AI Article Synopsis
- Alternaria is a global plant pathogen that causes citrus brown spot disease and produces harmful mycotoxins, negatively impacting the citrus industry.
- A new visual fluorescent assay that utilizes CRISPR-Cas12a and rolling circle amplification (RCA) has been developed for detecting Alternaria in citrus plants.
- This innovative method produces a clear red fluorescence in positive samples, making it easy to identify infected plants compared to negative samples, and has been tested for its effectiveness in both lab-cultured and real citrus samples.
Article Abstract
Alternaria is a plant pathogen that spreads globally and is prone to causing citrus brown spot disease and metabolizing mycotoxins, thus seriously hindering the development of this economic crop industry. Herein, a "label-free" and "turn on" visual fluorescent assay for citrus Alternaria based on CRISPR-Cas12a and rolling circle amplification (RCA) was described. Using ssDNA complementary to RCA primer as a trans-cleavage substrate for CRISPR-Cas12a, the two systems of CRISPR-Cas12a and RCA-amplified G-quadruplex were skillfully integrated. By using a portable light source for excitation, the positive sample produced obvious red fluorescence, while the negative sample remained almost colorless, making them easy to differentiate with the naked eye. In addition, the specificity was demonstrated by distinguishing Alternaria from other citrus disease related pathogens. Moreover, the practicality was verified by analyzing cultured Alternaria and Alternaria in actual citrus leaf and fruit samples. Therefore, this method may contribute to the on-site diagnosis of Alternaria.
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| http://dx.doi.org/10.1016/j.ijbiomac.2024.129983 | DOI Listing |
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