Attractant and repellent properties of Senna didymobotrya plant extracts to Amblyomma variegatum and Rhipicephalus appendiculatus.

The growing challenge of acaricide resistance and geographical range expansion of invasive tick species demands other interventions, like plant-based alternatives, for sustainable tick control. Leaves, flowers, seedpods, and twig branch extracts of Senna didymobotrya were analyzed using coupled gas chromatography mass spectrometry (GC-MS). Response of adult Amblyomma variegatum and Rhipicephalus appendiculatus to extracts was evaluated. The most attractive plant extract was fractionated and ticks' responses to its fractions assessed. Potential tick attractants in the attractive plant part extract and its fractions were identified by GC-MS analysis. Non- significant qualitative and quantitative differences were observed in the plant parts' extract composition (R = 0.6178). Flower extracts attracted both species, with a 0.1-fold higher attraction in A. variegatum compared to the standard attraction aggregation attachment pheromone (AAAP). Leaf and seedpod extracts repelled ticks at various concentrations. Bioassays after fractionating flower extracts identified hexane and ethyl acetate fractions as most attractive to A. variegatum (P < 0.001) and R. appendiculatus (P < 0.001), respectively. Chemical analysis of the most attractive extracts and fractions identified compounds, including documented acarine attractants, squalene and linoleic acid. A squalene and linoleic acid blend (1:1) at 1 mg/mL significantly attracted adult A. variegatum (P < 0.01) and R. appendiculatus (P < 0.001). The results of this study broaden comprehension of how ticks respond to plants in nature, and showcase the promising potential for integrating these insights into effective tick management programs.

Electrophysiological and Behavioral Responses of Cabbage Aphid (Brevicoryne brassicae) to Rosemary (Rosmarinus officinalis) Volatiles, a Potential push Plant for Vegetable push-pull Cropping System.

The cabbage aphid (Brevicoryne brassicae) is a major pest of kale (Brassica oleraceae var. acephala), an important vegetable that is grown worldwide due to its high nutritional and economic value. Brevicoryne brassicae poses a great challenge to B. oleraceae var. acephala production, causing significant direct and indirect yield losses. Farmers overly rely on synthetic insecticides to manage the pest with limited success owing to its high reproductive behavior and development of resistance. This necessitates a search for sustainable alternatives to mitigate these challenges. This study assessed behavioral responses of B. brassicae to odors from rosemary (Rosmarinus officinalis) and B. oleraceae var. acephala headspace volatiles in a Perspex four-arm olfactometer. We identified and quantified volatiles emitted by each of the two plants and those eliciting antennal response using coupled gas chromatography-mass spectrometry (GC-MS) and GC-electroantennograhic detection(GC-EAD), respectively. Our findings revealed that B. brassicae spent more time in the arms of the olfactometer that contained B. oleraceae var. acephala volatiles compared to the arm that held R. officinalis volatiles. Additionally, B. brassicae spent more time in the olfactometer arms with B. oleracea var. acephala compared to the arms holding B. oleracea var. acephala and R. officinalis enclosed together and clean air. GC-MS analysis revealed diverse and higher quantities of volatile compounds in R. officinalis compared to B. oleraceae var. acephala. GC-EAD analysis showed that antennae of B. brassicae detected Linalool, α-Terpineol, Verbenone, Geraniol, Camphor, and Borneol from the volatiles of R. officinalis, and Sabinene, γ-Terpinene, and ß-Caryophyllene from B. oleraceae var. acephala volatiles. Our findings demonstrate the potential of R. officinalis as a repellent plant against B. brassicae and could be utilized as a 'push' plant in an intercropping strategy against this pest.

Host-Parasitoid Phenology, Distribution, and Biological Control under Climate Change.

