Culture Methods Research Articles

Innovative Approaches for Sustainable Greenhouse Pest Management: Integrating Advanced Diagnostics and Preventive Strategies for Optimal Crop Health

Sustainable greenhouse pest management is critical for achieving good crop yields while reducing environmental impact. This paper investigates novel ways that incorporate improved diagnostic tools and preventive tactics into the framework of Integrated Pest Management (IPM) in greenhouse ecosystem. Understanding pest life cycles and utilizing cutting-edge tools such as DNA barcoding and enzyme-linked immunosorbent assays (ELISA) might help greenhouse managers achieve precise pest diagnosis and early responses. Preventive measures, such as tight cleanliness protocols, quarantine techniques, and the use of physical barriers, are critical in lowering pest incidence. Furthermore, biological controls, cultural methods such as crop rotation and intercropping, and the selective use of insecticides and biopesticides collectively help to ensure long-term pest management. This complete approach not only reduces insect damage but also creates a healthier, more resilient agricultural ecology. Furthermore, traditional diagnostic and preventive measures were also discussed in the article. The and recommendations are validated by recent studies and data, demonstrating the success of these integrated tactics in modern greenhouse operations.

The Coexistence of Klebsiella pneumoniae and Candida albicans Enhanced Biofilm Thickness but Induced Less Severe Neutrophil Responses and Less Inflammation in Pneumonia Mice Than K. pneumoniae Alone

Due to the possible coexistence of Klebsiella pneumoniae (KP) and Candida albicans (CA), strains of KP and CA with biofilm production properties clinically isolated from patients were tested. The production of biofilms from the combined organisms (KP+CA) was higher than the biofilms from each organism alone, as indicated by crystal violet and z-stack immunofluorescence. In parallel, the bacterial abundance in KP + CA was similar to KP, but the fungal abundance was higher than CA (culture method), implying that CA grows better in the presence of KP. Proteomic analysis was performed to compare KP + CA biofilm to KP biofilm alone. With isolated mouse neutrophils (thioglycolate induction), KP + CA biofilms induced less prominent responses than KP biofilms, as determined by (i) neutrophilic supernatant cytokines (ELISA) and (ii) neutrophil extracellular traps (NETs), using immunofluorescent images (neutrophil elastase, myeloperoxidase, and citrullinated histone 3), peptidyl arginine deiminase 4 (PAD4) expression, and cell-free DNA. Likewise, intratracheal KP + CA in C57BL/6 mice induces less severe pneumonia than KP alone, as indicated by organ injury (serum creatinine and alanine transaminase) (colorimetric assays), cytokines (ELISA), bronchoalveolar lavage fluid parameters (bacterial culture and neutrophil abundances using a hemocytometer), histology score (H&E stains), and NETs (immunofluorescence on the lung tissue). In conclusion, the biofilm biomass of KP + CA was mostly produced from CA with less potent neutrophil activation and less severe pneumonia than KP alone. Hence, fungi in the respiratory tract might benefit the host in some situations, despite the well-known adverse effects of fungi.

Overview of Brinjal (Solanum melongena L.) Pests and Their Management: A Review

Brinjal (eggplant or aubergine), a widely cultivated vegetable in tropical and subtropical regions, faces significant threats from a variety of insect pests. These pests cause substantial damage at different stages of crop growth, leading to reduced yields and compromised fruit quality. This review provides a comprehensive overview on the major insect pests that affect brinjal cultivation, focusing on their biology, damage mechanisms, and current management practices. Special emphasis is placed on integrated pest management (IPM) strategies, combining cultural, biological and chemical control methods to promote sustainable brinjal production.

