Latijnhouwers Maita, Ligterink Wilco, Vleeshouwers Vivianne G A A, van West Pieter, Govers Francine
Laboratory of Phytopathology, Wageningen University, Binnenhaven, The Netherlands.
Mol Microbiol. 2004 Feb;51(4):925-36. doi: 10.1046/j.1365-2958.2003.03893.x.
The heterotrimeric G-protein pathway is a ubiquitous eukaryotic signalling module that is known to regulate growth and differentiation in many plant pathogens. We previously identified Pigpa1, a gene encoding a G-protein alpha subunit from the potato late blight pathogen Phytophthora infestans. P. infestans belongs to the class oomycetes, a group of organisms in which signal transduction processes have not yet been studied at the molecular level. To elucidate the function of Pigpa1, PiGPA1-deficient mutants were obtained by homology-dependent gene silencing. The Pigpa1-silenced mutants produced zoospores that turned six to eight times more frequently, causing them to swim only short distances compared with wild type. Attraction to the surface, a phenomenon known as negative geotaxis, was impaired in the mutant zoospores, as well as autoaggregation and chemotaxis towards glutamic and aspartic acid. Zoospore production was reduced by 20-45% in different Pigpa1-silenced mutants. Transformants expressing constitutively active forms of PiGPA1, containing amino acid substitutions (R177H and Q203L), showed no obvious phenotypic differences from the wild-type strain. Infection efficiencies on potato leaves ranged from 3% to 14% in the Pigpa1-silenced mutants, compared with 77% in wild type, showing that virulence is severely impaired. The results prove that PiGPA1 is crucial for zoospore motility and for pathogenicity in an important oomycete plant pathogen.
异源三聚体G蛋白途径是一种普遍存在的真核信号传导模块,已知可调节许多植物病原体的生长和分化。我们之前鉴定出了Pigpa1,这是一个来自马铃薯晚疫病病原体致病疫霉的编码G蛋白α亚基的基因。致病疫霉属于卵菌纲,这是一类尚未在分子水平上研究信号转导过程的生物。为了阐明Pigpa1的功能,通过同源依赖性基因沉默获得了PiGPA1缺陷型突变体。Pigpa1沉默的突变体产生的游动孢子转向频率增加了6到8倍,导致它们与野生型相比只能短距离游动。突变体游动孢子对表面的吸引力(一种称为负趋地性的现象)、自动聚集以及对谷氨酸和天冬氨酸的趋化性均受损。不同的Pigpa1沉默突变体中,游动孢子的产生减少了20%至45%。表达含有氨基酸替代(R177H和Q203L)的组成型活性形式的PiGPA1的转化体与野生型菌株没有明显的表型差异。Pigpa1沉默突变体在马铃薯叶片上的感染效率为3%至14%,而野生型为77%,表明其毒力严重受损。结果证明,PiGPA1对于一种重要的卵菌植物病原体中的游动孢子运动性和致病性至关重要。