Search Results (165)

To identify microRNAs (miRNAs) responsive to powdery mildew (PM) infection and elucidate their regulatory roles in melon PM resistance, thereby laying a foundation for deciphering the underlying molecular mechanisms, we combined high-throughput sequencing with bioinformatics analysis to screen PM-responsive miRNAs and their target genes using PM-resistant and PM-susceptible melon genotypes. In total, 113 non-redundant miRNAs were identified in both genotypes, including 70 known and 43 novel miRNAs. Subsequent differential expression analysis revealed distinct miRNA responses to PM infection between resistant and susceptible genotypes. Upon PM inoculation, 13 miRNAs showed significant differential expression in both susceptible and resistant genotypes. In addition, four miRNAs, including miR164c, miR396a, miR398a and miRn39, displayed differential expression specifically in the susceptible genotype. Conversely, five miRNAs (miR167c, miR398b, miR399g, miR530a and miRn8) showed differential expression exclusively in the resistant genotype upon PM infection. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses demonstrated that these miRNAs mediate melon’s susceptibility or resistance to PM by modulating plant immune homeostasis, antioxidant metabolism, and pathogen-triggered cell death. Quantitative real-time PCR (qRT-PCR) validation confirmed a negative regulatory relationship between the expression of PM-responsive miRNAs and their predicted target genes. Collectively, our findings provide novel insights and candidate targets for further investigations into miRNA functions and regulatory mechanisms underlying melon PM resistance. Full article

Receptor-like cytoplasmic kinases (RLCKs) constitute a core family of signaling proteins that modulate diverse cellular activities and participate in multiple physiological and biochemical processes in plants. As a unique subclade of RLCKs, the receptor-like cytoplasmic kinases VII (RLCK VII) subfamily features a conserved kinase domain but lacks extracellular and transmembrane domains, and has been proven to exert crucial functions in plant immunity, growth and development, and yield formation. Background/Objectives: This study systematically identified and characterized RLCK VII family genes in the rapeseed (Brassica napus) genome, and explored their biological functions in plant resistance to Sclerotinia sclerotiorum. Methods: Multiple bioinformatics databases and online tools were employed to conduct a comprehensive analysis of the identified BnaRLCK VII genes, including their physicochemical properties, signal peptide characteristics, subcellular localization, phylogenetic relationships, gene structure, conserved motifs and domains, cis-regulatory elements, tissue-specific expression patterns, and gene duplication events. Transcriptome profiling (RNA-seq) and transient overexpression assays in Nicotiana tabacum were further performed to verify the function of BnaRLCK VII genes in S. sclerotiorum resistance. Results: A total of 173 RLCK VII family members were identified from the B. napus genome and designated as BnaRLCK VII genes. These genes were clustered into 10 distinct subgroups based on phylogenetic analysis. We further comprehensively analyzed their gene structures, conserved motifs, cis-acting elements, duplication patterns, and tissue expression profiles. RNA-seq analysis revealed that 31 BnaRLCK VII genes were significantly differentially expressed following S. sclerotiorum infection. Among these differentially expressed genes (DEGs), a homolog of Arabidopsis thaliana BIK1 was selected for functional validation and named BnaBIK1. Agrobacterium-mediated transient infiltration assays on tobacco leaves demonstrated that overexpression of BnaBIK1 could improve plant tolerance to S. sclerotiorum, which indicates that BnaBIK1 is a promising candidate gene for regulating rapeseed resistance against this pathogen. Conclusions: Collectively, these findings indicate that BnaRLCK VII family genes, particularly BnaBIK1, serve as key positive regulators of S. sclerotiorum resistance in B. napus. Full article

Plant pathogens pose a severe threat to global agricultural production, and their pathogenicity is closely linked to the biosynthesis of secondary metabolites. Basidiomycete within the family Ustilaginaceae represent significant plant pathogens, among which Ustilago maydis, as a model species, has been extensively studied for its secondary metabolites. However, the biosynthetic potential of other species within this family remains poorly understood. In this study, we conducted whole-genome bioinformatic analyses of 16 Ustilaginaceae species, including U. maydis, to systematically identify the distribution of biosynthetic gene clusters (BGCs), core gene domain compositions, and interspecies similarities. A total of 181 predicted BGCs were identified, averaging approximately 11 per species. BGCs for mannosylerythritol lipids (MELs), siderophores, and itaconic acid, as well as the melanin-associated genes pks1 and pks2, were widely distributed across most species. Conversely, an additional melanin biosynthetic gene cluster was found exclusively in U. maydis strain 521, indicating species-specific occurrence. Furthermore, this study identified a novel class of polyketide synthase (PKS) gene clusters with uncharacterized functions across 15 species, exhibiting high sequence and structural conservation between species. These findings reveal the rich metabolic diversity and species-specific biosynthetic potential of Ustilaginaceae, and by using U. maydis as a reference model, we highlight several BGCs (e.g., for MELs, siderophores, itaconic acid, and melanin) that are known to contribute to virulence or pathogenicity in plant hosts. This provides new insights into their pathogenic mechanisms. Full article

