resistant genes, and effective management. However, collecting samples from field populations without considering temporal resolution and calculating parameters related to historical gene flow may not capture contemporary genetic pattern and metapopulation dynamics of highly dispersive pests. Plutella xylostella (L.), the most widely distributed Lepidopteran pest that developed resistance to almost all current insecticides, migrates heterogeneously across space and time. To investigate its real-time genetic pattern and dynamics, we executed four samplings over two consecutive years across Southern China and Southeast Asia, and constructed population network based on contemporary gene flow. Across 48 populations, genetic structure analysis identified two differentiated insect swarms, of which the one with higher genetic variation was replaced by the other over time. We further inferred gene flow by estimation of kinship relationship and constructed migration network in each sampling time. Interestingly, we found mean migration distance at around 1,000 km. Such distance might have contributed to the formation of step-stone migration and migration circuit over large geographical scale. Probing network clustering across sampling times, we found a dynamic P. xylostella metapopulation with more active migration in spring than in winter, and identified a consistent pattern that some regions are sources (e.g., Yunnan in China, Myanmar and Vietnam) while several others are sinks (e.g., Guangdong and Fujian in China) over 2 years. Rapid turnover of insect swarms and highly dynamic metapopulation highlight the importance of temporal sampling and network analysis in investigation of source-sink relationships and thus effective pest management of P. xylostella, and other highly dispersive insect pests.

Many barriers to behavioural change exist when it comes to climate change action. A key element to overcoming some of these barriers is effective communication of complex scientific information. The use of visualizations, such as photographs or interactive maps, can increase knowledge dissemination, helping community members understand climatic and environmental changes. These techniques have been utilized in many disciplines but have not been widely embraced by climate change scholars. This paper discusses the utility of climate change data visualization as a tool for climate change knowledge mobilization. This paper draws on the case studying drivers of coastline change of Lake Ontario in the Town of Lincoln, Ontario, Canada. Historical aerial photographs were used to measure the rate of coastline change and visualize vulnerable sections of the coast. To better visualize the changes that occurred over time from a resident viewpoint, selected land-based historical photographs were replicated by taking new photographs at the same locations. These visualization tools can be useful to support the community in developing strategies to adapt to climate change by increasing understanding of the changes and knowledge through social learning. These tools can be generalized to other case studies dealing with community engagement in coastal adaptation efforts.

The diamondback moth, Plutella xylostella is a cosmopolitan agricultural pest. By analyzing 532 genomes from 114 populations around the world, the authors find evidence supporting a South American origin of this moth, and look for signatures of positive selection.

Although traditional agriculture carried out by ethnic groups is considered for its high biodiversity and important for food security and sovereignty, few studies have investigated the potential of these systems in the interest of promoting a sustainable agricultural development policy according to United Nations Sustainable Development Goals. Using the FAO's Sustainability Assessment of Food and Agriculture (SAFA) methodology, this study analyzed the sustainability of four traditional agricultural systems, three indigenous (Waorani, Shuar, and Kichwa) and one migrant settler populations in the Yasuní Biosphere Reserve (YBR) and identified synergies and trade-offs among the dimensions of sustainability. The results showed different dynamics in all dimensions of sustainability-specifically, trade-offs in the dimensions of good governance with environmental integrity and social well-being, economic resilience, and social well-being. It was identified that the differences in terms of sustainability are narrowing between the indigenous Shuar people's traditional agricultural systems and those of migrant settlers, which provides policymakers with specific information to design sustainable development policies and rescue traditional agricultural systems in the Amazon region.

Vitelline membrane proteins (VMPs) are the main proteins that form the inner shell (vitelline membrane layer) of insect eggs and are an integral part of egg formation and embryo development. Here, we characterized the molecular structure and expression patterns of the VMP26 gene and analyzed its reproductive functions in diamondback moth, Plutella xylostella (L.), a worldwide migratory pest of cruciferous plants. The PxVMP26 gene was shown to be a single exon gene that contained an open reading frame of 852 base pairs (bp) encoding 283 amino acids. Both qPCR and western blot analyses showed that PxVMP26 was specifically expressed in female adults and was significantly highly expressed in the ovary. Further anatomical analysis indicated that the expression level of PxVMP26 in the ovarian tube with an incomplete yolk was significantly higher than that in the ovarian tube with a complete yolk. CRISPR/Cas9-induced PxVMP26 knockout successfully created two homozygous strains with 8- and 46-bp frameshift mutations. The expression deficiency of the PxVMP26 protein was detected in the mutant strains using immunofluorescence and western blot. No significant difference was found in the number of eggs laid within three days between wild and mutant individuals, but there was a lower egg hatchability. The loss of the PxVMP26 gene changed the mean egg size, damaged the structure of the vitelline membrane, and increased the proportion of abnormal eggs due to water loss, resulting in egg collapse. This first analysis of the roles of the VMP gene in the oocyte formation and embryonic development of P. xylostella, using CRISPR/Cas9 technology, provides a basis for screening new genetic control targets of P. xylostella.