Climate change raises a serious threat to global entomofauna-the foundation of many ecosystems-by threatening species preservation and the ecosystem services they provide. Already, changes in climate-warming-are causing (i) sharp phenological mismatches among host-parasitoid systems by reducing the window of host susceptibility, leading to early emergence of either the host or its associated parasitoid and affecting mismatched species' fitness and abundance; (ii) shifting arthropods' expansion range towards higher altitudes, and therefore migratory pest infestations are more likely; and (iii) reducing biological control effectiveness by natural enemies, leading to potential pest outbreaks. Here, we provided an overview of the warming consequences on biodiversity and functionality of agroecosystems, highlighting the vital role that phenology plays in ecology. Also, we discussed how phenological mismatches would affect biological control efficacy, since an accurate description of stage differentiation (metamorphosis) of a pest and its associated natural enemy is crucial in order to know the exact time of the host susceptibility/suitability or stage when the parasitoids are able to optimize their parasitization or performance. Campaigns regarding landscape structure/heterogeneity, reduction of pesticides, and modelling approaches are urgently needed in order to safeguard populations of natural enemies in a future warmer world.

Immunological responses and gut microbial shifts in Phthorimaea absoluta exposed to Metarhizium anisopliae isolates under different temperature regimes.

The invasive tomato leaf miner, Phthorimaea absoluta, is conventionally controlled through chemical insecticides. However, the rise of insecticide resistance has necessitated sustainable and eco-friendly alternatives. Entomopathogenic fungi (EPF) have shown potential due to their ability to overcome resistance and have minimal impact on non-target organisms. Despite this potential, the precise physiological mechanisms by which EPF acts on insect pests remain poorly understood. To attain a comprehensive understanding of the complex physiological processes that drive the successful control of P. absoluta adults through EPF, we investigated the impacts of different Metarhizium anisopliae isolates (ICIPE 665, ICIPE 20, ICIPE 18) on the pest's survival, cellular immune responses, and gut microbiota under varying temperatures. The study unveiled that ICIPE 18 caused the highest mortality rate among P. absoluta moths, while ICIPE 20 exhibited the highest significant reduction in total hemocyte counts after 10 days at 25°C. Moreover, both isolates elicited notable shifts in P. absoluta's gut microbiota. Our findings revealed that ICIPE 18 and ICIPE 20 compromised the pest's defense and physiological functions, demonstrating their potential as biocontrol agents against P. absoluta in tomato production systems.

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.

Immune Responses and Transcriptomic Analysis of Nilaparvata lugens against Metarhizium anisopliae YTTR Mediated by Rice Ragged Stunt Virus.

Plant viruses and entomopathogenic fungi (EPF) can both elicit immune responses in insects. This study was designed to clarify whether plant viruses could affect the efficacy of EPF and explore the immune responses of brown planthopper (BPH), Nilaparvata lugens, in response to different pathogen infections. In this study, a strain of Metarhizium anisopliae YTTR with high pathogenicity against BPH was selected and explored whether rice ragged stunt virus (RRSV) could affect its lethality against BPH. RNA-seq was used to detect the inner responses of BPH in response to RRSV and M. anisopliae YTTR infection. Results showed that M. anisopliae YTTR has strong lethality against BPH (RRSV-carrying and RRSV-free). RRSV invasion did not affect the susceptibility of BPH against M. anisopliae YTTR at all concentrations. At 1 × 108 spores/mL, M. anisopliae YTTR caused a cumulative mortality of 80% to BPH at 7 days post-treatment. The largest numbers of differentially expressed genes (DEGs) was obtained in BPH treated with the two pathogens than in other single pathogen treatment. In addition, KEGG enrichment analysis showed that the DEGs were mostly enriched in immune and physiological mechanisms-related pathways. Both RRSV and M. anisopliae YTTR could induce the expression changes of immune-related genes. However, most of the immune genes had varying expression patterns in different treatment. Our findings demonstrated that RRSV invasion did not have any significant effect on the pathogenicity of M. anisopliae YTTR, while the co-infection of M. anisopliae YTTR and RRSV induced more immune and physiological mechanisms -related genes' responses. In addition, the presence of RRSV could render the interplay between BPH and M. anisopliae YTTR more intricate. These findings laid a basis for further elucidating the immune response mechanisms of RRSV-mediated BPH to M. anisopliae infection.

Comprehensive genome-wide identification and expression profiling of ADF gene family in Citrus sinensis, induced by endophytic colonization of Beauveria bassiana.

