Revisiting the intron hypothesis of QoI resistance in Phyllosticta ampelicida, the causal agent of grape black rot, and other Phyllosticta species

Author:

Horváth Áron N.1ORCID,Molnár Orsolya1ORCID,Németh Márk Z.1ORCID,Pintye Alexandra1ORCID,Dankó Tamás1,Spitzmüller Zsolt2,Váczy Zsuzsanna2,Váczy Kálmán Z.2ORCID,Onesti Giovanni3,Reis Pedro4ORCID,Rego Cecilia4ORCID,Bereczky Zsolt1,Kiss Levente15ORCID,Kovács Gábor M.16ORCID

Affiliation:

1. Centre for Agricultural Research, Plant Protection Institute, Hungarian Research Network (HUN‐REN) Budapest Hungary

2. Food and Wine Research Institute Eszterházy Károly Catholic University Eger Hungary

3. Department of Sustainable Crop Production Università Cattolica del Sacro Cuore Piacenza Italy

4. LEAF—Linking Landscape, Environment, Agriculture and Food Research Center, Associated Laboratory TERRA, Instituto Superior de Agronomia Universidade de Lisboa Lisbon Portugal

5. Centre for Crop Health University of Southern Queensland Toowoomba Queensland Australia

6. Department of Plant Anatomy, Institute of Biology Eötvös Loránd University Budapest Hungary

Abstract

AbstractChemical control of grape black rot, caused by Phyllosticta ampelicida, relies mainly on the use of demethylation inhibitors (DMIs) and quinone outside inhibitors (QoIs). The effectiveness of QoI fungicides is influenced by alternative respiration activity, and the exon/intron structure and point mutations in the target protein's gene, the cytochrome b (cytb) gene. Our study aims to investigate the QoI fungicide sensitivity of 48 P. ampelicida isolates in vitro by measuring EC50 and the molecular characteristics of the cytb gene and its mRNA in P. ampelicida and other Phyllosticta species. Mycelial growth tests revealed that the P. ampelicida isolates were sensitive to both azoxystrobin and trifloxystrobin; baseline EC50 values were 0.029 and 0.022 μg/mL, respectively. Addition of salicylhydroxamic acid (SHAM) resulted in lower EC50 values (0.024 and 0.017 μg/mL, respectively). None of the typical point mutations conferring resistance to QoIs in some fungi were detected. A group I intron was present right after the 143rd codon in the cytb gene in four of the six Phyllosticta species examined. The sequence and exon/intron structure of the cytb gene of Pampelicida isolated from Vitis vinifera is studied in detail and published here. Our results indicate a low risk of QoI resistance development via the G143A mutation in P. ampelicida.

Funder

Nemzeti Kutatási Fejlesztési és Innovációs Hivatal

European Regional Development Fund

Publisher

Wiley

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