Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Rozvojová 263, 16502 Praha 6, Czech Republic.
J Exp Bot. 2011 Mar;62(6):2107-16. doi: 10.1093/jxb/erq402. Epub 2011 Jan 3.
Recently, the octameric vesicle-tethering complex exocyst was found in plants and its importance for Arabidopsis morphogenesis was demonstrated. Exo70 exocyst subunits in plants, unlike in yeasts and mammals, are represented by a multigene family, comprising 23 members in Arabidopsis. For Exo70B2 and Exo70H1 paralogues, transcriptional up-regulation was confirmed on treatment with an elicitor peptide, elf18, derived from the bacterial elongation factor. Their ability to participate in the exocyst complex formation was inferred by the interaction of both the Exo70s with several other exocyst subunits using the yeast two-hybrid system. Arabidopsis plants mutated in these two genes were used to analyse their local reaction upon inoculation with Pseudomonas syringae pv. maculicola and the fungal pathogen Blumeria graminis f. sp. hordei. The Pseudomonas sensitivity test revealed enhanced susceptibility for the two exo70B2 and one H1 mutant lines. After Blumeria inoculation, an increase in the proportion of abnormal papilla formation, with an unusual wide halo made of vesicle-like structures, was found in exo70B2 mutants. Intracellular localization of both Exo70 proteins was studied following a GFP fusion assay and Agrobacterium-mediated transient expression of the constructs in Nicotiana benthamiana leaf epidermis. GFP-Exo70H1 localizes in the vesicle-like structures, while GFP-Exo70B2 is localized mainly in the cytoplasm. It is concluded that both Exo70B2 and Exo70H1 are involved in the response to pathogens, with Exo70B2 having a more important role in cell wall apposition formation related to plant defence.
最近,在植物中发现了八聚体囊泡拴系复合物外泌体(exocyst),并证明其对拟南芥形态发生的重要性。植物中的 Exo70 外泌体亚基与酵母和哺乳动物不同,由一个多基因家族代表,在拟南芥中包含 23 个成员。对于 Exo70B2 和 Exo70H1 同源物,在用细菌延伸因子衍生的激发肽 elf18 处理后,证实了其转录水平的上调。通过酵母双杂交系统,这两种 Exo70 与其他几种外泌体亚基的相互作用,推断它们能够参与外泌体复合物的形成。使用这两个基因发生突变的拟南芥植物来分析它们在接种丁香假单胞菌 pv. maculicola 和真菌病原体禾生球腔菌时的局部反应。对拟南芥的敏感性测试显示,两种 exo70B2 和一种 H1 突变系的敏感性增强。在接种禾生球腔菌后,在 exo70B2 突变体中发现异常的乳突形成比例增加,形成了异常宽的晕圈,由囊泡样结构组成。通过 GFP 融合测定和用农杆菌介导的 GFP 融合构建体在本氏烟叶片表皮中的瞬时表达,研究了两种 Exo70 蛋白的细胞内定位。GFP-Exo70H1 定位于囊泡样结构中,而 GFP-Exo70B2 主要定位于细胞质中。结论是,Exo70B2 和 Exo70H1 都参与了对病原体的反应,Exo70B2 在与植物防御相关的细胞壁附着形成中起着更重要的作用。