Roach Melissa J, Deyholos Michael K
Department of Biological Sciences, University of Alberta, Edmonton, Canada.
Mol Genet Genomics. 2007 Aug;278(2):149-65. doi: 10.1007/s00438-007-0241-1. Epub 2007 May 15.
To better understand the molecular processes associated with the development of the unusually long (> 30 mm) and strong bast fibre cells within the phloem of flax stems, we conducted a gene discovery experiment to identify transcripts enriched in fibre-bearing tissues, with the intention that these transcripts would serve as future targets for crop improvement and research in phloem development and cell wall deposition. We produced a library of 9,600 cDNA clones from the peels of flax stems, and selected tissue-specific cDNAs for sequencing based on two series of microarray experiments. In the first microarray series, we compared transcript abundance in stem-peels and leaves, and identified stem-enriched transcripts putatively involved in the processes of polysaccharide and cell wall metabolism. In the second microarray series, we compared gene expression in three segments of the vertical stem axis, which constituted a developmental series for phloem fibres and other cell types. The expression of specific LTP and AGP transcripts was particularly well-correlated with stem segments during either the elongation phase or cell-wall thickening phase of phloem fibre development, and the phloem-specific enrichment of these transcripts was confirmed by qRT-PCR. Transcripts representing multiple, distinct chitinases, beta-galactosidases, arabinogalactan proteins (AGP), and lipid transfer proteins (LTPs) were among the interesting transcripts enriched in specific stages of the developing stem. Considered together, the results of our analyses suggest similarity between the molecular mechanisms underlying phloem fibre development and the gelatinous fibres of tension wood in trees.
为了更好地理解与亚麻茎韧皮部中异常长(>30毫米)且坚韧的韧皮纤维细胞发育相关的分子过程,我们进行了一项基因发现实验,以鉴定在含纤维组织中富集的转录本,目的是这些转录本可作为未来作物改良以及韧皮部发育和细胞壁沉积研究的目标。我们从亚麻茎的外皮中构建了一个包含9600个cDNA克隆的文库,并基于两个系列的微阵列实验选择组织特异性cDNA进行测序。在第一个微阵列系列中,我们比较了茎皮和叶片中的转录本丰度,并鉴定了可能参与多糖和细胞壁代谢过程的茎富集转录本。在第二个微阵列系列中,我们比较了垂直茎轴三个区段中的基因表达,这三个区段构成了韧皮纤维和其他细胞类型的一个发育系列。在韧皮纤维发育的伸长阶段或细胞壁加厚阶段,特定的脂质转移蛋白(LTP)和阿拉伯半乳聚糖蛋白(AGP)转录本的表达与茎区段的相关性特别好,并且通过qRT-PCR证实了这些转录本在韧皮部中的特异性富集。代表多种不同几丁质酶、β-半乳糖苷酶、阿拉伯半乳聚糖蛋白(AGP)和脂质转移蛋白(LTP)的转录本是发育茎特定阶段中富集的有趣转录本之一。综合考虑,我们的分析结果表明韧皮纤维发育的分子机制与树木中张力木的胶质纤维之间存在相似性。