Suitable Host Research Articles

Pastoral grasses and legumes as potential host plants for fall armyworm Spodoptera frugiperda (J.E. Smith) development.

AbstractThe fall armyworm, Spodopterafrugiperda (J.E. Smith) (Lepidoptera: Noctuidae) is a highly polyphagous, migratory pest native to the tropical and subtropical region of Americas. Grass crops particularly corn have been most heavily impacted since S. frugiperda invaded Australia in 2020. In northern Western Australia, S. frugiperda is primarily a problem in dry season (May to October) crops when there are abundant host plants available in monocultures. During the wet season (November to April) it is not known what host plants may support larval growth and development. Accordingly, this present study undertook a no-choice feeding bioassay to assess larval and pupal development on several grasses and legumes that are present over the wet season in northern Western Australia to evaluate their potential as suitable host plants for S. frugiperda. Maize (control) and sorghum were the most suitable hosts for S. frugiperda development as they supported greater larval and pupal survival, heavier larvae and pupae, and shorter development period. Following maize and sorghum, pastoral grasses, particularly blue grass, and a weedy grass, barnyard grass, had comparable effects on larval and pupal development. Legume pastures (Cavalcade, butterfly pea, and siratro) were unsuitable hosts in this bioassay as very few larvae completed their development to adults. This study demonstrates the potential for rangeland pasture grasses and weedy grasses as wet season host plants for S. frugiperda. They provide the necessary green bridge for S. frugiperda populations to survive. It is recommended that pastoralists monitor their grass forage for S. frugiperda and consider management if significant reductions in biomass are detected.

Synergistic removal of Staphylococcus aureus biofilms by using a combination of phage Kayvirus rodi with the exopolysaccharide depolymerase Dpo7.

Bacteriophages have been shown to penetrate biofilms and replicate if they find suitable host cells. Therefore, these viruses appear to be a good option to tackle the biofilm problem and complement or even substitute more conventional antimicrobials. However, in order to successfully remove biofilms, in particular mature biofilms, phages may need to be administered along with other compounds. Phage-derived proteins, such as endolysins or depolymerases, offer a safer alternative to other compounds in the era of antibiotic resistance. This study examined the interactions between phage Kayvirus rodi with a polysaccharide depolymerase (Dpo7) from another phage (Rockefellervirus IPLA7) against biofilms formed by different Staphylococcus aureus strains, as determined by crystal violet staining, viable cell counts and microscopy analysis. Our results demonstrated that there was synergy between the two antimicrobials, with a more significant decreased in biomass and viable cell number with the combination treatment compared to the phage and enzyme alone. This observation was confirmed by microscopy analysis, which also showed that polysaccharide depolymerase treatment reduced, but did not eliminate extracellular matrix polysaccharides. Activity assays on mutant strains did not identify teichoic acids or PNAG/PIA as the exclusive target of Dpo7, suggesting that may be both are degraded by this enzyme. Phage adsorption to S. aureus cells was not significantly altered by incubation with Dpo7, indicating that the mechanism of the observed synergistic interaction is likely through loosening of the biofilm structure. This would allow easier access of the phage particles to their host cells and facilitate infection progression within the bacterial population.

Analysis of thermally-induced fracture of Callovo-Oxfordian claystone: From lab tests to field scale

Argillaceous rocks are a suitable host rock for the deep geological disposal of exothermic High-Level Radioactive Waste (HLW) and Spent Fuel (SF). Excess pore pressures develop in this type of rocks when subject to increased temperatures that, in some circumstances, may lead to the fracturing of the rock. The paper explores this phenomenon by means of coupled numerical analyses carried out within a fully coupled thermo-hydro-mechanical (THM) framework. The general THM formulation is described as well as the anisotropic elastic and anisotropic elastoviscoplastic constitutive laws employed. Thermal extension triaxial tests are simulated as a check on the performance of the numerical formulation and to provide calibration data for the tensile strength of the material. Selected results from a comprehensive set of three-dimensional analyses of a large-scale field heating test, designed to study the possibility of thermal-induced failure in the rock, are presented and discussed. The analyses reproduce satisfactorily the observed patterns of behaviour. The effects of the constitutive law, material parameters and the presence of the excavation damage zone (EDZ) around the main drift and around the heater boreholes are studied. In particular, their effects on the state of the stress in the heated area are examined in the context of the potential for thermal fracturing of the rock.

