Specific detection of intracellular symbiotic bacteria of aphids by oligonucleotide-probed in situ hybridization
Specific detection of intracellular symbiotic bacteria of aphids by oligonucleotide-probed in situ hybridization
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DOI：
10.1303/aez.33.461
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发表时间：
1998-08-01
影响因子：
1.3
通讯作者：
Sekiguchi, Y
中科院分区：
文献类型：
--
作者：
Fukatsu, T;Watanabe, K;Sekiguchi, Y
尝试利用针对细菌 rRNA 的寡核苷酸探针原位杂交来检测、可视化和表征蚜虫的细胞内共生细菌。利用与真细菌16S rRNA通用杂交的EUB338探针，在组织薄片上成功对各种蚜虫的初级和次级细胞内共生体进行了染色。当用GAM42A和BET42A探针分别靶向变形菌门的γ亚类和P亚类的23S rRNA进行原位杂交时，结果表明，松果松和Nippolachnus piri的次生共生体属于变形菌门的γ亚类，而根根四脉菌属于变形菌门的β亚类。变形菌。这是关于蚜虫次生细胞内共生体的系统发育亲和力的第一份报告。由于昆虫组织一般具有较强的自发荧光，因此使用生物素和地高辛标记探针的非荧光原位杂交更适合检测蚜虫组织切片中的共生细菌。
Oligonucleotide-probed in situ hybridization targeted to bacterial rRNAs was attempted to detect, visualize and characterize the intracellular symbiotic bacteria of aphids. Using the EUB338 probe that hybridizes universally with 16S rRNA of eubacteria, the primary and secondary intracellular symbionts of various aphids were successfully stained on tissue thin sections. When in situ hybridization was conducted with GAM42A and BET42A probes that are targeted to 23S rRNA of gamma- and P-subdivision of the Proteobacteria, respectively, it was shown that the secondary symbionts of Cinara pini and Nippolachnus piri belong to the gamma-subdivision of the Proteobacteria whereas that of Tetraneura radicicola is a member of the beta-subdivision of the Proteobacteria. This is the first report on the phylogenetic affinity of the secondary intracellular symbionts of aphids. Because insect tissues have strong autofluorescence in general, non-fluorescent in situ hybridization using biotin- and digoxigenin-labelled probes was more suitable for detecting symbiotic bacteria in aphid tissue sections.