The rice white tip nematode (RWTN) Aphelenchoides besseyi secretes effectors that manipulate host plant cells to facilitate successful parasitism and sustain infection.  Although the number of identified RWTN effectors remains limited, their mechanisms of interaction with host plants are largely unknown.  Profilins (PFNs) function as molecular hubs that regulate complex interaction networks.  To advance understanding of PFN3 in plant-parasitic nematodes, we identified an effector from A. besseyi, designated AbPFN3.  AbPFN3 is transcriptionally upregulated during the juvenile stage of the nematode, and in situ hybridization localized its expression to the esophageal glands.  Three rice (Oryza sativa) proteins, ADP/ATP carrier protein 1 (OsAAC1), B-cell receptor-associated protein 31 (OsBAP31) and Small Auxin Up RNA 50 (OsSAUR50), were identified as interactors of AbPFN3, with interactions occurring in distinct cellular compartments, including the endoplasmic reticulum, cytoplasm, and plasma membrane.  Transgenic analyses revealed that AbPFN3 expression significantly increased plant height and upregulated AAC1 and BAP31, while downregulating RGA2 and SAUR50.  This study characterizes AbPFN3 as a novel effector secreted by A. besseyi that interacts with multiple host proteins, highlighting its potential role in modulating host defense responses and cell development processes.

One of the main diseases that adversely impacts the global citrus industry is citrus bacterial canker (CBC), caused by the bacteria Xanthomonas citri subsp. citri (Xcc).  Response to CBC is a complex process, with both protein-DNA as well as protein–protein interactions for the regulatory network.  To detect such interactions in CBC resistant regulation, a citrus high-throughput screening system with 203 CBC-inducible transcription factors (TFs), were developed.  Screening the upstream regulators of target by yeast-one hybrid (Y1H) methods was also performed.  A regulatory module of CBC resistance was identified based on this system.  One TF (CsDOF5.8) was explored due to its interactions with the 1-kb promoter fragment of CsPrx25, a resistant gene of CBC involved in reactive oxygen species (ROS) homeostasis regulation.  Electrophoretic mobility shift assay (EMSA), dual-LUC assays, as well as transient overexpression of CsDOF5.8, further validated the interactions and transcriptional regulation.  The CsDOF5.8–CsPrx25 promoter interaction revealed a complex pathway that governs the regulation of CBC resistance via H₂O₂ homeostasis.  The high-throughput Y1H/Y2H screening system could be an efficient tool for studying regulatory pathways or network of CBC resistance regulation.  In addition, it could highlight the potential of these candidate genes as targets for efforts to breed CBC-resistant citrus varieties.

Mycobacterium avium subsp. paratuberculosis (MAP) causes paratuberculosis (pTB) in ruminants and may be linked to Crohn's disease in humans. Despite extensive MAP genomic data from various animals worldwide, there is a significant lack of such data and understanding of MAP pathogenicity in China. This study used whole-genome sequencing (WGS) and pathogenicity analysis in mice to examine virulence differences among six MAP field strains (designated NM10, LN219, HLJ37, HLJ160, XJ41, and XJ121) isolated from cattle and sheep in various regions of China affected by pTB. The WGS and pan-genome analysis revealed close genomic relatedness among the six MAP strains. However, strains LN219 and NM10 exhibited two and three hypervirulence factors, respectively, while the other four isolated strains each contained only one hypervirulence factor within their specific genomes. Moreover, AlphaFold predictions indicated that the nine amino acid deletions identified in the anti-anti-σ factor of strains LN219 and NM10 led to the lowest binding affinity in the anti-anti-σ factor_anti-σ factor complexes, relative to the other four Chinese strains and the K-10 strain. In addition, bacterial phenotype analyses and in vivo animal experiments have shown that the pathogenicity and virulence of the LN219 and NM10 strains were significantly elevated compared to the other four isolated strains. These factors may partially account for the differences in virulence observed among MAP strains circulating in China. Furthermore, identifying the genes in this bacterium that are associated with critical disease phenotypes can enable targeted functional experiments on these genes, thereby improving control strategies for pTB.

Brucellosis, caused primarily by Brucella abortus, Brucella melitensis, and Brucella suis, remains a critical global public health challenge, particularly in regions where these pathogens persist in livestock and wildlife reservoirs. Despite decades of control measures-including vaccination, test-and-removal programs, and biosecurity protocols-persistent human and animal cases highlight the limitations of existing diagnostic and intervention strategies. CRISPR-based diagnostics have emerged as a transformative tool, offering rapid, ultrasensitive, and field-deployable pathogen detection. Here, we present BOVDS (Brucella melitensis/abortus/suis-other pathogens-vaccine detection and differentiation system), an innovative CRISPR/Cas13a-based platform that integrates ultrahigh sensitivity (10 copies/µL), screening for 10 major abortifacient pathogens, and precise strain differentiation-overcoming key challenges in Brucella diagnostics. By incorporating mismatched spacer designs, BOVDS achieves robust discrimination between B. melitensis, B. abortus, and B. suis despite their high genomic conservation. Additionally, the platform enables differentiation between vaccine and wild-type strains, addressing critical gaps in vaccination monitoring and epidemiological surveillance. Uniting laboratory-level accuracy with on-farm practicality, BOVDS facilitates real-time outbreak management, targeted culling, and environmental decontamination, advancing One Health initiatives toward sustainable brucellosis prevention and control. This system sets a new benchmark for next-generation zoonotic disease diagnostics, with broad applicability in global public and veterinary health.