Fluorescent markers for the Spitzenkörper and exocytosis in Zymoseptoria tritici.
Fluorescent markers for the Spitzenkörper and exocytosis in Zymoseptoria tritici.
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DOI：
10.1016/j.fgb.2015.04.014
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发表时间：
2015-06
期刊：
影响因子：
--
通讯作者：
Steinberg G
中科院分区：
文献类型：
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作者：
Guo M;Kilaru S;Schuster M;Latz M;Steinberg G
我们建立Z。三个极性标记ZtSec 4、ZtMlc 1、ZtRab 11、ZtExo 70和ZtSpa 2。所有的标记都正确定位，标记Spitzenkörper和极性胞吐的位点。我们提供了5个抗羧菌素的传递载体，用于将所有标记整合到sdi 1位点。我们提供了5个潮霉素B抗性的转运载体，用于随机整合所有标记。真菌菌丝是高度极化的细胞，通过尖端生长侵入其基质。在植物病原真菌中，菌丝的生长对于宿主的入侵是必不可少的。这使得极性因子和分泌调节剂成为新型杀真菌剂的潜在新靶标。极化需要将分泌囊泡递送到顶端Spitzenkörper，然后在扩展的细胞尖端进行极化胞吐。在这里，我们引入荧光标记可视化顶端Spitzenkörper和顶端网站的小麦病原菌Zymoseptoria的菌丝胞吐。我们将绿色荧光蛋白融合到小GTdR ZtSec 4、肌球蛋白轻链ZtMlc 1和小GTdR ZtRab 11上，并将融合蛋白与染料FM 4 -64共定位于菌丝顶端，表明标记物标记了Z.三个月。此外，我们本地化GFP融合的外囊蛋白ZtExo 70，极性体蛋白ZtSpa 2。与子囊菌粗糙脉孢菌中的结果一致，这些标记物确实定位在菌丝顶端的质膜附近，并且仅部分与FM 4 -64共定位。因此，这些荧光标记物是有用的分子工具，允许Z中突变体的表型分析。三个月。这些工具将有助于开发新的研究途径，以控制小麦中的STB感染。
We establish Z. tritici polarity markers ZtSec4, ZtMlc1, ZtRab11, ZtExo70 and ZtSpa2. All markers localize correctly, labeling the Spitzenkörper and sites of polar exocytosis. We provide 5 carboxin-resistance conveying vectors for integration of all markers into the sdi1 locus. We provide 5 hygromycin B-resistance conveying vectors for random integration of all markers. Fungal hyphae are highly polarized cells that invade their substrate by tip growth. In plant pathogenic fungi, hyphal growth is essential for host invasion. This makes polarity factors and secretion regulators potential new targets for novel fungicides. Polarization requires delivery of secretory vesicles to the apical Spitzenkörper, followed by polarized exocytosis at the expanding cell tip. Here, we introduce fluorescent markers to visualize the apical Spitzenkörper and the apical site of exocytosis in hyphae of the wheat pathogen Zymoseptoria tritici. We fused green fluorescent protein to the small GTPase ZtSec4, the myosin light chain ZtMlc1 and the small GTPase ZtRab11 and co-localize the fusion proteins with the dye FM4-64 in the hyphal apex, suggesting that the markers label the hyphal Spitzenkörper in Z. tritici. In addition, we localize GFP-fusions to the exocyst protein ZtExo70, the polarisome protein ZtSpa2. Consistent with results in the ascomycete Neurospora crassa, these markers did localize near the plasma membrane at the hyphal tip and only partially co-localize with FM4-64. Thus, these fluorescent markers are useful molecular tools that allow phenotypic analysis of mutants in Z. tritici. These tools will help develop new avenues of research in our quest to control STB infection in wheat.