Endophytic entomopathogenic species are known to systematically colonize host plants and form symbiotic associations that benefit the plants they live with. The actin-depolymerizing factors (ADFs) are a group of gene family that regulate growth, development, and defense-related functions in plants. Systematic studies of ADF family at the genome-wide level and their expression in response to endophytic colonization are essential to understand its functions but are currently lacking in this field. 14ADF genes were identified and characterized in the Citrus sinensis genome. The ADF genes of C. sinensis were classified into five groups according to the phylogenetic analysis of plant ADFs. Additionally, the cis-acting analysis revealed that these genes play essential role in plant growth/development, phytohormone, and biotic and abiotic responses; and the expression analysis showed that the symbiotic interactions generate a significant expression regulation level of ADF genes in leaves, stems and roots, compared to controls; thus enhancing seedlings' growth. Additionally, the 3D structures of the ADF domain were highly conserved during evolution. These results will be helpful for further functional validation of ADFs candidate genes and provide important insights into the vegetative growth, development and stress tolerance of C. sinensis in responses to endophytic colonization by B. bassiana.

Interactions between the Entomopathogenic Fungus Metarhizium anisopliae ICIPE 20 and the Endoparasitoid Dolichogenidea gelechiidivoris, and Implications for Combined Biocontrol of Tuta absoluta.

The Integrated Pest Management (IPM) approach have been widely promoted and used for the management of native and invasive pests, while the use of various components of the IPM can have a synergetic, additive, or antagonistic effect on each other; this study evaluated the susceptibility of Dolichogenidea gelechiidivoris (Marsh) (Hymenoptera: Braconidae), to the Metarhizium anisopliae (Metschnikoff) ICIPE 20 through direct and indirect infection approaches. The effect of fungus on parasitoid longevity, survival of parasitized-larvae, preference of the parasitoid to fungal treated and untreated larvae, and percent parasitism of Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) under different infection scenarios were assessed. The direct application of dry conidia to the parasitoid prior to exposure to the host, reduced D. gelechiidivoris longevity, though the infected female wasps still yielded high parasitism (over 70%). Infecting the parasitized larvae at different ages led to a respective reduction of parasitoid emergence by 35% and 23% for infection at 1 and 5 days post-parasitisation. Exposure of healthy-D. gelechiidivoris adults to a plant-sprayed with fungus did not affect their longevity, and no discriminatory host selection was observed. The highest mortality (~80%) of T. absoluta was achieved when D. gelechiidivoris and M. anisopliae ICIPE 20 were used in combination, indicating an additive impact on the target pest; however, field validation can shed more light on this outcome.

Genome-Wide Identification and Characterization of Toll-like Receptors (TLRs) in Diaphorina citri and Their Expression Patterns Induced by the Endophyte Beauveria bassiana.

Toll-like receptors (TLRs) are pathogen recognition receptors (PRRs), which play key roles in helping the host immune system fight pathogen invasions. Systematic information on TLRs at the genome-wide level and expression profiling in response to endophytic colonization is very important to understand their functions but is currently lacking in this field. Here, a total of two TLR genes were identified and characterized in Diaphorina citri. The TLR genes of D. citri were clustered into five families according to the phylogenetic analysis of different species' TLRs. The domain organization analyses suggested that the TLRs were constituted of three important parts: a leucine-rich repeat (LRR) domain, a transmembrane region (TR) and a Toll/interleukin-1 receptor (TIR) domain. The mRNA expression levels of the two TLR genes (DcTOLL and DcTLR7) were highly regulated in both nymphs and adults of D. citri. These results elucidated the potentiated TLR gene expression in response to endophytically colonized plants. Furthermore, the 3D structures of the TIR domain were highly conserved during evolution. Collectively, these findings elucidate the crucial roles of TLRs in the immune response of D. citri to entomopathogens systematically established as endophytes, and provide fundamental knowledge for further understanding of the innate immunity of D. citri.

Genetic diversity of Diaphorina citri (Hemiptera: Liviidae) unravels phylogeographic structure and invasion history of eastern African populations.