Abstract Invasive species may change the life history strategies, distribution, genetic configuration and trophic interactions of native species. The diamondback moth, Plutella xylostella L., is an invasive herbivore attacking cultivated and wild brassica plants worldwide. Here we present phylogeographic analyses of P. xylostella and one of its major parasitoids, Cotesia vestalis, using mitochondrial markers, revealing the genetic diversity and evolutionary history of these two species. We find evidence that C. vestalis originated in Southwest China, then adapted to P. xylostella as a new host by ecological sorting as P. xylostella expanded its geographic range into this region. Associated with the expansion of P. xylostella, Wolbachia symbionts were introduced into local populations of the parasitoid through horizontal transfer from its newly associated host. Insights into the evolutionary history and phylogeographic system of the herbivore and its parasitoid provide an important basis for better understanding the impacts of biological invasion on genetic configuration of local species.

Abstract Background The diamondback moth (DBM), Plutella xylostella (L.), is a major pest of cruciferous crops worldwide. While the species has become a model for genomics, post-transcriptional mechanisms associated with development and sex determination have not been comprehensively studied and the lack of complete structure of mRNA transcripts limits further research. Results Here, we combined the methods of single-molecule long-read sequencing technology (IsoSeq) and RNA-seq to re-annotate the published DBM genome and present the genome-wide identification of alternative splicing (AS) associated with development and sex determination of DBM. In total, we identified ~ 13,900 genes (~ 77%) annotated in the DBM genome (version-2), resulting in the correction of 1586 wrongly annotated genes and identification of 78,000 previously unannotated transcripts. We also identified 1804 genes showing alternative splicing (AS) in each of the developmental stages and sexes, suggesting that AS events are ubiquitous in DBM. Comparative analyses showed that these AS events were rarely shared among developmental stages, indicating that they may play key specific roles in regulation of insect development. Further, we found 156 genes showing different AS events and expression patterns between males and females, linking them to potential functions in sex determination. Conclusion Overall, the P. xylostella transcriptome provides the significant information about regulatory alternative splicing events, which are shown to be involved in development and sex determination. Our work presents a solid foundation to better understand the mechanism of post-transcriptional regulation, and offers wider insights into insect development and sex determination.

An introduction to the special issue's book reviews

An introduction from the guest editors

The diamondback moth, Plutella xylostella, is a cosmopolitan pest and the first species to develop field resistance to toxins from the gram-positive bacterium Bacillus thuringiensis (Bt). Although previous work has suggested that mutations of ATP-binding cassette transporter subfamily C2 (ABCC2) or C3 (ABCC3) genes can confer Cry1Ac resistance, here we reveal that P. xylostella requires combined mutations in both PxABCC2 and PxABCC3 to achieve high-level Cry1Ac resistance, rather than simply a mutation of either gene. We identified natural mutations of PxABCC2 and PxABCC3 that concurrently occurred in a Cry1Ac-resistant strain (Cry1S1000) of P. xylostella, with a mutation (RA2) causing the mis-splicing of PxABCC2 and another mutation (RA3) leading to the premature termination of PxABCC3. Genetic linkage analysis showed that RA2 and RA3 were tightly linked to Cry1Ac resistance. Introgression of RA2 and RA3 enabled a susceptible strain (G88) of P. xylostella to obtain high resistance to Cry1Ac, confirming that these genes confer resistance. To further support the role of PxABCC2 and PxABCC3 in Cry1Ac resistance, frameshift mutations were introduced into PxABCC2 and PxABCC3 singly and in combination in the G88 strain with CRISPR/Cas9 mediated mutagenesis. Bioassays of CRISPR-based mutant strains, plus genetic complementation tests, demonstrated that the deletion of PxABCC2 or PxABCC3 alone provided < 4-fold tolerance to Cry1Ac, while disruption of both genes together conferred >8,000-fold resistance to Cry1Ac, suggesting the redundant/complementary roles of PxABCC2 and PxABCC3. This work advances our understanding of Bt resistance in P. xylostella by demonstrating mutations within both PxABCC2 and PxABCC3 genes are required for high-level Cry1Ac resistance.