Mitochondrial AOX1a and an H2O2 feed‐forward signalling loop regulate flooding tolerance in rice

SummaryFlooding is a widespread natural disaster that causes tremendous yield losses of global food production. Rice is the only cereal capable of growing in aquatic environments. Direct seeding by which seedlings grow underwater is an important cultivation method for reducing rice production cost. Hypoxic germination tolerance and root growth in waterlogged soil are key traits for rice adaptability to flooded environments. Alternative oxidase (AOX) is a non‐ATP‐producing terminal oxidase in the plant mitochondrial electron transport chain, but its role in hypoxia tolerance had been unclear. We have discovered that AOX1a is necessary and sufficient to promote germination/coleoptile elongation and root development in rice under flooding/hypoxia. Hypoxia enhances endogenous H2O2 accumulation, and H2O2 in turn activates an ensemble of regulatory genes including AOX1a to facilitate the conversion of deleterious reactive oxygen species to H2O2 in rice under hypoxia. We show that AOX1a and H2O2 act interdependently to coordinate three key downstream events, that is, glycolysis/fermentation for minimal ATP production, root aerenchyma development and lateral root emergence under hypoxia. Moreover, we reveal that ectopic AOX1a expression promotes vigorous root and plant growth, and increases grain yield under regular irrigation conditions. Our discoveries provide new insights into a unique sensor–second messenger pair in which AOX1a acts as the sensor perceiving low oxygen tension, while H2O2 accumulation serves as the second messenger triggering downstream root development in rice against hypoxia stress. This work also reveals AOX1a genetic manipulation and H2O2 pretreatment as potential targets for improving flooding tolerance in rice and other crops.

OBTAINING ASEPTIC CULTURE IN VITRO OF RHODIOLA SEMENOWII L. FROM VARIOUS TYPES OF EXPLANTS

In vitro cultivation of callus cells allows for the stable production of secondary metabolites, being an alternative to the use of medicinal plant biomass from natural populations. Standardization of cultivation protocols and technological processes for the synthesis of secondary metabolites in vitro conditions determines significant interest in the use of tissue culture methods (PTC – Plant Tissue Culture). The aim of the study was to obtain an aseptic culture of Rhodiola semenowii in vitro from various types of explants, as well as the effect of the hormonal composition of the nutrient medium on the induction of callusogenesis in the culture of isolated organs of R. semenowii. As a result of the research, the optimal mode of obtaining R. semenovii aseptic cultures from various explants was determined. It has been shown that the use of 70% EtOH in combination with 0.01% HgCl2 or 20% NaClO makes it possible to obtain aseptic cultures from any type of R. semenovii explants. It has been established that the type of explant is R. semenovii influenced both the frequency of callus formation and the beginning of its formation. The least optimal types of R. semenovii explants are stem and rhizome segments, the frequency of callusogenesis varied from 5.8% to 75.6%. The induction of callusogenesis also depended on the hormonal composition of the nutrient medium: the highest percentage of callus formation in all types of explants was on media containing 0.5mg L−1 thidiazuron (TDZ) + 1.0 mg L−1 1-naphthaleneacetic acid (NAA) and 0.5mg L−12,4D + 2.0 mg L−1 6-Benzylaminopurine (BAP).

Cultivation and Growth Dynamics of Capelin (Mallotus villosus) from Hatch to Adulthood

This study describes the first successful rearing of capelin from hatch to adulthood in a laboratory setting using intensive culture methods. Over the span of about two years, the capelin were reared in aquaculture tanks under a constant temperature of 7 °C. The capelin demonstrated a robust linear growth during their first year of life, with a mean length increment of 0.36 mm per day. Due to their accelerated growth, some of the capelin became sexually mature as early as one year post-hatch. The first year was characterized by a rapid increase in condition factor (CF) while the second year showed a plateau. The von Bertalanffy growth equation effectively described the two-year growth of the cultivated capelin, predicting an asymptotic length (L∞) of 18.4 cm, similar to the 18.6 cm median L∞ of wild Icelandic capelin (1981–2018 cohorts). The cultivated capelin were projected to reach this length in 2.6 years, compared to about 6 years for wild capelin. This study provides new insights into the growth dynamics of capelin and although the species is sensitive to handling, it demonstrates that intensive culture methods can be used to investigate biological aspects of this important forage species.

