Hewezi Tarek, Howe Peter, Maier Tom R, Hussey Richard S, Mitchum Melissa Goellner, Davis Eric L, Baum Thomas J
Department of Plant Pathology, Iowa State University, Ames, Iowa 50011, USA.
Plant Cell. 2008 Nov;20(11):3080-93. doi: 10.1105/tpc.108.063065. Epub 2008 Nov 11.
Plant-parasitic cyst nematodes secrete a complex of cell wall-digesting enzymes, which aid in root penetration and migration. The soybean cyst nematode Heterodera glycines also produces a cellulose binding protein (Hg CBP) secretory protein. To determine the function of CBP, an orthologous cDNA clone (Hs CBP) was isolated from the sugar beet cyst nematode Heterodera schachtii, which is able to infect Arabidopsis thaliana. CBP is expressed only in the early phases of feeding cell formation and not during the migratory phase. Transgenic Arabidopsis expressing Hs CBP developed longer roots and exhibited enhanced susceptibility to H. schachtii. A yeast two-hybrid screen identified Arabidopsis pectin methylesterase protein 3 (PME3) as strongly and specifically interacting with Hs CBP. Transgenic plants overexpressing PME3 also produced longer roots and exhibited increased susceptibility to H. schachtii, while a pme3 knockout mutant showed opposite phenotypes. Moreover, CBP overexpression increases PME3 activity in planta. Localization studies support the mode of action of PME3 as a cell wall-modifying enzyme. Expression of CBP in the pme3 knockout mutant revealed that PME3 is required but not the sole mechanism for CBP overexpression phenotype. These data indicate that CBP directly interacts with PME3 thereby activating and potentially targeting this enzyme to aid cyst nematode parasitism.
植物寄生性胞囊线虫分泌一系列细胞壁消化酶,有助于其穿透根部并在根内迁移。大豆胞囊线虫大豆异皮线虫还产生一种纤维素结合蛋白(Hg CBP)分泌蛋白。为了确定CBP的功能,从能够感染拟南芥的甜菜胞囊线虫甜菜异皮线虫中分离出一个直系同源cDNA克隆(Hs CBP)。CBP仅在取食细胞形成的早期阶段表达,而在迁移阶段不表达。表达Hs CBP的转基因拟南芥根系更长,对甜菜异皮线虫的易感性增强。酵母双杂交筛选鉴定出拟南芥果胶甲酯酶蛋白3(PME3)与Hs CBP强烈且特异性相互作用。过表达PME3的转基因植物根系也更长,对甜菜异皮线虫的易感性增加,而pme3基因敲除突变体则表现出相反的表型。此外,CBP过表达增加了植物体内PME3的活性。定位研究支持PME3作为细胞壁修饰酶的作用模式。在pme3基因敲除突变体中表达CBP表明,PME3是必需的,但不是CBP过表达表型的唯一机制。这些数据表明,CBP直接与PME3相互作用,从而激活并可能将该酶靶向以帮助胞囊线虫寄生。