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Tomato brown rugose fruit virus (ToBRFV) poses a significant threat to tomato production worldwide, prompting extensive research into its genetic diversity, evolutionary dynamics, and adaptive strategies. In this study, we conducted a comprehensive analysis of ToBRFV at the codon level, focusing on codon usage bias, selection pressures, and evolutionary patterns across multiple genes. Our analysis revealed distinct patterns of codon usage bias and selection pressures within the ToBRFV genome, with varying levels of genetic diversity and evolutionary constraints among different genes. We observed a transition/transversion bias of 2.07 across the entire ToBRFV genome, with the movement protein (MP) gene exhibiting the highest transition/transversion bias and SNP density, suggesting potential evolutionary pressures or a higher mutation rate in this gene. Furthermore, our study identified episodic positive selection primarily in the MP gene, highlighting specific codons subject to adaptive changes in response to host immune pressures or environmental factors. Comparative analysis of codon usage bias in the coat protein (CP) and RNA-dependent RNA polymerase (RdRp) genes revealed gene-specific patterns reflecting functional constraints and adaptation to the host's translational machinery. Our findings provide valuable insights into the molecular mechanisms driving ToBRFV evolution and adaptation, with implications for understanding viral pathogenesis, host-virus interactions, and the development of control strategies. Future research directions include further elucidating the functional significance of codon usage biases, exploring the role of episodic positive selection in viral adaptation, and leveraging these insights to inform the development of effective antiviral strategies and crop protection measures.

Nowadays, the emergence of some microbial species resistant to antibiotics, both gram-positive and gram-negative bacteria, is due to changes in molecular activities, biological processes and their cellular structure in order to survive. The aim of the gene network analysis for the drug-resistant Enterococcus faecium as gram-positive and Salmonella Typhimurium as gram-negative bacteria was to gain insights into the important interactions between hub genes involved in key molecular pathways associated with cellular adaptations and the comparison of survival mechanisms of these two bacteria exposed to ciprofloxacin. To identify the gene clusters and hub genes, the gene networks in drug-resistant E . faecium and S . Typhimurium were analyzed using Cytoscape. Subsequently, the putative regulatory elements were found by examining the promoter regions of the hub genes and their gene ontology (GO) was determined. In addition, the interaction between milRNAs and up-regulated genes was predicted. RcsC and D920_01853 have been identified as the most important of the hub genes in S . Typhimurium and E . faecium , respectively. The enrichment analysis of hub genes revealed the importance of efflux pumps, and different enzymatic and binding activities in both bacteria. However, E . faecium specifically increases phospholipid biosynthesis and isopentenyl diphosphate biosynthesis, whereas S . Typhimurium focuses on phosphorelay signal transduction, transcriptional regulation, and protein autophosphorylation. The similarities in the GO findings of the promoters suggest common pathways for survival and basic physiological functions of both bacteria, including peptidoglycan production, glucose transport and cellular homeostasis. The genes with the most interactions with milRNAs include dpiB , rcsC and kdpD in S . Typhimurium and EFAU004_01228 , EFAU004_02016 and EFAU004_00870 in E . faecium , respectively. The results showed that gram-positive and gram-negative bacteria have different mechanisms to survive under antibiotic stress. By deciphering their intricate adaptations, we can develop more effective therapeutic approaches and combat the challenges posed by multidrug-resistant bacteria.

Although the role of long non-coding RNAs (lncRNAs) in key biological processes in animals and plants has been confirmed for decades, their identification in fungi remains limited. In this study, we discovered and characterized lncRNAs in Aspergillus flavus in response to changes in water activity, CO 2 concentration, and temperature, and predicted their regulatory roles in cellular functions. A total of 472 lncRNAs were identified in the genome of A. flavus , consisting of 470 novel lncRNAs and 2 putative lncRNAs (EFT00053849670 and EFT00053849665). Our analysis of lncRNA expression revealed significant differential expression under stress conditions in A. flavus . Our findings indicate that lncRNAs in A. flavus , particularly down-regulated lncRNAs, may play pivotal regulatory roles in aflatoxin biosynthesis, respiratory activities, cellular survival, and metabolic maintenance under stress conditions. Additionally, we predicted that sense lncRNAs down-regulated by a temperature of 30 °C, osmotic stress, and CO 2 concentration might indirectly regulate proline metabolism. Furthermore, subcellular localization analysis revealed that up-and down-regulated lncRNAs are frequently localized in the nucleus under stress conditions, particularly at a water activity of 0.91, while most up-regulated lncRNAs may be located in the cytoplasm under high CO 2 concentration.

