Department of Microbiology and Plant Pathology, University of California, Riverside, CA, 92521, USA.
Mol Microbiol. 2018 Nov;110(4):562-575. doi: 10.1111/mmi.14114. Epub 2018 Oct 22.
MADS-box transcription factors play significant roles in eukaryotes, but have not yet been characterized in oomycetes. Here, we describe a MADS-box protein from Phytophthora infestans, which causes late blight of potato. P. infestans and most other oomycetes express a single MADS-box gene. PiMADS is not transcribed during vegetative growth, but is induced early during asexual sporulation. Its mRNA levels oscillate in response to light, which suppresses sporulation. The protein was not detected in nonsporulating mycelia, but was found in sporulating mycelia and spores. Both mRNA and protein levels decline upon spore germination. A similar expression pattern as well as nuclear localization was observed when the protein was expressed with a fluorescent tag from the native promoter. Gene silencing triggered by a construct expressing 478 nt of MADS sequences indicated that PiMADS is required for sporulation but not hyphal growth or plant colonization. A comparison of wild type to a silenced strain by RNA-seq indicated that PiMADS regulates about 3000 sporulation-associated genes, and acts before other genes previously shown to regulate sporulation. Analysis of the silenced strain also indicated that the native gene was not transcribed while the transgene was still expressed, which contradicts current models for homology-dependent silencing in oomycetes.
MADS 框转录因子在真核生物中发挥着重要作用,但在卵菌中尚未得到表征。在这里,我们描述了一种来自致病疫霉的 MADS 框蛋白,它会导致马铃薯晚疫病。致病疫霉和大多数其他卵菌只表达一个 MADS 框基因。PiMADS 在营养生长期间不转录,但在无性孢子形成的早期被诱导。其 mRNA 水平对光有波动响应,从而抑制孢子形成。该蛋白在不进行孢子形成的菌丝体中未被检测到,但在进行孢子形成的菌丝体和孢子中被发现。mRNA 和蛋白质水平在孢子萌发时下降。当使用来自天然启动子的荧光标记物表达该蛋白时,观察到类似的表达模式和核定位。由表达 478 个核苷酸的 MADS 序列的构建物触发的基因沉默表明,PiMADS 是孢子形成所必需的,但不是菌丝生长或植物定殖所必需的。野生型与沉默菌株的 RNA-seq 比较表明,PiMADS 调节大约 3000 个与孢子形成相关的基因,并且在以前显示调节孢子形成的其他基因之前起作用。对沉默菌株的分析还表明,当转基因仍在表达时,天然基因未被转录,这与卵菌中同源依赖性沉默的现有模型相矛盾。