Search Results (38)

Iron Age settlements in the Salento peninsula (Southern Italy, 8th–6th century BC) underwent fundamental transformations in social organization, marked by the emergence of local elites through trade development and intense contacts with the Greek world. This study examines organic residue assemblages from 99 ceramic sherds from one key Iron Age site to clarify the role of locally produced ceramics—both coarse ware containers and Japigian matt-painted vessels—in commensal and beverage production practices. Chromatographic analyses identified a wide variety of animal and plant by-products, including fats, oils, waxes, and resin compounds. Integrated phytolith and starch analysis revealed evidence consistent with fermentation processes, particularly through the identification of fungal remains and damaged starch granules suggesting brewing activities in a subset of vessels. Matt-painted pottery forms—characterized by conical rims, funnel-shaped necks, bowls, and jugs—show distinctive use-alteration patterns and residue profiles associated with fermented beverage consumption and preparation in approximately 26% of the analyzed assemblage. Integrating organic residue analysis, experimental archaeology, and microfossil investigation suggests the central role of locally produced pottery in Iron Age commensal activities and status display, though alternative interpretations for some biomarker profiles cannot be excluded. This multiproxy approach demonstrates functional differentiation and consumption practices, refining interpretations of vessel use and providing new insights into food economies and social life during the Iron Age in southern Italy. Full article

Excessive fertilizer use drives soil degradation and resource waste. This study investigates how arbuscular mycorrhizal fungi (AMF) formulations (powder vs. granular) optimize maize (Zea mays L.) yield, soil microbiome, and economic benefits under 50% and 75% fertilizer reduction. Field trials showed that the AMF powder formulation under 50% fertilizer reduction (AP50) increased maize yield by 14.67%. This increase was associated with rapid root colonization (85.3%), enhanced phosphorus availability, and the recruitment of beneficial fungi such as Mortierellomycota. Granular formulation at 75% reduction (AG75) achieved 7.18% yield gain via sustained symbiosis. Fungal communities exhibited greater sensitivity to fertilization than bacteria (Chao1, p = 0.0094), with AMF suppressing Fusarium by 42% while enriching functional taxa (Actinobacteria, Mortierellomycota). Economic analysis confirms that AP50 (30,435 CNY/ha) and AG75 (26,954 CNY/ha) yield higher net profits, where CNY denotes Chinese Yuan. Powder formulations maximize immediate benefits in medium- to low-fertility soils, whereas granules support long-term soil health in high-organic systems, providing a precision strategy for sustainable agriculture. Full article

The fungus Coleosporium zanthoxyli is the causal agent of leaf rust in Chinese prickly ash pepper (Zanthoxylum armatum ‘Hanyuan putaoqing’), seriously impacting its industrial development. However, little is currently known about the infection and pathogenesis of C. zanthoxyli on Z. armatum. In this study, the infection of Z. armatum by C. zanthoxyli was reported at histological and cytological levels by a fluorescence microscope and transmission electron microscopy (TEM) for the first time. Fluorescence microscopy with fluorophore Alexa 488 (WGA-FITC) stained samples revealed that the infection process comprised three distinct stages: penetration (0–1 days post inoculation, dpi), parasitic growth (3–5 dpi), and sporulation (≥7 dpi). The number of haustoria increased during the osmotic and parasitic periods and then decreased; the length of hyphae also increased rapidly and then decreased. TEM analysis during these stages demonstrated that as disease severity increased, chloroplasts and mitochondria enlarged significantly, accompanied by a marked accumulation of starch granules and osmiophilic granules. At later stages, the nuclei became irregular, the grana lamellae were blurred, and the lamellar structure was arranged disorderly, and leaf tissues were extensively colonized by fungal hyphae and haustoria, leading to cellular necrosis and distorted cell walls. Notably, the sporulation phase was characterized by dense rust spore clusters covering the leaf surface. These findings provide critical insights into the ultrastructural changes induced by C. zanthoxyli during infection, elucidating key mechanisms of rust-induced damage in Chinese prickly ash and identifying the parasitic phase as a critical window for control strategies. This study lays a foundation for further research on rust pathogenesis and the development of Chinese prickly ash targeted control strategies. Full article