Case report: A case of blood culture-negative Bartonella quintana endocarditis: blood mNGS is an efficient method for early diagnosis

Bartonella quintana is one of the main causes of blood culture-negative endocarditis, and routine blood culture and serological methods are difficult to achieve early diagnosis. We report a case of blood culture-negative Bartonella quintana endocarditis from southwestern Chongqing. The patient was a 67-year-old male scavenger who presented with heart failure without fever as the main clinical manifestation upon admission. He stated having had contact with stray cats in the past 2 months. The combination of clinical symptoms, echocardiography, and blood mNGS testing confirmed the infection of Bartonella quintana.

Potential Impact of Rapid Molecular Microbiologic Diagnosis for Mechanically Ventilated Children in Intensive Care With Suspected Pneumonia

Background: Lower respiratory tract infections (LRTIs) remain a leading cause of community-acquired and nosocomial infection in children and a common indication for antimicrobial use and intensive care admission. Determining the causative pathogen for LRTIs is difficult and traditional culture-based methods are labor- and time-intensive. Emerging molecular diagnostic tools may identify pathogens and detect antimicrobial resistance more quickly, to enable earlier targeted antimicrobial therapy. Methods: This is a single-center, prospective observational laboratory study evaluating the use of the Biofire FilmArray pneumonia panel (FA-PP) (BioFire Diagnostics, Salt Lake City, UT) for bronchoalveolar lavage specimens from mechanically ventilated children admitted with suspected or presumed pneumonia. We aimed to determine its feasibility and utility for identifying pathogens, antimicrobial resistance and its potential influence on antibiotic prescribing. Results: We analyzed 50 samples taken from 41 children with a median age of 6 months. Positive agreement between culture and FA-PP was 83% and negative agreement was 76%. Agreement between FA-PP (mecA/C or MREJ) and culture was high for methicillin-resistant Staphylococcus aureus. In 3 cases, extended-spectrum beta-lactamase-producing Gram-negative organisms were detected by culture and not FA-PP. Hypothetically, FA-PP results would have affected antimicrobial prescribing in approximately half the cases (24, 48%). Conclusions: FA-PP is a useful adjunct to traditional culture methods in mechanically ventilated children with LRTIs and may influence clinical decision-making regarding antibiotic escalation or stewardship.

Improvement of Transplanting Rice Yield and Nitrogen Use Efficiency by Increasing Planting Density in Northeast China Under the Optimal Nitrogen Split-Fertilizer Applications

There are few studies on how nitrogen (N) fertilizer application rates and transplanting densities impact rice yield, root distribution, and N use efficiency in the cold regions of Northeast China. This research involved a two-year field trial utilizing Jinongda 667 as the material. In 2021, three N split-fertilizer applications—T1 (6:3:1), T2 (5:3:2), T3 (4:3:3)—and two transplanting densities—D1 (30 cm × 13.3 cm) and D2 (30 cm × 20 cm)—were compared with the conventional cultivation mode (T0: 175 kg N hm−2, 6:3:1), whereby the N application mode most suitable for increasing density was explored. In 2022, four N application levels—0 (N0), 125 (N1), 150 (N2), and 175 (N3) kg N hm−2—were assessed under the same density treatment to analyze the yield, resource utilization efficiency, and root traits of Jinongda 667. The results indicated that when the transplanting density was 30 cm × 13.3 cm, the application of 5:3:2 fertilizer was more conducive to improving rice yield. Increasing planting density under reduced N input significantly enhanced both rice yield and N use efficiency. In contrast to the conventional cultivation method (D2N3), the treatment of increased planting density (D1N2) under reduced N input led to a 21.2% rise in the number of panicles per square meter and an 8.6% boost in rice yield. Furthermore, increasing planting density under reduced N input significantly enhanced the agronomic efficiency of N fertilizer, the apparent utilization rate, and the N harvest index. It also boosted the SPAD value, photosynthetic rate, and the utilization efficiency of light and N resources in rice. However, it was noted that root enzyme activity decreased. This study demonstrated that increasing planting density, combined with the N application mode of 5:3:2 and an N application rate of 150 kg hm−2, maximized resource utilization efficiency, optimized root absorption capacity, and resulted in higher yields.

