2025/06/05 by Lindsey P. Burbank, Elizabeth Deyett, Nancy Her +4 · 1 voice
Agricultural and Biological Sciences · #Bacteria #Biology #Botany #Cocoa and Sweet Potato Agronomy #Computational biology #Gene #Genetics #Genome #Identification (biology) #Mycorrhizal Fungi and Plant Interactions #Phytoplasmas and Hemiptera pathogens #Transposable element #Virulence #Xylella fastidiosa
paper · doi:10.1094/phytofr-10-24-0111-r
published in PhytoFrontiers™ 5(3), 480-490
openalex publication_date 2025/06/05 · openalex created_date 2025/06/06 · openalex updated_date 2026/06/11
In bacterial genetics, large-scale screening approaches, such as sequencing transposon mutant pools, can be highly effective for identifying and characterizing genes with unknown functions. In the plant pathogen Xylella fastidiosa, this approach is challenging due to the fastidious nature of this bacterial species and its niche-specific growth in the plant xylem tissue. The purpose of this study was to explore the feasibility of transposon sequencing (Tnseq) for identification of virulence genes in X. fastidiosa, with the hypothesis that this would uncover genes or pathways not previously associated with plant infection. Predicted essential genes were compared after X. fastidiosa strain M23 was grown in culture and in planta using two known susceptible host species (grapevine and almond). After growth in planta, several gene categories were predicted as essential, including hemagglutinins, tRNAs, toxin-antitoxin systems, and prophage genes. Three candidate genes (XfasM230359, XfasM230360, and XfasM230972) were chosen for further validation by making targeted deletion mutants. Deletion mutants exhibited reduced disease in grapevines but normal growth and aggregation phenotypes in culture. Overall, the Tnseq approach has some practical limitations due to the nature of the X. fastidiosa pathosystem, and significant bottleneck effects of inoculation, but was still able to identify genes contributing to disease in plants. Recommendations for future Tnseq studies in X. fastidiosa are discussed based on the challenges and results of this work. [Formula: see text] The author(s) have dedicated the work to the public domain under the Creative Commons CC0 “No Rights Reserved” license by waiving all of his or her rights to the work worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law, 2025.