Ranik Martin, Creux Nicky M, Myburg Alexander A
Department of Genetics, Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria, Pretoria, 0002, South Africa.
Tree Physiol. 2006 Mar;26(3):365-75. doi: 10.1093/treephys/26.3.365.
Despite the availability of high-throughput transcript profiling technology, little is known about tissue-specific gene expression patterns in the wood-forming tissues of Eucalyptus plantation tree species. We used cDNA-amplified fragment length polymorphism (AFLP) analysis in combination with infrared fragment detection and semi-automated band quantification to profile gene expression in a 6-year-old, fast- growing Eucalyptus tree. The expression profiles of 6385 transcript-derived fragments (TDFs) were analyzed across four major woody tissues (mature xylem, immature xylem, phloem and cork) collected from two stem positions, to provide a global view of transcript abundance and variability in the Eucalyptus stem. About 21% of the TDFs were differentially expressed and could be grouped into clusters representing co- expressed genes. A total of 71 TDFs representing different gene clusters were isolated and characterized. These included genes implicated in cell fate, signal transduction and cell wall biosynthesis, processes closely associated with xylogenesis. Analysis of the expression levels of selected TDFs by quantitative RT-PCR corroborated the TDF quantification and confirmed that cDNA-AFLP analysis is a highly efficient and accurate tool for transcript profiling and gene discovery in wood-forming tissues of tree species.
尽管已有高通量转录谱分析技术,但对于桉树人工林树种木质形成组织中的组织特异性基因表达模式仍知之甚少。我们将cDNA扩增片段长度多态性(AFLP)分析与红外片段检测及半自动条带定量相结合,对一棵6年生的速生桉树的基因表达进行了分析。分析了从两个茎部位置采集的四种主要木质组织(成熟木质部、未成熟木质部、韧皮部和木栓)中6385个转录本衍生片段(TDF)的表达谱,以全面了解桉树茎中转录本丰度和变异性。约21%的TDF存在差异表达,可分为代表共表达基因的簇。共分离并鉴定了71个代表不同基因簇的TDF。其中包括与细胞命运、信号转导和细胞壁生物合成相关的基因,这些过程与木质部形成密切相关。通过定量RT-PCR对选定TDF的表达水平进行分析,证实了TDF定量结果,并确认cDNA-AFLP分析是一种用于树种木质形成组织中转录谱分析和基因发现的高效且准确的工具。