The vitellogenin receptor (VgR) belongs to the low-density lipoprotein receptor (LDLR) gene superfamily and plays an indispensable role in Vg transport, yolk deposition, and oocyte development. For this reason, it has become a promising target for pest control. The involvement of VgR in Vg transport and reproductive functions remains unclear in diamondback moths, Plutella xylostella (L.), a destructive pest of cruciferous crops. Here, we cloned and identified the complete cDNA sequence of P. xylostella VgR, which encoded 1805 amino acid residues and contained four conserved domains of LDLR superfamily. PxVgR was mainly expressed in female adults, more specifically in the ovary. PxVgR protein also showed the similar expression profile with the PxVgR transcript. CRISPR/Cas9-mediated PxVgR knockout created a homozygous mutant of P. xylostella with 5-bp-nucleotide deletion in the PxVgR. The expression deficiency of PxVgR protein was detected in the ovaries and eggs of mutant individuals. Vg protein was still detected in the eggs of the mutant individuals, but with a decreased expression level. However, PxVg transcripts were not significantly affected by the PxVgR knockout. Knockout of PxVgR resulted in shorter ovarioles of newly emerged females. No significant difference was detected between wild and mutant individuals in terms of the number of eggs laid in the first 3 days after mating. The loss of PxVgR gene resulted in smaller and whiter eggs and lower egg hatching rate. This study represents the first report on the functions of VgR in Vg transport, ovary development, oviposition, and embryonic development of P. xylostella using CRISPR/Cas9 technology. This study lays the foundation for understanding molecular mechanisms of P. xylostella reproduction, and for making use of VgR as a potential genetic-based molecular target for better control of the P. xylostella.

Environmental and climatic changes are disproportionately felt in coastal communities, where drivers of coastline change are complicated with continued development. This study analyzed the coastline change of Lake Ontario in the Town of Lincoln, Ontario, Canada, using a mixed-methods two-phased approach that is novel to the study area. The first phase of the methodology included a coastline change analysis using historical aerial photographs in a geographic information system to identify the most vulnerable sections of the coastline. To better understand the calculated changes, the second phase explored the roles of select climatic and non-climatic drivers of coastline change, such as historic storms and land use changes. The results indicated that four main areas of Lincoln’s coast were more vulnerable, with rates of erosion between −0.32 and −0.66 m/yr between 1934 and 2018. Sections of coastline that had less erosion included those that were more heavily vegetated, attempted a cooperative protection approach, or utilized revetment stones in areas without steep banks. This methodology can help municipalities understand coastline change in a more holistic way to increase their adaptive capacity and allows for the creation of useful visualizations that better communicate to residents and town staff the level of vulnerability of their coasts.

Coastal communities are increasingly vulnerable to climate change and its effects may push coastal ecosystems to undergo irreversible changes. This is especially true for shorebirds with the loss of biodiversity and resource-rich habitats to rest, refuel, and breed. To protect these species, it is critical to conduct research related to nature-based Solutions (NbS). Through a scoping review of scientific literature, this paper initially identified 85 articles with various ecosystem-based adaptation (EbA) strategies that could help conserve shorebird populations and promote ecotourism. Of these 85 articles, 28 articles had EbA strategies that were examined, with some like coral reefs and mangroves eliminated as they were inappropriate for this region. The scoping review identified four major EbA strategies for the Greater Niagara Region with living shorelines and beach nourishment being the most suitable, especially when combined. These strategies were then evaluated against the eight core principles of nature-based solutions protecting shorebird as well as human wellbeing. Living shoreline strategy was the only one that met all eight NbS principles. As the coastline of the region greatly varies in substrate and development, further research will be needed to decide which EbA strategies would be appropriate for each specific area to ensure their efficacy.

