Detection and identification of Bogia coconut syndrome phytoplasma from seed-associated tissues and seedlings of coconut (Cocos nucifera) and betel nut (Areca catechu).

Evidence for seed transmission of phytoplasmas has grown in several pathosystems including coconut (Cocos nucifera). Bogia coconut syndrome (BCS) is a disease associated with the lethal yellowing syndrome associated with the presence of 'Candidatus Phytoplasma noviguineense' that affects coconut, betel nut (Areca catechu) and bananas (Musa spp.) in Papua New Guinea. Coconut and betel nut drupes were sampled from BCS-infected areas in Papua New Guinea, dissected, the extracted nucleic acid was used in polymerase chain reaction (PCR), and loop mediated isothermal amplification (LAMP) used to check for presence of phytoplasma DNA. In a second study, drupes of both plant species were collected from multiple field sites and grown in insect-proof cages. Leaf samples taken at 6 months were also tested with PCR and LAMP. The studies of dissected coconut drupes detected phytoplasma DNA in several tissues including the embryo. Drupes from betel nut tested negative. Among the seedlings, evidence of possible seed transmission was found in both plant species. The results demonstrate the presence of 'Ca. P. noviguineense' in coconut drupes and seedlings, and in seedlings of betel nut; factors that need to be considered in ongoing management and containment efforts.

Thermal acclimation uncovers a simple genetic basis of adaptation to high temperature in a cosmopolitan pest.

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.

Harnessing artificial intelligence for analysing the impacts of nectar and pollen feeding in conservation biological control.

Plant-derived foods, such as nectar and pollen, have garnered substantial research attention due to their potential to support natural enemies of pests. This review is a pioneering exploration of the potential for artificial intelligence approaches to provide insights into the factors that drive the success of conservation biological control (CBC). Nectar and pollen were confirmed as key plant food resources for natural enemies. These have been widely used across differing crop systems and provided by a wide range of CBC interventions, such as field margin flower strips. The combined use of parasitoids and predators is revealed as more successful than either of these guilds alone. CBC success was greater in field crops than in vine and berry crops, whilst interventions using dicotyledonous species that produce nectar in addition to pollen were more successful than using grassy interventions.

Developmental Shifts in the Microbiome of a Cosmopolitan Pest: Unraveling the Role of Wolbachia and Dominant Bacteria.

Wolbachia bacteria (phylum Proteobacteria) are ubiquitous intracellular parasites of diverse invertebrates. In insects, coevolution has forged mutualistic associations with Wolbachia species, influencing reproduction, immunity, development, pathogen resistance, and overall fitness. However, the impact of Wolbachia on other microbial associates within the insect microbiome, which are crucial for host fitness, remains less explored. The diamondback moth (Plutella xylostella), a major pest of cruciferous vegetables worldwide, harbors the dominant Wolbachia strain plutWB1, known to distort its sex ratio. This study investigated the bacterial community diversity and dynamics across different developmental life stages and Wolbachia infection states in P. xylostella using high-throughput 16S rDNA amplicon sequencing. Proteobacteria and Firmicutes dominated the P. xylostella microbiome regardless of life stage or Wolbachia infection. However, the relative abundance of dominant genera, including an unclassified genus of Enterobacteriaceae, Wolbachia, Carnobacterium, and Delftia tsuruhatensis, displayed significant stage-specific variations. While significant differences in bacterial diversity and composition were observed across life stages, Wolbachia infection had no substantial impact on overall diversity. Nonetheless, relative abundances of specific genera differed between infection states. Notably, Wolbachia exhibited a stable, high relative abundance across all stages and negatively correlated with an unclassified genus of Enterobacteriaceae, Delftia tsuruhatensis, and Carnobacterium. Our findings provide a foundational understanding of the complex interplay between the host, Wolbachia, and the associated microbiome in P. xylostella, paving the way for a deeper understanding of their complex interactions and potential implications for pest control strategies.

Field evaluation of electrophysiologically-active dung volatiles as chemical lures for trapping of dung beetles.

