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本文利用MTT染色法和点接生物测定法对不同温度干燥处理36h后的葡萄生单轴霉孢子囊存活力和致病力进行了检测,并应用单因素试验和正交试验对其孢子囊进行了MTT染色条件的优化。结果表明:葡萄生单轴霉孢子囊MTT染色的最佳条件为温度36℃、MTT浓度0.05%、染色48h,孢子囊染色率可达83.0%。20℃恒温干燥处理36h显著提高了孢子囊存活力,葡萄生单轴霉孢子囊的蓝色染色率为79.3%,叶片点接发病率为98.9%,显著高于对照的蓝色染色率52.0%和发病率62.7%。MTT染色得到的孢子囊蓝色染色率与点接生物测定法得出的发病率存在很好的线性关系y=1.276 1x-1.939 1,R2=0.996 1,孢子囊的蓝色染色率与叶片点接发病率呈正相关。本研究表明MTT染色法可以用于葡萄生单轴霉孢子囊存活力的快速和准确检测。
In this paper, viability and virulence of Plasmopara viticola were measured by MTT staining and point-to-point bioassay after 36h drying at different temperatures. The sporocysts were tested by single factor test and orthogonal experiment MTT staining conditions optimization. The results showed that the optimal conditions for MTT staining of Plasmopara viticola were 36 ℃, MTT concentration 0.05%, staining 48h, and the staining rate of sporocysts up to 83.0%. Drying at 20 ℃ for 36h significantly enhanced the spore capsule viability. The blue staining rate of Plasmopara viticola was 79.3%, the incidence of leaf spot incidence was 98.9%, significantly higher than that of the control (52.0% And the incidence of 62.7%. There was a good linear relationship between the blue staining rate of sporangia and the incidence rate of spotted bioassay by MTT staining y = 1.276 1x-1.939 1, R2 = 0.996 1, the blue staining rate of sporangium and leaf spot Then the incidence was positively correlated. This study shows that MTT staining can be used for the rapid and accurate detection of Plasmopara viticola sporulation.