AI Article Synopsis
- Fusarium head blight (FHB) in wheat reduces crop yield and introduces harmful mycotoxins, necessitating early and accurate detection methods.
- A new CRISPR-Cas12a-based nucleic acid assay offers a rapid diagnostic solution by specifically targeting genetic sequences associated with the disease, achieving detection levels as low as 1 fg/μL of DNA.
- This assay distinguishes FHB from other fungi using a dual recognition process, enhancing the applicability of CRISPR technology in diagnosing crop diseases.
Article Abstract
Fusarium head blight (FHB) of wheat, mainly caused by () infection, reduces crop yield and contaminates grain with mycotoxins. We report a clustered regularly interspaced short palindromic repeats (CRISPR)-Cas12a-based nucleic acid assay for an early and rapid diagnosis of wheat FHB. Guide RNA (gRNA) was screened for highly specific recognition of polymerase chain reaction (PCR) amplicon of the internal transcribed spacer (ITS) region and the transcription elongation factor 1α () of . The trans-activation of Cas12a protein cleaves the single-stranded DNA probes with the terminal fluorophore and quencher groups, thus allowing us to report the presence of ITS and of . Owing to the dual recognition process through PCR primers and gRNA hybridization, the approach realized specific discrimination of from other pathogenic fungi. It also allowed us to detect as low as 1 fg/μL total DNA from , which is sufficient to diagnose a 4 day infection. CRISPR-Cas12a-based nucleic acid assay promises the molecular diagnosis of crop diseases and broadens the application of CRISPR tools.
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| http://dx.doi.org/10.1021/acs.jafc.1c08391 | DOI Listing |
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