Accurate and Robust Wide‐Range Luminescent Microthermometer Based on ALD‐Encapsulated Ga2O3:Cr DBR Microcavities

AbstractThe high spatial resolution and contactless optical readout capabilities of luminescence thermometry offer significant advantages in numerous fields, including biomedicine, space exploration and optoelectronics. In addition, robust, reproducible, and accurate temperature measurements are essential in these areas. The ultra‐wide band gap semiconductor material Ga2O3 is a suitable host for optical sensing in harsh environments due to its high stability. In this work, the thermometric operation of Ga2O3:Cr‐based microcavities are evaluated. They are designed as follows: Ga2O3:Cr microwires are encapsulated in multilayers fabricated by atomic layer deposition (ALD), which act as both Bragg reflectors and protective layers for the thermometric sensor. Prior to the ALD encapsulation step, focused ion beam carved trenches at the microwire ends are necessary to accommodate the multilayer coating. The structural and optical properties of the devices are assessed experimentally, analytically and by simulations. The developed microthermometers can be easily calibrated using a cubic polynomial for the temperature‐dependent resonant peak position shift. A better than 0.5 °C temperature resolution and accuracy for temperatures above −80 °C is demonstrated. Additionally, the devices show robustness against excitation laser densities of at least 34 W mm−2, can operate at temperatures up to 600 °C and remain functional in liquids.

Infection of human organoids supports an intestinal niche for Chlamydia trachomatis.

Several reports suggest that intestinal tissue may be a natural niche for Chlamydia trachomatis infection and a reservoir for persistent infections in the human body. Due to the human specificity of the pathogen and the lack of suitable host models, there is limited knowledge on this topic. In our study, we modelled the course of the chlamydial infection in human primary gastrointestinal (GI) epithelial cells originating from patient-derived organoids. We show that GI cells are resistant to apical infection and C. trachomatis needs access to the basolateral membrane to establish an infection. Transmission electron microscopy analysis reveals the presence of both normal as well as aberrant chlamydial developmental forms in the infected cells, suggesting a possible cell-type specific nature of the infection. Furthermore, we show that the plasmid-encoded Pgp3 is an important virulence factor for the infection of human GI cells. This is the first report of C. trachomatis infection in human primary intestinal epithelial cells supporting a possible niche for chlamydial infection in the human intestinal tissue.

Production of Reverse Transcriptase and DNA Polymerase in Bacterial Expression Systems.

DNA amplification and reverse transcription enzymes have proven to be invaluable in fast and reliable diagnostics and research applications because of their processivity, specificity, and robustness. Our study focused on the production of mutant Taq DNA polymerase and mutant M-MLV reverse transcriptase in the expression hosts Vibrio natriegens and Escherichia coli under various expression conditions. We also examined nonspecific extracellular production in V. natriegens. Intracellularly, M-MLV was produced in V. natriegens at the level of 11% of the total cell proteins (TCPs) compared with 16% of TCPs in E. coli. We obtained a soluble protein that accounted for 11% of the enzyme produced in V. natriegens and 22% of the enzyme produced in E. coli. Taq pol was produced intracellularly in V. natriegens at the level of 30% of TCPs compared with 26% of TCPs in E. coli. However, Taq pol was almost non-soluble in E. coli, whereas in V. natriegens, we obtained a soluble protein that accounted for 23% of the produced enzyme. We detected substantial extracellular production of Taq pol. Thus, V. natriegens is a suitable alternative host with the potential for production of recombinant proteins.