Growth Response of Kappaphycus Alvarezii at Different Depth in the Waters of Batu Bao, East Nusa Tenggara

ABSTRACT Seaweed is one of the important fisheries commodities with significant economic value. Kappaphycus alvarezii, commonly known as Eucheuma cottoni in the market, is a type of seaweed that produces kappa carrageenan. This research aims to determine the effective planting depth for the growth, morphological and histological changes, and the environmental parameters influencing the growth of K. alvarezii. The cultivation method used in this research is the longline method, and it employs a Completely Randomized Design consisting of four treatments: 15cm, 30cm, 45cm, and 60cm, with each treatment replicated five times. The data were analyzed using analysis of variance, and if significant differences were found, Duncan's post hoc test was conducted. The research results indicate that an increase in planting depth has a significant impact on specific growth rate (SGR) and average growth rate (AGR). Deeper planting depths result in slower growth. The best depth for the growth, morphology, and histology of K. alvarezii during the study was observed at a depth of 15 cm. Environmental parameters such as temperature, salinity, water flow rate, water clarity, and pH have a positive influence on the growth of K. alvarezii.

Multiple skin abscesses due to Nocardia neocaledoniensis: a case report and literature review

N. neocaledoniensis is a very rare infectious pathogen that causes human disease, particularly in immunocompromised individuals. In this case report, we describe the successful diagnosis of N. neocaledoniensis in a patient confirmed by mNGS and the treatment of multiple skin abscesses due to N. neocaledoniensis infection. mNGS is an important diagnostic method complementary to routine bacterial culture and identification methods, especially for rare, novel, co-infected pathogens, and pathogens that are difficult to culture. This report may provide a reference for the clinical treatment and diagnosis of N. neocaledoniensis infection in humans.

Advancements in Organoid Culture Technologies: Current Trends and Innovations.

Organoids have emerged as valuable tools in investigating disease mechanisms, drug efficacy, and personalized medicine due to their capacity to recapitulate crucial aspects of tissue physiology, including cell-cell interactions, heterogeneity, microenvironmental cues, and drug responses. Despite their broad applicability across various research domains, conventional organoid culture methods are plagued by several limitations that hinder research progress. These limitations include the inability to faithfully recreate tissue microenvironments, immune contexts, and vascular systems. Fortunately, ongoing advancements in organoid culture techniques are addressing these shortcomings. In this review, we provide a comprehensive overview of current mainstream organoid culture protocols. By evaluating these protocols, researchers can identify the most suitable experimental methods, thereby optimizing resource allocation and experimental outcomes.

Wastewater-associated plastispheres: A hidden habitat for microbial pathogens?

Wastewater treatment plants (WWTPs) receive wastewater from various sources. Despite wastewater treatment aiming to remove contaminants, microplastics persist. Plastic surfaces are quickly colonized by microbial biofilm ("plastispheres"). Plastisphere communities are suggested to promote the spread and survival of potential human pathogens, suggesting that the transfer of plastispheres from wastewater to the environment could pose a risk to human and environmental health. The study aimed to identify pathogens in wastewater plastispheres, specifically food-borne pathogens, in addition to characterizing the taxonomic diversity and composition of the wastewater plastispheres. Plastispheres that accumulated on polypropylene (PP), polyvinyl chloride (PVC), and high-density polyethylene propylene (HDPE) surfaces exposed to raw and treated wastewater were analyzed via cultivation methods, quantitative reverse transcription PCR (RT‒qPCR) and 16S rRNA amplicon sequencing. RT‒qPCR revealed the presence of potential foodborne pathogenic bacteria and viruses, such as Listeria monocytogenes, Escherichia coli, norovirus, and adenovirus. Viable isolates of the emerging pathogenic species Klebsiella pneumoniae and Acinetobacter spp. were identified in the plastispheres from raw and treated wastewater, indicating that potential pathogenic bacteria might survive in the plastispheres during the wastewater treatment. These findings underscore the potential of plastispheres to harbor and disseminate pathogenic species, posing challenges to water reuse initiatives. The taxonomic diversity and composition of the plastispheres, as explored through 16S rRNA amplicon sequencing, were significantly influenced by the wastewater environment and the duration of time the plastic spent in the wastewater. In contrast, the specific plastic material did not influence the bacterial composition, while the bacterial diversity was affected. Without efficient wastewater treatment and proper plastic waste management, wastewater could act as a source of transferring plastic-associated pathogens into the food chain and possibly pose a threat to human health. Continued research and innovation are essential to improve the removal of microplastics and associated pathogenic microorganisms in wastewater.

