Identification of Arabidopsis thaliana phloem RNAs provides a search criterion for phloem-based transcripts hidden in complex datasets of microarray experiments
Identification of Arabidopsis thaliana phloem RNAs provides a search criterion for phloem-based transcripts hidden in complex datasets of microarray experiments
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DOI：
10.1111/j.1365-313x.2008.03555.x
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发表时间：
2008-09-01
期刊：
影响因子：
7.2
通讯作者：
Hedrich, Rainer
中科院分区：
文献类型：
--
作者：
Deeken, Rosalia;Ache, Peter;Hedrich, Rainer
韧皮部移动信号在植物营养、发育和通讯中起着重要作用。在后一种情况下，韧皮部移动RNA与植物组织之间的信号传导有关。在本研究中，我们着重于模式植物拟南芥茎部韧皮部中的转录本的鉴定。为了分离韧皮部薄壁细胞和伴细胞筛元件复合物中表达的转录物，我们使用激光显微切割结合激光压力催化（LMPC）。筛选分子中的移动的转录物分离自叶韧皮部分泌物。在优化取样和固定后，从两种来源中分离出高质量的RNA。本文所述的RNA扩增程序的修改非常适合于产生用于微阵列实验的足够产量和质量的RNA。与LMPC衍生的韧皮部组织或韧皮部汁液RNA杂交的微阵列允许区分韧皮部表达的和移动的转录物种类。使用这组韧皮部转录本，并将其与来自光，激素和营养处理实验数据库的微阵列进行比较，我们确定了韧皮部衍生的RNA作为移动的，潜在的长距离信号。因此，我们的数据集提供了一个搜索标准的韧皮部为基础的信号隐藏在复杂的数据集的微阵列实验。这些全面的韧皮部转录谱的可用性将有利于反向遗传研究和正向遗传筛选韧皮部和长距离RNA信号突变体。
Phloem-mobile signals play a major role in plant nutrition, development and communication. In the latter context, phloem-mobile RNAs have been associated with signalling between plant tissues. In this study, we focused on the identification of transcripts in the shoot phloem of the model plant Arabidopsis thaliana. To isolate transcripts expressed in phloem parenchyma cells and in companion cell-sieve element complexes, we used laser microdissection coupled to laser pressure catapulting (LMPC). Mobile transcripts in sieve elements were isolated from leaf phloem exudates. After optimization of sampling and fixation, RNA of high quality was isolated from both sources. The modifications to the RNA amplification procedure described here were well suited to production of RNA of sufficient yield and quality for microarray experiments. Microarrays hybridized with LMPC-derived phloem tissue or phloem sap RNA allowed differentiation between phloem-expressed and mobile transcript species. Using this set of phloem transcripts and comparing them with microarrays derived from databases of light, hormone and nutrient treatment experiments, we identified phloem-derived RNAs as mobile, potential long-distance signals. Our dataset thus provides a search criterion for phloem-based signals hidden in the complex datasets of microarray experiments. The availability of these comprehensive phloem transcript profiles will facilitate reverse-genetic studies and forward-genetic screens for phloem and long-distance RNA signalling mutants.