Dung beetles are economically important beneficial insects that process dung. To locate this source, they use volatile organic compounds (VOCs). The objectives of the study were to evaluate the attractiveness of ten electrophysiologically-active dung volatiles (phenol, skatole, indole, p-cresol, butanone, butyric acid, eucalyptol, dimethyl sulphide, dimethyl disulphide, and toluene) to dung beetles in the field and to investigate how the composition of volatile blends influences efficacy as lures for use in traps. Six combinations of the compounds were compared with field collected cattle dung bait and a negative control, across three seasons. Both dung and synthetic baits captured all exotic dung beetle species present in the study area. A six-compound mix (M1), comprising major dung volatiles, served as an attractive chemical mixture. The addition of dimethyl sulphide, dimethyl disulphide (M2) and toluene (M4) enhanced attractancy of M1 for dung beetles, while eucalyptol (M3) decreased the attractancy. The degree of attraction by various dung beetle species to synthetic baits varied, but baits proved to be effective, especially for summer trapping. The trap design used in this study presented a convenient and practical way to sample dung beetle and other associated scarabs from open pastures. The attraction of introduced dung beetle species to synthetic baits is documented here for the first time in Australia. In addition, necrophagous Omorgus sp. is reported here for the first time to be attracted to synthetic baits. They showed a significant attraction to the mixture containing dimethyl sulphide and dimethyl disulphide (M2). The current study represents a promising first step towards formulating a synthetic chemical lure for dung beetles, offering a consistent, standardised, and bio-secure trapping method compared to use of naturally occurring dung baits, especially as a multi-species lure.

Gut bacteria mediated adaptation of diamondback moth, Plutella xylostella, to secondary metabolites of host plants.

IMPORTANCE: In 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.

Characterisation of Antennal Sensilla and Electroantennography Responses of the Dung Beetles Bubas bison, Onitis aygulus and Geotrupes spiniger (Coleoptera: Scarabaeoidea) to Dung Volatile Organic Compounds.

Locating sporadically distributed food resources and mate finding are strongly aided by volatile cues for most insects, including dung beetles. However, there is limited information on the olfactory ecology of dung beetles. We conducted a scanning electron microscopy study on the morphology and distribution of the antennal sensilla of three introduced dung beetle species in Australia: Geotrupes spiniger (Coleoptera: Geotrupidae), Bubas bison and Onitis aygulus (Coleoptera: Scarabaeidae). Three main morphological types of antennal sensilla were identified: sensilla trichodea (ST), sensilla basiconica (SB) and sensilla chaetica (SCh). Distinct variations of SB distribution were observed in B. bison and G. spiniger and on different lamellar surfaces in both sexes of all three species. Sexual dimorphism in antennal sensilla distribution or their abundance was not evident. To complement the morphological characterisation of sensilla, electroantennography (EAG) was carried out to construct EAG response profiles of the three species to selected dung volatiles. An initial study revealed that antennae of all species were sensitive to a mix of phenol, skatole, indole, p-cresol, butanone and butyric acid, common components of livestock dung headspace. In addition to these six compounds, dimethyl sulfide, dimethyl disulfide, eucalyptol and toluene were tested for antennal activity. All compounds evoked measurable EAG responses, confirming antennal sensitivity. Geotrupes spiniger exhibited significant responses to all the compounds compared to the control, whereas B. bison and O. aygulus only responded to a subset of compounds. A comparison of relative EAG amplitudes revealed highly significant responses to p-cresol in G. spiniger and to skatole in B. bison. Geotrupes spiniger displayed differential responses to all the compounds. Pooled EAG data suggest highly significant differences in responses among the three species and among compounds. Our findings suggest that a blend of volatiles may offer potential for the trapping of dung beetles, thereby avoiding the use of dung baits that are inconvenient, inconsistent and may pose a threat to farm biosecurity.

Gut microbiota assemblages of generalist predators are driven by local- and landscape-scale factors.

