Establishment of a Taqman-based approach to monitor Fusarium spp. Airbone spores.

Abstract

Aerobiological studies provide important information about the biological particles present in the air. Monitoring the presence of airborne fungal spores can help farmers to prevent the onset of fungal diseases that may affect both quantity and quality of crops. The fungi Fusarium spp. are among the most important phytopathogenic fungal communities with high impact at regional level by affecting important cultures such as almond, tomato, maize and cereals. The establishment of an approach that would enable farmers to early react upon the possibility of a Fusarium spp. infection will lead to a better control of the diseases associated to those fungi. Currently, to monitor Fusarium spp. airborne spores it is followed the Hirst-type methodology, which is based on spore’s identification and quantification by optical microscope; a hard and time consuming process due to the spore’s small size and colorless wall. In this context, the development of an alternative methodology that enable to get accurate and reliable results in a faster way will be of high interest. A Taqman specific assay for Fusarium spp. detection and quantification was previously established for a different purpose [1] and was here applied as a molecular-based tool to detect airborne Fusarium spp. spores. To collect the biological particles from the atmosphere a Burkard 7-Day Volumetric Spore Trap, and the Hirst associated methodology, was used as the methodology recommended by the European Aerobiology Society (EAS) and International Association for Aerobiology (IAA) [2]. As proof-of-concept, the analysis was focused on samples weekly collect, from 1st October - 31st December 2018 (14 weeks) at the station of Portuguese Aerobiology Network (RPA – SPAIC) (38° 34’ N; 7° 54’ W). Genomic DNA (gDNA) was extracted from collected biological particles adhered to the melinex tape following the CTAB protocol [3] with some modifications. Considering the results achieved, we consider the Taqman-specific assay as an alternative methodology for monitoring Fusarium spp. airborne spores.