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ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » Molecular Plant Pathology Laboratory » Research » Publications at this Location » Publication #422145
Research Project: Omics-Based Approach to Detection, Identification, and Systematics of Plant Pathogenic Phytoplasmas and Spiroplasmas

Location: Molecular Plant Pathology Laboratory

Title: CRISPR/Cas-based detection of Spiroplasma citri for citrus stubborn disease management

Author
item SHIH, JUSTIN - Orise Fellow
item Wei, Wei

Submitted to: Phytopathogenic Mollicutes
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/26/2024
Publication Date: 3/5/2025
Citation: Shih, J., Wei, W. 2025. CRISPR/Cas-based detection of Spiroplasma citri for citrus stubborn disease management. Phytopathogenic Mollicutes. 15(1):161-162. https://doi.org/10.5958/2249-4677.2025.00083.6.
DOI: https://doi.org/10.5958/2249-4677.2025.00083.6

Interpretive Summary: Spiroplasma citri is a small, plant-pathogenic bacterium that infects citrus plants, causing Citrus Stubborn Disease, a significant threat to citrus production worldwide. To address this challenge, ARS scientists in Beltsville, Maryland, have developed a new detection method using CRISPR/Cas technology, a cutting-edge system for precisely detecting, editing, and controlling DNA sequences. By using specially designed "guides" that target the bacterium’s unique DNA sequence, the new approach can identify even the trace amounts of S. citri in just two hours, providing a fast and accurate method for early detection. To improve accessibility, efforts are underway to create simple, portable test strips that can be used directly in the field without requiring specialized equipment. The new detection tool enables faster detection and early intervention, benefiting citrus farmers, inspectors, biosecurity agencies, and researchers. The new system helps prevent outbreaks, reduce crop losses, limit disease spread, and protect citrus production and trade.

Technical Abstract: Spiroplasma citri, the causal agent of Citrus Stubborn Disease, remains a challenging plant pathogen to control, causing substantial economic loss in global citrus production. Rapid and efficient detection assays are essential for timely and effective mitigation interventions. Our research focused on developing a CRISPR/Cas-based detection system using the DETECTR method to create a point-of-care assay for S. citri. Preliminary results demonstrated the successful design of RPA primers, crRNAs, and a protocol for detecting S. citri at a concentration as low as 1 fM in two hours. Further optimization will be performed to enhance field applicability, including developing lateral flow strips and enabling direct, equipment-free detection in the field.