Calcofluor White M2R 与Sytox Green 双重染色法鉴别蜜蜂微孢子虫
Detection and identification of Nosema ceranae by dual fluorescent staining with Calcofluor White M2R and Sytox Green
秦浩然1, 2**，李继莲1，和绍禹2***，吴杰
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作者单位：1. 农业部授粉昆虫生物学重点实验室中国农业科学院蜜蜂研究所北京100093；2. 云南农业大学 东方蜜蜂研究所昆明650201
中文关键词：东方蜜蜂微孢子虫，荧光染色，Calcofluor White M2R，Sytox Green
英文关键词：Nosema ceranae，fluorescent staining，Calcofluor White M2R，Sytox Green
中文摘要：东方蜜蜂微孢子虫 (Nosema ceranae)是一种广泛寄生于东方蜜蜂 Apis cerana，西方蜜蜂 Apis mellifera和熊蜂 Bombus Latreille上的寄生虫，对蜜蜂和熊蜂的危害较大，进而影响养蜂业的发展。本实验采用荧光染色试剂 Calcofluor White M2R与核酸染料 Sytox Green双重染法来鉴别蜜蜂或熊蜂体内的N． ceranae及孢子的存活状态。结果得出，在荧光显微镜下可见死孢子被染上黄绿色荧光，活的呈现蓝白色荧光，而寄主细胞、细菌、病毒等不被染色。这是一种快速有效鉴别N． ceranae及其死活的方法，从而判定蜜蜂或熊蜂体内的微孢子虫在是否具有侵染活性，对微孢子虫的研究及药物防治具有重要作用。
英文摘要： Nosema ceranae is a microsporidian parasite of Apis cerana，Apis mellifera and bumblebees that has an adverse effect on pollination，especially of fruits and vegetables，and apiculture． In this study，Calcofluor White M2R and Sytox Green stains were used to discriminate between live and deadN． ceranae． The results show that deadN． ceranae spores had yellow-green fluorescence whereas live spores fluoresced white－blue． This method allows easy detection and identification of live and deadN． ceranae and should therefore contribute to further research onN． ceranae and itstreatment．