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B. cinerea was isolated from decayed tomato fruit and incubated on potato dextrose agar medium (PDA). Spore suspensions of the strain were prepared by flooding the 7-d-old culture dishes incubated at 25¡À1 ◦C with 0.05% Tween-80 solution. The spore suspension was adjusted to 1¡Á106 conidia/mL with a hemocytometer. The inoculations were carried out in a sterilized laminar flow cabinet 1, 3, 6, 9 and 12 d after fruit were treated. Ten fruit per replicate and three replicates per treatment were surface-sterilized by immersion in 70% ethanol for 1 min, rinsed three times in sterile distilled water, and then wounded with a sterile nail at three points (4mm deep¡Á2mm wide) on the equator of each fruit. Then 10uL of spore suspension was injected into each wound site and the fruit incubated at 25¡À1 ◦C, 85¨C90% RH. Disease symptoms on each fruit were recorded on the 3rd day after inoculation (when the symptoms were visible and countable).Lesion size = 3.14¡Á(lesion diameter/2)2. [ Last edited by ÎôÄê²ÐÃÎ on 2011-3-21 at 22:15 ] |
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