The gut microbiomes of arthropods have significant impact on key physiological functions such as nutrition, reproduction, behavior, and health. Spiders are diverse and numerically dominant predators in crop fields where they are potentially important regulators of pests. Harnessing spiders to control agricultural pests is likely to be supported by an understanding of their gut microbiomes, and the environmental drivers shaping microbiome assemblages. This study aimed to deciphering the gut microbiome assembly of these invertebrate predators and elucidating potential implications of key environmental constraints in this process. Here, we used high-throughput sequencing to examine for the first time how the assemblages of bacteria in the gut of spiders are shaped by environmental variables. Local drivers of microbiome composition were globally-relevant input use system (organic production vs. conventional practice), and crop identity (Chinese cabbage vs. cauliflower). Landscape-scale factors, proportion of forest and grassland, compositional diversity, and habitat edge density, also strongly affected gut microbiota. Specific bacterial taxa were enriched in gut of spiders sampled from different settings and seasons. These findings provide a comprehensive insight into composition and plasticity of spider gut microbiota. Understanding the temporal responses of specific microbiota could lead to innovative strategies development for boosting biological control services of predators.

A very long-chain fatty acid enzyme gene, PxHacd2 affects the temperature adaptability of a cosmopolitan insect by altering epidermal permeability.

Temperature fluctuations pose challenges to poikilotherms, such as insects, especially under climate change conditions. Very long-chain fatty acids (VLCFAs) form important structural components of membranes and epidermal surfaces, so play important roles in adaptation to temperature stress in plants. It has been unclear whether VLCFAs are involved in epidermis formation and thermal resistance in insects. In this study, we focused on the 3-hydroxy acyl-CoA dehydratase 2 (Hacd2), an important enzyme in the synthesis pathway of VLCFAs, in a cosmopolitan pest, the diamondback moth, Plutella xylostella. Hacd2 was cloned from P. xylostella and the relative expression pattern was identified. Epidermal permeability increased with the decreased VLCFAs in the Hacd2-deficient P. xylostella strain, which was constructed by using the CRISPR/Cas9 system. Survival and fecundity of the Hacd2-deficient strain was significantly lower than that of the wildtype strain when subject to desiccating environmental stress. Hacd2 mediates thermal adaptability in P. xylostella by changing epidermal permeability so is likely to be key to its remaining a major pest species under predicted climate change conditions.

Continent-wide evidence that landscape context can mediate the effects of local habitats on in-field abundance of pests and natural enemies.

Landscape-scale factors known to influence in-field abundance of pest herbivores and their natural enemies, but little is known about effects that operate through the shorter-range influences exerted by habitats immediately adjacent to crop fields.This study first compared the abundance of brassica insect pests and their natural enemy arthropods in 24 spatially independent brassica vegetable fields across southern Australia. An 'edge effect' index was used to compare the abundance of each taxon in the field center with abundance in areas of the crop adjacent to differing habitats. Then, three landscape properties: landscape composition, edge density, and connectivity of diverse crop and non-crop habitats were analyzed at five scales up to 5 km from these focal field centers to assess longer-range influences on arthropod abundances in field centers and on the edge effects.Edge effect of adjacent woody vegetation promoted ladybirds and reduced diamondback moth and whiteflies. Conversely, the presence of crops and pastures immediately adjacent to focal crop fields reduced whiteflies and aphids but with no effect on natural enemies.Effect of landscape composition and connectivity on arthropod abundance at field center found promotion of aphids (cabbage aphid and green peach aphid) by woodland in the landscape.Effect of landscape properties on the edge effects of adjacent habitats was contrasting; strengthened (landscape composition and edge density on edge effect of crops, pasture and woody vegetation in reducing diamondback moth and whiteflies) as well as weakened (edge density and landscape connectivity on edge effect of crops, pasture and woody vegetation in reducing diamondback moth on diamondback moth, whiteflies and aphids, and promoting ladybirds). Synthesis and applications: Findings of this geographically extensive study help define the level of pest risk associated with sites as well as suggest potential interventions such as establishment or restorations of woody vegetation adjacent to crop fields that could reduce risk.

DNA metabarcoding of gut contents reveals key habitat and seasonal drivers of trophic networks involving generalist predators in agricultural landscapes.