The structure of the Cryptocotyle lingua miracidium (Digenea: Opisthorchiidae)

In the course of Digenea evolution the transitions to a passive strategy of infection of the first intermediate host occurred multiple times. In these cases, miracidia – ciliated larvae – relinquished their role as active seekers for a suitable snail host and began relying on the chance of being accidently ingested by it. The structural changes resulting from these transitions are diverse, but the majority of them are reductive. In this study we present our ultrastructural survey on the opisthorchioid Cryptocotyle lingua miracidium. The transition to passivity led to extensive reduction in almost all somatic elements of the larva: only four epithelial plates cover its body; just a single nervous cell, a single protonephridium and a single longitudinally-oriented muscle cell have been found in this miracidium. Several undifferentiated cells comprise the germinal material of the larva. The greatest part of its body is occupied by three glandular elements. The position of these glands is consistent within Opisthorchioidea and strikingly corresponds to that of some of the optically studied miracidia of Plagiorchioidea. This similarity is suspicious, because the C. lingua larva appeared to differ vastly from other ultrastructurally described passive miracidia. Here we create a basis for the future comparison of the miracidia of Opisthorchioidea and Plagiorchioidea, expecting the ultrastructural data on the latter to be published someday.

Development of a subunit vaccine against the cholangiocarcinoma causing Opisthorchis viverrini: a computational approach.

Opisthorchis viverrini is the etiological agent of the disease opisthorchiasis and related cholangiocarcinoma (CCA). It infects fish-eating mammals and more than 10 million people in Southeast Asia suffered from opisthorchiasis with a high fatality rate. The only effective drug against this parasite is Praziquantel, which has significant side effects. Due to the lack of appropriate treatment options and the high death rate, there is a dire need to develop novel therapies against this pathogen. In this study, we designed a multi-epitope chimeric vaccine design against O. viverrini by using immunoinformatics approaches. Non-allergenic and immunogenic MHC-1, MHC-2, and B cell epitopes of three candidate proteins thioredoxin peroxidase (Ov-TPx-1), cathepsin F1 (Ov-CF-1) and calreticulin (Ov-CALR) of O. viverrini, were predicted to construct a potent multiepitope vaccine. The coverage of the HLA-alleles of these selected epitopes was determined globally. Four vaccine constructs made by different adjuvants and linkers were evaluated in the context of their physicochemical properties, antigenicity, and allergenicity. Protein-protein docking and MD simulation found that vaccines 3 was more stable and had a higher binding affinity for TLR2 and TLR4 immune receptors. In-silico restriction cloning of vaccine model led to the formation of plasmid constructs for expression in a suitable host. Finally, the immune simulation showed strong immunological reactions to the engineered vaccine. These findings suggest that the final vaccine construct has the potential to be validated by in vivo and in vitro experiments to confirm its efficacy against the CCA causing O. viverrini.

Response of growth and physiological enzyme activities in Eriogyna pyretorum to various host plants.

Morphological attributes and chemical composition of host plants shape growth and development of phytophagous insects via influences on their behavior and physiological processes. This research delves into the relationship between Eriogyna pyretorum and various host plants through studuying how feeding on different host tree species affect growth, development, and physiological enzyme activities. We examined E. pyretorum response to three distinct host plants: Camphora officinarum, Liquidambar formosana and Pterocarya stenoptera. Notably, larvae feeding on C. officinarum and L. formosana displayed accelerated development, increased pupal length, and higher survival rates compared to those on P. stenoptera. This underlines the pivotal role of host plant selection in shaping the E. pyretorum's life cycle. The activities of a-amylase, lipase and protective enzymes were the highest in larvae fed on the most suitable host L.formosana which indicated that the increase of these enzyme activities was closely related to growth and development. Furthermore, our investigation revealed a relationship between enzymatic activities and host plants. Digestive enzymes, protective enzymes, and detoxifying enzymes exhibited substantial variations contingent upon the ingested host plant. Moreover, the total phenolics content in the host plant leaves manifested a noteworthy positive correlation with catalase and lipase activities. In contrast, a marked negative correlation emerged with glutathione S-transferase and α-amylase activities. The total developmental duration of larvae exhibited a significant positive correlation with the activities of GST and CarE. The survival rate of larvae showed a significant positive correlation with CYP450. These observations underscore the insect's remarkable adaptability in orchestrating metabolic processes in accordance with available nutritional resources. This study highlights the interplay between E.pyretorum and its host plants, offering novel insights into how different vegetation types influence growth, development, and physiological responses. These findings contribute to a deeper comprehension of insect-plant interactions, with potential applications in pest management and ecological conservation.

