College of Horticulture, Northwest A&F University, Yangling, Shaanxi, 712100, People's Republic of China.
Protoplasma. 2011 Apr;248(2):415-23. doi: 10.1007/s00709-010-0162-4. Epub 2010 May 29.
Vitis pseudoreticulata glyoxal oxidase (VpGLOX) was previously isolated from the Chinese wild vine V. pseudoreticulata accession "Baihe-35-1" during a screen for genes that are upregulated in response to infection with grapevine powdery mildew (Erysiphe necator, PM). In the present study, a possible function of VpGLOX for defense against PM was investigated using Agrobacterium-mediated transient expression. After optimizing agro-infiltration, VpGLOX was transiently overexpressed in leaves of either PM-susceptible (accession "6-12-2") or PM-resistant (accession "6-12-6") plants. The efficiency of transfection was verified using a β-glucuronidase (GUS) reporter and was found to comprise most leaf areas regardless of the initial leaf position. Upon infection with E. necator, clear differences were observed with respect to hyphal development between agro-infiltrated leaves and control groups of both, the susceptible and the resistant, genotypes. The expression of VpGLOX was followed by real-time polymerase chain reaction in both genotypes. Whereas in the susceptible host ("6-12-2") expression was found to increase only in transfected leaves and remained transient, in the resistant host ("6-12-6"), a second peak appeared later in transfected leaves, probably representing the response of the endogenous VpGLOX. The data support the interpretation that VpGLOX is sufficient to confer resistance to E. necator.
葡萄卷叶相关氧化酶(VpGLOX)是从中国野生葡萄品种‘白河-35-1’中分离出来的,在筛选白粉病(葡萄球腔菌,PM)感染后上调表达的基因时发现。本研究通过农杆菌介导的瞬时表达来研究 VpGLOX 防御 PM 的可能功能。在优化了农杆菌浸润条件后,VpGLOX 在 PM 敏感品种( accession "6-12-2")和 PM 抗性品种( accession "6-12-6")叶片中瞬时过表达。使用β-葡萄糖醛酸酶(GUS)报告基因验证了转染效率,发现无论初始叶片位置如何,转染效率都涵盖了大部分叶片区域。在 E. necator 感染后,观察到农杆菌浸润叶片与敏感和抗性基因型对照组叶片之间的菌丝发育存在明显差异。在这两种基因型中,通过实时聚合酶链反应(PCR)跟踪 VpGLOX 的表达。在易感宿主(“6-12-2”)中,仅在转染叶片中发现表达增加且呈瞬时表达,而在抗性宿主(“6-12-6”)中,转染叶片中出现了第二个峰值,可能代表内源性 VpGLOX 的反应。数据支持以下结论:VpGLOX 足以赋予宿主对白粉病的抗性。