Collaborative Research: Creating Synthetic Lichen to Elucidate how Morphology Impacts Mutualistic Exchanges in Microbial Communities.

合作研究：创造合成地衣来阐明形态学如何影响微生物群落的互惠交换。

• 批准号: 2334681
• 负责人: Daniel Ducat
• 金额: $ 27.31万
• 依托单位: Michigan State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-02-01 至 2027-01-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2334681&HistoricalAwards=false
• 关键词: Collaborative; Research; Creating; Synthetic; Lichen

Complex communities of microbes colonize most surfaces on the planet including rocks, leaves, and our organs. Their activities and interactions on these surfaces have major impacts on human, plant, animal, and ecosystem health. However, the impact of the 3D structure of these microbial communities on their capacity to exchange resources remains poorly understood. This has limited the capacity both to predict the growth rate and resilience of natural microbes in changing environments, as well as to engineer synthetic microbiomes for industrial applications. This work uses synthetic biology to engineer industrial microbes to recapitulate aspects of lichen symbiosis, where structure plays a central role in microbial exchange. This approach facilitates generating novel insights into the contribution of morphology to symbiosis and powerful new tools for engineering the growth and communication between microbes. The synthetic lichen created through this work is a powerful new platform to study symbiosis, as well as a novel photosynthetic biomanufacturing platform. Additionally, this project uses synthetic lichen as both a tool and a metaphor to create art that can communicate the ideas underpinning this work to undergraduate students and the broader public. In this project a synthetic lichen is created from free living cyanobacteria as photobiont and filamentous fungi as mycobiont, to explore how the physical coupling created by morphology impacts the physiological outcomes of metabolic exchanges. The morphological patterning of the fungi is implemented using a previously validated synthetic cell-cell signaling system consisting of an Indole 3-Acetic Acid (auxin) biosynthesis pathway and a library of auxin-inducible Cas9-based transcription factors (HACRs) with a range of sensitivities. The cyanobacteria is engineered to generate auxin-based patterning cues and HACRs are used to connect these signals to the expression of genes regulating morphology in the fungus, to generate a lichen-like morphology. In parallel, morphological features of the cyanobacterium are modulated by utilizing either a filamentous or single-celled strain, or established pathways for converting rod-shaped unicellular cyanobacteria into elongated filaments of varying lengths. These microbes are also engineered to perform altruistic metabolic behaviors, namely the secretion of sucrose and extracellular polysaccharide by the photobiont and mycobiont respectively, resulting in symbioses that mimic the relationship observed in lichens. This project elucidates how the different degrees of physical coupling that morphology generates between microbes impact the metabolic exchanges which drive consortia-level physiology.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

微生物的复杂群落在地球上定居大多数表面，包括岩石，叶子和我们的器官。他们在这些表面上的活动和互动对人，植物，动物和生态系统健康产生了重大影响。但是，这些微生物群落的3D结构对交换资源的能力的影响仍然很少。这限制了预测自然微生物在不断变化的环境中的增长率和弹性的能力，以及用于工业应用的工程师合成微生物组。这项工作使用合成生物学来设计工业微生物，以概括地衣共生的各个方面，其中结构在微生物交换中起着核心作用。这种方法促进了对形态对共生和强大的新工具的贡献的新见解，以设计微生物之间的生长和交流。通过这项工作创建的合成地衣是研究共生的强大新平台，以及一个新型的光合生物制造平台。此外，该项目将合成地衣既是工具又是隐喻来创造艺术，这些艺术可以传达这项工作的想法，将这项工作与本科生和更广泛的公众交流。在这个项目中，从自由活着的蓝细菌作为光叶族和丝状真菌作为mycobiont创建了合成地衣，以探索形态产生的物理耦合如何影响代谢交流的生理结果。真菌的形态模式是使用先前验证的合成细胞 - 细胞信号系统实施的，该系统由吲哚3-乙酸（Auxin）生物合成途径和生长素诱导的基于CAS9的转录因子（HACR）组成。蓝细菌经过设计，用于产生基于生长素的图案提示，HACR用于将这些信号与调节真菌中形态的基因的表达联系起来，以产生类似地衣的形态。同时，通过利用丝状或单细胞菌株或已建立的途径来调节蓝细菌的形态特征，将杆状的单细胞蓝细菌转化为长度不同的细丝。这些微生物还经过设计用于执行利他的代谢行为，即分别由光硫酸和霉菌分别分别分泌蔗糖和细胞外多糖，从而使象征性地模仿了在苔藓中观察到的关系。该项目阐明了形态在微生物之间产生的不同程度的物理耦合如何影响代谢交易所，这些交流促进了财团级的生理学。该奖项反映了NSF的法定任务，并被认为是值得通过基金会的知识分子和更广泛影响的审查标准来通过评估来通过评估来获得支持的。