Spatial dissection of the Arabidopsis thaliana transcriptional response to downy mildew using Fluorescence Activated Cell Sorting.
Spatial dissection of the Arabidopsis thaliana transcriptional response to downy mildew using Fluorescence Activated Cell Sorting.
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DOI：
10.3389/fpls.2015.00527
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发表时间：
2015
影响因子：
5.6
通讯作者：
Gifford ML
中科院分区：
文献类型：
--
作者：
Coker TL;Cevik V;Beynon JL;Gifford ML
基因表达的变化是植物对感染反应的重要组成部分。在过去的十年中，全叶表达谱在识别基因和过程中发挥了宝贵的作用，有助于模式植物拟南芥和各种病原体之间的相互作用。然而，对于一些病原体，例如由活体营养型卵菌病原体阿拉伯透明霜霉（Hyaloperonospora arabidopsidis，Hpa）引起的霜霉病，全叶分析可能无法捕获完整的拟南芥属响应，包括叶内未感染细胞以及感染细胞的响应。在感染细胞中发生的高度定位的表达变化可以被未感染细胞的相对丰度稀释。此外，不同性质的局部和全身Hpa反应可能会合并。为了解决这个问题，我们应用荧光激活细胞分选（FACS），通常用于分析植物非生物反应的技术，植物病原体相互作用的研究。我们分离historiated（Hpa近端）和非historiated（Hpa远端）细胞感染的幼苗样品，使用流式细胞仪，并测量全球基因表达。当与未感染的对照相比，278转录被确定为显着差异表达，其中绝大多数是差异表达特异性在Hpa近端细胞。通过将我们的数据与以前的整个器官研究进行比较，我们发现了许多高度局部调节的基因，这些基因可能与Hpa反应有关，并且首次使用我们灵敏的FACS技术发现。
Changes in gene expression form a crucial part of the plant response to infection. In the last decade, whole-leaf expression profiling has played a valuable role in identifying genes and processes that contribute to the interactions between the model plant Arabidopsis thaliana and a diverse range of pathogens. However, with some pathogens such as downy mildew caused by the biotrophic oomycete pathogen Hyaloperonospora arabidopsidis (Hpa), whole-leaf profiling may fail to capture the complete Arabidopsis response encompassing responses of non-infected as well as infected cells within the leaf. Highly localized expression changes that occur in infected cells may be diluted by the comparative abundance of non-infected cells. Furthermore, local and systemic Hpa responses of a differing nature may become conflated. To address this we applied the technique of Fluorescence Activated Cell Sorting (FACS), typically used for analyzing plant abiotic responses, to the study of plant-pathogen interactions. We isolated haustoriated (Hpa-proximal) and non-haustoriated (Hpa-distal) cells from infected seedling samples using FACS, and measured global gene expression. When compared with an uninfected control, 278 transcripts were identified as significantly differentially expressed, the vast majority of which were differentially expressed specifically in Hpa-proximal cells. By comparing our data to previous, whole organ studies, we discovered many highly locally regulated genes that can be implicated as novel in the Hpa response, and that were uncovered for the first time using our sensitive FACS technique.
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