Abstract Background Structural variation among genomes is now viewed to be as important as single nucleoid polymorphisms in influencing the phenotype and evolution of a species. Segmental duplication (SD) is defined as segments of DNA with homologous sequence. Results Here, we performed a systematic analysis of segmental duplications (SDs) among five lepidopteran reference genomes (Plutella xylostella, Danaus plexippus, Bombyx mori, Manduca sexta and Heliconius melpomene) to understand their potential impact on the evolution of these species. We find that the SDs content differed substantially among species, ranging from 1.2% of the genome in B. mori to 15.2% in H. melpomene. Most SDs formed very high identity (similarity higher than 90%) blocks but had very few large blocks. Comparative analysis showed that most of the SDs arose after the divergence of each linage and we found that P. xylostella and H. melpomene showed more duplications than other species, suggesting they might be able to tolerate extensive levels of variation in their genomes. Conserved ancestral and species specific SD events were assessed, revealing multiple examples of the gain, loss or maintenance of SDs over time. SDs content analysis showed that most of the genes embedded in SDs regions belonged to species-specific SDs (“Unique” SDs). Functional analysis of these genes suggested their potential roles in the lineage-specific evolution. SDs and flanking regions often contained transposable elements (TEs) and this association suggested some involvement in SDs formation. Further studies on comparison of gene expression level between SDs and non-SDs showed that the expression level of genes embedded in SDs was significantly lower, suggesting that structure changes in the genomes are involved in gene expression differences in species. Conclusions The results showed that most of the SDs were “unique SDs”, which originated after species formation. Functional analysis suggested that SDs might play different roles in different species. Our results provide a valuable resource beyond the genetic mutation to explore the genome structure for future Lepidoptera research.

Ecdysteroids play an essential role in controlling insect development and reproduction. Their pathway is regulated by a group of enzymes called Halloween gene proteins. The relationship between the Halloween genes and ecdysteroid synthesis has yet to be clearly understood in diamondback moth, Plutella xylostella (L.), a worldwide Lepidoptera pest attacking cruciferous crops and wild plants. In this study, complete sequences for six Halloween genes, neverland (nvd), shroud (sro), spook (spo), phantom (phm), disembodied (dib), shadow (sad), and shade (shd), were identified. Phylogenetic analysis revealed a strong conservation in insects, including Halloween genes of P. xylostella that was clustered with all other Lepidoptera species. Three Halloween genes, dib, sad, and shd were highly expressed in the adult stage, while nvd and spo were highly expressed in the egg and pupal stages, respectively. Five Halloween genes were highly expressed specifically in the prothorax, which is the major site of ecdysone production. However, shd was expressed predominantly in the fat body to convert ecdysone into 20-hydroxyecdysone. RNAi-based knockdown of sad, which is involved in the last step of ecdysone biosynthesis, significantly reduced the 20E titer and resulted in a longer developmental duration and lower pupation of fourth-instar larvae, as well as caused shorter ovarioles and fewer fully developed eggs of P. xylostella. Furthermore, after the knockdown of sad, the expression levels of Vg and VgR genes were significantly decreased by 77.1 and 53.0%. Meanwhile, the number of eggs laid after 3 days was significantly reduced in sad knockdown females. These results suggest that Halloween genes may play a critical role in the biosynthesis of ecdysteroids and be involved in the development and reproduction of P. xylostella. Our work provides a solid basis for understanding the functional importance of these genes, which will help to screening potential genes for pest management of P. xylostella.

The establishment of an expression quantification system that can be easily applied for the comparison of microRNAs (miRNAs) from biological samples is an important step toward understanding functional mechanisms in organisms. However, there is lack of attention on the selection of reference genes for miRNA expression profiling in insect herbivores. Here, we explored the candidate reference genes in a notorious pest of cruciferous crops, Plutella xylostella, for normalization of miRNA expression in developmental stages and tissues and in response to a change of food source from artificial diet to host plant Arabidopsis thaliana. We first compared the expression levels and stability of eight small RNAs using qRT-PCR, and found that miR11 was the most suitable reference gene for expression quantification of the miRNAs. We then confirmed this finding using digital droplet PCR and further validated with a well-studied cross-kingdom miRNA derived from A. thaliana (ath-miR159a). However, none of the reference genes was applicable for all experimental conditions, and multiple reference genes were sometimes required within the same experiment. Our work provides a method for the selection of reference genes for quantification of plant-derived miRNAs, which paves the way for unveiling their roles in the insect-plant coevolution.

