.5 mL was added and incubated for 5 min at root temperature (RT). The suspension of released nuclei was filtered through a 50-μm CellTris disposable filter to remove cell debris. Stained nuclei were analyzed by the Partec PAI Flow Cytometer using
been shown to survive in the soil, on plant debris, and in fumigated field soil, the main source of inoculum is assumed to originate on infected strawberry plant material from the nursery ( Eastburn and Gubler, 1990 ). However, the pathogen may be
30 h. After fixing, flower buds were transferred to 70% ethanol and stored at 4 °C. Flower buds were washed in sterile distilled water and anthers removed. Anthers were squashed in 1% acetocarmine stain on glass slides, debris removed, coverslip
small water drops or tiny specks of debris. However, because all anomalies were much smaller (usually less than 500 pixels) than the 10 actual berries (usually greater than 8000 pixels), the noise was easily filtered and the 10 individual data lines
tissue samples (≈5 mg) were immersed in nuclei extraction buffer (Partec, Munster, Germany) and macerated with a razor blade to release nuclei. The suspension was incubated for 5 min and then passed through 20-μm nylon mesh to remove debris. The nuclei in
). Inoculated and noninoculated plants were grown for 2 months. Next, an aliquot of the root was taken with a core sampler (23 mm diameter and 110 mm length) from 26 plants of each treatment. Roots were washed with tap water to remove soil debris and then
®; Partec, Münster, Germany) for 1 to 2 min at 25 °C. The preparation was then filtered using Partec CellTrics® disposable filters to remove debris. Nuclei were stained with 1.5 mL 4′, 6-diamidino-2-phenylindole staining buffer and incubated again at 25 °C
collection, the seeds were manually cleaned of excess floral parts and other debris with a screen and stored in sealed plastic bags under ambient laboratory conditions (22.8 °C, 27.5% relative humidity) for 2 to 4 months before undergoing testing. Pre
. The experiment was replicated three times. Each sample was homogenized in 1.0 mL of 80% (v/v) acetone using a pestle in an Eppendorf tube. After removing cellular debris by centrifugation, the supernatant containing chlorophyll was diluted 1:10 (v
mowed to 3.8 cm with a rotary mower, debris was removed, and the seedbed received two additional passes with a vertical mower set to a depth of ≈1.3 cm. Perennial ryegrass (Manhattan IV perennial ryegrass; Pure Seed Testing, Inc., Hubbard, OR) and tall