The genus Cunninghamella is widely distributed, primarily saprotrophic, occasionally endophytic and phytopathogenic. Analysis based on the internal transcribed spacer (ITS), the large subunit (LSU) of ribosomal DNA, and translation elongation factor 1 alpha (TEF1α), along with morphological comparisons, resulted in a discovery of eight new species. Molecular phylogenetic analyses placed each of these new species within well-supported clades. Cunninghamella crassior sp. nov., with short and thick spines, and C. fusca sp. nov. with brown sporangiola, are sister clades to each other. C. diffundens sp. nov., containing dispersed granules in sporangiola, is closely related to C. irregularis Zhao. C. tuberculata sp. nov., producing sporangiola with nodule-like protrusions. C. fulvicolor sp. nov., sister to C. irregularis, forms yellowish-brown pigmented colonies. C. guttulata sp. nov., with teardrop-shaped sporangiola, and C. inaequalis sp. nov., with uneven sporangiola, are both closely related to C. regularis Zhao. C. monosporangiola sp. nov., characterized by only one sporangiolum on some vesicles, is sister to C. verrucosa Zhao. This study represents the eleventh installment in a series investigating early-diverging fungal diversity in China and expands the number of accepted species in Cunninghamella to 39. Full article

This study explores the effects of Tricholoma matsutake-derived insoluble dietary fiber (TMIDF) on the pasting behavior, structural properties, and in vitro digestibility of rice flour. The incorporation of 5% TMIDF significantly increased the peak viscosity (from 2573.21 to 2814.52 mPa·s) by competitively adsorbing water and forming a dense transient network, while simultaneously reducing the final viscosity (from 1998.27 to 1886.18 mPa·s) by inhibiting amylose recrystallization. Multi-scale structural analyses revealed that TMIDF enhanced V-type crystallinity and limited enzyme access via a porous fibrous matrix. Fourier-transform infrared spectroscopy and low-field nuclear magnetic resonance analyses confirmed that hydrogen bonding and water redistribution were key interaction mechanisms. TMIDF significantly lowered in vitro starch digestibility and increased resistant starch content by 16% (from 14.36% to 30.94%) through synergistic effects, including physical encapsulation of starch granules, formation of enzyme-resistant amylose-lipid complexes, and α-amylase inhibition (31.08%). These results demonstrate that TMIDF possesses a unique multi-tiered modulation mechanism, involving structural optimization, enzyme suppression, and diffusion control, which collectively surpasses the functional performance of conventional plant-derived insoluble dietary fibers. This research establishes a theoretical basis for applying fungal insoluble dietary fibers to develop low glycemic index functional foods, highlighting their dual role in improving processing performance and nutritional quality. Full article

Objective: The study aimed to optimize protocols for the joint extraction of deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) from 0.025 × 10⁶ CFU of Candida albicans, targeting to overcome the challenges in the extraction of these genetic materials. Materials and methods: From this, treated silicon carbide (SiC) granules were added to fungal samples from methods 1, 2, and 3 obtained from aliquots of BHI or Sabouraud medium to cause cell lysis and enable the isolation of these macromolecules by phenol and chloroform. The concentration and integrity of the extracted nucleic acids were analyzed, respectively, by spectrophotometry using the A₂₆₀/A₂₈₀ ratios and 1% agarose gel electrophoresis. Results: Therefore, method 3 is the one that most comprises samples considered pure of both DNA and RNA, simultaneously. Furthermore, the presence of intact RNAs corresponding to the base pair size such as 5.8 S rRNA and tRNA was verified during electrophoresis, considering the particularities of RNA, which makes it very unstable and easily degraded. Conclusions: Thus, it results in a faster and simpler method in addition to obtain promising results using minimal amounts of biological sample and offering a valuable alternative for small laboratories to work with molecular biology. Full article