In Chinese tea plantations, yellow sticky cards and light traps are increasingly used to control insect pests, especially the tea green leafhopper Empoasca onukii. In this study, a 16-week open-field experiment with daily weather monitoring was designed to test the responses of tea green leafhopper, parasitoids and spiders to yellow sticky cards and three light traps with different wavelengths (covered with sticky cards). An exclosure experiment was also designed to further test the influence of the three light systems (without sticky card) on the same species. The results showed that all three light emitting diode (LED) light traps (white, green and yellow) and yellow sticky cards attracted many more E. onukii male adults than females during the course of the open field experiment, with less than 25% of trapped adults being females. Parasitoids and spiders were also attracted by these systems. Weather variables, especially rainfall, influenced the trapping efficiency. In the exclosure experiment, the population of leafhoppers in the yellow sticky card treatment did not decline significantly, but the number of spiders significantly decreased. The green and white light treatments without sticky cards showed a significant control of E. onukii and no obvious harm to spiders. These results suggest that yellow sticky cards and light traps have limited capacity to control tea green leafhoppers. However, light, especially green light, may be a promising population control measure for tea green leafhoppers, not as killing agents in the traps, but rather as a behavioral control system.

Evolutionary and ecological forces are important factors that shape gut microbial profiles in hosts, which can help insects adapt to different environments through modulating their metabolites. However, little is known about how gut microbes and metabolites are altered when lepidopteran pest species switch hosts. In the present study, using 16S-rDNA sequencing and mass spectrometry-based metabolomics, we analyzed the gut microbiota and metabolites of three populations of Plutella xylostella: one feeding on radish (PxR) and two feeding on peas (PxP; with PxP-1 and PxP-17 being the first and 17th generations after host shift from radish to peas, respectively). We found that the diversity of gut microbes in PxP-17 was significantly lower than those in PxR and PxP-1, which indicates a distinct change in gut microbiota after host shift. Kyoto Encyclopedia of Genes and Genomes analysis revealed that the functions of energy metabolism, signal transduction, and xenobiotics biodegradation and metabolism were increased in PxP-17, suggesting their potential roles in host adaptation. Metabolic profiling showed a significant difference in the abundance of gut metabolites between PxR and PxP-17, and significant correlations of gut bacteria with gut metabolites. These findings shed light on the interaction among plants, herbivores, and symbionts, and advance our understanding of host adaptation associated with gut bacteria and metabolic activities in P. xylostella.

Amaranthus hybridus is a noxious weed in Ontario, with demonstrated allelopathic properties that can lead to decreased agricultural production. We tested the germination and growth of five cover crop species exposed to A. hybridus extracts, and to dried or fresh materials in soil. A germination index was calculated, and the dry weight of plant organs were measured to quantify responses to treatments. All species had reduced germination (≤29%) in 100% extract. Trifolium pratense had significant root weight reductions in extract (52%) and dried (72%) treatments, whereas shoot weight only decreased (48%) in dried treatment. Medicago sativa shoot weight decreased (52%) in 20g fresh treatment, while root weight decreased (62%) in dried treatment. Shoot weight of Raphanus sativus increased (32%) at mid-extract concentrations, while root weight increased (33%) only with dried treatment; however, both its shoot and root weight decreased (>40%) in fresh treatment. Only the shoot weight of Lolium multiflorum increased (41% in 75% extract and 55% in dried treatment). Both Cichorium intybus shoot and root weights decreased (~50%) in fresh treatment. Crop responses to A. hybridus are complex, and material and species-dependant. Further testing in the field may provide a more comprehensive understanding of how to improve the management of A. hybridus.

Tea plantations have used many synthetic chemicals to ensure performance and control of pests. This has led to increased contamination of soils and reduced tea growth. We assessed the levels of heavy metals, including Cd, Cr, Pb, Cu, Ni, Zn, Hg, As, and pesticide residues, such as HCHs, biphenyl chrysanthemum ester, methamidophos, imidacloprid, permethrin, in the soil of tea plantations of Taiwan, Tibet, Guangdong, and Fujian. The Potential Ecological Risk Index and the Nemerow comprehensive pollution index were used to analyze the data. The results showed that risk indices in Tibet, Guangdong and Fuzhou were considered as moderate ecological harm level. Ecological risk assessment index of Anxi organic and Anxi conventional tea gardens suggested a “low” risk level. The Nemerow comprehensive pollution indices for soil pesticide residues in the tea plantations of Taiwan, Tibet, Anxi organic and Anxi conventional were considered mild. Guangdong and Fuzhou had values suggesting “slight pollution” levels. According to National Soil Environmental Quality Standard (GB15618-1995), soil in tea plantations in Taiwan, Tibet, and Anxi conventional matched the national first grade of soil quality and those from Guangdong, Fuzhou, and Anxi organic tea garden matched the national second grade.

