Abstract
Marssonina coronaria is considered a threat to organic apple production in Central Europe. Since the application of fungicides is limited in organic production, breeding for resistance seems to be a promising strategy to manage the disease in the future. In this study, an artificial inoculation method similar to procedures used in apple scab greenhouse screenings was developed using German strains of the pathogen M. coronaria for evaluating 110 Malus domestica cultivars. The strains were morphologically and molecularly characterized and confirmed as M. coronaria. Symptom development was significantly influenced by incubation method, conidia concentration and point of inoculation, but not by leaf position and different water types used for inoculation. The success of inoculation and spread of the fungus on infected leaves were confirmed by conventional PCR. Moreover, there was a difference in development of disease symptoms between inoculations with conidia from in vitro grown strains (in vitro inoculum) and conidia from diseased leaves from the orchard (field inoculum) in a time dependent manner. These differences which were first found with the susceptible cultivar ‘Topaz’ after artificial inoculation with in vitro and field inoculum were confirmed on 20 more apple cultivars inoculated with different in vitro grown strains. In summary, all tested cultivars, including 21 which are scab-resistant, developed symptoms of this disease. Results from three years of investigation indicate a decrease in virulence of M. coronaria strains, when cultivated on artificial culture (growth)-media. Hence field inoculum is recommended for artificial greenhouse screenings for the evaluation of disease resistance in Malus genetic resources.
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Acknowledgements
The authors acknowledge Thomas Nothnagel fore pictures and assistance during the microscopic preparation of samples. We also acknowledge the KOB Bavendorf and Jan Hinrichs-Berger for the delivery of symptomatic leaves. Special thanks to Sabine Bartsch for excellent technical assistance and, to the staff of the experimental orchard at JKI. This work was funded by the Julius Kühn-Institut.
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Table S1
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Table S2
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Figure S1
Inoculation trial with M .coronaria in the greenhouse. (JPG 3358 kb)
Figure S2
Growth characterization ofM. coronariaon different cultural media. Strain FU0021 was characterized on three cultural media (MA, PDA, PCDA) and water agar as control. The growth of M. coronaria mycelial bodies was evaluated in two experiments. Five mycelial bodies each were transferred to five petri dishes containing MA, PCDA, PDA or water agar and cultivated with permanent light in a growth chamber (CLF Plant Climatics, Donau-Wörth, Germany) or kept in the dark. Each colony was photographed with a camera (ColorView III, Olympus Soft Imaging Solutions GmbH) integrated into a stereo microscope (Stemi SV11, Zeiss). The area on the media of each mycelial body was measured using the software Cell^D (Version 2.7, Olympus Soft Imaging Solutions GmbH) after 0, 7 and 14 days growth at a temperature of 20 °C. - MA-malt agar; PCDA-potato carrot dextrose agar; PDA - potato dextrose agar (PNG 163 kb)
Figure S3
Cross section of an apple leave after inoculation withM. coronaria. Discs from symptomatic areas of infected leaves were incubated at 90 °C for 10 min in Ethanol (99%) to release chlorophyll. The procedure was performed in a ThermoMixer in 1.5 ml Eppendorf tubes (Eppendorf AG, Hamburg, Germany). After bleaching, the leaf discs were embedded in Technovit 7100 (Heraeus Kulzer GmbH, Wehrheim, Germany) following the standard procedure of the kit. The leaf discs were sliced with an automated microtome and stained with aniline blue. (PNG 2906 kb)
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Wöhner, T., Girichev, V., Radatz, S. et al. Evaluation of Malus gene bank resources with German strains of Marssonina coronaria using a greenhouse-based screening method. Eur J Plant Pathol 153, 743–757 (2019). https://doi.org/10.1007/s10658-018-1588-3
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DOI: https://doi.org/10.1007/s10658-018-1588-3