Q150r plus rotary pumped coater

Surfaces of inoculated citrus leaves both treated with chemicals and untreated were observed with a scanning electron microscope to see fine structures of D. citri. Inoculated parts of leaves were cut to sizes of 5 × 5 mm² and fixed with a 2% glutaraldehyde in cacodylic buffer (pH 7.2) at 4°C for 2 h. Samples were washed with cacodylic buffer three times (10 min each). Post fixation was carried out with 1% osmium tetroxide in cacodylic buffer at 4°C for 2 h. These fixed samples were washed with distilled water two times for 10 min each. Dehydration was performed with 30%, 50%, 70%, 90%, and 100% ethanol for 30 min each followed by treatment with 100% twice for 30 min each. Samples were dried with a critical point dryer (CPD 030, BAL-Tec, Los Angeles, CA, USA) and coated with platinum by sputter coater-platinum (Q150R Plus – Rotary Pumped Coater, Quorum Technologies Ltd., Sussex, UK) at 20 mA for 90 s. Coated samples were observed with a field emission scanning electron microscope (FE-SEM Mira3, Tescan Ltd., Brno, Czech Republic).

Kiwifruit surfaces of ‘Zesy002’ pre-treated with agrichemicals and then inoculated with B. dothidea were observed at 1, 2, and 3 days after inoculation using a scanning electron microscope (FE-SEM Mira3, Tescan Ltd., Brno, Czech Republic).
The inoculated kiwifruit were cut to 0.4 × 0.6 mm² using a sterile blade. Fixation, dehydration and embedding of the fruit were performed according to Hayat (1989). The leaf samples were fixed in 2% (v/v) glutaraldehyde in 0.05 M phosphate buffer (pH 7.4) for 2 h. After washing with phosphate buffer for 10 min three times each, post fixation was performed in 2% (v/v) osmium tetroxide in phosphate buffer for 2 h at room temperature. After washing three times, the samples were dehydrated through an alcohol series (30%, 50%, 70%, 90%, and 100% two times for 30 min each). The samples were gently dried using a critical point drier (EM CPD 300, Leica, Jena, Germany). Samples were mounted on metallic stubs, gold-coated (~100 Å) with a sputter coater (Q150R Plus – Rotary Pumped Coater, Quorum Technologies Ltd., Sussex, UK) and viewed under the FE-SEM at 20 kV.

Fine structure of C. orbiculare on the surface of cucumber leaves
pre-treated with bio-sulfur was observed with a scanning electron microscope at 1 and
3 days after inoculation. Leaves were cut with a razor into a size of
1 × 3 mm². Samples were fixed with 2% glutaraldehyde in 0.1 M sodium cacodylic
buffer (pH 7.2) for 2 h. After fixation, samples were washed with the same buffer for
10 min three times. To remove moisture from samples, 30, 50, 70, 80, 90, 95, and 100% of
ethanol series were used for treatment (10 min for each treatment, three treatments for
each ethanol step). Finally, 100% acetone was used for treatment (30 min each time for a
total of two times).
Each sample was dried with a critical point dryer (CPD 030; BAL-Tec, Los Angeles, CA) and
coated with a platinum using sputter coater-platinum (Q150R Plus – Rotary Pumped Coater,
Quorum technologies Ltd., Sussex, UK) at 20 mA for 90 s. Samples were observed with a
field emission scanning electron microscope (FE-SEM Mira3, Tescan Ltd., Brno, Czech
Republic).