ppa12001-sup-0001-FigS1.tifimage/tif3834KFigure S1. Effectiveness of triticale seedling resistance to Microdochium nivale is dependent on plant genotype and duration of plant hardening. Two cultivars of triticale, Magnat (a) and Hewo (b), were subjected to hardening periods of 0, 14, 21, 28, 42 and 98 days at 4°C. Plants were inoculated with mycelium and incubated at 4°C for 21 days. Control (c) uninfected plants were hardened 28 days and incubated in similar conditions to those of the infected plants, except for the fungal treatment. Then plants were cut at 4 cm of height over the soil level and kept in optimal conditions. Representative examples of plants after 10 days of regrowth were selected from five independent experiments.
ppa12001-sup-0002-FigS2.tifimage/tif2652KFigure S2. Genotype-dependent changes of plant cell walls under cold determine hydrophobic properties of the triticale leaf surface. Two cultivars of triticale, Magnat (M20, M4) and Hewo (H20, H4), were subjected to a hardening period of 28 days at 4°C (M4, H4). Control plants grew at 20°C (H20, M20). Leaf surfaces were untreated, preincubated for 24 h with freshly prepared fungal homogenate or for 24 h with redistilled water as a control. Droplets (7 μL) of redistilled water or fungal homogenate were applied to the surfaces of detached leaves of the seedlings under test, and mounted on double-sided tape to the edge of a ruler. Imaging of the leaf surfaces and droplets was performed with a microscope. The distance between bars corresponds to 1 mm.
ppa12001-sup-0003-FigS3.tifimage/tif6890KFigure S3. Cell wall permeability is a key factor of the cold-induced resistance. Plants were hardened 0–98 days at 4°C and then incubated for 24 h with fungal extract. Unhardened control plants (c) were incubated with redistilled water. Leaf cell wall permeability for DAPI was measured 10 and 60 min after staining for Hewo and Magnat. Microscopic analyses were performed under UV light. Images show representative examples of leaves selected from 20 biological repeats. Magnification ×100.
ppa12001-sup-0004-FigS4.tifimage/tif4163KFigure S4. Cell wall strengthening with phenolic compounds results from cold hardening. Two cultivars of triticale, Magnat and Hewo, were subjected to a hardening period (28 days at 4°C. Control plants grew at 20°C. Phenolic substances were visualized by autofluorescence of cell walls (arrows) under UV light. Nuclei were stained with DAPI. Bar, 20 μm. Numeric data expresses the mean concentration of total phenolic substances (μg g−1 dry weight) determined in leaves spectrophotometrically (Singleton & Rossi, 1965) and standard error (n = 10).

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