The Asian citrus psyllid (Diaphorina citri Kuwayama) is a key pest of Citrus sp. worldwide, as it acts as a vector for Candidatus Liberibacter asiaticus, the bacterial pathogen that causes citrus Huanglongbing. Diaphorina citri has been reported in Kenya, Tanzania, and more recently in Ethiopia. This study assessed the genetic diversity and phylogeographic structure of the pest to gain insights into the potential sources of its introduction into Africa. Population structure and differentiation of D. citri populations from China, Ethiopia, Kenya, Tanzania, and the USA were assessed using 10 microsatellite loci. Additionally, five new complete mitogenomes of D. citri collected in China, Ethiopia, Kenya, Tanzania, and the USA were analyzed in the context of publicly available sequences. Genotype data grouped the D. citri populations from Kenya and Tanzania in one cluster, and those from Ethiopia formed a separate cluster. The two genetic clusters inferred from genotype data were congruent with mitochondrial sequence data. The mitogenomes from Kenya/Tanzania/China had 99.0% similarity, and the Ethiopia/USA had 99.9% similarity. In conclusion, D. citri populations in eastern Africa have different sources, as the Kenyan and Tanzanian populations probably originated from southeastern Asia, while the Ethiopian population most probably originated from the Americas.

Comparative microbiome analysis of Diaphorina citri and its associated parasitoids Tamarixia radiata and Diaphorencyrtus aligarhensis reveals Wolbachia as a dominant endosymbiont.

Microbiome analysis in a host-parasitoid interaction network was conducted to compare the taxonomic composition of bacterial communities of Diaphornia citri, Tamarixia radiata, and Diaphorencyrtus aligarhensis. The comparative analysis revealed differences in the composition and diversity of the symbiont populations across the host and its associated parasitoids. Proteobacteria was the most dominant phylum, representing 67.80% of the total bacterial community, while Candidatus Profftella armature and Wolbachia were the dominant genera across the host and parasitoids. There were clear differences observed in alpha and beta diversity of microbiota through the host and its associated parasitoids. The function prediction of bacterial communities and Pearson correlation analysis showed that specific bacterial communities displayed positive correlations with the carbohydrate metabolism pathway. Furthermore, when symbiotic bacteria were eliminated using a broad-spectrum antibiotic, tetracycline hydrochloride, the parasitoids' median survival time and longevity were significantly reduced. We confirmed the physiological effects of symbiotic bacteria on the fitness of parasitoids and demonstrated the effect of antibiotics in decreasing the food intake and measurement of amino acids in the hemolymph. This study sheds light on basic information about the mutualism between parasitoids and bacteria, which may be a potential source for biocontrol strategies for citrus psyllid, especially D. citri.

Effectiveness of Entomopathogenic Fungi on Immature Stages and Feeding Performance of Fall Armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae) Larvae.

Maize is a major staple crop in China, and the sustainable productivity of this primary crop has been recently threatened by fall armyworm (FAW), Spodoptera frugiperda, invasion. The five fungal isolates, Aspergillus sp. BM-3 and SE-2-1, Cladosporium tenuissimum SE-10, Penicillium citrinum CTD-24, and Beauveria bassiana ZK-5 were assessed for their efficacy in causing mortality against first to sixth instar eggs and neonate larvae seven days post-treatment, and their effects on the feeding performance of sixth instar S. frugiperda larvae at 48 h post-treatment at three concentrations (1 × 106, 1 × 107, and 1 × 108 conidia mL-1) were also assessed. The six instar S. frugiperda larvae were not susceptible to the five tested fungal isolates. However, B. bassiana ZK-5 caused the highest egg mortality of 40, 70, and 85.6% at 1 × 106, 1 × 107, and 1 × 108 conidia mL-1, respectively, followed by P. citrinum CTD-24 (30.6, 50, and 75.6%) and C. tenuissimum SE-10 (25.6, 40, and 55.6%). In addition, B. bassiana ZK-5 caused the highest neonate mortality of 54.3% at 1 × 108 conidia mL-1. B. bassiana ZK-5 and P. citrinum CTD-24 caused cumulative mortality, including 93.3 and 83.3% mortality of eggs and neonates, respectively, at 1 × 108 conidia mL-1. Furthermore, B. bassiana ZK-5 reduced the feeding efficacy of first to third instar S. frugiperda larvae by 66.7 to 78.6%, while P. citrinum CTD-24 and C. tenuissimum SE-10 reduced larval feeding by 48.3 to 57.1% at 1 × 108 conidia mL-1. However, these fungal isolates were less potent in reducing the feeding activity of fourth to sixth instar S. frugiperda larvae (>46% with B. bassiana at 48 h post-treatment). The tested fungal isolates could play an essential role as microbial biopesticides in suppressing the S. frugiperda population in China after further investigations on their efficacy are obtained in the field.