The development of insecticide resistance in insect pests is a worldwide concern and elucidating the underlying mechanisms is critical for effective crop protection. Recent studies have indicated potential links between insect gut microbiota and insecticide resistance and these may apply to the diamondback moth, Plutella xylostella (L.), a globally and economically important pest of cruciferous crops. We isolated Enterococcus sp. (Firmicutes), Enterobacter sp. (Proteobacteria), and Serratia sp. (Proteobacteria) from the guts of P. xylostella and analyzed the effects on, and underlying mechanisms of insecticide resistance. Enterococcus sp. enhanced resistance to the widely used insecticide, chlorpyrifos, in P. xylostella, while in contrast, Serratia sp. decreased resistance and Enterobacter sp. and all strains of heat-killed bacteria had no effect. Importantly, the direct degradation of chlorpyrifos in vitro was consistent among the three strains of bacteria. We found that Enterococcus sp., vitamin C, and acetylsalicylic acid enhanced insecticide resistance in P. xylostella and had similar effects on expression of P. xylostella antimicrobial peptides. Expression of cecropin was down-regulated by the two compounds, while gloverin was up-regulated. Bacteria that were not associated with insecticide resistance induced contrasting gene expression profiles to Enterococcus sp. and the compounds. Our studies confirmed that gut bacteria play an important role in P. xylostella insecticide resistance, but the main mechanism is not direct detoxification of insecticides by gut bacteria. We also suggest that the influence of gut bacteria on insecticide resistance may depend on effects on the immune system. Our work advances understanding of the evolution of insecticide resistance in this key pest and highlights directions for research into insecticide resistance in other insect pest species.

Herbivore specialists adapt to feed on a specific group of host plants by evolving various mechanisms to respond to plant defenses. Insects also possess complex gut microbiotas but their potential role in adaptation is poorly understood. Our previous study of the genome of diamondback moth, Plutella xylostella, revealed an intrinsic capacity to detoxify plant defense compounds, which is an important factor in its success as a pest. Here we expand on that work with a complete taxonomic and functional profile of the P. xylostella gut microbiota obtained by metagenomic sequencing. Gene enrichment in the metagenome, accompanied by functional identification, revealed an important role of specific gut bacteria in the breakdown of plant cell walls, detoxification of plant phenolics, and synthesis of amino acids. Microbes participating in these pathways mainly belonged to three highly abundant bacteria: Enterobacter cloacae, Enterobacter asburiae, and Carnobacterium maltaromaticum. Results show that while the gut microbial community may be complex, a small number of functionally active species can be disproportionally important. The presence of specific enzymes in the microbiota community, such as supporting amino acid synthesis, digestion and detoxification functions, demonstrates the beneficial interactions between P. xylostella and its gut microbiota. These interactions can be potential targets for manipulation to provide novel pest management approaches.

Insect behaviors are often influenced by light conditions including photoperiod, light intensity, and wavelength. Understanding pest insect responses to changing light conditions may help with developing alternative strategies for pest control. Little is known about the behavioral responses of leafhoppers (Hemiptera: Cicadellidae) to light conditions. The behavior of the tea green leafhopper, Empoasca onukii Matsuda, was examined when exposed to different light photoperiods or wavelengths. Observations included the frequency of locomotion and cleaning activities, and the duration of time spent searching. The results suggested that under normal photoperiod both female and male adults were generally more active in darkness (i.e., at night) than in light. In continuous darkness (DD), the locomotion and cleaning events in Period 1 (7:00-19:00) were significantly increased, when compared to the leafhoppers under normal photoperiod (LD). Leafhoppers, especially females, changed their behavioral patterns to a two day cycle under DD. Under continuous illumination (continuous quartz lamp light, yellow light at night, and green light at night), the activities of locomotion, cleaning, and searching were significantly suppressed during the night (19:00-7:00) and locomotion activities of both females and males were significantly increased during the day (7:00-19:00), suggesting a shift in circadian rhythm. Our work suggests that changes in light conditions, including photoperiod and wavelength, can influence behavioral activities of leafhoppers, potentially affecting other life history traits such as reproduction and development, and may serve as a method for leafhopper behavioral control.

