University of Tübingen, Interfaculty Institute of Cell Biology, Section Animal Genetics, Auf der Morgenstelle 15, 72076, Tübingen, Germany.
School of Pharmaceutical Science and Technology, Tianjin University, Tianjin, 300072, China.
BMC Res Notes. 2021 Sep 8;14(1):351. doi: 10.1186/s13104-021-05766-7.
As in most organisms, the surface of the fruit fly Drosophila melanogaster is associated with bacteria. To examine whether this association depends on cuticle quality, we isolated and quantified surface bacteria in normal and melanized flies applying a new and simple protocol.
On wild flies maintained in the laboratory, we identified two persistently culturable species as Lactobacillus plantarum and Acetobacter pomorum by 16S rDNA sequencing. For quantification, we showered single flies for DNA extraction avoiding the rectum to prevent contamination from the gut. In quantitative PCR analyses, we determined the relative abundance of these two species in surface wash samples. On average, we found 17-times more A. pomorum than L. plantarum. To tentatively study the importance of the cuticle for the interaction of the surface with these bacteria, applying Crispr/Cas9 gene editing in the initial wild flies, we generated flies mutant for the ebony gene needed for cuticle melanisation and determined the L. plantarum to A. pomorum ratio on these flies. We found that the ratio between the two bacterial species reversed on ebony flies. We hypothesize that the cuticle chemistry is crucial for surface bacteria composition. This finding may inspire future studies on cuticle-microbiome interactions.
与大多数生物一样,果蝇的体表与细菌有关。为了研究这种关联是否依赖于表皮质量,我们应用一种新的简单方案,从正常和黑化的果蝇中分离和定量了体表细菌。
通过 16S rDNA 测序,我们在实验室饲养的野生果蝇中鉴定出两种可持久培养的物种,即植物乳杆菌和醋酸杆菌。为了定量分析,我们对单只果蝇进行喷淋以提取 DNA,避免直肠污染肠道。在定量 PCR 分析中,我们确定了这两种物种在体表冲洗样本中的相对丰度。平均而言,我们发现醋酸杆菌的丰度是植物乳杆菌的 17 倍。为了初步研究表皮对与这些细菌表面相互作用的重要性,我们在最初的野生果蝇中应用了 Crispr/Cas9 基因编辑,生成了需要黑化表皮的 ebony 基因突变果蝇,并确定了这些果蝇上植物乳杆菌和醋酸杆菌的比例。我们发现这两种细菌的比例在 ebony 果蝇上发生了反转。我们假设表皮化学性质对体表细菌组成至关重要。这一发现可能为未来研究表皮-微生物组相互作用提供启示。