Lamp - a modern tool for the detection of fungal infections in the vineyard

Lisa M. FRISCH, Magdalena A. MANN, Rudi F. VOGEL, Ludwig, NIESSEN
Technical University of Munich, Germany

Email contact: lisa.frisch@tum.de

AIM: Loop-mediated isothermal amplification (LAMP) ^[1] is a modern technology for fast and sensitive amplification of specific DNA sequences under isothermal conditions. Its simple handling and no need for dedicated equipment together with an evaluation of the amplification event by in-tube detection make this method advantageous and economically affordable for on-site investigations in the industry. In this study, the applicability of such assays for the detection of fungal infections in grape, soil, and must samples was tested and optimized.

METHODS: 88 grape, 42 soil, and 15 must samples from different vineyards in Europe collected during the harvest 2020 were tested with LAMP assays optimized for the specific detection of Botrytis (B.) cinerea ^[2] responsible for Botrytis bunch rot, the gushing-inducing fungus Penicillium (P.) oxalicum ^[3], and with a newly developed LAMP assay for the detection of the mycotoxin-producing and gushing-inducing fungus P. expansum ^[4,5].

RESULTS: The optimized LAMP assay for the detection of B. cinerea revealed positive samples in all tested vineyards. For P. oxalicum, 6% of grape samples showed positive results while soil and must were tested negative. P. expansum was only found in Germany with 28% of grape, 10% of soil, and 13% of must samples revealing positive results.

CONCLUSIONS: The application of LAMP assays for the detection of fungal infections prior to the occurrence of visual mold symptoms by testing samples from vineyards is particularly beneficial. A specific and sensitive detection can be performed within 60 minutes of incubation and results can be monitored by naked eye inspection at day light. A simple sample preparation and the use of simple equipment like a water bath make LAMP a powerful tool for on-site investigations in the winemaking industry.

SUPPORT: AiF 19952 N.

References:

1. Notomi, T. (2000). Loop-mediated isothermal amplification of DNA. Nucleic Acids Res, 28 (12).
2. Tomlinson, J. A., Dickinson, M. J., Boonham, N. (2010). Detection of Botrytis cinerea by loop-mediated isothermal amplification. Lett Appl Microbiol, 51(6).
3. Vogt, E. I., Kupfer, V. M., Bechtner, J. D., Frisch, L. M., Vogel, R. F., Niessen, L. (2017). Detection of Penicillium oxalicum in grapes with a loop-mediated isothermal amplification (LAMP) assay. J Microbiol Biotechnol Food Sci, 7(3).
4. Frisch, L. M., Mann, M. A., Marek, D. N., Niessen, L. (2021). Development and optimization of a loop-mediated isothermal amplification (LAMP) assay for the species-specific detection of Penicillium expansum. Food Microbiol, 95.
5. Frisch, L. M., Mann, M. A., Marek, D. N., Baudrexl, M., Vogel, R. F., Niessen, L. (2021). Studies on the gushing potential of Penicillium expansum. Food Res Int, 139.