Histidine triad (HIT) family proteins contain a conserved histidine triad motif and play key roles in fungal metabolism and pathogenicity. This study focused on VD9136, a member of the HIT family in Verticillium dahliae, aiming to elucidate its biological function and mechanism underlying its role in cotton pathogenesis. A systematic investigation of the VD9136 gene in V. dahliae was conducted using bioinformatics analysis, gene knockout, genetic complementation, and pathogenicity assays. The results showed that VD9136 protein consists of 136 amino acids and is a stable, neutral, and weakly hydrophilic protein that lacks transmembrane domains and signal peptides; it is localized to the extracellular space via a non-classical secretion pathway. Its secondary structure is predominantly composed of α-helices and random coils. Phylogenetic analysis revealed that VD9136 is closely related to VliHIT, a homologous protein from V. longisporum, the pathogen responsible for Verticillium wilt in rapeseed. The promoter region of VD9136 contains multiple cis-acting elements, including light-responsive, hormone-responsive, and stress-responsive elements, indicating that its transcription may be regulated by multiple signaling pathways. VD9136 was significantly upregulated during the early stage of cotton infection (6–24 h post-inoculation). Pathogenicity assays demonstrated that V. dahliae knockout mutants lacking VD9136 exhibited a significant reduction in virulence, as evidenced by a lower disease index, decreased fungal biomass within plant tissues, and attenuated vascular browning in cotton plants. The pathogenic phenotype was successfully restored in genetic complementation strains. This study identified VD9136 as a key regulatory factor in the pathogenic process of V. dahliae, and its loss of function reduces the pathogenicity of V. dahliae. The findings provide a theoretical basis for elucidating the pathogenic mechanism of cotton Verticillium wilt and for developing corresponding prevention and control strategies. Full article

The CFEM (Common in Fungal Extracellular Membrane) domain defines a family of cysteine-rich proteins unique to fungi, playing pivotal roles in host–pathogen interactions. However, the repertoire and functions of CFEM proteins in the broad-host-range necrotrophic pathogen Sclerotinia sclerotiorum remain largely unexplored. Through genome-wide bioinformatic analysis, we identified 13 CFEM-containing proteins (SsCFEM1–13) in S. sclerotiorum. Characterization revealed substantial diversity in their physicochemical properties, domain architecture, and predicted subcellular localization. Ten proteins possess a secretion signal, with six predicted to be GPI-anchored and three classified as high-confidence effectors. Members lacking transmembrane domains were predicted to adopt the conserved CFEM “helical-basket” fold. Phylogenetic analysis grouped SsCFEMs into two distinct clades and indicated a complex evolutionary history involving both conserved ancestry and lineage-specific expansion. Transcriptomic profiling showed that most genes were upregulated during early infection of various host plants, with SsCFEM8 exhibiting particularly strong and consistent induction. Crucially, transient expression assays in Nicotiana benthamiana revealed that several SsCFEM proteins, notably SsCFEM4 and SsCFEM9, function as cell death suppressors, validating their predicted effector roles and identifying key virulence candidates. This study provides the first comprehensive catalog and functional prediction of the CFEM protein family in S. sclerotiorum, establishing a foundation for future mechanistic studies on their roles in the pathogenesis of this devastating fungal pathogen. Full article

