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RESEARCH-PGR: Sieve Tube Proteomics - Unraveling the Physiology and Cell Biology of an Arcane Cell Type

RESEARCH-PGR：筛管蛋白质组学 - 揭示神秘细胞类型的生理学和细胞生物学

基本信息

批准号：
1940827
负责人：
Michael Knoblauch
金额：
$ 70万
依托单位：
Washington State University
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-01 至 2024-01-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1940827&HistoricalAwards=false
关键词：
RESEARCH PGR Sieve Tube Proteomics

项目摘要

The food consumed by humans is produced in plants by photosynthesis, either directly (think of potatoes) or indirectly (meat from plant-eating animals). Photosynthesis occurs in the green parts of plants, but, with exceptions such as lettuce, hungry humans are more interested in non-photosynthesizing storage organs of plants including grain, tubers, and fruits. Plants transfer the products of photosynthesis (sugars, chemically speaking) from their green parts to storage organs via a network of microscopic pipes, the so-called sieve tubes, in the phloem tissue. Due to their extreme sensitivity, the function of live sieve tubes is notoriously hard to study, and our understanding of sieve tube transport and its regulation has remained lamentably poor. This is unfortunate, especially since numerous phloem-sucking pests, for instance aphids, exploit the continuous stream of energy-rich substances in the sieve tubes. What often is worse are the many pathogenic viruses and bacteria that hitch rides in this stream to infest the entire plant, starting from a single point of entry. As these factors cause massive crop losses every year, the critical function of phloem in plant performance translates into an economic significance that can hardly be overstated. This project will make important contributions to comprehension of fundamental mechanisms of sieve tube function, which will facilitate the design of treatments and preventive measures targeting pests and pathogens. Proteins potentially involved in sieve tube physiology will be identified by a large-scale biochemical approach (‘proteomics’), and then further analyzed one by one through directed modification of the genes that encode them. With regard to Broader Impacts, the project will engage public interest with an artistic exhibition that incorporates large format photographs of phloem cells displayed in the terminal of the Pullman/Moscow regional airport and will also continue updating a “Phloem” webpage. In addition the PI will work with the Office of Multicultural Student Services at Washington State University to recruit minority undergraduate students to work in the lab.Vascular systems allow organisms to distribute resources internally by bulk flow, thus overcoming size limitations set by diffusion. The development of vascular tissues including sieve tubes drove the evolution of large land plants (tracheophytes) which caused a major increase in the productivity of terrestrial ecosystems. This productivity supports life on earth in its various forms. The largest photoautotroph marine organisms, kelps, have convergently evolved transporting sieve tubes, highlighting the significance of this transport and communication system for large photosynthetic and sessile life forms. This project aims at elucidating open questions in the cell biology and physiology of sieve tubes. State-of-the-art proteomics approaches will be applied to identify proteins potentially involved in transport regulation, by comparing the proteomes of vascular tissue and isolated sieve elements. Candidate proteins will be fluorescently tagged by modifications of their encoding genes, and the subcellular localization of the proteins in transporting tubes will be determined. Advanced bioimaging tools will be applied to assign sieve tube proteins to specific organelles or membrane systems, to generate 3D models of sieve tube organelle interactions, and to characterize the responses of protein distribution to stress. The insights gained are expected to provide essential information for the generation of more robust and pest-resistant crops.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.

人类消耗的食物是通过光合作用在植物中产生的，直接（想想土豆）或间接（食草动物的肉）。光合作用发生在植物的绿色部分，但是，除了莴苣等例外，饥饿的人类对植物的非光合储存器官更感兴趣，包括谷物，块茎和水果。植物将光合作用的产物（从化学上讲是糖）从绿色部分通过韧皮部组织中的一个微观管道网络（所谓的筛管）转移到储存器官。由于它们的极端敏感性，活筛管的功能是出了名的难以研究，我们对筛管运输及其调节的理解仍然很差。这是不幸的，特别是因为许多韧皮部吸吮害虫，如蚜虫，利用筛管中连续不断的富含能量的物质。更糟糕的是，许多致病病毒和细菌从一个入口开始，在这条溪流中搭便车，感染整个植物。由于这些因素每年造成大量的作物损失，韧皮部在植物性能中的关键功能转化为经济意义，这一点几乎不被夸大。该项目将为理解筛管功能的基本机制做出重要贡献，这将有助于设计针对害虫和病原体的治疗和预防措施。可能参与筛管生理学的蛋白质将通过大规模生物化学方法（“蛋白质组学”）进行鉴定，然后通过编码它们的基因的定向修饰逐一进行进一步分析。关于更广泛的影响，该项目将通过一个艺术展览吸引公众的兴趣，该展览将展出普尔曼/莫斯科地区机场航站楼中展示的韧皮部细胞的大幅照片，并将继续更新“韧皮部”网页。此外，PI将与华盛顿州立大学的多元文化学生服务办公室合作，招募少数民族本科生在实验室工作。血管系统允许生物体通过大量流动在内部分配资源，从而克服扩散所设定的尺寸限制。包括筛管在内的维管组织的发展推动了大型陆生植物（维管植物）的进化，从而导致陆地生态系统生产力的大幅提高。这种生产力以各种形式支持着地球上的生命。最大的光合自养海洋生物，海带，收敛进化运输筛管，突出了大型光合和固着生命形式的运输和通信系统的意义。本计画旨在阐明筛管的细胞生物学与生理学中尚未解决的问题。国家的最先进的蛋白质组学的方法将被应用到确定蛋白质可能参与运输调节，通过比较蛋白质组的血管组织和分离的筛元件。候选蛋白质将通过其编码基因的修饰进行荧光标记，并确定蛋白质在转运管中的亚细胞定位。先进的生物成像工具将被应用于分配筛管蛋白质到特定的细胞器或膜系统，以生成筛管细胞器相互作用的3D模型，并表征蛋白质分布对压力的响应。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。