Isaria cateniannulata is a very important and virulent entomopathogenic fungus that infects many insect pest species. Although I. cateniannulata is commonly exposed to extreme environmental temperature conditions, little is known about its molecular response mechanism to temperature stress. Here, we sequenced and de novo assembled the transcriptome of I. cateniannulata in response to high and low temperature stresses using Illumina RNA-Seq technology. Our assembly encompassed 17,514 unigenes (mean length = 1,197 bp), in which 11,445 unigenes (65.34%) showed significant similarities to known sequences in NCBI non-redundant protein sequences (Nr) database. Using digital gene expression analysis, 4,483 differentially expressed genes (DEGs) were identified after heat treatment, including 2,905 up-regulated genes and 1,578 down-regulated genes. Under cold stress, 1,927 DEGs were identified, including 1,245 up-regulated genes and 682 down-regulated genes. The expression patterns of 18 randomly selected candidate DEGs resulting from quantitative real-time PCR (qRT-PCR) were consistent with their transcriptome analysis results. Although DEGs were involved in many pathways, we focused on the genes that were involved in endocytosis: In heat stress, the pathway of clathrin-dependent endocytosis (CDE) was active; however at low temperature stresses, the pathway of clathrin-independent endocytosis (CIE) was active. Besides, four categories of DEGs acting as temperature sensors were observed, including cell-wall-major-components-metabolism-related (CWMCMR) genes, heat shock protein (Hsp) genes, intracellular-compatible-solutes-metabolism-related (ICSMR) genes and glutathione S-transferase (GST). These results enhance our understanding of the molecular mechanisms of I. cateniannulata in response to temperature stresses and provide a valuable resource for the future investigations.

Plant microRNAs (miRNAs) have recently been reported to be involved in the cross-kingdom regulation of specific cellular and physiological processes in animals. However, little of this phenomenon is known for the communication between host plant and insect herbivore. In this study, the plant-derived miRNAs in the hemolymph of a cruciferous specialist Plutella xylostella were identified by small RNAs sequencing. A total of 39 miRNAs with typical characteristics of plant miRNAs were detected, of which 24 had read counts ≥ 2 in each library. Three plant-derived miRNAs with the highest read counts were validated, and all of them were predicted to target the hemocyanin domains-containing genes of P. xylostella. The luciferase assays in the Drosophila S2 cell demonstrated that miR159a and novel-7703-5p could target BJHSP1 and PPO2 respectively, possibly in an incomplete complementary pairing mode. We further found that treatment with agomir-7703-5p significantly influenced the pupal development and egg-hatching rate when reared on the artificial diet. The developments of both pupae and adults were severely affected upon their transfer to Arabidopsis thaliana, but this might be independent of the cross-kingdom regulation of the three plant-derived miRNAs on their target genes in P. xylostella, based on expression analysis. Taken together, our work reveals that the plant-derived miRNAs could break the barrier of the insect mid-gut to enter the circulatory system, and potentially regulate the development of P. xylostella. Our findings provide new insights into the co-evolution of insect herbivore and host plant, and novel direction for pest control using plant-derived miRNAs.

Argonaute (Ago) protein family plays a key role in the RNA interference (RNAi) process in different insects including Lepidopteran. However, the role of Ago proteins in the RNAi pathway of Plutella xylostella is still unknown. We cloned an Argonaute3 gene in P. xylostella (PxAgo3) with the complete coding sequence of 2832 bp. The encoded protein had 935 amino acids with an expected molecular weight of 108.9 kDa and an isoelectric point of 9.29. It contained a PAZ (PIWI/Argonaute/Zwile) domain and PIWI (P-element-induced whimpy testes) domain. PxAgo3 was classified into the Piwi subfamily of Ago proteins with a high similarity of 93.0% with Bombyx mori Ago3 (BmAgo3). The suppression of PxAgo3 by dsPxAgo3 was observed 3 h after treatment and was maintained until 24 h. Knockdown of PxAgo3 decreased the suppression level of PxActin by dsPxActin in P. xylostella cells, while overexpression of PxAgo3 increased the RNAi efficiency. Our results suggest that PxAgo3 play a key role in the double stranded RNA (dsRNA)-regulated RNAi pathway in P. xylostella.