Cys2/His2-type zinc finger transcription factors (C2H2 TFs) constitute one of the largest and most functionally diverse transcription factor families in plants, playing core regulatory roles in multiple aspects of plant growth, development, and stress adaptation. Based on literature data from databases including PubMed (1995–April 2026) and integrated with bioinformatics analyses, this review provides a comprehensive overview of this family. We first summarize the structural characteristics and classification systems of C2H2 TFs, and elucidate their evolutionary dynamics from lower plants to angiosperms. Regarding their impact on plant organ development, beyond key biological processes, this review details the molecular mechanisms of C2H2 TFs in floral organ morphogenesis (e.g., petal, sepal, stamen, and ovule development), pollen fertility maintenance, and flowering time regulation. Concurrently, we systematically analyze their functional pathways in responses to abiotic stresses (drought, high salinity, low temperature, aluminum toxicity, etc.) and biotic stresses (pathogens, pests), clarifying the molecular networks through which they coordinate reactive oxygen species (ROS) homeostasis, stomatal movement, and osmotic regulation by modulating hormone signaling pathways such as ABA, SA, and JA. Furthermore, this review discusses major limitations of current research, including knowledge gaps concerning functional redundancy, pseudogenization phenomena, and cell type-specific regulation. We also provide perspectives on future research directions leveraging cutting-edge technologies such as CRISPR gene editing, single-cell sequencing, and multi-omics integration, as well as their application prospects in crop stress resistance breeding and quality improvement. This review provides ideas for in-depth research on the regulatory network and related functions of C2H2 TFs, and offers reference value for improving plant traits, enhancing plant resistance, and increasing the production of plant secondary metabolites. Full article

Phytopathogens threaten natural ecosystems and global food security. Horticultural trade is the main long-distance pathway that causes the spread of these organisms and disease outbreaks worldwide. Most inspections for disease symptoms are conducted visually, but this is insufficient given the large number of plants and the prevalence of asymptomatic infections. Therefore, there is increasing interest in the use of high-throughput sequencing (HTS) and environmental DNA (eDNA) for plant health surveillance. Many studies have used these technologies to detect phytopathogens, but fewer have done so in horticultural settings. Furthermore, much work has focused on the molecular and bioinformatic approaches for this work, with relatively little attention given to sample collection. This systematic review therefore provides an overview of the available sampling methods and their target plant pathogens, with a particular focus on the utility of these sampling methods in horticultural nurseries. It highlights some striking gaps in the literature and opportunities for further research: for example, the detection of bacterial phytopathogens using eDNA has received little attention despite having considerable potential as a surveillance and/or diagnostic tool. Full article

Quorum-sensing (QS) systems play a crucial role in regulating virulence, biofilm formation, and antibiotic resistance in clinically relevant microbes. This review explores the potential of QS systems as targets for developing novel plant-based therapeutic strategies using bioinformatics, aimed at combating highly pathogenic bacteria: uropathogenic Escherichia coli (UPEC) and Pseudomonas aeruginosa. We examine the key components and molecular pathways of QS systems in these microbes, including autoinducer synthases, receptors, and regulatory proteins. In UPEC, we discuss the LuxS-dependent autoinducer (AI)-2 system, while for P. aeruginosa, we analyze the more complex interconnected Las, Rhl, and PQS circuits. We highlight how these systems control the expression of virulence factors and contribute to biofilm formation, emphasizing their importance in pathogenesis. Furthermore, we explore bioinformatics approaches for identifying and characterizing QS components, i.e., by predicting protein structures and interactions. The potential of in silico screening for QS inhibitors is also discussed, along with challenges and opportunities in targeting QS systems for therapeutic interventions. By integrating microbiological, molecular, and computational perspectives, this review aims to provide insights into the application of bioinformatics in understanding and targeting QS systems in these clinically significant pathogens. The goal is to facilitate the development of novel anti-virulence approaches in search of novel antibiotics that could complement or replace traditional antibiotic treatments, addressing the growing concern of antimicrobial resistance in these clinically relevant microbes. Full article

Fungal diseases continue to limit global crop production and drive major economic losses. Conventional diagnostic and control approaches depend on time-consuming culture-based methods and broad-spectrum chemicals, which offer limited precision. Advances in molecular identification have changed this landscape. PCR, qPCR, LAMP, sequencing and portable platforms enable rapid and species-level detection directly from plant tissue. These tools feed into RNA-based control strategies, where knowledge of pathogen genomes and sRNA exchange enables targeted suppression of essential fungal genes. Host-induced and spray-induced gene silencing provide selective control without the long-term environmental costs associated with chemical use. CRISPR/Cas9 based tools now refine both diagnostics and resistance development, and bioinformatics improves target gene selection. Rising integration of artificial intelligence indicates a future in which disease detection, prediction and management connect in near real time. The major challenge lies in limited field validation and the narrow range of fungal species with complete molecular datasets, yet coordinated multi-site trials and expansion of annotated genomic resources can enable wider implementation. The combined use of molecular diagnostics and RNA-based strategies marks a shift from disease reaction to disease prevention and moves crop protection towards a precise, sustainable and responsive management system. This review synthesizes the information related to current molecular identification tools and RNA-based management strategies, and evaluates how their integration supports precise and sustainable approaches for fungal disease control under diverse environmental settings. Full article