Effect of male parental gamma irradiation on host suitability of its F1 progeny of a lepidopteran tropical pest, Spodoptera litura (Fabr.) towards development and virulence of entomopathogenic nematodes, Steinernema thermophilum

The suitability of F1 progeny insect larvae of the irradiated male parent, Spodoptera litura (Fabr.) for infective juveniles (IJs) of entomopathogenic nematodes (EPN), Steinernema thermophilum was assessed to comprehend the feasibility of combining EPNs with nuclear pest control tactic. As compared to the control, the IJs induced faster host mortality with reduced proliferation in F1 host larvae. IJs derived from F1 host larvae exhibited almost similar proliferation capacity on normal hosts as in control. Further, the molecular basis of EPNs induced mortality in F1 host larvae was evaluated. Dual stress of EPN infection and irradiation induced downregulation of the relative mRNA expression of antimicrobial genes and upregulated expression of antioxidative genes. A pronounced effect of EPNs in association with irradiation stress was apparent on host mortality. Radiation induced sterile F1 insect larvae of S. litura acted as a reasonably suitable host for EPNs and also provided the environment for developing viable EPNs for their potential use as biocontrol agents.

Prebiotic Chemistry and Sepiolite: A Density Functional Theory Approach

In this study, the molecular interactions of sepiolite, a biocompatibility clay mineral with known as biomaterial, and purine and pyrimidine molecules forming the bases of DNA and RNA molecules were modeled by Density Functional Theory. In addition to the geometry optimization, interaction energy, bond critical point, and electrostatic potential calculations revealed that essential molecules for our source of life interact with the basal surface of the clay. For example, the best interaction energies between bases / sepiolite were found to be -127.47, -121.35 kJ / mol for guanine and cytosine, respectively. Looking at the modeling results, one of the most important factors affecting the interaction energies is H-bond. In order to reveal this, bond critical points analyze were performed and it was computed that a large amount of intermolecular interaction energies came from H-bonds. For example, it has been calculated that close to 70% of the total energy in the guanine/TOT model is from H-bonds. Besides, this value of the cytosine/TOT model was found to be around 72%. The most effective indexes in these two models are 145 and 135, and the H-bond energies were recorded as -22.41 and 31.41 kJ/mol, respectively. Considering all the analyzes run, it can be concluded that the basal surfaces of the sepiolite are a suitable host for the nitrogenous bases, which are the main sources of life.

Lupeol a predisposing factor in grey mould (Amphobotrys ricini [N.F. Buchw.]) Hennebert disease pathogenesis in castor (Ricinus communis L.)

AbstractTo understand the castor wax layer's multifaceted contribution to Amphobotrys ricini pathogenesis, the hydrophobic components of cuticular wax were analysed from waxy and non‐waxy castor genotypes. Gas chromatography–mass spectrometry (GC–MS) technology enabled the stable detection and quantitative determination of various fatty acids and terpenoids. The investigation revealed a significant presence of triterpenoid compound ‘lupeol’, accounting for approximately 53.6% of the wax composition in the waxy genotype (DCH‐519), which was absent in non‐waxy genotype (ICS‐324). On exposure to lupeol, about 93.3% of conidia germinated leading to rapid mycelium growth and sporulation of A. ricini. SEM analysis of waxy and non‐waxy genotypes infected with A. ricini confirmed faster germination and production of longer germ tubes on waxy genotype compared with non‐waxy genotype, which may likely due to early recognition of the suitable host in the presence of lupeol, ultimately aiding in speedy germination and growth of the pathogen setting pace for pathogenesis.