Improved visualisation of ACP-engineered osteoblastic spheroids: a comparative study of contrast-enhanced micro-CT and traditional imaging techniques

This study investigates osteoblastic cell spheroid cultivation methods, exploring flat-bottom, U-bottom, and rotary flask techniques with and without amorphous calcium phosphate (ACP) supplementation to replicate the 3D bone tissue microenvironment. ACP particles derived from eggshell waste exhibit enhanced osteogenic activity in 3D models. However, representative imaging of intricate 3D tissue-engineered constructs poses challenges in conventional imaging techniques due to notable scattering and absorption effects in light microscopy, and hence limited penetration depth. We investigated contrast-enhanced micro-CT as a methodological approach for comprehensive morphological 3D-analysis of thein-vitromodel and compared the technique with confocal laser scanning microscopy, scanning electron microscopy and classical histology. Phosphotungstic acid and iodine-based contrast agents were employed for micro-CT imaging in laboratory and synchrotron micro-CT imaging. Results revealed spheroid shape variations and structural integrity influenced by cultivation methods and ACP particles. The study underscores the advantage of 3D spheroid models over traditional 2D cultures in mimicking bone tissue architecture and cellular interactions, emphasising the growing demand for novel imaging techniques to visualise 3D tissue-engineered models. Contrast-enhanced micro-CT emerges as a promising non-invasive imaging method for tissue-engineered constructs containing ACP particles, offering insights into sample morphology, enabling virtual histology before further analysis.

An Etiology of Sputum Culture Contamination and their Drugs Susceptibility Patterns among TB-suspected Patients at Epicentre Mbarara Research Centre, Uganda

Aims: Contamination in TB cultures is problematic as it allows the growth of non-target bacterial or fungal species in sputum samples, potentially concealing M. tuberculosis. This study aimed to determine the prevalence, characterization, and drug susceptibility patterns of contaminants in sputum cultures from TB-suspected patients. Methods: A laboratory-based cross-sectional study was conducted from October 2023 to June 2024 at the Epicentre Mbarara Research Centre, involving 81 sputum samples from TB-suspected patients. Conventional decontamination procedures were used, and both solid Lowenstein Jensen (LJ) and liquid Mycobacterium Growth Indicator Tube (MGIT) culture methods were applied. Standard culture techniques and physicochemical analyses were employed. Bacteria were incubated at 37°C in O2 and Co2 incubators, with growth observed within 24 to 48 hours and up to three days for yeast contaminants. Sabouraud Dextrose Agar cultures were monitored for two weeks for filamentous fungi. Antibiograms for bacterial isolates were determined using the Kirby-Bauer, while broth microdilution was used for fungal isolates. Results: Of 81 samples, 14(17.28%) were contaminated. The most common bacterial contaminants were Staphylococcus aureus, Bacillus subtilis, Streptococcus viridans, Staphylococcus epidermidis, and Escherichia coli. Fungal contaminants included Aspergillus flavus, Penicillium species, Candida albicans, Cryptococcus neoformans, and non-albican Candida species. High resistance to erythromycin (73%) and tetracycline (50%) was noted among bacterial isolates, with sensitivity to vancomycin (67%), gentamycin (60%), and chloramphenicol (58%). Yeast isolates showed (87.5%) susceptibility to itraconazole and (75%) to amphotericin B but (50%) resistance to 5-flucytosine. Mold isolates exhibited (100%) susceptibility to itraconazole, amphotericin B, and 5-flucytosine but complete resistance to caspofungin. Conclusion: The level of contamination in TB cultures was noted, and Staphylococcus aureus and Candida albicans were observed as the most common contaminants. Bacterial isolates resisted erythromycin and tetracycline, while fungal isolates were generally susceptible to itraconazole and amphotericin B. We recommend Strengthening decontamination and monitoring common resistant contaminants.

