Collaborative Research: EDGE FGT: Functional Genomic Tools for Parasitic Nematodes and their Bacterial Symbionts

合作研究：EDGE FGT：寄生线虫及其细菌共生体的功能基因组工具

• 批准号: 2128266
• 负责人: Heidi Goodrich-Blair
• 金额: $ 60万
• 依托单位: University of Tennessee Knoxville
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-15 至 2025-07-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2128266&HistoricalAwards=false
• 关键词: Collaborative; Research; EDGE; FGT; Functional

All animals, including humans live in association with microbes and parasites that can promote health and cause disease. The mechanisms by which animals communicate with microbes and parasites to block, initiate, maintain, and dissolve such associations are just beginning to be uncovered. Because these mechanisms are often conserved across biology, they can be investigated using model animals such as roundworms, or nematodes, which associate with microbes and are themselves parasites. This project will develop new experimental tools in roundworms that have developed partnerships, or symbioses, with specific bacteria. Together these roundworms and their bacterial partners infect and kill insects, using them as a food source. Developing new tools to study this elegant animal-bacterium system will help expand our understanding how animals and bacteria form partnerships, and how they work together to parasitize other animals. The tools and knowledge gained in this project will be rapidly shared with the community of researchers involved in drug discovery, agricultural control of crop pests, and in the study of parasitism, infectious disease, beneficial microbiome function, and fundamental cell, molecular, developmental, and evolutionary biology. As part of this project, undergraduates will be involved in a discovery-based microbiology lab where they will practice isolating and identifying new insect-killing roundworms from the environment. Young people (K-12) and educators will be engaged through collaboration with the Science Journal for Kids, where a basic curriculum and summary of key research findings will be developed for classroom use. Entomopathogenic nematodes in the genera Heterorhabditis and Steinernema are mutualistically associated with bacteria in the genera Photorhabdus and Xenorhabdus, respectively. The nematode-bacterium symbiotic pair obligately parasitizes insects as a nutrient source for reproduction and has utility as a biological control agent for agricultural insect pests and as a source of novel compounds. The entomopathogenic pair and their insect hosts are models to understand fundamental biological principles, including the evolution and molecular and cellular basis of mutualistic and antagonistic organismal interactions. Numerous features make this system an excellent experimental model including ease and cost of husbandry, fast generation time, and optical transparency. Although genetic techniques have been developed for representative bacterial symbiotic partners and insect hosts, to date there have been no broadly adopted, reliable genetic modification tools in either Steinernema or Heterorhabditis nematodes. This inability to interrogate nematode gene function has hobbled full use of this system to yield much-needed insights into parasitism, animal microbiome interactions, and other areas. Here we propose to capitalize on a recently isolated Steinernema nematode that has promising characteristics for development of genetic tools, including hermaphroditic reproduction, amenability to long-term freezing, healthy development on agar bacterial lawns, resilience to microinjection, and significant pathogenicity to lab insects and agricultural pests. By fully sequencing the genomes and developing genetic techniques and tools for both nematode and symbiont, including CRISPR-Cas9 genome editing in the nematode, and creating an arrayed mutant library for the bacterium, our team will help realize the full potential of this elegant animal-microbe model system.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.

所有动物，包括人类，都与可促进健康并引起疾病的微生物和寄生虫生活在一起。动物与微生物和寄生虫沟通以阻止、启动、维持和消除这种联系的机制才刚刚开始被揭示。由于这些机制在生物学中通常是保守的，因此可以使用模型动物（例如蛔虫或线虫）来研究它们，这些动物与微生物相关，本身就是寄生虫。该项目将在与特定细菌建立伙伴关系或共生关系的蛔虫中开发新的实验工具。这些蛔虫和它们的细菌伙伴一起感染并杀死昆虫，并将它们作为食物来源。开发新工具来研究这种优雅的动物细菌系统将有助于扩大我们对动物和细菌如何形成伙伴关系以及它们如何共同寄生其他动物的理解。该项目中获得的工具和知识将与参与药物发现、作物害虫农业防治以及寄生、传染病、有益微生物组功能以及基础细胞、分子、发育和进化生物学研究的研究人员群体迅速共享。作为该项目的一部分，本科生将参与一个基于发现的微生物学实验室，在那里他们将练习从环境中分离和识别新的杀虫蛔虫。年轻人（K-12）和教育工作者将通过与《儿童科学杂志》合作来参与其中，该杂志将开发供课堂使用的基本课程和关键研究成果摘要。异杆菌属和斯氏线虫属的昆虫病原线虫分别与光杆菌属和致病杆菌属的细菌互惠相关。线虫-细菌共生对专门寄生于昆虫作为繁殖的营养源，并可用作农业害虫的生物防治剂和新型化合物的来源。昆虫病原对及其昆虫宿主是理解基本生物学原理的模型，包括互利和拮抗有机体相互作用的进化以及分子和细胞基础。该系统具有众多特性，使其成为出色的实验模型，包括饲养的简便性和成本、快速的生成时间和光学透明度。尽管已经针对代表性细菌共生伙伴和昆虫宿主开发了遗传技术，但迄今为止，在斯氏线虫或异杆线虫中还没有广泛采用的、可靠的遗传修饰工具。由于无法询问线虫基因功能，因此无法充分利用该系统来获得对寄生、动物微生物组相互作用和其他领域急需的见解。在这里，我们建议利用最近分离的斯氏线虫，该线虫具有用于开发遗传工具的有希望的特性，包括雌雄同体繁殖、适合长期冷冻、在琼脂菌苔上健康发育、对显微注射的抵抗力以及对实验室昆虫和农业害虫的显着致病性。通过对基因组进行全面测序并开发用于线虫和共生体的遗传技术和工具，包括在线虫中的 CRISPR-Cas9 基因组编辑，以及为细菌创建阵列突变体库，我们的团队将帮助实现这种优雅的动物微生物模型系统的全部潜力。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力优势和能力进行评估，被认为值得支持。 更广泛的影响审查标准。

项目成果

Conjugation and transposon mutagenesis of Xenorhabdus griffiniae HGB2511, the bacterial symbiont of the nematode Steinernema hermaphroditum (India).

• DOI: 10.17912/micropub.biology.000772
• 发表时间: 2023
• 期刊: microPublication biology
• 作者: Alani, Omar S;Cao, Mengyi;Goodrich-Blair, Heidi;Heppert, Jennifer K
• 通讯作者: Heppert, Jennifer K