tomato_6421_infected_with_cg
Single-spore cultures of Colletotrichum gloeosporioides (Penz.) Cg-14 isolate were obtained from a decayed avocado fruit (Wattad et al., 1994). The isolate was thoroughly analyzed in tomato infections (Alkan et al., 2013b). Cultures were grown at 27°C in the dark, and maintained on M3S plates. Conidia were harvested with 10 mL of sterile distilled water supplemented with 0.01% (v/v) Tween 80 (Sigma-Aldrich, Copenhagen, Denmark). Cells were visualized with a model BX60F-3 microscope and a model SZ-60 stereoscope (both from Olympus America Inc., Melville, NY, USA) and counted with a hemocytometer (Brand, GMBH, Werheim, Germany).  Tomato (Solanum lycopersicum) plants were grown in the greenhouse under standard conditions. Transgenic tomatoes SlPH are impaired of the PH ortholog controls fruit acidity. Fruit juice of the silenced plants had nearly a 2 pH units increase compared with the non-silenced plants, together with reduced % acid.  Tomato fruits were inoculated by placing 5 μL of a conidial suspension containing 106 spores mL-1 on each of four 2-mm-deep, 2-mm-diameter wounds spaced evenly in a circle around the upper part of the stem end of 8 fruits. Following inoculation, the fruits were incubated for 3 days at 25°C in covered plastic containers containing wet paper towels. Samples of healthy and infected tissues were collected and frozen with liquid nitrogen for subsequent RNA analysis.
tomato_6421_infected_with_cg