Colorimetric loop-mediated isothermal amplification (LAMP) detection of Fusarium solani in black pepper (Piper nigrum L.) for rapid diagnosis of yellowing disease

Yellowing disease caused by Fusarium solani significantly impacts black pepper, leading to yield reduction and economic losses. A simple, rapid, and reliable diagnostic method is essential for timely intervention and management. This study aims to develop a colorimetric loop-mediated isothermal amplification (LAMP) assay for diagnostic detection of F. solani. Specific LAMP primers were designed targeting the translation elongation factor-1 alpha (EF-1α) gene. Optimized reaction conditions were 65 °C for 60 min, achieving a DNA detection limit of 100 pg. Positive reactions were confirmed by a colour change and ladder-like patterns on the agarose gel. LAMP assay showed specificity for F. solani in comparison to other fungal species. It is a simpler and faster alternative to PCR, providing visible results in 60 min. The assay showed 100% specificity and accuracy in detecting F. solani in artificially infected plants. This method enables rapid assessment of disease pressure and facilitates timely management interventions.

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