Microbotryum lychnidis-dioicae is an obligate fungal species colonizing the plant host, Silene latifolia. The fungus synthesizes and secretes effector proteins into the plant host during infection to manipulate the host for completion of the fungal lifecycle. The goal of this study was to continue functional characterization of such M. lychnidis-dioicae effectors. Here, we identified three putative effectors and their putative host-plant target proteins. MVLG_02245 is highly upregulated in M. lychnidis-dioicae during infection; yeast two-hybrid analysis suggests it targets a tubulin α-1 chain protein ortholog in the host, Silene latifolia. A potential plant protein interacting with MVLG_06175 was identified as CASP-like protein 2C1 (CASPL2C1), which facilitates the polymerization of the Casparian strip at the endodermal cells. Proteins interacting with MVLG_05122 were identified as CSN5a or 5b, involved in protein turnover. Fluorescently labelled MVLG_06175 and MVLG_05122 were expressed in the heterologous plant, Arabidopsis thaliana. MVLG_06175 formed clustered granules at the tips of trichomes on leaves and in root caps, while MVLG_05122 formed a band structure at the base of leaf trichomes. Plants expressing MVLG_05122 alone were more resistant to infection with Fusarium oxysporum. These results indicate that the fungus might affect the formation of the Casparian strip in the roots and the development of trichomes during infection as well as alter plant innate immunity. Full article

Mast cells (MCs) are abundant at sites exposed to the external environment and pathogens. Local activation of these cells, either directly via pathogen recognition or indirectly via interaction with other activated immune cells and results in the release of pre-stored mediators in MC granules. The release of these pre-stored mediators helps to enhance pathogen clearance. While MCs are well known for their protective role against parasites, there is also significant evidence in the literature demonstrating their ability to respond to viral, bacterial, and fungal infections. Vitamin D is a fat-soluble vitamin and hormone that plays a vital role in regulating calcium and phosphorus metabolism to maintain skeletal homeostasis. Emerging evidence suggests that vitamin D also has immunomodulatory properties on both the innate and adaptive immune systems, making it a critical regulator of immune homeostasis. Vitamin D binds to its receptor, called the vitamin D receptor (VDR), which is present in almost all immune system cells. The literature suggests that a vitamin D deficiency can activate MCs, and vitamin D is necessary for MC stabilization. This manuscript explores the potential of vitamin D to regulate MC activity and combat pathogens, with a focus on its ability to fight viruses. Full article

Candida auris, a multidrug-resistant fungal pathogen, significantly threatens global public health. Recent studies have identified melanin production, a key virulence factor in many pathogenic fungi that protects against external threats like reactive oxygen species, in C. auris. However, the melanin regulation mechanism remains elusive. This study explores the role of the Ras/cAMP/PKA signaling pathway in C. auris melanization. It reveals that the catalytic subunits Tpk1 and Tpk2 of protein kinase A (PKA) are essential, whereas Ras1, Gpr1, Gpa2, and Cyr1 are not. Under melanin-promoting conditions, the tpk1Δ tpk2Δ strain formed melanin granules in the supernatant akin to the wild-type strain but failed to adhere them properly to the cell wall. This discrepancy is likely due to a decreased expression of chitin-synthesis-related genes. Our findings also show that Tpk1 primarily drives melanization, with Tpk2 having a lesser impact. To corroborate this, we found that C. auris must deploy Tpk1-dependent melanin deposition as a defensive mechanism against antioxidant exposure. Moreover, we confirmed that deletion mutants of multicopper oxidase and ferroxidase genes, previously assumed to influence C. auris melanization, do not directly contribute to the process. Overall, this study sheds light on the role of PKA in C. auris melanization and enhances our understanding of the pathogenicity mechanisms of this emerging fungal pathogen. Full article