Cold atmospheric plasma (CAP), specifically cold air glow discharge plasma (CAGDP), offers a novel approach to enhancing plant defense against viral infections. This study investigates the effects of CAGDP on alternative splicing (AS) and transcriptome-wide gene expression in tomato seedlings infected with Tomato brown rugose fruit virus (ToBRFV). Using high-throughput RNA sequencing and bioinformatics analyses via CLC Genomics Workbench, custom Python scripts, and functional enrichment tools including STRING database and KEGG REST API, we identified significant AS changes, predominantly exon skipping and intron retention, in chromosomes associated with disease resistance. These splicing alterations were linked to key biological processes such as metabolic pathways, catalytic activity, and hormone signaling. Moreover, integration of miRNA–mRNA networks predicted by psRNATarget and visualized in Cytoscape revealed a complex regulatory system involving both transcriptional and post-transcriptional mechanisms. The identification of 19 differentially expressed AS genes highlights the coordinated reprogramming induced by CAGDP to activate antiviral defenses. These findings highlight cold plasma technology as a promising, eco-friendly tool to reprogram plant molecular pathways and enhance resistance to viral pathogens.

This study investigates the efficacy of Trichoderma spp. and Bacillus spp., as well as their gamma radiation-induced mutants, as potential biological control agents against Meloidogyne javanica (Mj) in tomato plants. The research encompasses in vitro assays, greenhouse trials, and molecular identification methodologies to comprehensively evaluate the biocontrol potential of these agents. In vitro assessments reveal significant nematicidal activity, with Bacillus spp. demonstrating notable effectiveness in inhibiting nematode egg hatching (16–45%) and inducing second-stage juvenile (J2) mortality (30–46%). Greenhouse trials further confirm the efficacy of mutant isolates, particularly when combined with chitosan, in reducing nematode-induced damage to tomato plants. The combination of mutant isolates with chitosan reduces the reproduction factor (RF) of root-knot nematodes by 94%. By optimizing soil infection conditions with nematodes and modifying the application of the effective compound, the RF of nematodes decreases by 65–76%. Molecular identification identifies B. velezensis and T. harzianum as promising candidates, exhibiting significant nematicidal activity. Overall, the study underscores the potential of combined biocontrol approaches for nematode management in agricultural settings. However, further research is essential to evaluate practical applications and long-term efficacy. These findings contribute to the development of sustainable alternatives to chemical nematicides, with potential implications for agricultural practices and crop protection strategies.

Plant viruses pose a significant threat to global agriculture and require efficient tools for their timely detection. We present AutoPVPrimer, an innovative pipeline that integrates artificial intelligence (AI) and machine learning to accelerate the development of plant virus primers. The pipeline uses Biopython to automatically retrieve different genomic sequences from the NCBI database to increase the robustness of the subsequent primer design. The design_primers_with_tuning module uses a random forest classifier that optimizes parameters and provides flexibility for different experimental conditions. Quality control measures, including the evaluation of poly-X content and melting temperature, increase primer reliability. Unique to AutoPVPrimer is the visualize_primer_dimer module, which supports the visual evaluation of primer dimers—a feature missing in other tools. Primer specificity is validated via primer BLAST, which contributes to the overall efficiency of the pipeline. AutoPVPrimer has been successfully applied to the tomato mosaic virus, proving its adaptability and efficiency. The modular design allows customization by the user and extends the applicability to different plant viruses and experimental scenarios. The pipeline represents a significant advance in primer design and provides researchers with an effective tool to accelerate molecular biology experiments. Future developments aim to extend compatibility and incorporate user feedback to consolidate AutoPVPrimer as an innovative contribution to the bioinformatics toolbox and a promising resource for the advancement of plant virology research.

Tomato plants face biotic challenges like infections by cucumber mosaic virus (CMV), a member of the Cucumovirus genus in the Bromoviridae family, as well as beneficial interactions, such as colonization by the symbiotic fungus Funneliformis mosseae , which belongs to the Glomeraceae family. While this symbiosis boosts nutrient uptake and stress tolerance, viral infections can reduce yield and quality. Understanding how tomatoes manage these interactions is vital for enhancing crop productivity. To explore the molecular mechanisms behind these interactions, this study focuses on long non-coding RNAs (lncRNAs), which play crucial roles in gene regulation, stress response, and plant metabolic pathways. Tomato RNA-seq data were analyzed to identify lncRNAs and their interactions with microRNAs (miRNAs) through de novo assembly, mapping, expression analysis, and localization prediction. In this study, 3210 lncRNAs were identified from 12 SRA datasets of tomato plants, including control, CMV-infected, F. mosseae -colonized, and co-infected samples. Among these, 3194 were novel lncRNAs and 16 were conserved. Expression analysis revealed significant differential expression patterns across treatments. Pathway analysis indicated that these lncRNAs are involved in key metabolic processes, such as carbon metabolism, amino acid biosynthesis, and secondary metabolite production, suggesting their role in enhancing disease resistance. Furthermore, we predicted interactions between identified lncRNAs and miRNAs, including miR160a, miR166a/b, miR167a, miR171a/b/c, miR1917, miR1918, and miR395a/b, thereby highlighting potential regulatory networks that could modulate stress responses. The subcellular localization of identified lncRNAs revealed a predominance in the cytoplasm, implying their involvement in post-transcriptional regulation. This study accentuates the significance of lncRNAs in tomato plant defense mechanisms and provides a foundation for future research focused on enriching resistance to viral infections and boosting stress resilience.

