Department of Fundamental Microbiology, University of Lausanne, Lausanne, Switzerland.
PLoS Biol. 2024 Mar 5;22(3):e3002523. doi: 10.1371/journal.pbio.3002523. eCollection 2024 Mar.
The honey bee is a powerful model system to probe host-gut microbiota interactions, and an important pollinator species for natural ecosystems and for agriculture. While bacterial biosensors can provide critical insight into the complex interplay occurring between a host and its associated microbiota, the lack of methods to noninvasively sample the gut content, and the limited genetic tools to engineer symbionts, have so far hindered their development in honey bees. Here, we built a versatile molecular tool kit to genetically modify symbionts and reported for the first time in the honey bee a technique to sample their feces. We reprogrammed the native bee gut bacterium Snodgrassella alvi as a biosensor for IPTG, with engineered cells that stably colonize the gut of honey bees and report exposure to the molecules in a dose-dependent manner through the expression of a fluorescent protein. We showed that fluorescence readout can be measured in the gut tissues or noninvasively in the feces. These tools and techniques will enable rapid building of engineered bacteria to answer fundamental questions in host-gut microbiota research.
蜜蜂是研究宿主-肠道微生物群相互作用的有力模式生物，也是自然生态系统和农业的重要传粉媒介。虽然细菌生物传感器可以为宿主与其相关微生物群之间发生的复杂相互作用提供重要的见解，但缺乏非侵入性采样肠道内容物的方法，以及用于工程共生体的有限遗传工具，迄今为止，它们在蜜蜂中的发展受到了阻碍。在这里，我们构建了一个多功能的分子工具包，用于遗传修饰共生体，并首次在蜜蜂中报告了一种采样其粪便的技术。我们将本土蜜蜂肠道细菌 Snodgrassella alvi 重新编程为 IPTG 的生物传感器，经过工程改造的细胞可以稳定地定植于蜜蜂肠道，并通过表达荧光蛋白以剂量依赖的方式报告暴露于这些分子。我们表明，荧光读数可以在肠道组织中或非侵入性地在粪便中测量。这些工具和技术将使构建工程细菌能够快速回答宿主-肠道微生物群研究中的基本问题。
