Department of Biology, University of Massachusetts, Amherst, Massachusetts.
Curr Protoc. 2021 Dec;1(12):e309. doi: 10.1002/cpz1.309.
The chytrid fungus Batrachochytrium dendrobatidis (Bd) is a causative agent of chytridiomycosis, a skin disease associated with amphibian population declines around the world. Despite the major impact Bd is having on global ecosystems, much of Bd's basic biology remains unstudied. In addition to revealing mechanisms driving the spread of chytridiomycosis, studying Bd can shed light on the evolution of key fungal traits because chytrid fungi, including Bd, diverged before the radiation of the Dikaryotic fungi (multicellular fungi and yeast). Studying Bd in the laboratory is, therefore, of growing interest to a wide range of scientists, ranging from herpetologists and disease ecologists to molecular, cell, and evolutionary biologists. This protocol describes how to maintain developmentally synchronized liquid cultures of Bd for use in the laboratory, how to grow Bd on solid media, as well as cryopreservation and revival of frozen stocks. © 2021 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Reviving cryopreserved Bd cultures Basic Protocol 2: Establishing synchronized liquid cultures of Bd Basic Protocol 3: Regular maintenance of synchronous Bd in liquid culture Alternate Protocol 1: Regular maintenance of asynchronous Bd in liquid culture Basic Protocol 4: Regular maintenance of synchronous Bd on solid medium Alternate Protocol 2: Starting a culture on solid medium from a liquid culture Basic Protocol 5: Cryopreservation of Bd.
蛙壶菌（Batrachochytrium dendrobatidis，Bd）是一种致病真菌，会引发蛙壶菌病，这种疾病与世界各地的蛙类种群减少有关。尽管 Bd 对全球生态系统造成了重大影响，但它的许多基本生物学特性仍未得到研究。除了揭示导致蛙壶菌病传播的机制外，研究 Bd 还可以阐明关键真菌特征的进化，因为包括 Bd 在内的壶菌在二型真菌（多细胞真菌和酵母）辐射之前就已经分化了。因此，研究 Bd 引起了广泛科学家的兴趣，包括两栖动物学家、疾病生态学家、分子、细胞和进化生物学家。本方案描述了如何维持发育同步的 Bd 液体培养物，以及如何在固体培养基上培养 Bd，以及冷冻储存和复苏冷冻库存。© 2021 作者。Wiley Periodicals LLC 出版的《当代协议》。基本方案 1：复苏冷冻保存的 Bd 培养物基本方案 2：建立 Bd 的同步液体培养物基本方案 3：同步 Bd 在液体培养基中的常规维持备用方案 1：液体中异步 Bd 的常规维持备用方案 2：从液体培养物开始在固体培养基上培养基本方案 4：同步 Bd 在固体培养基上的常规维持备用方案 2：从液体培养物开始在固体培养基上培养基本方案 5：Bd 的冷冻保存。
