Potato late blight is caused by the oomycete Phytophthora infestans, the most destructive pathogen affecting cultivated potato (Solanum tuberosum L.). Under favorable conditions, P. infestans can infect leaves and tubers and severely spread in field if left uncontrolled. The use of resistant varieties, when available, along with effective detection tools can be considered sustainable strategies to control the disease compared to chemical control. The aim of the present work was to set up a potato seedling assay in order to monitor the colonization by P. infestans and test different molecular techniques for the detection of the pathogen. For this purpose, 2-week-old seedlings of different varieties were sprayed with a sporangial suspension of the pathogen and kept under controlled conditions in a phytotron for 10 days. At different stages of the infection leaf samples were collected and analyzed by a targeted PCR approach using P. infestans ribosomal ITS specific primers and by real-time qPCR using housekeeping genes primers for the quantification of the plant/pathogen DNA ratio. Both approaches provided a method to detect P. infestans in relation to the infection stage and presence/absence of symptoms. Future implementation of this assay will involve the use of Nanopore sequencing so that conventional qPCR will validate both reliability and sensitivity of this next generation sequencing approach as a potential tool for late blight diagnostics.

Molecular approaches to detect Phytophthora infestans from different varieties of potato seedlings artificially inoculated / Fratini, ROSITA SILVANA; Beccaccioli, Marzia; Faino, Luigi; Reverberi, Massimo. - (2022). (Intervento presentato al convegno 21st Annual Meeting of the Oomycete Molecular Genetics Network tenutosi a Brno; Repubblica Ceca).

Molecular approaches to detect Phytophthora infestans from different varieties of potato seedlings artificially inoculated

Rosita Silvana Fratini
;
Marzia Beccaccioli;Luigi Faino;Massimo Reverberi
2022

Abstract

Potato late blight is caused by the oomycete Phytophthora infestans, the most destructive pathogen affecting cultivated potato (Solanum tuberosum L.). Under favorable conditions, P. infestans can infect leaves and tubers and severely spread in field if left uncontrolled. The use of resistant varieties, when available, along with effective detection tools can be considered sustainable strategies to control the disease compared to chemical control. The aim of the present work was to set up a potato seedling assay in order to monitor the colonization by P. infestans and test different molecular techniques for the detection of the pathogen. For this purpose, 2-week-old seedlings of different varieties were sprayed with a sporangial suspension of the pathogen and kept under controlled conditions in a phytotron for 10 days. At different stages of the infection leaf samples were collected and analyzed by a targeted PCR approach using P. infestans ribosomal ITS specific primers and by real-time qPCR using housekeeping genes primers for the quantification of the plant/pathogen DNA ratio. Both approaches provided a method to detect P. infestans in relation to the infection stage and presence/absence of symptoms. Future implementation of this assay will involve the use of Nanopore sequencing so that conventional qPCR will validate both reliability and sensitivity of this next generation sequencing approach as a potential tool for late blight diagnostics.
2022
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1725474
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