Hippe S
Institut für Biology III, Rheinisch-Westfälische-Technische Hochschule, Aachen, Federal Republic of Germany.
Histochemistry. 1987;87(4):309-15. doi: 10.1007/BF00492583.
The intracellular localization of the sterol-biosynthesis-inhibiting (SBI) fungicide (3H)triadimenol A is investigated in vitro in the fungus Ustilago avenae. For this purpose low temperature preparation techniques (shock freezing, freeze substitution, embedding in Lowicryl HM20) are combined with conventional electron microscopic (EM) autoradiography. In particular the suitability of Lowicryl HM20 embedded specimens for EM autoradiography with regard to the finestructure preservation is shown. For the localization of (3H)triadimenol the filamentous grain development as well as the application of the gold latensification method resulting in the appearance of spherical silver grains is tested. Fungicide sensitive wild type sporidia of U. avenae are compared with fungicide resistant cells of the mutant r8. A quantitative analysis of the autoradiographs of the wild type developed according to the gold latensification process shows a relatively homogeneous distribution of silver grains over the entire cell. On the other hand, the resistant mutant is characterized by an accumulation of silver deposits over the vacuoles as compared with the lower density of grains over the cell walls and cytoplasm. The data are discussed in the context of possible resistance mechanisms against SBI-fungicides.
在体外对燕麦黑粉菌进行研究，以确定甾醇生物合成抑制（SBI）杀菌剂（3H）三唑醇A在细胞内的定位。为此，将低温制备技术（冲击冷冻、冷冻置换、包埋于Lowicryl HM20中）与传统电子显微镜（EM）放射自显影相结合。特别展示了包埋于Lowicryl HM20中的标本对于EM放射自显影在超微结构保存方面的适用性。对于（3H）三唑醇的定位，测试了丝状颗粒的形成以及应用金强化法导致球形银颗粒的出现。将燕麦黑粉菌对杀菌剂敏感的野生型孢子与突变体r8的抗杀菌剂细胞进行比较。根据金强化过程显影的野生型放射自显影片的定量分析表明，银颗粒在整个细胞中分布相对均匀。另一方面，与细胞壁和细胞质上较低的颗粒密度相比，抗性突变体的特征是液泡上有银沉积物的积累。结合对SBI杀菌剂可能的抗性机制对这些数据进行了讨论。