Pseudostellaria heterophylla (Miq.) Pax is a popular clinical herb and nutritious health food. However, leaf spot disease caused by fungal pathogens frequently occurs and seriously influences the growth, quality, and yield of P. heterophylla. In this work, the field control roles of difenoconazole, chitosan, and their combination in the leaf spot disease in P. heterophylla and their effects on the disease resistance, photosynthetic capacity, medicinal quality, and root yield of P. heterophylla are investigated. The results manifest that 37% difenoconazole water-dispersible granule (WDG) with 5000-time + chitosan 500-time dilution liquid had a superior control capacity on leaf spot disease with the control effects of 91.17%~88.19% at 15~30 days after the last spraying, which significantly (p < 0.05) exceeded that of 37% difenoconazole WDG 3000-time dilution liquid and was significantly (p < 0.01) higher than that of 37% difenoconazole WDG 5000-time dilution liquid, chitosan 500-time dilution liquid, or chitosan 1000-time dilution liquid. Simultaneously, this combination could more effectively enhance the disease resistance, photosynthetic capacity, medicinal quality, and tuberous root yield of P. heterophylla compared to when these elements were applied alone, as well as effectively reduce difenoconazole application. This study emphasizes that chitosan combined with a low dosage of difenoconazole can be proposed as a green, efficient, and alternative formula for controlling leaf spot disease in P. heterophylla and enhancing its resistance, photosynthesis, quality, and yield. Full article

In the starch processing industry including the food and pharmaceutical industries, α-amylase is an important enzyme that hydrolyses the α-1,4 glycosidic bonds in starch, producing shorter maltooligosaccharides. In plants, starch molecules are organised in granules that are very compact and rigid. The level of starch granule rigidity affects resistance towards enzymatic hydrolysis, resulting in inefficient starch degradation by industrially available α-amylases. In an approach to enhance starch hydrolysis, the domain architecture of a Glycoside Hydrolase (GH) family 13 α-amylase from Aspergillus niger was engineered. In all fungal GH13 α-amylases that carry a carbohydrate binding domain (CBM), these modules are of the CBM20 family and are located at the C-terminus of the α-amylase domain. To explore the role of the domain order, a new GH13 gene encoding an N-terminal CBM20 domain was designed and found to be fully functional. The starch binding capacity and enzymatic activity of N-terminal CBM20 α-amylase was found to be superior to that of native GH13 without CBM20. Based on the kinetic parameters, the engineered N-terminal CBM20 variant displayed surpassing activity rates compared to the C-terminal CBM20 version for the degradation on a wide range of starches, including the more resistant raw potato starch for which it exhibits a two-fold higher Vmax underscoring the potential of domain engineering for these carbohydrate active enzymes. Full article

Neutrophils are a vital component of the innate immune system and play an essential function in the recognition and clearance of bacterial and fungal pathogens. There is great interest in understanding mechanisms of neutrophil dysfunction in the setting of disease and deciphering potential side effects of immunomodulatory drugs on neutrophil function. We developed a high throughput flow cytometry-based assay for detecting changes to four canonical neutrophil functions following biological or chemical triggers. Our assay detects neutrophil phagocytosis, reactive oxygen species (ROS) generation, ectodomain shedding, and secondary granule release in a single reaction mixture. By selecting fluorescent markers with minimal spectral overlap, we merge four detection assays into one microtiter plate-based assay. We demonstrate the response to the fungal pathogen, Candida albicans and validate the assay’s dynamic range using the inflammatory cytokines G-CSF, GM-CSF, TNFα, and IFNγ. All four cytokines increased ectodomain shedding and phagocytosis to a similar degree while GM-CSF and TNFα were more active in degranulation when compared to IFNγ and G-CSF. We further demonstrated the impact of small molecule inhibitors such as kinase inhibition downstream of Dectin-1, a critical lectin receptor responsible for fungal cell wall recognition. Bruton’s tyrosine kinase (Btk), Spleen tyrosine kinase (Syk), and Src kinase inhibition suppressed all four measured neutrophil functions but all functions were restored with lipopolysaccharide co-stimulation. This new assay allows for multiple comparisons of effector functions and permits identification of distinct subpopulations of neutrophils with a spectrum of activity. Our assay also offers the potential for studying the intended and off-target effects of immunomodulatory drugs on neutrophil responses. Full article

