Toh Su San, Treves David S, Barati Michelle T, Perlin Michael H
Department of Biology and Program on Disease Evolution, University of Louisville, Louisville, KY, 40292, USA.
Indiana University Southeast, New Albany, IN, USA.
Arch Microbiol. 2016 Oct;198(8):813-25. doi: 10.1007/s00203-016-1244-2. Epub 2016 May 23.
Microbotryum lychnidis-dioicae is a member of a species complex infecting host plants in the Caryophyllaceae. It is used as a model system in many areas of research, but attempts to make this organism tractable for reverse genetic approaches have not been fruitful. Here, we exploited the recently obtained genome sequence and transcriptome analysis to inform our design of constructs for use in Agrobacterium-mediated transformation techniques currently available for other fungi. Reproducible transformation was demonstrated at the genomic, transcriptional and functional levels. Moreover, these initial proof-of-principle experiments provide evidence that supports the findings from initial global transcriptome analysis regarding expression from the respective promoters under different growth conditions of the fungus. The technique thus provides for the first time the ability to stably introduce transgenes and over-express target M. lychnidis-dioicae genes.
微小麦角菌是一种感染石竹科宿主植物的复合种。它在许多研究领域被用作模型系统，但使这种生物体适用于反向遗传学方法的尝试尚未取得成果。在这里，我们利用最近获得的基因组序列和转录组分析来指导构建体的设计，这些构建体可用于目前适用于其他真菌的农杆菌介导转化技术。在基因组、转录和功能水平上都证明了可重复的转化。此外，这些初步的原理验证实验提供了证据，支持了最初的全局转录组分析关于真菌在不同生长条件下各自启动子表达的发现。该技术首次提供了稳定导入转基因并过表达目标微小麦角菌基因的能力。