BackgroundInsect midgut microbiota is important in host nutrition, development and immune response. Recent studies indicate possible links between insect gut microbiota and resistance to biological and chemical toxins. Studies of this phenomenon and symbionts in general have been hampered by difficulties in culture-based approach. In the present study, DNA sequencing was used to examine the midgut microbiota of diamondback moth (DBM), Plutella xylostella (L.), a destructive pest that attacks cruciferous crops worldwide. Its ability to develop resistance to many types of synthetic insecticide and even Bacillus thuringiensis toxins makes it an important species to study.Methodology/principal findingsBacteria of the DBM larval midgut in a susceptible and two insecticide (chlorpyrifos and fipronil) resistant lines were examined by Illumina sequencing sampled from an insect generation that was not exposed to insecticide. This revealed that more than 97% of the bacteria were from three orders: Enterobacteriales, Vibrionales and Lactobacillales. Both insecticide-resistant lines had more Lactobacillales and the much scarcer taxa Pseudomonadales and Xanthomonadales with fewer Enterobacteriales compared with the susceptible strain. Consistent with this, a second study observed an increase in the proportion of Lactobacillales in the midgut of DBM individuals from a generation treated with insecticides.Conclusions/significanceThis is the first report of high-throughput DNA sequencing of the entire microbiota of DBM. It reveals differences related to inter- and intra-generational exposure to insecticides. Differences in the midgut microbiota among susceptible and insecticide-resistant lines are independent of insecticide exposure in the sampled generations. While this is consistent with the hypothesis that Lactobacillales or other scarcer taxa play a role in conferring DBM insecticide resistance, further studies are necessary to rule out other possibilities. Findings constitute the basis for future molecular work on the functions of insect midgut microbiota taxa and their possible role in conferring host resistance to toxins.

Water has often been the source of crises and their frequency will intensify due to climate change impacts. The Niagara River Watershed provides an ideal case to study water crises as it is an international transboundary system (Canada-United States) and has both historical and current challenges associated with water quantity and quality, which resonates broadly in water basins throughout the world. The aim of this study was to understand how stakeholders perceive ecosystems and the relationship with preferences for governance approaches in the context of water governance. An online survey instrument was employed to assess perceptions of the system in terms of resilience (engineering, ecological, social-ecological, or epistemic), preferences for governance approaches (state, citizen, market, and hybrid forms), and the most pressing issues in the watershed. Responses showed that, despite demographic differences and adherence to different resilience perspectives, support was strongest for governance approaches that focused on state or state-citizen hybrid forms. The validity of the resilience typology as a grouping variable is discussed. The roles of institutional constraints, pragmatism in governance approach preferences, and the influence of multiple crises are explored in relation to the context of the study site, as well as to water governance scholarship more broadly.

A shift appears to be occurring in thinking about flooding, from a resistance-based approach to one of resilience. Accordingly, how stakeholders in flood-prone regions perceive the system and its governance are salient questions. This study queried stakeholders’ internal representations of ecosystems (resistance- or resilience-based), preferences for governance actors and mechanisms for flooding, and the relationship between them in five different regions of the world. The influence of personal experience on these variables was also assessed. Most respondents aligned themselves with a resilience-based approach in relation to system connectedness and response to disturbance; however, respondents were almost evenly split between resistance- and resilience-based approaches when considering system management. Responses generally were considered to hold for other disturbances as well. There was no clear relationship between internal representations and preferences for governance actors or mechanisms. Respondents generally favoured actor combinations that included governments and mechanism combinations that included regulations and policies. Those who had personal experience with flooding tended to align themselves with a resilience-based internal representation of system management, but personal experience showed no clear relationship with governance preferences. The findings support an evolutionary perspective of flood management where emerging paradigms enhance preceding ones, and prompt a critical discussion about the universality of resilience as a framing construct.

This paper aims to present an approach that was developed as part of the Community-University Alliance Research (CURA) entitled “Coastal Communities Challenges of the Estuary and the Gulf of St. Lawrence under a Changing Climate” (CURA-CCC). We describe the process of capacity building we used in several coastal communities in Eastern Canada to enhance their resilience. We use the examples of the municipalities of Maria and Bonaventure in Quebec to illustrate this approach, which is based on participatory action research. The co-construction of adaptation strategies that emerged from this work is based on an exploratory model examining the hazards and their effects on these communities, considering the uncertainty of knowledge and capacities of the actors to act. These coastal habitats are subject to climate change hazards including storm surges causing coastal erosion, floods and river overflows. It is important for these communities to develop a better understanding of these issues when developing long-term collective solutions that can go beyond their local territories. In fact, we believe that such an approach could potentially be used in several other coastal rural areas of eastern Canada.