BACKGROUND: Understanding the networks of trophic interactions into which generalist predators are embedded is key to assessing their ecological role of in trophic networks and the biological control services they provide. The advent of affordable DNA metabarcoding approaches greatly facilitates quantitative understanding of trophic networks and their response to environmental drivers. Here, we examine how key environmental gradients interact to shape predation by Lycosidae in highly dynamic vegetable growing systems in China. RESULTS: For the sampled Lycosidae, crop identity, pesticide use and seasons shape the abundance of prey detected in spider guts. For the taxonomic richness of prey, local- and landscape-scale factors gradients were more influential. Multivariate ordinations confirm that these crop-abundant spiders dynamically adjust their diet to reflect environmental constraints and seasonal availability to prey. CONCLUSION: Plasticity in diet composition is likely to account for the persistence of spiders in relatively ephemeral brassica crops. Our findings provide further insights into the optimization of habitat management for predator-based biological control practices. © 2022 Society of Chemical Industry.

Contrasting Volatilomes of Livestock Dung Drive Preference of the Dung Beetle Bubas bison (Coleoptera: Scarabaeidae).

Volatile cues can play a significant role in the location and discrimination of food resources by insects. Dung beetles have been reported to discriminate among dung types produced by different species, thereby exhibiting behavioral preferences. However, the role of volatile organic compounds (VOCs) in dung localization and preference remains largely unexplored in dung beetles. Here we performed several studies: firstly, cage olfactometer bioassays were performed to evaluate the behavioral responses of Bubas bison (Coleoptera: Scarabaeidae) to VOCs emanating from fresh horse, sheep, and cattle dung; secondly, concurrent volatilome analysis was performed to characterize volatilomes of these dung types. Bubas bison adults exhibited greater attraction to horse dung and less attraction to cattle dung, and they preferred dung from horses fed a pasture-based diet over dung from those fed lucerne hay. Volatilomes of the corresponding dung samples from each livestock species contained a diverse group of alkanes, alkenes, alkynes, alcohols, aldehydes, ketones, esters, phenols, and sulfurous compounds, but the composition and abundance of annotated VOCs varied with dung type and livestock diet. The volatilome of horse dung was the most chemically diverse. Results from a third study evaluating electroantennogram response and supplementary olfactometry provided strong evidence that indole, butyric acid, butanone, p-cresol, skatole, and phenol, as well as toluene, are involved in the attraction of B. bison to dung, with a mixture of these components significantly more attractive than individual constituents.

Large-scale genome-wide study reveals climate adaptive variability in a cosmopolitan pest.

Understanding the genetic basis of climatic adaptation is essential for predicting species' responses to climate change. However, intraspecific variation of these responses arising from local adaptation remains ambiguous for most species. Here, we analyze genomic data from diamondback moth (Plutella xylostella) collected from 75 sites spanning six continents to reveal that climate-associated adaptive variation exhibits a roughly latitudinal pattern. By developing an eco-genetic index that combines genetic variation and physiological responses, we predict that most P. xylostella populations have high tolerance to projected future climates. Using genome editing, a key gene, PxCad, emerged from our analysis as functionally temperature responsive. Our results demonstrate that P. xylostella is largely capable of tolerating future climates in most of the world and will remain a global pest beyond 2050. This work improves our understanding of adaptive variation along environmental gradients, and advances pest forecasting by highlighting the genetic basis for local climate adaptation.

Raspberry ketone diet supplement reduces attraction of sterile male Queensland fruit fly to cuelure by altering expression of chemoreceptor genes.

Sterile male Queensland fruit fly, Bactrocera tryoni (Froggatt), fed as immature adults on the plant compound raspberry ketone (RK), show a reduced attraction to cuelure, a synthetic analogue of RK used as an attractant in Male Annihilation Technique. We hypothesized the reduced attraction of RK-fed adult males to cuelure may be a consequence of altered expression of chemoreception genes. A Y-tube olfactometer assay with RK-fed and RK-unfed sterile B. tryoni males tested the subsequent behavioural response to cuelure. Behavioral assays confirmed a significant decrease in attraction of RK-fed sterile males to cuelure. RK-fed, non-responders (to cue-lure) and RK-unfed, responders (to cue-lure) males were sampled and gene expression compared by de novo RNA-seq analysis. A total of 269 genes in fly heads were differentially expressed between replicated groups of RK-fed, cuelure non-responders and RK-unfed, cuelure responders. Among them, 218 genes including 4 chemoreceptor genes were up regulated and 51 genes were down regulated in RK-fed, cuelure non-responders. De novo assembly generated many genes with unknown functions and no significant BLAST hits to homologues in other species. The enriched and suppressed genes reported here, shed light on the transcriptional changes that affect the dynamics of insect responses to chemical stimuli.

