Rapid Fluorescence-Based Screening for Wolbachia Endosymbionts in Drosophila Germ Line and Somatic Tissues
Rapid Fluorescence-Based Screening for Wolbachia Endosymbionts in Drosophila Germ Line and Somatic Tissues
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DOI：
10.1128/aem.00215-11
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发表时间：
2011-07-01
影响因子：
4.4
通讯作者：
Sullivan, William
中科院分区：
文献类型：
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作者：
Casper-Lindley, Catharina;Kimura, Scott;Sullivan, William
沃尔巴克氏体是一种广泛存在于节肢动物和线虫体内的细菌内共生体。敏感的PCR为基础的方法的出现，极大地促进了沃尔巴克氏体感染的个人和人群感染水平的分析识别。在这里，一个互补的视觉荧光为基础的沃尔巴克氏体筛选方法进行说明。通过使用荧光染料Syto-11，可以在包括卵巢在内的各种果蝇组织中有效地检测Wolbachia。Syto-11还能使其他昆虫体内的沃尔巴克氏体染色。由于沃尔巴克氏体是通过母系生殖系遗传的，因此细菌驻留在受感染种群中苍蝇的卵巢中。这种染色方法的一个优点是，它告知沃尔巴克氏体滴度以及其组织和细胞分布。使用该方法，确定了加州中部两个地点的昆虫种群的感染状况，并分离出具有异常低或高的沃尔巴克氏体滴度的变体。此外，还鉴定了一种卵巢管包含感染和未感染卵室的变体。还分析了Cardinium和Spiroplasma感染的昆虫的Syto-11染色。
Wolbachia is a globally distributed bacterial endosymbiont present in arthropods and nematodes. The advent of sensitive PCR-based approaches has greatly facilitated the identification of Wolbachia-infected individuals and analysis of population infection levels. Here, a complementary visual fluorescence-based Wolbachia screening approach is described. Through the use of the fluorescent dye Syto-11, Wolbachia can be efficiently detected in various Drosophila tissues, including ovaries. Syto-11 also stains Wolbachia in other insects. Because Wolbachia is inherited through the maternal germ line, bacteria reside in the ovaries of flies in infected populations. An advantage of this staining approach is that it informs about Wolbachia titer as well as its tissue and cellular distribution. Using this method, the infection status of insect populations in two central California locations was determined, and variants with unusually low or high Wolbachia titers were isolated. In addition, a variant with ovarioles containing both infected and uninfected egg chambers was identified. Syto-11 staining of Cardinium- and Spiroplasma-infected insects was also analyzed.