Background: Plant growth promoting bacteria (PGPB) are non-pathogenic bacteria that enhance plant growth through several mechanisms such as nutrient mobilization, phytohormones production, defense against phytopathogens, and alleviation of plant stress. Hence, these bacteria are used as ecologic biofertilizers to diminish the use of agrochemicals. Nevertheless, some PGPR strains can harbor antibiotic resistance determinants and the possibility of spreading them upon releasing these bacteria is an environmental concern. Objectives: The objectives of this work are as follows: (1) evaluating the antibiotic resistance in a collection of PGPB, and (2) prospecting antibiotic resistance genes in the genomes of PGPB in order to predict the risk for antibiotic resistance dissemination. Methods: The resistance towards 12 antibiotics in a collection of 20 PGPB (10 Gram-positive and 10 Gram-negative strains) has been evaluated using disk diffusion in agar, broth microdilution, and agar dilution tests. In addition, the whole genomes of six strains have been sequenced in order to find the correlation between the resistance levels and AMR genes by using bioinformatic tools. Results: The results indicated a wide range of halo diameters, but in general Gram-negatives showed higher resistance compared to Gram-positives. The four most resistant strains and the two more susceptible strains were selected for further analysis and sequencing the whole genomes. The resistant strains were identified as Achromobacter spanius N6, Leclercia adecarboxylata H17, Priestia aryabhattai strain MHA1, and Bacillus cereus N25. The susceptible strains were identified as Pantoea sp. S3 and Priestia megaterium MS4. Mining antibiotic resistance genes in the genomes confirmed the existence of resistance determinants responsible for the phenotypic behavior, indicating the potential of genomics for predicting antibiotic resistance in PGPB. However, there was not an exact correspondence between the presence of the genes and the level of resistance, suggesting the existence of additional regulatory mechanisms. Conclusions: The information obtained by genomics must be complemented experimentally by tests for antibiotic resistance determination. In this regard, it is necessary to develop a global antibiotic resistance database for PGPB, due to the difficulty of interpretation of the antibiotic susceptibility tests after comparing the experimental results with those tabulated for clinical species. Full article

Phytomycobiomes refer to the fungal consortia that inhabit plant tissues and the rhizosphere. Their documented functions include nutrient mobilization, carbon retention, stress mitigation and pathogen suppression, although measurable effects often depend on plant and soil conditions. In this review, we examine the current evidence for their ecological relevance and assess the molecular approaches most commonly used to characterize them. Arbuscular Mycorrhizal (AM) fungi, endophytes and saprotrophic taxa indicate measurable gains in nutrient acquisition, disease resistance and soil aggregation, although long-term consistency is rarely evaluated. Each function appears to have an explicit mechanistic attribution, with direct links between fungal groups, enzymatic pathways and measurable ecosystem outcomes. Several sequencing-based techniques are available, yet none offer complete accuracy. Internal Transcribed Spacer (ITS) amplicon surveys provide rapid taxonomic coverage but suffer from primer bias; shotgun metagenomics offers functional insight but at significant financial cost; and quantitative polymerase chain reaction (qPCR) assays remain useful for targeted quantification, whereas long-read technologies show promise but still lack widespread adoption. The field faces a number of unresolved constraints, including limited knowledge of host range, inconsistent performance under fluctuating environmental conditions and the absence of a standardized bioinformatic pipeline. Despite these limitations, we regard phytomycobiomes as viable candidates for replacing or reducing synthetic inputs, provided their application is guided by context-specific evidence rather than broad generalization. Full article

In this study, we report bioinformatics analysis of the endoglucanase GH9 gene family in tomato (Solanum lycopersicum L.) using the SL5.0 genome, confirming the presence of 19 members that clustered into classes A, B, and C. To explore their potential role in plant–microbe interactions, we determined the transcriptional regulation of 10 SlGH9 gene members in tomato leaves and roots during interactions with the mutualistic root mycorrhizal fungus Rhizophagus irregularis and the foliar pathogen Sclerotinia sclerotiorum. The upregulation of several SlGH9 genes in the leaves of mycorrhizal plants suggests that they are involved in cellulose remodeling and biosynthesis rather than its degradation. This would be consistent with the observed increase in foliar area. On the other hand, downregulation of some SlGH9 genes in leaves of pathogen-infected mycorrhizal plants suggests that these genes may play a role in the enhanced resistance observed by reducing cellulose degradation, thereby maintaining cell wall integrity. The potential involvement of endoglucanase genes in expansive growth (foliar area) and in defense in mycorrhizal and pathogen-infected plants may reflect a growth–defense trade-off. Full article