Across lower eukaryotes, the transcription factor Crz1 is dephosphorylated by calcineurin, which facilitates Crz1 translocation to the nucleus to regulate gene expression. In the fungal pathogen Cryptococcus neoformans, calcineurin–Crz1 signaling maintains calcium homeostasis, thermotolerance, cell wall integrity, and morphogenesis. How Crz1 distinguishes different stressors and differentially regulates cellular responses is poorly understood. Through monitoring Crz1 subcellular localization over time, we found that Crz1 transiently localizes to granules after exposure to high temperature or calcium. These granules also host the phosphatase calcineurin and Pub1, a ribonucleoprotein stress granule marker, suggesting a role of stress granules in modulating calcineurin–Crz1 signaling. Additionally, we constructed and analyzed an array of Crz1 truncation mutants. We identified the intrinsically disordered regions in Crz1 contribute to proper stress granule localization, nuclear localization, and function. Our results provide the groundwork for further determination of the mechanisms behind the complex regulation of Crz1. Full article

Neutrophil extracellular traps (NETs) are net-like structures released from neutrophils. NETs predominantly contain cell-free deoxyribonucleic acid (DNA) decorated with histones and neutrophil granule proteins. Numerous extrinsic and intrinsic stimuli can induce the formation of NETs such as pathogens, cytokines, immune complexes, microcrystals, antibodies, and other physiological stimuli. The mechanism of NETosis induction can either be ROS-dependent or independent based on the catalase producing activity of the pathogen. NADPH is the source of ROS production, which in turn depends on the upregulation of Ca2+ production in the cytoplasm. ROS-independent induction of NETosis is regulated through toll-like receptors (TLRs). Besides capturing and eliminating pathogens, NETs also aggravate the inflammatory response and thus act as a double-edged sword. Currently, there are growing reports of NETosis induction during bacterial and fungal ocular infections leading to different pathologies, but there is no direct report suggesting its role during herpes simplex virus (HSV) infection. There are innumerable independent reports showing that the major effectors of NETosis are also directly affected by HSV infection, and thus, there is a strong possibility that HSV interacts with these facilitators that can either result in virally mediated modulation of NETosis or NETosis-mediated suppression of ocular HSV infection. This review focuses on the mechanism of NETs formation during different ocular pathologies, with its prime focus on highlighting their potential implications during HSV ocular infections and acting as prospective targets for the treatment of ocular diseases. Full article

Airborne fungi are ubiquitous in the environment and are commonly associated with airway inflammatory diseases. The innate immune defense system eliminates most inhaled fungi. However, some influence the development of chronic rhinosinusitis. Fungal CRS is thought of as not a common disease, and its incidence increases over time. Fungi are present in CRS patients and in healthy sinonasal mucosa. Although the immunological mechanisms have not been entirely explained, CRS patients may exhibit different immune responses than healthy people against airborne fungi. Fungi can induce Th1 and Th2 immune responses. In CRS, Th2-related immune responses against fungi are associated with pattern recognition receptors in nasal epithelial cells, the production of inflammatory cytokines and chemokines from nasal epithelial cells, and interaction with innate type 2 cells, lymphocytes, and inflammatory cells. Fungi also interact with neutrophils and eosinophils and induce neutrophil extracellular traps (NETs) and eosinophil extracellular traps (EETs). NETs and EETs are associated with antifungal properties and aggravation of chronic inflammation in CRS by releasing intracellular granule proteins. Fungal and bacterial biofilms are commonly found in CRS and may support chronic and recalcitrant CRS infection. The fungal–bacterial interaction in the sinonasal mucosa could affect the survival and virulence of fungi and bacteria and host immune responses. The interaction between the mycobiome and microbiome may also influence the host immune response, impacting local inflammation and chronicity. Although the exact immunopathologic role of fungi in the pathogenesis of CRS is not completely understood, they contribute to the development of sinonasal inflammatory responses in CRS. Full article