Doering-Saad C, Newbury H J, Couldridge C E, Bale J S, Pritchard J
School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
J Exp Bot. 2006;57(12):3183-93. doi: 10.1093/jxb/erl082. Epub 2006 Aug 25.
The aim of this study was to identify genes that are expressed in the phloem. Increased knowledge of phloem regulation will contribute to our understanding of its many roles, from transport of solutes to information about interactions with pathogens. A cDNA library constructed from phloem-enriched sap exuding from cut Ricinus communis (L.) hypocotyls was sequenced. To assess contamination from other tissues, two libraries were constructed: one using the first 15 min of exudation and the other from sap collected after 120 min of exudation had elapsed. Of 1012 clones sequenced, 158 unique transcripts were identified. The presence of marker molecules such as profilin, the low occurrence of chloroplast-related mRNAs, and the sieve element localization of constituent mRNA using in situ hybridization were consistent with a phloem origin of the sap. Functional analysis of the cDNAs revealed classifications including ribosomal function, interaction with the environment, transport, DNA/RNA binding, and protein turnover. An analysis of the closest Arabidopsis thaliana (L.) homologue for each clone indicated that genes involved in cell localization, protein synthesis, tissue localization, organ localization, organ differentiation, and cell fate were represented at twice the level occurring in the whole Arabidopsis genome. The transcripts found in this phloem-enriched library are discussed in the context of phloem function and the relationship between the companion cell and sieve element.
本研究的目的是鉴定在韧皮部表达的基因。对韧皮部调控的深入了解将有助于我们理解其众多作用,从溶质运输到与病原体相互作用的信息传递。对从蓖麻(Ricinus communis (L.))下胚轴切口处渗出的富含韧皮部的汁液构建的cDNA文库进行了测序。为了评估来自其他组织的污染,构建了两个文库:一个使用渗出的前15分钟的汁液,另一个使用渗出120分钟后收集的汁液。在测序的1012个克隆中,鉴定出158个独特的转录本。诸如肌动蛋白结合蛋白等标记分子的存在、叶绿体相关mRNA的低发生率以及使用原位杂交对组成mRNA的筛管分子定位,均与汁液的韧皮部来源一致。对cDNA的功能分析揭示了包括核糖体功能、与环境相互作用、运输、DNA/RNA结合以及蛋白质周转等分类。对每个克隆的拟南芥(Arabidopsis thaliana (L.))最相近同源物的分析表明,参与细胞定位、蛋白质合成、组织定位、器官定位、器官分化和细胞命运的基因在文库中的出现水平是整个拟南芥基因组中的两倍。本文在韧皮部功能以及伴胞与筛管分子之间关系的背景下,对在这个富含韧皮部的文库中发现的转录本进行了讨论。