Collaborative Research - Role of Pulsatile Ca^2+ in Controlling Polarized Hyphal Tip Growth

知识价值。所有生物都有识别和适应环境的复杂方法，因此它们能够生存或避免恶劣条件，开发新的食物来源，甚至引起疾病。在测序、解码和修改DNA的方法上取得的一些最新进展，以及在统一方法中使用强大显微镜的技术改进，现在可以在活细胞内监测目标遗传变化的微妙异常影响。这项研究使用真菌作为模型系统来确定细胞如何能够感知其微观环境的变化，并在随后的一系列改变细胞行为的事件中招募高度特化的蛋白质，这些蛋白质与钙离子结合。遗传方法将用于全面破坏真菌模式生物中所有已知的关键钙相互作用蛋白，并附加特殊标签，以监测活细胞内钙的变化和/或多种钙相互作用和相关蛋白的运动。这种方法将导致对哪些靶向蛋白质允许细胞以快速和定向的方式生长的全面理解。该项目的科学影响不仅仅是更好地理解导致小麦严重疾病的模式真菌镰刀菌（Fusarium graminearum）的钙信号生物学。它将为研究钙信号的进化创造一个新的工具箱，钙信号是一种高度保守的古老分子语言，几乎所有生物都使用它将外部环境信号转化为所需的细胞变化。该项目汇集了一组具有互补背景的科学家，包括；生物化学、细胞生物学、细胞学、进化生物学、真菌生物学、信息学、数学和遗传学。更广泛的影响。该项目的广泛和跨学科范围将为本科生、研究生和博士后提供一个理想的环境，以实践团队为基础的综合问题解决方案。面对日益复杂的生物学问题，对这种教育的需求不断增加。本项目支持的博士后将在不久的将来通过已建立的指导计划指导其发展自己的项目。通过为期一周的实践研讨会和全面的公开网站，其他人将获得所有资源并接受培训，以实施开发的工具。钙信号的发现和显微镜技术将在研究生课程和网络工具中介绍。本科生（包括少数民族学生）将通过NSF REU和EPSCoR项目以及宾夕法尼亚州立大学和特拉华大学的其他现有学分和外展项目参与该项目。这些努力将帮助参与的学生获得广阔的视野和新技术，以开展学术和工业环境。该真菌模型将揭示动物和植物中其他类似生长形式（即神经元、出芽和裂变酵母、上皮细胞、根毛、花粉管）中钙信号通路的共同成分，并揭示其进化适应和功能多样性的潜在独特属性。真菌Ca2+信号的独特方面可能被用于控制疾病。

Intellectual merit. All organisms have sophisticated ways for recognizing and adapting to their environment so they can survive or avoid harsh conditions, exploit new food sources or even cause disease. Several recent advances in methods for sequencing, decoding and modifying DNA and technological improvements in powerful microscopes when used in a unified approach now enable subtle abnormal effects of targeted genetic changes to be monitored inside living cells. This research uses fungi as a model system for determining how cells are able to sense changes in their microscopic environment and recruit highly specialized proteins that bind to calcium ions in a subsequent cascade of events that change the cells behavior. Genetic methods will be used to comprehensively disrupt all known key calcium interacting proteins in a fungal model organism and attach special tags that allow monitoring of calcium changes and/or movement of multiple calcium interacting and related proteins to be tracked inside of living cells. This approach will result in a comprehensive understanding of which targeted proteins allow cells to grow in a rapid and directed fashion. The scientific impacts of this project go beyond better understanding the biology of calcium signaling in the model fungus Fusarium graminearum which causes a serious disease in wheat. It will create a novel toolbox for studying the evolution of calcium signaling, a highly conserved ancient molecular language used by in virtually all organisms for converting external environmental signals into needed cellular changes. The project brings together a group of scientists with complementary backgrounds including; biochemistry, cell biology, cytology, evolutionary biology, fungal biology, informatics, math, and genetics.Broader impacts. The broad and interdisciplinary scope of this project will provide an ideal environment for practicing team-based integrative problem solving with undergraduate, graduate, and postdoctoral students. Faced with the rapidly growing complexity of biological questions, the need for such education continues to increase. A postdoc supported by this project will be mentored to develop her own program in the near future through established mentoring programs. Through hands-on week long workshops and comprehensive publicly available websites, others will have access to all resources and trained to implement the developed tools. The calcium signaling discoveries and microscopy techniques developed here will be introduced in graduate level courses and web-based tools. Undergraduate students (including minority students) will be involved in this project through NSF REU and EPSCoR programs as well as other existing credit and outreach programs at both Penn State and University of Delaware. These efforts will help the participating students gain broad perspectives and new technologies to carry forth into academic and industrial settings. This fungus model will reveal shared components of calcium signaling pathways in other similar growth forms (i.e. neurons, budding and fission yeast, epithelial cells, root hairs, pollen tubes) in animals and plants, and also expose unique attributes potentially relating to their evolutionary adaptation and functional diversity. Unique aspects of fungal Ca2+ signaling may be exploited for controlling disease.