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应用镰刀菌(Fusarium)纯毒素脱氧雪腐镰刀菌烯醇Deoxynivalenol(简称DON)加入小麦叶肉原生质体悬浮液,设30,50,100ppm三个DON毒素浓度处理及空白对照,在18—20℃和26—28℃两种温度下,1—7小时不同时间取样。经二乙酸荧光素(Fluorescein dlacetate)染色,于荧光显微镜下观察各处理原生质体存活率。并分别与对照存活率进行成对数据t测验。结果表明,在18—20℃6小时后取样观察,50ppm毒素对不同抗性小麦品种原生质体反应差异最显著。首次发现毒素能显著提高抗病品种的原生质体活力。如抗病品种望水白和苏麦3号的原生质体处理存活率分别比对照高24.1和9.7个百分点。但DON毒素对感病品种原生质体则有显著伤害作用,使其质膜严重受损,原生质处逸,最终导致原生质体死亡。如感病品种徐州21和绵阳11的原生质体存活率分别比其对照低20.8和20.6个百分点。中抗和中感品种原生质体处理和对照之间,存活率均无显著差异。以不同小麦品种原生质体对DON毒素的敏感性差异为依据,建立小麦品种抗赤霉病性生物测定法,经用该方法(DON毒素法)测试的14个小麦品种抗赤性与其田间抗赤性基本相符。
The Fusarium detoxification deoxynivalenol (DON) was added to the wheat mesophyll protoplast suspension. Three DON concentrations of 30, 50 and 100 ppm were treated with the blank control. At 18-20 ℃ and 26 -28 ℃ two kinds of temperature, 1-7 hours sampling at different times. After being stained with Fluorescein dlacetate, the viability of protoplasts of each treatment was observed under a fluorescence microscope. And paired data t test with control survival rate respectively. The results showed that at the temperature of 18-20 ℃ 6 hours after sampling, 50ppm toxin had the most significant difference in protoplast response of different resistant wheat varieties. The discovery of toxins for the first time significantly increased the viability of protoplasts of resistant varieties. For example, the protoplast survival rates of resistant variety Wangshuibai and Sumai 3 were respectively 24.1 and 9.7% higher than that of the control. However, DON toxin had significant damage to the protoplasts of susceptible varieties, severely damaged the plasma membrane, protoplasm yi at the end and eventually led to the death of protoplasts. For example, the survival rate of protoplasts of susceptible cultivars Xuzhou 21 and Mianyang 11 were 20.8 and 20.6 percentage points lower than their control respectively. There was no significant difference in the survival rate between the protoplasts of Zhonggan and Zhonggan cultivars and the control. Based on the sensitivity of protoplasts of different wheat varieties to DON toxin, the bioassay of wheat cultivars against Fusarium head blight was established. The resistance of 14 wheat cultivars tested by this method (DON toxin method) The basic match.