Microtubule-organization during fungal invasion into barley epidermal cells

植物寄生真菌在克服植物先天免疫方面发挥着多种策略。在生物营养真菌的情况下，寄生虫还操纵宿主细胞功能，以在完整的活宿主细胞中建立和维持感染结构。这涉及对植物机制的敌意控制，以控制亚细胞结构和功能。我们发现宿主小RAC/ROP g蛋白（大麦RACB）和微管是真菌成功或失败建立寄生的关键问题。在这个项目中，我们旨在填补草RAC/ROP蛋白与植物-真菌相互作用中微管组织之间的知识空白。为此，我们将利用最近公布的大麦基因组序列信息。我们将利用这些信息来鉴定组织微管细胞骨架和控制生物营养性子囊菌真菌Blumeria graminis f.sp渗透成功的RAC/ROP执行蛋白。hordei。候选蛋白（RIPs和RICs）因其与RAC/ROPs和/或微管相互作用的蛋白结构域保守而被确定。我们将在酵母和植物中展示RICs或rip与RACB的相互作用。本研究采用瞬时诱导基因沉默和过表达的方法，对大麦表皮细胞进行了蛋白功能的研究。hordei。荧光标记版本的RICs和rip被短暂表达，以研究依赖于真菌攻击和/或依赖于上游RAC/ROPs的异位激活的亚细胞定位。活细胞成像将显示受RAC/ROPs以及RICs或rip的功能和丰度影响的MT组织。这将证明大麦的RICs和rip在RAC/ROPs下游和真菌渗透过程中影响MT阵列的潜力。

Plant parasitic fungi exert diverse strategies for overcoming plant innate immunity. In the case of biotrophic fungi, the parasite additionally manipulates host cellular function for establishment and maintenance of infection structures in intact living host cells. This involves hostile control of plant mechanisms to control subcellular architecture and function. We identified host small RAC/ROP G-proteins (barley RACB) and microtubules to be key to the question of fungal success or failure in establishing parasitism. In this project, we aim at filling the knowledge gap between grass RAC/ROP proteins and microtubule organization in plant-fungus interaction. To this end, we are going to make use of the recently published sequence information of the barley genome. We will use this information for identification of RAC/ROP executor proteins which organize the microtubule cytoskeleton and control penetration success of the biotrophic ascomycete fungus Blumeria graminis f.sp. hordei. Candidate proteins (RIPs and RICs) were identified because of their protein domains conserved for interaction with RAC/ROPs and/or with microtubules. We are going to demonstrate interaction of RICs or RIPs with RACB in yeast and in planta. We study protein function by transient induced gene silencing and over-expression in barley epidermal cells followed by challenge inoculation with conidia of Blumeria graminis f.sp. hordei. Fluorescent-tagged versions of RICs and RIPs are transiently expressed to study subcellular localization dependent on fungal attack and/or depending on ectopic activation of upstream RAC/ROPs. Live cell imaging will illustrate MT organization influenced by function and abundance of RAC/ROPs as well as RICs or RIPs. This will demonstrate the potential of barley RICs and RIPs to influence MT arrays downstream of RAC/ROPs and during fungal penetration.