PCR diagnostic methods for Ascosphaera infections in bees
PCR diagnostic methods for Ascosphaera infections in bees
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DOI：
10.1016/j.jip.2005.08.004
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发表时间：
2005-10-01
影响因子：
3.4
通讯作者：
Skinner, JS
中科院分区：
文献类型：
--
作者：
James, RR;Skinner, JS
子囊壳属真菌是影响蜜蜂幼虫生存力的主要疾病--白垩病的病原体。由于缺乏可区分的特征，基于形态特征的单个囊腔菌属物种的鉴定一直很困难。大多数鉴定是基于子囊、孢子球和分生孢子的大小和形状。不幸的是，这些结构的大小发生了很多重叠，一些Ascosphaera物种不会在体外产生有性结构。我们报告了一种快速可靠的诊断方法，用于确定Ascosphaera感染切叶蜂（切叶蜂）使用PCR标记，采用属特异性引物Ascosphaera，和物种特异性引物已知与切叶蜂属。使用这些方法，可以直接对蜜蜂进行物种鉴定，包括渐近个体。此外，PCR标记物可以检测在单个宿主中的多个囊腔菌属物种的共感染。我们还确定了一个标志Ascosphaera apis，在蜜蜂inellifera，蜜蜂白垩病的主要原因。我们的诊断方法消除了对培养样品的需要，并可用于处理大量的现场收集的蜜蜂幼虫。(c)2005年爱思唯尔公司All rights reserved.
Fungi in the genus Ascosphaera are the causative agents of chalkbrood, a major disease affecting bee larval viability. Identification of individual Ascosphaera species based on morphological features has been difficult due to a lack of distinguishing characteristics. Most identifications are based on the size and shape of the ascomata, spore balls and conidia. Unfortunately, much overlap occurs in the size of these structures, and some Ascosphaera species will not produce sexual structures in vitro. We report a quick and reliable diagnostic method for identifying Ascosphaera infections in Megachile bees (leafcutting bees) using PCR markers that employ genus-specific primers for Ascosphaera, and species-specific primers for species known to be associated with Megachile spp. Using these methods, species identifications can be performed directly on bees, including asymptornatic individuals. Furthermore, the PCR markers can detect co-infections of multiple Ascosphaera species in a single host. We also identified a marker for Ascosphaera apis, the predominant cause of chalkbrood in Apis inellifera, the honey bee. Our diagnostic methods eliminate the need for culturing samples, and could be used to process a large number of field collected bee larvae. (c) 2005 Elsevier Inc. All rights reserved.