Risk assessment of Phlyctinus callosus for the EU.

Following a request from the European Commission, the EFSA Panel on Plant Health performed a quantitative risk assessment for the EU of Phlyctinus callosus (Coleoptera: Curculionidae), a polyphagous pest occurring in Australia, New Zealand and South Africa. The current risk assessment focused on potential pathways for entry, the climatic conditions allowing establishment, the expected spread capacity and the impact considering a time horizon of 10 years (2023-2032). The Panel identified the import of apples, cut flowers and table grapes as the most relevant entry pathways. Over the next 10 years, an annual median estimate of approximately 49.5 (90% certainty range, CR, ranging from 4.0 to 881.2) potential P. callosus founder populations are expected. When the probability of establishment is considered and climatic indicators are used to define the areas in the EU where establishment is possible, the model estimated a median of 1 founder population every 1.3 years (90% CR: 1 every 30.8 years to 23.3 per year) in the scenario where the areas are defined by the union of all the climatic indicators and 1 founder population every 11.9 years (90% CR: 1 every 256.6 years to 2.5 per year) in the scenario where establishment is possible only in the areas defined by the climatic indicator of minimum soil temperature. The estimated number of founder populations per year is mostly driven by the probability of establishment in the rural areas, infestation rate in table grapes and the probability of transfer to a suitable host in the rural area. The risk of entry for cut flowers and apples is substantially lower than the risk from the table grapes. If such founder populations were to establish, P. callosus is estimated to spread by natural dispersal and common agricultural practices at a rate of 15.5 m/year (90% CR 5.1-46.8 m/year) after a lag phase of 4.0 years (90% CR 1.3-8.7 years). The impact, expressed as percentage loss of the production directly attributable to P. callosus in the areas where establishment is possible and assuming farmers do not apply specific control measures was estimated at 0.5% (90% CR 0.01%-2.8%) for cut flowers/foliage, 5.2% (90% CR 2.2%-11.7%) for apples and 2% (90% CR 1.3%-5.2%) for table grapes. Options for risk reduction are discussed, but their effectiveness is not quantified.

Barite Replacement as a Key Factor in the Genesis of Sediment-Hosted Zn-Pb±Ba and Barite-Sulfide Deposits: Ore Fluids and Isotope (S and Sr) Signatures from Sediment-Hosted Zn-Pb±Ba Deposits of Iran

Iran hosts more than 350 Precambrian to Cenozoic sediment-hosted Zn-Pb±Ba and barite-sulfide deposits, including shale-hosted massive sulfide (SHMS, also called SEDEX) and Irish-type and Mississippi Valley-type (MVT) mineralization, and barite is a common mineral in these deposits. In the SHMS deposits, barite is typically found as fine-grained disseminations in thin laminae. In these deposits, the sulfide laminae often occur as diagenetic replacements and as bands containing authigenic and diagenetic barite and pyrite framboids. In the Irish-type Zn-Pb-Ba and stratabound barite-sulfide deposits, barite exhibits various textures, including fine-grained disseminated barite, banded zebra textures, veins, and massive barite lenses. In some of the giant Irish-type deposits, as well as in the stratabound barite-sulfide mineralization, the main stratabound sulfide ore is developed within a barite envelope and is characterized by the replacement of barite and pyrite by chalcopyrite, galena, and sphalerite. In the MVT deposits, the formation of barite is often related to dolomitization, and sulfide mineralization involves the replacement of the dolomitized carbonate rocks, as well as associated barite. Fluid inclusion studies on the Irish-type deposits indicate that the temperatures and salinities of the sulfide-forming fluids are higher compared to those of the barite-forming fluids. Fluid inclusion analyses of coarse-grained barites from Irish and MVT deposits reveal their hydrothermal origin. The δ3⁴S values of sulfide minerals (pyrite, sphalerite, and galena) in Irish-type deposits exhibit a broad range of low values (mostly −28 to +5‰), primarily revealing a process of bacterial sulfate reduction (BSR). However, the textures (replacement, colloform, and banded) and more positive sulfur isotope values (+1 to +36‰) in the SHMS Zn-Pb deposits suggest that bacterial sulfate reduction (BSR) plays a less significant role. We suggest that thermochemical sulfate reduction (TSR) connected to the direct replacement of barite plays a more relevant role in providing sulfur for the sulfide mineralization in the SHMS, barite-sulfide, and MVT deposits. Based on the textual evidence, sulfur isotopic data, and fluid inclusion studies, barite has been identified as a key controller for the subsequent Zn-Pb mineralization by providing a suitable host and significant sulfur contribution in the sediment-hosted Zn-Pb and stratabound barite-sulfide deposits. This implies that diagenetic barite might be a precursor to all types of sediment-hosted Zn-Pb mineralization.

