Mousami Poudel, n/a
Graduate Research Assistant
University of Florida
Gainesville, Florida, United States
Apekshya Parajuli, n/a
Graduate Research Assistant
UNIVERSITY OF FLORIDA
GAINESVILLE, Florida, United States
Gerald V. Minsavage, n/a
Biological Scientist
UNIVERSITY OF FLORIDA
GAINESVILLE, Florida, United States
Samuel F. Hutton, PhD
Associate Professor
University of Florida
Wimuama, Florida, United States
Jeffrey B. Jones, PhD (he/him/his)
Distinguished Professor
University of Florida
Gainesville, FL, USA
Anuj Sharma, PhD
Assistant professor
University of Florida
Wimauma, Florida, United States
Bacterial leaf spot (BLS) caused by Xanthomonas is a severe disease of tomato (Solanum lycopersicum) and pepper (Capsicum annuum) plants. The deployment of resistant cultivars offers a sustainable approach to managing BLS. In pepper, mutations in the Bs6 and Bs8 susceptibility genes have been linked to resistance, leading to the reduced bacterial growth and absence of disease symptoms. The objective of this study is to generate BLS-resistant tomatoes by knocking out Bs6 and Bs8 homologs using CRISPR/Cas9 gene editing. We identified three putative homologs of Bs6 candidates and one homolog of Bs8 in tomato and designed CRISPR/Cas9 constructs targeting conserved regions of each homolog for mutagenesis. Screening of transformant lines using PCR and Sanger sequencing identified several disruptive insertion and deletion events in all targeted genes. Preliminary disease assays on mutant lines suggest a reduction in bacterial growth, supporting the role for these genes in susceptibility. Future work will focus on validating disease resistance in multiple generations in greenhouse and field conditions. The expected impact of this study is the development of durable BLS-resistant tomato lines, reducing reliance on chemical control methods and contributing to more sustainable disease management strategies.