Kounatidis Ilias, Crotti Elena, Sapountzis Panagiotis, Sacchi Luciano, Rizzi Aurora, Chouaia Bessem, Bandi Claudio, Alma Alberto, Daffonchio Daniele, Mavragani-Tsipidou Penelope, Bourtzis Kostas
Department of Genetics, Development and Molecular Biology, School of Biology, Faculty of Sciences, Aristotle University of Thessaloniki, 54124 Thessalonici, Greece.
Appl Environ Microbiol. 2009 May;75(10):3281-8. doi: 10.1128/AEM.02933-08. Epub 2009 Mar 20.
Following cultivation-dependent and -independent techniques, we investigated the microbiota associated with Bactrocera oleae, one of the major agricultural pests in olive-producing countries. Bacterial 16S rRNA gene libraries and ultrastructural analyses revealed the presence of several bacterial taxa associated with this insect, among which Acetobacter tropicalis was predominant. The recent increased detection of acetic acid bacteria as symbionts of other insect model organisms, such as Anopheles stephensi (G. Favia et al., Proc. Natl. Acad. Sci. USA 104:9047-9051, 2007) or Drosophila melanogaster (C. R. Cox and M. S. Gilmore, Infect. Immun. 75:1565-1576, 2007), prompted us to investigate the association established between A. tropicalis and B. oleae. Using an A. tropicalis-specific PCR assay, the symbiont was detected in all insects tested originating from laboratory stocks or field-collected from different locations in Greece. This acetic acid bacterium was successfully established in cell-free medium, and typing analyses, carried out on a collection of isolates, revealed that different A. tropicalis strains are present in fly populations. The capability to colonize and lodge in the digestive system of both larvae and adults and in Malpighian tubules of adults was demonstrated by using a strain labeled with a green fluorescent protein.
采用依赖培养和不依赖培养的技术后,我们研究了与油橄榄实蝇相关的微生物群,油橄榄实蝇是橄榄生产国的主要农业害虫之一。细菌16S rRNA基因文库和超微结构分析揭示了与这种昆虫相关的几种细菌类群,其中热带醋杆菌占主导地位。最近,作为其他昆虫模式生物(如斯蒂芬按蚊(G. Favia等人,《美国国家科学院院刊》104:9047 - 9051,2007年)或黑腹果蝇(C. R. Cox和M. S. Gilmore,《感染与免疫》75:1565 - 1576,2007年))的共生菌,醋酸菌的检测频率增加,这促使我们研究热带醋杆菌与油橄榄实蝇之间建立的关联。使用热带醋杆菌特异性PCR检测方法,在所有测试的昆虫中都检测到了这种共生菌,这些昆虫来自实验室种群或从希腊不同地点野外采集。这种醋酸菌在无细胞培养基中成功培养,对一系列分离株进行的分型分析表明,果蝇种群中存在不同的热带醋杆菌菌株。通过使用标记有绿色荧光蛋白的菌株,证明了其在幼虫和成虫的消化系统以及成虫的马氏管中定殖和存活的能力。