Ecology and geography of Cache Valley virus assessed using ecological niche modeling

BackgroundCache Valley virus (CVV) is an understudied Orthobunyavirus with a high spillover transmission potential due to its wide geographical distribution and large number of associated hosts and vectors. Although CVV is known to be widely distributed throughout North America, no studies have explored its geography or employed computational methods to explore the mammal and mosquito species likely participating in the CVV sylvatic cycle.MethodsWe used a literature review and online databases to compile locality data for CVV and its potential vectors and hosts. We linked location data points with climatic data via ecological niche modeling to estimate the geographical range of CVV and hotspots of transmission risk. We used background similarity tests to identify likely CVV mosquito vectors and mammal hosts to detect ecological signals from CVV sylvatic transmission.ResultsCVV distribution maps revealed a widespread potential viral occurrence throughout North America. Ecological niche models identified areas with climate, vectors, and hosts suitable to maintain CVV transmission. Our background similarity tests identified Aedes vexans, Culiseta inornata, and Culex tarsalis as the most likely vectors and Odocoileus virginianus (white-tailed deer) as the most likely host sustaining sylvatic transmission.ConclusionsCVV has a continental-level, widespread transmission potential. Large areas of North America have suitable climate, vectors, and hosts for CVV emergence, establishment, and spread. We identified geographical hotspots that have no confirmed CVV reports to date and, in view of CVV misdiagnosis or underreporting, can guide future surveillance to specific localities and species.Graphical

Broadening the application of Yarrowia lipolytica synthetic biology tools to explore the potential of Yarrowia clade diversity.

Yeasts have established themselves as prominent microbial cell factories, and the availability of synthetic biology tools has led to breakthroughs in the rapid development of industrial chassis strains. The selection of a suitable microbial host is critical in metabolic engineering applications, but it has been largely limited to a few well-defined strains. However, there is growing consideration for evaluating strain diversity, as a wide range of specific traits and phenotypes have been reported even within a specific yeast genus or species. Moreover, with the advent of synthetic biology tools, non-type strains can now be easily and swiftly reshaped. The yeast Yarrowia lipolytica has been extensively studied for various applications such as fuels, chemicals, and food. Additionally, other members of the Yarrowia clade are currently being evaluated for their industrial potential. In this study, we demonstrate the versatility of synthetic biology tools originally developed for Y. lipolytica by repurposing them for engineering other yeasts belonging to the Yarrowia clade. Leveraging the Golden Gate Y. lipolytica tool kit, we successfully expressed fluorescent proteins as well as the carotenoid pathway in at least five members of the clade, serving as proof of concept. This research lays the foundation for conducting more comprehensive investigations into the uncharacterized strains within the Yarrowia clade and exploring their potential applications in biotechnology.