Fusarium head blight (FHB) negatively affects wheat yield and quality worldwide. As wheat varieties differ in terms of their resistance to FHB, the identification of FHB-resistant genes is of great importance for the genetic improvement for FHB resistance in wheat breeding. Although sugar transporter proteins (STPs) play vital roles in plant–pathogen interactions, the functions of STP genes in wheat FHB resistance remain poorly understood. In this study, bioinformatics analyses were conducted to identify novel STP genes and characterize their expression profiles in wheat. We confirmed the presence of the 81 TaSTP genes previously reported and identified one additional member, designated as TaSTP6-2D. Based on RNA-seq profiles, 50 TaSTP genes that showed differential expression under biotic or abiotic stress were selected to explore the potential function in the resistance to Fusarium head blight. RT-qPCR analysis revealed that 11 TaSTP genes (TaSTP1-2D, TaSTP3-2A, TaSTP3-2B, TaSTP6-2A, TaSTP6-2B, TaSTP13-4B, TaSTP13-4D, TaSTP19-4A, TaSTP26-5A, TaSTP28-3A and TaSTP28-3D) were differential expressed following the treatment with chitin, Fusarium graminearum or deoxynivalenol. Among them, TaSTP26-5A showed a 28-fold upregulation to chitin in “Yangmai 158” compared to a 6-fold change in “Fielder”. These findings establish a foundation for understanding the function of TaSTP genes in FHB resistance and provide potential genetic targets for improving disease resistance in wheat. Full article

Metagenomic investigation of gut microbiome is a comprehensive and rapid technique for the analysis and diagnosis of numerous diseases. The gut microbiome is an intricate ecosystem, coordinated by the interaction of various microbes and the metabolites produced by them, which helps in developing and sustaining immunity and homeostasis. A healthy gut microbiome is driven by different factors, such as nutrition, lifestyle, etc. The current study examines the association of diet to gut microbiome dysbiosis and its role in various disease conditions. Gut microbiome data was collected from 73 patients and tested at BioAro Inc. lab, using shotgun metagenomics through next generation sequencing. It was then analyzed and compared with data from 20 healthy subjects from HMP database. An in-house bioinformatics pipeline (PanOmiQ) and Pathogen Fast Identifier were utilized for secondary analysis, while tertiary analysis was accomplished using R software. Results showed a higher number of opportunistic pathogen microorganisms in the gut microbiome of subjects consuming a meat diet, as compared to those consuming a plant diet. These opportunistic pathogens included Ruminococcus torques (>3.34%), Ruminococcus gnavus (>2.22%), and Clostridium symbiosum (>1.87%). The study also found a higher relative abundance of these pathogens in cancer patients, as compared to healthy subjects. We also observed a highly significant (p < 0.0001) correlation of a meat diet with obesity in comparison to the subjects on a plant diet and the healthy subjects. Our findings suggest that patients following a plant diet have a lower relative abundance of pathogens that are associated with cancer and obesity. These findings provide critical insight into how we can use shotgun metagenomics to study the composition and diversity of the gut microbiome and the effects of a diet on the gut microbiome and its role in metabolic diseases. This is the first report investigating gut microbiota using shotgun metagenomics, correlating with different diseases and diet followed, which might impact the presence of opportunistic pathogens or keystones species. Additionally, it can provide valuable insights to physicians and dietetic practitioners for providing personalized treatment or customizing a diet plan. Full article

Iron is a limiting factor for plant and microbial growth because, in soil environments, it is predominantly present as oxyhydroxide minerals, rendering it unavailable to plants and microorganisms. Siderophores are chelating agents secreted to solubilize iron and facilitate its uptake. To understand the evolutionary and ecological dynamics of microbial communities, as well as the evolution of pathogens within hosts, it is essential to study the genes shared between microorganisms for environmental adaptation and survival. In this study, we conducted microbiological assays to evaluate the effect of the siderophore produced by Rouxiella badensis strain SER3 on the mycelial growth of fungal pathogens such as Fusarium brachygibbosum 4BF. Using spectrophotometric techniques and bioinformatics tools, we identified desferrioxamine E (nocardamine) in the culture supernatant, and the corresponding biosynthetic gene cluster in the SER3 genome was confirmed through antiSMASH analysis and synteny comparisons. Gene expression analysis by RT-PCR showed differential expression of biosynthetic precursors when strain SER3 was grown alone or in interaction with fungal pathogen. Finally, scanning electron microscopy revealed structural damage to F. brachygibbosum hyphae during co-culture with strain SER3. These results suggest that the production of desferrioxamine E may act as a biocontrol mechanism employed by R. badensis SER3 against F. brachygibbosum. Full article