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Rapid Detection of Monilinia fructicola and Monilinia laxa on Peach and Nectarine using Loop-Mediated Isothermal Amplification.

Authors
  • Ortega, Sara Franco1
  • Del Pilar Bustos López, Maria1, 2
  • Nari, Luca3
  • Boonham, Neil4
  • Gullino, Maria Lodovica1, 2
  • Spadaro, Davide1, 2
  • 1 Centre of Competence for the Innovation in the Agro-environmental Sector-AGROINNOVA, University of Turin, via Paolo Braccini 2, I-10095 Grugliasco, TO, Italy. , (Italy)
  • 2 Department of Agricultural, Forestry and Food Sciences, University of Torino, via Paolo Braccini 2,I-10095 Grugliasco, TO, Italy. , (Italy)
  • 3 AGRION, Fondazione per la Ricerca l'Innovazione e lo Sviluppo Tecnologico dell'Agricoltura Piemontese, 12030 Manta (Cn), Italy. , (Italy)
  • 4 FERA Sand Hutton, York, United Kingdom. , (United Kingdom)
Type
Published Article
Journal
Plant Disease
Publisher
Scientific Societies
Publication Date
Sep 01, 2019
Volume
103
Issue
9
Pages
2305–2314
Identifiers
DOI: 10.1094/PDIS-01-19-0035-RE
PMID: 31306092
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Monilinia laxa and M. fructicola are two causal agents of brown rot, one of the most important diseases in stone fruit. Two species cause blight on blossoms and twigs and brown rot on fruit in pre- and postharvest. Both species are distributed worldwide in North and South America, Australia, and Japan. In Europe, M. laxa is endemic, while M. fructicola was introduced in 2001 and it is now widespread in several countries. Currently, both species coexist in European stone fruit orchards. Monilinia spp. overwinter in cankers and mummified fruit. Mummy monitoring during winter permits growers to understand which species of Monilinia will be prevalent in an orchard during the following season, permitting planning of an appropriate crop protection. Traditionally, the identification has been carried out using morphological features and even with polymerase chain reaction (PCR)-based assays that requires time and well-equipped laboratories. In this study, two isothermal-based methods were designed to identify these pathogens in a faster way than using traditional methods. The loop-mediated amplification (LAMP) assays were validated on some isolates of Monilinia spp. coming from the mummy monitoring according to the international European and Mediterranean Plant Protection Organization standard (PM7/98), taking into account specificity, sensitivity, repeatability, and reproducibility. The sensitivity of both assays was checked by monitoring (at different time points) two nectarine varieties artificially inoculated and stored at two different temperatures. The reliability of both LAMP assays against the quantification of the inoculum was compared with previously published quantitative PCR assays. Both LAMP methods were able to detect a low number of cells. These LAMP methods could be a useful tool for monitoring brown rot causal agents in the field and during postharvest.

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