Economic-ecological assessment of multi-crop small-scale model farms in the Republic of Armenia (piloting results from Gegharqunik region)

Background: Biogenic elements such as nitrogen (N), phosphorus (P), and potassium (K) are major pollutants of Lake Sevan's water, causing eutrophication and deterioration of water quality. Untreated wastewater, sewage, agriculture, fish farming, and other activities are significant sources of these pollutants. Vegetable cultivation, heavily reliant on mineral fertilizers and chemicals, and livestock breeding contribute substantially to nitrogen pollution in the lake. However, improved vegetable growing practices, including small-scale agriculture using organic fertilizers, natural soil improvers, and eco-friendly bio-liquids as growth stimulants, can enhance the socio-economic conditions for farmers in the Lake Sevan basin. These practices promote environmentally friendly, water-saving land-use methods, reducing nitrogen leaching into the lake. Objective: This study aims to demonstrate the socio-economic and environmental benefits of applying innovative agro-technology to small-scale farming in the Lake Sevan watershed (Gegharkunik region). Specifically, the study objectives are to determine the quality characteristics of crops grown in small-scale farms and compare these quality indicators between yields obtained through the proposed technology and traditional cultivation methods. Methods: Field trials of multi-crop systems were conducted on 0.25 ha land plots in four variations across four different locations in the Lake Sevan watershed. The trials were evaluated for various parameters. Nitrate content, dry matter, starch, vitamin C, and sugar levels were determined in potatoes, cabbage, broccoli, beets, and green beans, while fat content was measured in green beans and green peas. Results: Economic efficiency calculations revealed that utilizing the proposed innovative agro-technology in a multi-crop small-scale farm (0.25 ha) yielded a 20-25% increase compared to traditional cultivation with synthetic fertilizers, accompanied by notable environmental benefits. Conclusions: The study demonstrates that small-scale agricultural practices employing the proposed agro-technology have significant socioeconomic and environmental impacts on local farmers. Keywords: Cultivation of non-traditional crops, organic farming, small-scale farming, activities, economic and ecological evaluation, and food quality characteristics.

Chemical and Physical Aspects of Soil Health Resulting from Long-Term No-Till Management

The aim of this study was to compare the long-term effects of conventional tillage (CT) and no-till (NT) systems on the main soil properties that determine soil health. The research was conducted in a field experiment established in 1975 in Chylice, central Poland, at the WULS-SGGW Experimental Station Skierniewice. Soil samples collected from 0–10 and 10–20 cm of the mollic horizon of the Phaeozem were analysed for total organic carbon (TOC) content, fractional composition of SOM and spectroscopic properties of humin, soil structural stability, soil water retention characteristics and soil water repellency (SWR). The results showed that NT practice almost doubled the TOC in the 0–10 cm layer. However, optical parameters of humin indicated that NT management promoted the formation of humin with a lower molecular weight and lower degree of condensation of aromatic structures. In the NT 0–10 cm layer, a significant increase in the number of water-resistant macroaggregates was found. In the 0–10 cm layer, the water capacity increased by 9%, 18%, 22% and 26% compared to CT at (certain soil suction) pF values of 0.0, 2.0, 3.0 and 4.2, respectively. SWR occurs regardless of the cultivation method at a soil moisture equivalent to pF 4.2, and the greatest range of SWR was found in the NT 0–10 cm layer.

