Genetic and Molecular Dissection of Hyphal Anastomosis

丝状真菌通过尖端延伸、分枝和菌丝融合生长，形成菌丝网络，构成真菌个体。一些真菌菌落是微小的，而另一些则可以达到几英亩的大小。尽管对丝状真菌的尖端生长和分支进行了广泛的研究，但对菌丝融合的机制或菌丝网络形成的功能知之甚少。以往利用活细胞成像和共聚焦显微镜对丝状真菌粗神经孢子菌菌丝融合过程的研究表明，这是一个复杂而精心调控的生物过程，对真菌个体具有明显的影响。菌丝吻合是增加菌丝胞浆流动和相互联系的一种方式，这可能在菌丝内交流中起重要作用，影响菌丝模式的形成和发育过程。丝状真菌中的菌丝融合与其他生物中的细胞融合事件类似，如卵子和精子之间的受精事件，以及导致合胞体的体细胞融合事件，如肌肉分化过程中的成肌细胞融合和骨形成过程中的破骨细胞融合。丝状真菌菌丝吻合突变体的鉴定将提供信息和工具，通过遗传和分子表征信号方面和融合过程的机制，除了阐明真菌个体无法形成菌丝网络的表型后果。由于粗神经孢子虫具有活细胞成像技术的可追踪性，全基因组序列信息，遗传和分子技术的易用性以及神经孢子虫群落的积极相互作用，因此将对粗神经孢子虫进行研究。目的是通过结合活细胞成像，遗传分析和生化工具来剖析菌丝融合的过程，阐明自信号，极化和融合的机制。更广泛的影响：不同层次的学生将参与这个项目，他们将学习遗传学、细胞成像、基因组学和分子生物学等各种技术。来自弱势群体的学生将通过暑期实习项目获得支持。

Filamentous fungi grow by tip extension, branching and hyphal fusion to form a hyphal network that makes up a fungal individual. Some fungal colonies are microscopic, while others can be acres in size. Although tip growth and branching have been extensively studied in filamentous fungi, little is known about the mechanism of hyphal fusion or the function of the formation of a hyphal network. Previous studies on the hyphal fusion process by live cell imaging and confocal microscopy in the filamentous fungus, Neurospora crassa, have revealed a complex and carefully regulated biological process with obvious consequences to the fungal individual. Hyphal anastomosis is a way to increase cytoplasmic flow and interconnectedness of hypha, which may be important in intra-hyphal communication that influences hyphal pattern formation and developmental processes. Hyphal fusion in filamentous fungi is comparable to cell fusion events in other organisms, such as fertilization events between egg and sperm and somatic cell fusion events that result in syncytia, such as myoblast fusion during muscle differentiation and between osteoclasts during bone formation. The identification of hyphal anastomosis mutants in filamentous fungi will provide information and tools by which to genetically and molecularly characterize the signaling aspects and the machinery of the fusion process, in addition to illuminating the phenotypic consequences to the fungal individual of the inability to form a hyphal network. The studies will be performed on Neurospora crassa because of its tractability for live cell imaging techniques, full genome sequence information, the ease of genetic and molecular techniques and the positive interactions of the Neurospora community. The goal is to dissect the process of hyphal fusion by using a combination of live cell imaging, genetic analysis and biochemical tools to elucidate the mechanisms of self-signaling, polarization and fusion. Broader Impacts: Students at various levels will be involved in this project and they will learn a variety of techniques in genetics, cell imaging, genomics and molecular biology. Students from underrepresented groups will be supported through a summer internship program.