A Handy and Visual Detection Method for Potato Late Blight Utilizing Multi-Copy Specific Sequences and RPA-LFD...

Area of Science:

• Plant Pathology
• Molecular Biology
• Biotechnology

Background:

• Potato late blight, caused by Phytophthora infestans, is a major threat to global potato production.
• Early and accurate detection of P. infestans is crucial for effective disease management.

Purpose of the Study:

• To identify a novel DNA sequence for Phytophthora infestans detection.
• To develop a rapid, sensitive, and user-friendly diagnostic method for potato late blight.

Main Methods:

• Bioinformatic analysis to identify the SSPI1 (Specific Sequence of Phytophthora infestans 1) DNA sequence.
• Development of a Recombinase Polymerase Amplification (RPA) assay combined with Lateral Flow Dipstick (LFD) technology targeting SSPI1.
• Comparative analysis of SSPI1 with PiYPT1 and PiITS using PCR and RPA-LFD assays.

Main Results:

• The SSPI1 sequence demonstrated higher sensitivity and specificity compared to existing markers (PiYPT1 and PiITS).
• The developed RPA-LFD assay is rapid (35 min), sensitive (11 copies/μl, 100 pg genomic DNA), and requires no specialized equipment.
• The assay successfully detected P. infestans in early infection stages (24 h post-inoculation) and asymptomatic leaves under field conditions.

Conclusions:

• The novel SSPI1 sequence and the developed RPA-LFD assay provide a highly sensitive, specific, and practical tool for early and rapid detection of potato late blight.
• This method offers a significant advancement for disease surveillance and management in potato cultivation.
• The user-friendly nature of the assay facilitates its application in diverse field settings.