Experimental determination of maximum shear stress in Mobius® Breez perfusion microbioreactors and comparative analysis with stirred tank bioreactors

Perfusion processes have experienced increased popularity in recent years due to their ability to sustain high cell densities and productivities in biopharmaceutical production, offering advantages over traditional batch and fed-batch cultivation methods. The Mobius® Breez microbioreactor significantly reduces experimental effort by downsizing the classical volume of perfusion bioreactors to the mL range and thus represents a valuable tool for process development. However, miniaturization has raised questions regarding comparability with traditional bioreactors in terms of the physical environment, such as hydrodynamic shear stress. Therefore, the maximum hydrodynamic shear stress, cultivation performance, and membrane-wall contact were evaluated to elucidate the system's behavior. Findings reveal two distinct operational conditions, distinguished by the presence or absence of membrane-wall contact, resulting in varying levels of hydrodynamic stress. Conditions lacking membrane contact demonstrate stress levels within safe operating thresholds for CHO cells, while those involving membrane contact exceed these thresholds, potentially leading to cell damage. Through the identification of critical frequencies of membrane motion, this study offers insights for optimizing microbioreactor operation and enhancing overall bioprocess efficiency.

Establishing an Interactive Sequence Database for Shiitake Cultivar Identification.

Shiitake mushrooms (Lentinula edodes) hold significant cultural and economic value, particularly in Asia where they are extensively cultivated. The diversification of shiitake cultivars, driven by the need to adapt to various climatic conditions and cultivation methods, has resulted in over 200 distinct cultivars. Reliable identification of these cultivars is crucial for breeding, intellectual property protection, and effective genetic resource management. Traditional morphological methods are inadequate due to their subjectivity and labor-intensive nature. This study leverages nanopore high-throughput sequencing to comprehensively analyze the rDNA regions (SSU, ITS, LSU, IGS) of 41 shiitake strains from Taiwan's Bioresource Collection and Research Center (BCRC), comprising 5 wild strains, 33 commercial strains, and 3 wild-commercial hybrids. Our results identified the IGS1 region as the most variable and suitable for cultivar differentiation. Consequently, we developed an interactive online database (https://github.com/Raingel/ShiitakeIGS1) that integrates 317 IGS1 sequences from Taiwan, Japan, and China. This platform allows users to upload their IGS1 sequences and identify similar cultivars through a user-friendly interface, enhancing the precision and efficiency of shiitake cultivar identification.

Wood microbiome variation and interactions with fungal symbionts of invasive ambrosia beetles

The microbiomes of plants can modulate the impacts of pests, including through interactions with microbiomes of pathogen vectors such as ambrosia beetles. While physical and chemical traits of plant hosts are known to affect beetle-carried microbes, how beetle and host microbiomes interact is seldom explored. We aimed to determine whether wood-inhabiting endophytes mediate host susceptibility to Fusarium dieback, an emergent tree disease complex that includes ambrosia beetle vectors. We studied three competent host tree species (Persea americana, Salix spp., Platanus racemosa) common in disease hot spots in agricultural and wildland habitats. Using culturing methods, we compared wood microbiomes of 319 attacked and 133 non-attacked trees across a network of 47 beetle-infested and 41 non-infested plots in southern California, USA. We conducted 1,148 in vitro assays to evaluate antagonism by wood-inhabiting endophytic fungi (60 species) and bacteria (40 species) to the Fusarium pathogens, finding 15 fungal and 11 bacterial species with clear antagonism to the pathogen. Such wood endophytes may potentially protect tree hosts as biological control agents. Antagonistic microbes were more common in attacked trees than in non-attacked trees, suggesting the abundance of antagonistic fungi and bacteria in the wood is insufficient to determine host susceptibility to attack or they are enriched in attacked trees. Wood-inhabiting microbial communities were consistently different between cultivated P. americana and wildland tree species, as there were some differences based on host attack status. Differences between attacked and non-attacked trees were reflected in different microbial consortia rather than the abundance of individual, antagonistic microbial species.