Model Application of Entomopathogenic Fungi as Alternatives to Chemical Pesticides: Prospects, Challenges, and Insights for Next-Generation Sustainable Agriculture.

In the past few decades, the control of pests and diseases of cultivated plants using natural and biological measures has drawn increasing attention in the quest to reduce the level of dependence on chemical products for agricultural production. The use of living organisms, predators, parasitoids, and microorganisms, such as viruses, bacteria, and fungi, has proven to be a viable and sustainable pest management technique. Among the aforementioned, fungi, most importantly the insect-pathogenic species, have been in use for more than 150years. These include the most popular strains belonging to the genera Beauveria, Metarhizium, Isaria, Hirsutella, and Lecanicillium. Their application is usually through an inundative approach, which inherently involves exposure of the fungal spores to unfavorable humidity, temperature, and solar radiation conditions. These abiotic factors reduce the persistence and efficacy of these insect-pathogenic fungi. Despite these limitations, over 170 strains have been formulated as mycopesticides and are available for commercial use. In the last few decades, numerous studies have suggested that these species of entomopathogenic fungi (EPF) offer far more benefits and have broader ecological functions than hitherto presumed. For instance, aside from their roles as insect killers, it has been well established that they also colonize various host plants and, hence, provide other benefits including plant pathogen antagonism and plant growth promotion and serve as sources of novel bioactive compounds and secondary metabolites, etc. In this light, the potential of EPF as alternatives or perhaps as supplements to chemical pesticides in plant protection is discussed in this review. The paper highlights the numerous benefits associated with endophytic fungal entomopathogen and host plant associations, the mechanisms involved in mediating plant defense against pests and pathogens, and the general limitations to the use of EPF in plant protection. A deeper understanding of these plant host-fungus-insect relationships could help unveil the hidden potentials of fungal endophytes, which would consequently increase the level of acceptance and adoption by users as an integral part of pest management programs and as a suitable alternative to chemical inputs toward sustainable crop production.

General Limitations to Endophytic Entomopathogenic Fungi Use as Plant Growth Promoters, Pests and Pathogens Biocontrol Agents.

The multiple roles of fungal entomopathogens in host plants' growth promotion, pest and pathogen management have drawn huge attention for investigation. Endophytic species are known to influence various activities of their associated host plants, and the endophyte-colonized plants have been demonstrated to gain huge benefits from these symbiotic associations. The potential application of fungal endophytes as alternative to inorganic fertilizers for crop improvement has often been proposed. Similarly, various strains of insect pathogenic fungi have been formulated for use as mycopesticides and have been suggested as long-term replacement for the synthetic pesticides that are commonly in use. The numerous concerns about the negative effects of synthetic chemical pesticides have also driven attention towards developing eco-friendly pest management techniques. However, several factors have been underlined to be militating the successful adoption of entomopathogenic fungi and fungal endophytes as plant promoting, pests and diseases control bio-agents. The difficulties in isolation and characterization of novel strains, negative effects of geographical location, vegetation type and human disturbance on fungal entomopathogens, are among the numerous setbacks that have been documented. Although, the latest advances in biotechnology and microbial studies have provided means of overcoming many of these problems. For instance, studies have suggested measures for mitigating the negative effects of biotic and abiotic stressors on entomopathogenic fungi in inundative application on the field, or when applied in the form of fungal endophytes. In spite of these efforts, more studies are needed to be done to achieve the goal of improving the overall effectiveness and increase in the level of acceptance of entomopathogenic fungi and their products as an integral part of the integrated pest management programs, as well as potential adoption as an alternative to inorganic fertilizers and pesticides.

Mechanism of Action of Endophytic Fungi Hypocrea lixii and Beauveria bassiana in Phaseolus vulgaris as Biopesticides against Pea Leafminer and Fall Armyworm.

