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A flax fibre proteome: identification of proteins enriched in bast fibres

机译:亚麻纤维蛋白质组学:鉴定富含韧皮纤维的蛋白质

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Background Bast fibres from the phloem tissues of flax are scientifically interesting and economically useful due in part to a dynamic system of secondary cell wall deposition. To better understand the molecular mechanisms underlying the process of cell wall development in flax, we extracted proteins from individually dissected phloem fibres (i.e. individual cells) at an early stage of secondary cell wall development, and compared these extracts to protein extracts from surrounding, non-fibre cells of the cortex, using fluorescent (DiGE) labels and 2D-gel electrophoresis, with identities assigned to some proteins by mass spectrometry. Results The abundance of many proteins in fibres was notably different from the surrounding non-fibre cells of the cortex, with approximately 13% of the 1,850 detectable spots being significantly (> 1.5 fold, p ≤ 0.05) enriched in fibres. Following mass spectrometry, we assigned identity to 114 spots, of which 51 were significantly enriched in fibres. We observed that a K+ channel subunit, annexins, porins, secretory pathway components, β-amylase, β-galactosidase and pectin and galactan biosynthetic enzymes were among the most highly enriched proteins detected in developing flax fibres, with many of these proteins showing electrophoretic patterns consistent with post-translational modifications. Conclusion The fibre-enriched proteins we identified are consistent with the dynamic process of secondary wall deposition previously suggested by histological and biochemical analyses, and particularly the importance of galactans and the secretory pathway in this process. The apparent abundance of β-amylase suggests that starch may be an unappreciated source of materials for cell wall biogenesis in flax bast fibres. Furthermore, our observations confirm previous reports that correlate accumulation proteins such as annexins, and specific heat shock proteins with secondary cell wall deposition.
机译:背景技术来自亚麻韧皮部组织的韧皮纤维在科学上是有趣的,并且在经济上是有用的,部分原因是二次细胞壁沉积的动态系统。为了更好地了解亚麻细胞壁发育过程的分子机制,我们在次级细胞壁发育的早期从单独解剖的韧皮部纤维(即单个细胞)中提取了蛋白质,并将这些提取物与周围非使用荧光(DiGE)标记和2D凝胶电泳,对皮层中的纤维细胞进行识别,并通过质谱将其身份分配给某些蛋白质。结果纤维中许多蛋白质的丰度与皮层周围的非纤维细胞明显不同,在1,850个可检测点中约有13%显着(> 1.5倍,p≤0.05)富含纤维。质谱分析后,我们为114个斑点指定了身份,其中51个斑点明显富含纤维。我们观察到,在发育中的亚麻纤维中,K + 通道亚基,膜联蛋白,孔蛋白,分泌途径成分,β-淀粉酶,β-半乳糖苷酶和果胶和半乳聚糖生物合成酶是最富集的蛋白质,这些蛋白质中的许多蛋白质显示出与翻译后修饰一致的电泳模式。结论我们确定的富含纤维蛋白与组织学和生化分析先前提出的次生壁沉积的动态过程一致,尤其是半乳聚糖和分泌途径在此过程中的重要性。 β-淀粉酶的表观丰度表明,淀粉可能是亚麻韧皮纤维中细胞壁生物发生的未知物质来源。此外,我们的观察结果证实了先前的报道,这些报道将积累蛋白(例如膜联蛋白)和特定的热激蛋白与次级细胞壁沉积相关联。

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