A DNA-based real-time PCR assay for robust growth quantification of the bacterial pathogen Pseudomonas syringae on Arabidopsis thaliana.
A DNA-based real-time PCR assay for robust growth quantification of the bacterial pathogen Pseudomonas syringae on Arabidopsis thaliana.
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DOI：
10.1186/s13007-016-0149-z
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发表时间：
2016
期刊：
影响因子：
5.1
通讯作者：
Somssich IE
中科院分区：
文献类型：
--
作者：
Ross A;Somssich IE
假单胞菌与拟南芥的相互作用是研究各种细菌-宿主相互关系最常用的系统之一。目前，大多数研究是基于病原体的生长定量来表征抗性或毒力靶标。然而，用于确定植物中细菌增殖的标准可用方法是费力的并且具有若干限制。在这里，我们提出了一种替代的强大的方法，这是基于细菌DNA的定量实时PCR。我们直接将该测定法与常规使用的平板计数法进行比较，以获得许多充分描述的拟南芥突变体中的细菌滴度。这些研究表明，基于DNA的技术是高度可靠和可比的。此外，该技术易于应用，稳健，是常规实验或更大规模分析的理想选择。本文的在线版本（doi：10.1186/s13007-016-0149-z）包含补充材料，可供授权用户使用。
The interaction of Pseudomonas syringae with Arabidopsis is one of the most commonly used systems to study various bacterial—host interrelationships. Currently, most studies are based on the growth quantification of the pathogen to characterize resistance or virulence targets. However, the standard available method for determining bacterial proliferation in planta is laborious and has several limitations. Here we present an alternative robust approach, which is based on the quantification of bacterial DNA by real-time PCR. We directly compared this assay with the routinely used plate counting method to access bacterial titers in a number of well described Arabidopsis mutants. These studies showed that the DNA-based technique is highly reliable and comparable. Moreover, the technique is easily applicable, robust, and ideal for routine experiments or for larger scale analyses. The online version of this article (doi:10.1186/s13007-016-0149-z) contains supplementary material, which is available to authorized users.
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影响因子：
11.6
作者：
WHALEN, MC;INNES, RW;STASKAWICZ, BJ
通讯作者：
STASKAWICZ, BJ
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