Endophytic fungal isolates Hypocrea lixii F3ST1 and Beauveria bassiana G1LU3 were evaluated for their potential to endophytically colonize and induce active compounds in Phaseolus vulgaris, as a defense mechanism against pea leafminer (Liriomyza huidobrensis) and fall armyworm (Spodoptera frugiperda). Endophytic colonization was achieved through seed inoculation with the volatile emissions from P. vulgaris plants being analyzed using GC-MS. The crude extracts of P. vulgaris obtained using methanol and dichloromethane were assayed against leafminer and fall armyworm larvae using leaf dipping and topical application, respectively. The two isolates successfully colonized the entire host plant (roots, stems, and leaves) with significant variation (p < 0.001) between fungal isolates and the controls. The results showed qualitative differences in the volatile profiles between the control plants, endophytically colonized and insect-damaged plants attributed to fungal inoculation and leafminer damage. The crude methanol extracts significantly reduced the percentage pupation of 2nd instar leafminer larvae (p < 0.001) and adult-flies emergence (p < 0.05). The survival of the 1st instar fall armyworm larvae was also significantly reduced (p < 0.001) compared to the controls. This study demonstrated the high potential of endophytic fungi H. lixii and B. bassiana in inducing mainly specific defense compounds in the common bean P. vulgaris that can be used against pea leafminer and fall armyworm.

Characterization of mycotoxins from entomopathogenic fungi (Cordyceps fumosorosea) and their toxic effects to the development of asian citrus psyllid reared on healthy and diseased citrus plants.

Entomopathogenic fungi (EPF) produce multiple mycotoxins, which play an essential role in improving fungal pathogenesis and virulence. To characterize various mycotoxins from the crude methanol extract of Cordyceps fumosorosea, a major EPF against various insect pests, we performed ultra-performance liquid chromatography coupled to quadrupole time of flight mass spectrometer (UPLC-QTOF MS) technique, and all compounds were identified through molecular mass and formulae. Bassianolide was assessed against the nymphs and adults of Diaphorina citri reared on healthy and Huánglóngbìng (HLB)-diseased Citrus spp. Plants under laboratory conditions. Overall, 17 compounds were identified from the fungal extract and categorized into three groups, i.e. (1) alkaloids (Isariotins A-C), (2) peptides (Bassianolide, Beauverolides, Beauvericin A, Isaridins and Destruxin E) and (3) polyketide (Tenuipyrone). The detected beauverolides (B, C, F, I, Ja) from C. fumosorosea were novel mycotoxins, and their detection intensity was the highest in the fungal extract. Furthermore, bassianolide caused more than 70% and 80% mortality of D. citri nymphs and adults after two days of application, respectively. After three days of chemical application, all nymphal and adult populations of D. citri were killed by bassianolide. However, the mortality rates of both populations, nymphs and adults, were higher on HLB-diseased plants as compared to healthy plants.

Citronellal perception and transmission by Anopheles gambiae s.s. (Diptera: Culicidae) females.

Anopheles gambiae s.s. is a key vector of Plasmodium parasites. Repellents, which may be a promising alternative to pesticides used to control malaria mosquitoes. Although citronellal is a known mosquito repellent, its repellency characteristics are largely unknown. Determining the specific odorant-binding proteins (OBPs) and odorant receptors (ORs) that detect and transfer the citronellal molecule in A. gambiae s.s. will help to define the mode of action of this compound. In this research, we assessed the repellent activity of citronellal in A. gambiae s.s. using a Y-tube olfactory meter, screened candidate citronellal-binding OBPs and ORs using reverse molecular docking, clarified the binding properties of predicted proteins for citronellal using fluorescence competition binding assay. Results showed that citronellal had a dosage effect on repelling A. gambiae s.s.. The 50% repellent rate was determined to be 4.02 nmol. Results of simulated molecular docking showed that the only proteins that bound tightly with citronellal were AgamOBP4 and AgamORC7. Fluorescence competitive binding assays confirmed the simulations. This research determined that citronellal was captured by AgamOBP4 and transmitted to AgamORC7 in A. gambiae s.s.. Our study will be beneficial in the further understanding the repellent mechanism of citronellal against A. gambiae s.s..

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.