Molecular gut content analysis indicates the inter- and intra-guild predation patterns of spiders in conventionally managed vegetable fields.

Inter- and intra-guild interactions are important in the coexistence of predators and their prey, especially in highly disturbed vegetable cropping systems with sporadic food resources. Assessing the dietary range of a predator taxon characterized by diverse foraging behavior using conventional approaches, such as visual observation and conventional molecular approaches for prey detection, has serious logistical problems. In this study, we assessed the prey compositions and compare the dietary spectrum of a functionally diverge group of predators-spiders-to characterize their trophic interactions and assess biological control potential in Brassica vegetable fields. We used high-throughput sequencing (HTS) and biotic interaction networks to precisely annotate the predation spectrum and highlight the predator-predator and predator-prey interactions. The prey taxa in the gut of all spider families were mainly enriched with insects (including dipterans, coleopterans, orthopterans, hemipterans, and lepidopterans) with lower proportions of arachnids (such as Araneae) along with a wide range of other prey factions. Despite the generalist foraging behavior of spiders, the community structure analysis and interaction networks highlighted the overrepresentation of particular prey taxa in the gut of each spider family, as well as showing the extent of interfamily predation by spiders. Identifying the diverse trophic niche proportions underpins the importance of spiders as predators of pests in highly disturbed agroecosystems. More specifically, combining HTS with advanced ecological community analysis reveals the preferences and biological control potential of particular spider taxa (such as Salticidae against lepidopterans and Pisauridae against dipterans), and so provides a valuable evidence base for targeted conservation biological control efforts in complex trophic networks.

Beneficial insects are associated with botanically rich margins with trees on small farms.

Beneficial insect communities on farms are influenced by site- and landscape-level factors, with pollinator and natural enemy populations often associated with semi-natural habitat remnants. They provide ecosystem services essential for all agroecosystems. For smallholders, natural pest regulation may be the only affordable and available option to manage pests. We evaluated the beneficial insect community on smallholder bean farms (Phaseolus vulgaris L.) and its relationship with the plant communities in field margins, including margin trees that are not associated with forest fragments. Using traps, botanical surveys and transect walks, we analysed the relationship between the floral diversity/composition of naturally regenerating field margins, and the beneficial insect abundance/diversity on smallholder farms, and the relationship with crop yield. More flower visits by potential pollinators and increased natural enemy abundance measures in fields with higher plant, and particularly tree, species richness, and these fields also saw improved crop yields. Many of the flower visitors to beans and potential natural enemy guilds also made use of non-crop plants, including pesticidal and medicinal plant species. Selective encouragement of plants delivering multiple benefits to farms can contribute to an ecological intensification approach. However, caution must be employed, as many plants in these systems are introduced species.

Conservation biological control research is strongly uneven across trophic levels and economic measures.

Conservation biological control suppresses pests by promoting established rather than inoculative or mass released natural enemies. Research in this approach has expanded rapidly this century but uptake remains limited. Why? Most of the 150 peer reviewed papers reporting field experiments include results on natural enemies and/or pests. Only a minority report effects on crop damage levels or yield, and very few consider economic consequences. This is despite evidence for potential benefits across this full spectrum of response variables. We argue that the limited scope of work to date constrains the development of a compelling evidence base to demonstrate the field effectiveness of conservation biological control, hampering its uptake so encourage researchers to include the assessment of economic impact in future studies of conservation biological control. © 2020 Society of Chemical Industry.

Insect pollination is important in a smallholder bean farming system.

