Two species of parasitic fungi from the phylum Chytridiomycota (chytrids) are annihilating global amphibian populations. These chytrid species-Batrachochytrium dendrobatidis and B. salamandrivorans-have high rates of mortality and transmission. Upon establishing infection in amphibians, chytrids rapidly multiply within the skin and disrupt their hosts' vital homeostasis mechanisms. Current disease models suggest that chytrid fungi locate and infect their hosts during a motile, unicellular 'zoospore' life stage. Moreover, other chytrid species parasitize organisms from across the tree of life, making future epidemics in new hosts a likely possibility. Efforts to mitigate the damage and spread of chytrid disease have been stymied by the lack of knowledge about basic chytrid biology and tools with which to test molecular hypotheses about disease mechanisms. To overcome this bottleneck, we have developed high-efficiency delivery of molecular payloads into chytrid zoospores using electroporation. Our electroporation protocols result in payload delivery to between 75-97% of living cells of three species: B. dendrobatidis, B. salamandrivorans, and a non-pathogenic relative, S. punctatus. This method lays the foundation for molecular genetic tools needed to establish ecological mitigation strategies and answer broader questions in evolutionary and cell biology.

中文翻译：

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高效真菌电穿孔

斜纹夜蛾门中的两种寄生真菌正在消灭全球的两栖动物种群。这些chytrid物种-Batrachochytrium dendrobatidis和B. salamandrivorans-死亡率和传播率很高。在两栖动物中建立感染后，糜蛋白酶在皮肤内迅速繁殖并破坏宿主的重要稳态机制。当前的疾病模型表明，壶菌属真菌在运动的单细胞“孢子”生命阶段定位并感染它们的宿主。此外，其他壶菌属物种寄生了整个生命树中的生物，使得新宿主中未来的流行病成为可能。由于缺乏关于基本壶菌生物学知识和用于检验关于疾病机制的分子假设的工具的知识，减轻了壶菌病的破坏和传播的努力已陷入困境。为了克服这一瓶颈，我们开发了使用电穿孔技术将分子有效载荷高效传递到糜菌游动孢子中的方法。B. dendrobatidis，B. salamandrivorans，和非病原性相对，S.毛虫。该方法为建立生态减缓策略并回答进化和细胞生物学中更广泛的问题所需的分子遗传工具奠定了基础。