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Analysis of xylem sap proteins from Brassica napus

机译：甘蓝型油菜木质部汁液蛋白的分析

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Background Substance transport in higher land plants is mediated by vascular bundles, consisting of phloem and xylem strands that interconnect all plant organs. While the phloem mainly allocates photoassimilates, the role of the xylem is the transport of water and inorganic nutrients from roots to all aerial plant parts. Only recently it was noticed that in addition to mineral salts, xylem sap contains organic nutrients and even proteins. Although these proteins might have important impact on the performance of above-ground organs, only a few of them have been identified so far and their physiological functions are still unclear. Results We used root-pressure xylem exudate, collected from cut Brassica napus stems, to extract total proteins. These protein preparations were then separated by high-resolution two-dimensional gel electrophoresis (2-DE). After individual tryptic digests of the most abundant coomassie-stained protein spots, partial peptide sequence information was deduced from tandem mass spectrometric (MS/MS) fragmentation spectra and subsequently used for protein identifications by database searches. This approach resulted in the identification of 69 proteins. These identifications include different proteins potentially involved in defence-related reactions and cell wall metabolism. Conclusion This study provides a comprehensive overview of the most abundant proteins present in xylem sap of Brassica napus . A number of 69 proteins could be identified from which many previously were not known to be localized to this compartment in any other plant species. Since Brassica napus , a close relative of the fully sequenced model plant Arabidopsis thaliana , was used as the experimental system, our results provide a large number of candidate proteins for directed molecular and biochemical analyses of the physiological functions of the xylem under different environmental and developmental conditions. This approach will allow exploiting many of the already established functional genomic resources, like i.e. the large mutant collections, that are available for Arabidopsis.
机译：背景高地植物中的物质运输是由维管束介导的，维管束由韧皮部和木质部链组成，这些韧皮部和木质部链相互连接所有植物器官。韧皮部主要分配光同化物，而木质部的作用是将水和无机养分从根部运输到植物的所有空中部分。直到最近才注意到，除木质盐外，木质部树液还含有有机营养物甚至蛋白质。尽管这些蛋白质可能对地上器官的性能产生重要影响，但到目前为止，仅鉴定了其中的几种，其生理功能仍不清楚。结果我们使用了从甘蓝型油菜切成的茎上收集的根压木质部分泌物提取总蛋白。然后通过高分辨率二维凝胶电泳（2-DE）分离这些蛋白质制剂。在对胰蛋白酶消化最丰富的考马斯蛋白染色的蛋白斑点进行单独的胰蛋白酶消化后，从串联质谱（MS / MS）碎片质谱图推导了部分肽序列信息，然后通过数据库搜索将其用于蛋白鉴定。这种方法导致鉴定出69种蛋白质。这些鉴定包括可能与防御相关反应和细胞壁代谢有关的不同蛋白质。结论本研究对甘蓝型油菜木质部汁液中最丰富的蛋白质进行了全面的综述。可以鉴定出69种蛋白质，其中许多先前未知的蛋白质都未知于任何其他植物物种中。由于将全测序模型植物拟南芥的近亲Brassica napus用作实验系统，因此我们的结果提供了大量候选蛋白，用于针对不同环境和发育条件下木质部的生理功能进行定向分子和生化分析。条件。这种方法将允许开发许多已经建立的功能基因组资源，例如可用于拟南芥的大型突变体集合。

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