CRISPR-Based Biosensors for Plant Disease Diagnostics: Innovations, Applications, and Future Prospects

Abstract

Timely, accurate plant pathogen diagnostics are critical for mitigating crop losses and enhancing food security. Conventional methods (PCR, ELISA, LAMP) are effective but constrained by laboratory infrastructure, trained personnel, and multi-hour workflows, limiting field deployment. CRISPR–Cas biosensors—leveraging programmable target recognition and Cas12/Cas13 collateral cleavage—offer rapid, sensitive, and portable nucleic-acid detection. This review synthesizes plant-focused advances (2021–2025), covering molecular mechanisms; integration with isothermal amplification; and readouts spanning fluorescence, lateral-flow strips, and electrochemical sensing. We consolidate validated applications across viral, bacterial, and fungal pathogens, highlighting time-to-result (≈15–45 min), low per-test cost, and suitability for resource-limited settings. We also examine practical barriers, including plant matrix inhibitors, reagent stability, contamination control in amplificationcoupled assays, and fragmented regulatory pathways for agricultural diagnostics. Emerging solutions—microfluidic cartridges, lyophilized reagents, multiplexed panels using orthogonal Cas enzymes, and AI-assisted crRNA design—are accelerating translation toward ―sample-in, answer-out‖ platforms. We position CRISPR biosensors within precision agriculture and mobile surveillance ecosystems, emphasizing needs for standardized validation and field-level interoperability. Collectively, these developments indicate that CRISPR diagnostics are poised to become core components of decentralized, real-time plant health monitoring.