Higher tree species richness and diversity in urban areas than in forests: Implications for host availability for invasive tree pests and pathogens

Urban and forest trees provide valuable ecosystem services. However, they are increasingly threatened by invasive forest pests and pathogens. Trees in urban areas are often the first potential hosts non-native tree-feeding insects and tree pathogens (“pests”) encounter after introduction in a novel region. If the trees encountered are suitable hosts, these pests can establish and become invasive – eventually also in surrounding forests. Here, we compared tree species and genus composition between urban areas and surrounding forests and examined the implications for host availability for forest pests and potential effects on invasibility. We compiled and standardised 26 urban tree inventories, containing ∼ 500.000 individual trees. We used multivariate analyses to compare urban tree composition with forest tree composition from forests surrounding each municipality (10 km radius), derived from the Swiss National Forest Inventory. With > 1300 different tree species, species richness of urban trees was 17 times higher than species richness in surrounding forests. Linear models and multivariate analyses revealed that host availability for forest quarantine pests is significantly higher in urban areas than in forests, with large differences in host suitability for different quarantine pests between urban and forest tree assemblages. This indicates that differences in species composition in urban and forest trees can result in increased host availability, possibly facilitating the establishment of quarantine forest pests.

Foraging behavior of Ganaspis brasiliensis in response to temporal dynamics of volatile release by the fruit–Drosophila suzukii complex

The lineage G1 of Ganaspis brasiliensis is a larval parasitoid of the worldwide pest Drosophila suzukii and one of its most effective natural enemies in the native area. Because of its high degree of host specificity, G. brasiliensis G1 is considered a suitable species for introduction in areas invaded by D. suzukii following a classical biological control approach. Indeed, the release of the parasitoid is currently implemented in the USA and Italy. G1 females attack only host larvae developing in ripening fresh fruits on the plant and not larvae that develop in decaying fruits. To date, virtually no information is available on the cues regulating the foraging behavior of G1. In this study, we therefore aimed to find out whether chemical cues are exploited by G1 females to: (i) locate host fruits; (ii) locate suitable host larvae within infested fruit; (iii) discriminate between infested fresh fruits and infested rotting ones. We used a model system composed of blueberries and D. suzukii tested in two-choice olfactometer bioassays (with D. suzukii-infested fruits, healthy fruits, and pure air as odor targets), followed by the collection and the characterization of volatile organic compounds (VOCs) released by the tested targets. The results showed a clear time-dependent choice made by G1 females of infested versus healthy fruits related to the concomitant development of D. suzukii larvae and fruit degradation. Attraction to infested fruits was recorded during the early stages of infestation, followed by a repellent phase coinciding with fruits largely degraded by larval feeding. We found that the attractiveness of G. brasiliensis G1 towards fruits infested by young larvae was associated with the detection of VOCs released by the infested blueberries, and host’s cuticular hydrocarbons. Conversely, the repellence of older and deteriorated fruits hosting developed D. suzukii larvae was associated with the detection of a fermentation compound produced by microorganisms likely carried inside the fruit by the flies. The response of G1 females to the temporal dynamics of chemical cues emitted by the fruit–host larvae complex was consistent with the high degree of specificity of the parasitoid towards the ripening host fruits and towards D. suzukii.

First Report of Powdery Mildew on Cardamine violifolia in China.

