Progress in a LAMP-PCR (Loop-Mediated Isothermal Amplification) assay for detecting the Sigatoka fungus in Musa sp. plants (#2184)

Authors

Fernández-Alvarado, Karla G.

Ballestas-Casallas, Yamil A.

Villalba Acevedo, Jorge L.

Chavarro Mesa, Edisson

Abstract

The cultivation of bananas and plantains ranks fourth in global food importance after rice, wheat, and corn. The fungus Mycosphaerella sp., known as Sigatoka, is a pathogen that mainly affects these crops. In Colombia, black and yellow Sigatoka are among the nine most significant diseases impacting Musa sp. production. This study aimed to characterize the fungal microbiota associated with Musa sp. using microbiological and molecular methodologies to identify the phytopathogenic fungus. The molecular technique LAMP-PCR (Loop-mediated Isothermal Amplification Polymerase Chain Reaction) was proposed for nucleic acid amplification at a constant temperature. DNA extraction was performed using the "Wizard® Genomic DNA Purification Kit" (PROMEGA), following the manufacturer's instructions. The LAMP-PCR assay was conducted with the "WarmStart® LAMP Kit (DNA & RNA)" (New England Biolabs). Microbiological processing allowed for the isolation of pure and viable cultures, essential for studying target microorganisms. The LAMP-PCR technique proved to be effective for detecting and amplifying specific genetic material, providing rapid and accurate results. The study continues the standardization process to optimize this test and implement it routinely for Sigatoka pathogen identification.