BACKGROUND: Many crops are dependent on pollination by insects. Habitat management in agricultural landscapes can support pollinator services and even augment crop production. Common bean (Phaseolus vulgaris L.) is an important legume for the livelihoods of smallholder farmers in many low-income countries, particularly so in East Africa. While this crop is autogamous, it is frequently visited by pollinating insects that could improve yields. However, the value of pollination services to common beans (Kariasii) yield is not known. METHODS: We carried out pollinator-exclusion experiments to determine the contribution of insect pollinators to bean yields. We also carried out a fluorescent-dye experiment to evaluate the role of field margins as refuge for flower-visitors. RESULTS: Significantly higher yields, based on pods per plant and seeds per pod, were recorded from open-pollinated and hand-pollinated flowers compared to plants from which pollinators had been excluded indicating that flower visitors contribute significantly to bean yields. Similarly, open and hand-pollinated plants recorded the highest mean seed weight. Extrapolation of yield data to field scale indicated a potential increase per hectare from 681 kg in self-pollinated beans to 1,478 kg in open-pollinated beans indicating that flower visitors contributed significantly to crop yield of beans. Our marking study indicated that flower-visiting insects including bees, flies and lepidopterans moved from the field margin flowers into the bean crop. Overall, these results show that insect pollinators are important for optimising bean yields and an important food security consideration on smallholder farms. Field margin vegetation also provides habitat for flower-visiting insects that pollinate beans. Hence, non-crop habitats merit further research focusing on establishing which field margin species are most important and their capacity to support other ecosystem services such as natural pest regulation or even pests.

Local management and landscape structure determine the assemblage patterns of spiders in vegetable fields.

Both field- and landscape-scale factors can influence the predator communities of agricultural pests, but the relative importance and interactions between these scales are poorly understood. Focusing on spiders, an important taxon for providing biological control, we tested the influence of field- and landscape-scale factors on structuring the spider communities in a highly dynamic brassica agroecosystem. We found that local factors (pesticide-use and crop type) and forested landscape significantly influenced the abundance and species richness of spiders, whilst grassland patches significantly affected the spider species richness. Correlation results demonstrated that assemblage patterns of most spider families positively responded to the interplay between local factors and forest patches in the landscape. The spiders abundance was greatest in cauliflower crops surrounded with forest and grassland patches in landscape. Similarly, ordination analyses revealed that organic fields of cauliflower in forested landscapes had a strong positive association with the abundance and species richness of spiders. In contrast, insecticide and synthetic fertilizer-treated fields of Chinese cabbage in landscapes with little non-crop habitat reduced the abundance and species richness of spiders. Our results highlight the extent of interaction between local- and landscape-scale factors, help explain recently reported inconsistent effects of landscape factors on conservation biological control.

Fungal Endophyte Communities of Crucifer Crops Are Seasonally Dynamic and Structured by Plant Identity, Plant Tissue and Environmental Factors.

Endophytic fungi are important in diverse plant functions but knowledge of the factors that shape assemblages of these symbionts is lacking. Here, using a culture-dependent approach, we report 4,178 endophytic fungal isolates representing 16 orders isolated from stems, roots and leaves of three cruciferous plant species, Chinese cabbage (Brassica rapa L.), radish (Raphanus sativus L.) and white cabbage (B. olerocea L.), collected from 21 focal fields with different landscape contexts and pesticide uses during four seasons (summer, autumn, winter and spring). The colonization rate of fungi was found to be most strongly affected by season, plant identity and plant tissue. The colonization was highest during autumn, followed by summer, spring and lowest during winter. The colonization was highest in B. olerocea (53.2%), followed by B. rapa (42.6%), and lowest in R. sativus (35.0%). The colonization was highest in stems (51.9%) in all plant types, followed by leaves (42.4%) and roots (37.5%). Hypocreales was the dominant order (33.3% of all the isolates), followed by Glomerellales (26.5%), Eurotiales (12.1%), Pleosporales (9.8%) and Capnodiales (6.0%). Fungal endophyte abundance (number of isolates) followed the same pattern as colonization rate, while species richness varied with season and host plant tissue. Ordination analyses showed that the abundance and richness of Hypocreales, Eurotiales and Sordariales were associated with plant roots, while Capnodiales, Pleosporales and Trichosphaeriales were associated with spring. Other environmental factors, elevation, and the proportions of grassland, forest, orchard and waterbodies in the surrounding landscape also exerted effects within some categories of other main effects or for certain fungal taxa. Our results indicate that while fungal endophyte communities of crucifer crops vary strongly with the season, they are also strongly structured by plant identity and plant tissue, to a lesser extent by pesticide use and only weakly by landscape composition. The understanding of the ecological roles of fungal endophytes could contribute to habitat management and consequently improve crop pest management.