Cardamine violifolia, also called Cardamine hupingshanensis, is an economically important medicinal plant renowned for accumulating selenium (Guo et al., 2022). Selenium is an essential trace element with anti-oxidant, anti-inflammatory, anti-cancer, and immune regulatory functions. In July 2023, an outbreak of powdery mildew was detected, infecting the leaves of numerous C. violifolia plants in Enshi (30°11'5.27''N; 109°48'48.45''E), Hubei Province, China. This disease caused severe damage to plant leaves and stems, starting as individual spots and merging into a large mold that covers the entire leaf. It affected nearly 25% all C. violifolia plants, resulting in significant yield loss, disruption of normal metabolism, and premature aging. The lower leaf blades and underside of the leaves were particularly vulnerable. The affected leaves were collected and subjected to morphological diagnostic analysis (Mori et al., 2000) (Fig. S1). The powdery mildew species aggressively spread throughout the leaves, pedicels, and pods, persisting until present and often covering the entire surface. The conidiophores were upright, cylindrical, composed of 3 to 4 cells, and measured 92.3 ± 12.9 × 9.2 ± 0.6 μm (n = 30). Conidial pedicels had 21.6 ± 3.4 μm (n = 50) long cylindrical podocytes. The monoconidia were columnar or barrel-columnar, 30.60-55.59 × 9.11-20.00 μm in size. Conidia lacked an obvious cellulose body. The bud tubes formed from the end of conidia, and papillary appressoria developed on the epiphytic mycelia. ITS region sequences were amplified using the specific powdery mildew universal primers ITS1 (5'-TCCGTAGGTGAACCTGCGG-3'), PM6 (5'-GYCRCYCTGTCGCGAG-3') for partial sequences of 18S and 28S ribosomal DNA genes (Takamatsu et al., 2001). The sequence was deposited in the GenBank under the accession number OR506156 and aligned with available sequences on NCBI, which were 99.2%(528/532) identical to the E. cruciferarum (MT309701, MF192845, and KY660929) sequences (Fig. S2). The ITS sequence from GenBank was used to conduct maximum likelihood phylogenetic analysis using MEGA 11.0. The analysis results showed both the strain and E. cruciferarum clustered on the same branch. To confirm Koch's postulates, pathogenicity testing was carried out using an illuminating incubator. Infected leaves were attached to healthy leaves of C. violifolia seedlings (n=8). All the plants were incubated under 25℃ and >80% relative humidity. After one month, all inoculated plants presented the same symptoms as those initially observed in the field. Morphological and molecular analysis confirmed the isolated fungi's identity as the same pathogen. Therefore, C. violifolia is a suitable host for E. cruciferarum in China. The growers must be informed of these findings to prevent serious economic losses caused by this pathogenic white powder and to prepare for proper management practices. To our knowledge, this is the first report of E. cruciferarum infecting C. violifolia in China.

Bi-directionalized promoter systems allow methanol-free production of hard-to-express peroxygenases with Komagataella Phaffii

BackgroundHeme-incorporating peroxygenases are responsible for electron transport in a multitude of organisms. Yet their application in biocatalysis is hindered due to their challenging recombinant production. Previous studies suggest Komagataella phaffi to be a suitable production host for heme-containing enzymes. In addition, co-expression of helper proteins has been shown to aid protein folding in yeast. In order to facilitate recombinant protein expression for an unspecific peroxygenase (AnoUPO), we aimed to apply a bi-directionalized expression strategy with Komagataella phaffii.ResultsIn initial screenings, co-expression of protein disulfide isomerase was found to aid the correct folding of the expressed unspecific peroxygenase in K. phaffi. A multitude of different bi-directionalized promoter combinations was screened. The clone with the most promising promoter combination was scaled up to bioreactor cultivations and compared to a mono-directional construct (expressing only the peroxygenase). The strains were screened for the target enzyme productivity in a dynamic matter, investigating both derepression and mixed feeding (methanol-glycerol) for induction. Set-points from bioreactor screenings, resulting in the highest peroxygenase productivity, for derepressed and methanol-based induction were chosen to conduct dedicated peroxygenase production runs and were analyzed with RT-qPCR. Results demonstrated that methanol-free cultivation is superior over mixed feeding in regard to cell-specific enzyme productivity. RT-qPCR analysis confirmed that mixed feeding resulted in high stress for the host cells, impeding high productivity. Moreover, the bi-directionalized construct resulted in a much higher specific enzymatic activity over the mono-directional expression system.ConclusionsIn this study, we demonstrate a methanol-free bioreactor production strategy for an unspecific peroxygenase, yet not shown in literature. Hence, bi-directionalized assisted protein expression in K. phaffii, cultivated under derepressed conditions, is indicated to be an effective production strategy for heme-containing oxidoreductases. This very production strategy might be opening up further opportunities for biocatalysis.