Microinjection of Urediospore Germlings of Uromyces appendiculatus

Ary Corrêa Jr., Harvey C. Hoch
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引用次数: 21

Abstract

Corrêa, A., Jr., and Hoch, H. C. 1993. Microinjection of urediospore germlings of Uromyces appendiculatus. Experimental Mycology 17, 253-273. Urediospore germlings of Uromyces appendiculatus were microinjected with a variety of materials to assess the applicability of microinjection methodologies for studying fungal cell growth and development. Until now, hyphal cells have not been successfully microinjected without first being pretreated with osmotica to lower the turgor pressures of the cells. We describe the development of a hydraulicly actuated system and protocols for successfully microinjecting Uromyces cells on a routine basis. Synthetic fluorocarbon and silicone fluids, various fluorophore-conjugated dextrans, rhodamine-conjugated phalloidin, rhodamine 123, cyclic AMP, and cytochalasin E were all microinjected into living urediospore germlings that grew or responded in ways consistent with expected pharmacological effects of the injected material. Germlings that were microinjected with synthetic fluids maintained the ability to sense and respond to inductive topographies for the formation of appressoria. Materials were injected at volumes of ≤0.5 to ≤400 fl. The results of this investigation indicate that fungal cells can be successfully microinjected with a variety of materials and, importantly, continue to function as normal cells.

尾尾尾尾菌脲孢子胚的显微注射
Corrêa, A., Jr.和Hoch, H. C. 1993。尾尾尾尾尾菌无孢子胚的显微注射。真菌学通报,17(3):593 - 593。用不同的材料对尾尾尿霉菌的尿孢子进行显微注射,以评估显微注射方法在研究真菌细胞生长发育中的适用性。到目前为止,如果不先用渗透剂预处理以降低细胞的膨胀压力,菌丝细胞还没有成功地进行微注射。我们描述了一个液压驱动系统的发展和方案成功微注射尿霉菌细胞的常规基础。合成氟碳和硅酮液体、各种荧光团共轭右旋糖聚糖、罗丹明共轭phalloidin、罗丹明123、环AMP和细胞chalasin E都被微量注射到活的脲孢子胚芽中,这些胚芽的生长或反应方式与所注射物质的预期药理作用一致。微注射合成液体的胚芽保持了对附着胞形成的感应地形的感知和响应能力。材料以≤0.5至≤400 fl的体积注射。本研究结果表明,真菌细胞可以成功地微注射各种材料,并且重要的是,继续像正常细胞一样发挥功能。
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