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缺少动力蛋白轻链 1 的烟草疫霉转化体产生无鞭毛游动孢子。

Phytophthora nicotianae transformants lacking dynein light chain 1 produce non-flagellate zoospores.

机构信息

Plant Science Division, Research School of Biology, The Australian National University, Canberra, ACT 2601, Australia.

出版信息

Fungal Genet Biol. 2010 Aug;47(8):663-71. doi: 10.1016/j.fgb.2010.04.008. Epub 2010 May 6.

Abstract

Biflagellate zoospores of the highly destructive plant pathogens in the genus Phytophthora are responsible for the initiation of infection of host plants. Zoospore motility is a critical component of the infection process because it allows zoospores to actively target suitable infection sites on potential hosts. Flagellar assembly and function in eukaryotes depends on a number of dynein-based molecular motors that facilitate retrograde intraflagellar transport and sliding of adjacent microtubule doublets in the flagellar axonemes. Dynein light chain 1 (DLC1) is one of a number of proteins in the dynein outer arm multiprotein complex. It is a 22 kDa leucine-rich repeat protein that binds to the catalytic motor domain of the dynein gamma heavy chain. We report the cloning and characterization of DLC1 homologues in Phytophthora cinnamomi and Phytophthora nicotianae (PcDLC1 and PnDLC1). PcDLC1 and PnDLC1 are single copy genes that are more highly expressed in sporulating hyphae than in vegetative hyphae, zoospores or germinated cysts. Polyclonal antibodies raised against PnDLC1 locallized PnDLC1 along the length of the flagella of P. nicotianae zoospores. RNAi-mediated silencing of PnDLC1 expression yielded transformants that released non-flagellate, non-motile zoospores from their sporangia. Our observations indicate that zoospore motility is not required for zoospore release from P. nicotianae sporangia or for breakage of the evanescent vesicle into which zoospores are initially discharged.

摘要

双鞭毛游动孢子是植物病原菌疫霉属中高度破坏性的病原体,负责引发宿主植物的感染。游动孢子的运动能力是感染过程中的一个关键组成部分,因为它使游动孢子能够主动寻找潜在宿主上合适的感染部位。真核生物的鞭毛组装和功能依赖于许多基于动力蛋白的分子马达,这些马达促进了鞭毛轴丝内的逆行鞭毛内运输和相邻微管二联体的滑动。动力蛋白轻链 1(DLC1)是动力蛋白外臂多蛋白复合物中的许多蛋白质之一。它是一种 22 kDa 的亮氨酸丰富重复蛋白,与动力蛋白γ重链的催化马达结构域结合。我们报道了疫霉属和烟草疫霉属中 DLC1 同源物的克隆和特性(PcDLC1 和 PnDLC1)。PcDLC1 和 PnDLC1 是单拷贝基因,在孢子形成菌丝中比在营养菌丝、游动孢子或萌发的胞囊中的表达水平更高。针对 PnDLC1 制备的多克隆抗体将 PnDLC1 定位在烟草疫霉游动孢子的鞭毛上。针对 PnDLC1 的 RNAi 介导的沉默导致转化体从其孢子囊中释放出无鞭毛、无动力的游动孢子。我们的观察表明,游动孢子的运动能力对于烟草疫霉孢子囊中释放游动孢子或使最初排出的暂态囊泡破裂并不是必需的。

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