中国农业科学2012，Vol.45Issue(15)：3075-3082,8.
微孔板法检测番茄灰霉病菌对杀菌剂的敏感性
Microtiter Method to Test the Sensitivity of Botrytis cinerea to Fungicides
摘要
Abstract
[Objective] The objective of this study is to establish a rapid and efficient method for testing the sensitivity of B. cinerea to fungicides by using the enzyme-labelling measuring instrument. [Method] With AOD value as an evaluation index, the conditions of microtiter method to test the sensitivity of B. cinerea to fungicides were optimized. The effect of six widely-used fungicides (boscalid, azoxystrobin, diethofencarb, thiophanate-methyl, iprodione and difenoconazole) against B. cinerea spore germination and mycelial growth was tested, and the results were compared with the conclusions of spore germination method and mycelium growth rate method. [Result] The conditions of microtiter method were optimized. The determination wavelength was 630 nm, the medium was PDE, the concentration of conidial suspension was 8×105-1.6×106 spores/mL, the optimum culture conditions were under shaking culture (150 r/min) at 21℃ in the dark, the text time for the inhibition of spore germination was 24 h after adding fungicide and conidial suspension in the wells, and the testing time for the inhibition of mycelial growth was 12-24 h after adding fungicide (conidial suspension in wells was incubated for 12 h before adding fungicide). The conclusion was almost consistent with that from conventional spore germination method or mycelium growth rate method. Fifty-seven isolates of B. cinerea from three regions of Shandong Province were tested by this method, and the mean EC50 values were 1.9311 and 5.6488 μg·mL-1 for inhibition of conidial germination and mycelial growth, respectively. [Conclusion] Microtiter method combines the advantages of both conventional methods together, it has characteristics of fast testing speed, high measuring accuracy and good reproducibility, so it possesses better practicability.关键词
微孔板法/灰霉病菌/敏感性/测试条件Key words
microtiter method/ Botrytis cinerea/ sensitivity/ testing conditions引用本文复制引用
杨晓楠,王猛,申瑞平,刘峰..微孔板法检测番茄灰霉病菌对杀菌剂的敏感性[J].中国农业科学,2012,45(15):3075-3082,8.基金项目
国家公益性行业(农业)科研专项(200903033) （农业）
