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以中黄13号大豆种子为材料,取生长8天的健康子叶,在其表面挖出约2×2mm的伤口,将50μl不同浓度的茉莉酸溶液和1mM的硝酸银溶液滴在伤口处,黑暗中(25℃)保温不同时间后取样,经甲醇提取,用高压液相色谱(HPLC)测定其中异戊烯化异黄酮glyceollins及其前体glycinol的累积情况。结果显示,健康子叶中没有glyceollins和glycinol的累积,1mM硝酸银比茉莉酸具有更强的诱导glyceollins和glycinol累积的能力,glyceollins的累积与茉莉酸浓度呈负相关。在各种处理中,glyceollinⅠ、Ⅱ、Ⅲ所占比例分别为:50%~80%、10%~40%和10%~20%,但是glyceollin三种异构体的比例随保温时间的延长而有所变化,glyceollinⅠ比例逐渐下降,glyceollinⅡ所占比例逐渐升高,在1mM硝酸银处理的子叶中表现更为明显。
Using healthy soybean cotyledons of Zhonghuang No.13 for 8 days, dredged about 2 × 2mm wounds on its surface. Fifty microliters of different concentrations of jasmonic acid solution and 1mM silver nitrate solution were dropped on the wounds. Darkness (25 ℃) for different time. The samples were extracted with methanol, and the accumulation of glyceolol and its precursor glycinol was determined by high pressure liquid chromatography (HPLC). The results showed that there was no accumulation of glyceolins and glycinol in healthy cotyledons. 1mM silver nitrate showed stronger ability of inducing glyceolol and glycinol accumulation than jasmonic acid, and the accumulation of glyceollins was negatively correlated with the concentration of jasmonic acid. The ratios of glyceollin I, II and III were 50% ~ 80%, 10% ~ 40% and 10% ~ 20% in all treatments, but the proportion of glyceollin isoforms was increased with the incubation time Some changes, glyceollin Ⅰ gradually decreased the proportion of glyceollin Ⅱ gradually increased, in 1mM silver nitrate treated cotyledons in the more obvious.