Potato ( Solanum tuberosum ) is a staple food crop that supports global food security, ranking as the world’s third most important food crop after rice and wheat in terms of human consumption, and it is threatened by Potato virus Y (PVY), which causes severe yield losses. This study integrates bioinformatics analysis and experimental approaches to elucidate molecular defense mechanisms against PVY infection. Using transcriptomic data from PVY-infected potato plants, we constructed protein-protein interaction (PPI) networks and identified hub genes central to defense responses. The qPCR validation showed that three hub genes ( NAD1 , NAD2 , NAD3 ) were upregulated in resistant Sante plants but downregulated in susceptible Agria. Among these, NAD2 showed a striking 5.58-fold increase in Sante, highlighting its critical role in stress signaling and antiviral defense. Network analysis revealed interactions with microRNAs (miRNAs), including stu-miR8015-5p and stu-miR396-5p, suggesting complex regulatory networks. Codon usage bias analysis highlighted adaptive codon preferences optimized for translational efficiency, supporting potential strategies like codon deoptimization to impair viral fitness. Promoter motif analysis identified stress-responsive cis -regulatory elements linked to abscisic acid signaling, critical for antiviral responses. This comprehensive study establishes a framework for targeting hub genes and miRNAs to engineer PVY-resistant cultivars, thereby offering a sustainable solution.

Tea plant necrotic ring blotch virus (TPNRBV) has emerged as a significant threat to tea plantations, primarily in China. Since 2020, similar symptoms have been observed in tea plants in northern Iran, raising concerns about the spread of this viral infection. In this study, we conducted an extensive investigation involving approximately 70 samples collected from both symptomatic and asymptomatic tea plants. Using reverse transcription-polymerase chain reaction with specially designed primers, we successfully amplified DNA fragments from 26 samples, confirming the presence of TPNRBV. Subsequent sequencing of these fragments revealed various segments of the TPNRBV genome. Our phylogenetic analysis revealed that the Iranian TPNRBV isolates formed a distinct sub-cluster alongside Chinese isolates, distinguishing them from Japanese isolates. These finding sheds light on the genetic diversity and relationships of TPNRBV across different regions. Additionally, we explored the potential modes of TPNRBV transmission. Mechanical transmission experiments confirmed the ability of the virus to infect Nicotiana rustica and Chenopodium quinoa seedlings, highlighting the risk of mechanical spread within tea plantations. Moreover, we investigated seed transmission and found evidence of TPNRBV in various parts of tea seeds, suggesting the possibility of seed-borne transmission. Overall, this comprehensive study enhances our understanding of the biological and molecular characteristics of TPNRBV, an emerging threat to global tea production. Our findings provide valuable insights into the virus’s transmission dynamics and genetic diversity, which are essential for developing effective management strategies to mitigate its impact on tea cultivation worldwide.

Tomato brown rugose fruit virus (ToBRFV) is a highly destructive and rapidly spreading tobamovirus that poses a serious threat to global tomato production. While low-dose gamma irradiation has emerged as a promising non-chemical strategy to enhance host resistance, the molecular mechanisms and transcriptomic reprogramming underlying this induced resistance remain largely unexplored. In this study, we employed a transcriptome-wide RNA sequencing approach to elucidate the specific gene expression networks and defense pathways activated in ToBRFV-infected tomato plants in response to low-dose gamma irradiation, addressing a critical gap in our understanding of host-virus interactions under irradiation priming. Naturally infected tomato seeds were exposed to an optimized gamma dose of 15 Gy, and transcriptomic profiles of irradiated plants were compared with those of non-irradiated infected controls. RNA-Seq analysis identified 469 differentially expressed genes (DEGs), including 157 upregulated and 312 downregulated transcripts (FDR < 0.05), indicating that gamma irradiation induces extensive transcriptional reprogramming. Functional enrichment analyses revealed significant activation of pathways related to metabolic reorganization, antioxidant defense, plant hormone signal transduction, secondary metabolite biosynthesis, and MAPK signaling. Notably, key defense-associated genes encoding peroxidases, protein kinases, and tetratricopeptide repeat (TPR) domain-containing proteins were strongly upregulated, suggesting enhanced reactive oxygen species (ROS) detoxification, stress signal amplification, and potential restriction of viral replication. In contrast, several growth- and development-related transcription factors and heat shock proteins were markedly downregulated, reflecting a shift in resource allocation toward defense responses. Quantitative RT-PCR validation of selected hormone-related genes confirmed the reliability of the RNA-Seq data and highlighted the coordinated involvement of auxin, ethylene, and gibberellin signaling in stress adaptation. Collectively, our results provide novel molecular evidence defining how low-dose gamma irradiation primes endogenous defense networks to reduce viral accumulation. This study offers new insights into the host-mediated transcriptional regulation of resistance against ToBRFV, establishing a foundation for future functional studies on irradiation-induced immunity.