An efficient method for visualization and growth of fluorescent Xanthomonas oryzae pv. oryzae in planta
An efficient method for visualization and growth of fluorescent Xanthomonas oryzae pv. oryzae in planta
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DOI：
10.1186/1471-2180-8-164
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发表时间：
2008-09-30
期刊：
影响因子：
4.2
通讯作者：
Ronald, Pamela C.
中科院分区：
文献类型：
--
作者：
Han, Sang-Wook;Park, Chang-Jin;Ronald, Pamela C.
背景：米黄单胞菌 pv。米曲霉是白叶枯病的病原体，是水稻的严重病原体。在这里，我们描述了一种荧光标记系统，用于研究米 X. oryzae pv 的毒力和致病性。结果：荧光 X. oryzae pv。表达绿色荧光蛋白 (GFP) 的米曲霉菲律宾小种 6 菌株 (PXO99(GFP)) 是使用载体 pPneo-gfp 中新霉素启动子控制下的 gfp 基因生成的。 PXO99(GFP)菌株表现出与野生型对照菌株PXO99相同的毒力和无毒力特性。使用荧光显微镜，直接观察水稻木质部导管中的细菌增殖和定植。使用荧光测定法和酶联免疫吸附测定 (ELISA) 评估细菌生长的准确和快速测定。结论：我们的结果表明荧光标记系统可用于评估细菌感染和监测植物中的细菌增殖。
Background: Xanthomonas oryzae pv. oryzae, the causal agent of bacterial blight disease, is a serious pathogen of rice. Here we describe a fluorescent marker system to study virulence and pathogenicity of X. oryzae pv. oryzae.Results: A fluorescent X. oryzae pv. oryzae Philippine race 6 strain expressing green fluorescent protein (GFP) (PXO99(GFP)) was generated using the gfp gene under the control of the neomycin promoter in the vector, pPneo-gfp. The PXO99(GFP)strain displayed identical virulence and avirulence properties as the wild type control strain, PXO99. Using fluorescent microscopy, bacterial multiplication and colonization were directly observed in rice xylem vessels. Accurate and rapid determination of bacterial growth was assessed using fluoremetry and an Enzyme- Linked ImmunoSorbant Assay (ELISA).Conclusion: Our results indicate that the fluorescent marker system is useful for assessing bacterial infection and monitoring bacterial multiplication in planta.