Barros Eugenia, van Staden Carol-Ann, Lezar Sabine
CSIR-Biosciences, Meiring Naude Road, Brummeria, Pretoria,South Africa.
BMC Biotechnol. 2009 May 27;9:51. doi: 10.1186/1472-6750-9-51.
Fast-growing Eucalyptus grandis trees are one of the most efficient producers of wood in South Africa. The most serious problem affecting the quality and yield of solid wood products is the occurrence of end splitting in logs. Selection of E. grandis planting stock that exhibit preferred wood qualities is thus a priority of the South African forestry industry. We used microarray-based DNA-amplified fragment length polymorphism (AFLP) analysis in combination with expression profiling to develop fingerprints and profile gene expression of wood-forming tissue of seven different E. grandis trees.
A 1578-probe cDNA microarray was constructed by arraying 768 cDNA-AFLP clones and 810 cDNA library clones from seven individual E. grandis trees onto silanised slides. The results revealed that 32% of the spotted fragments showed distinct expression patterns (with a fold change of at least 1.4 or -1.4 and a p value of 0.01) could be grouped into clusters representing co-expressed genes. Evaluation of the binary distribution of cDNA-AFLP fragments on the array showed that the individual genotypes could be discriminated.
A simple, yet general method was developed for genotyping and expression profiling of wood-forming tissue of E. grandis trees differing in their splitting characteristics and in their lignin contents. Evaluation of gene expression profiles and the binary distribution of cDNA-AFLP fragments on the chip suggest that the prototype chip developed could be useful for transcript profiling and for the identification of Eucalyptus trees with preferred wood quality traits in commercial breeding programmes.
速生巨桉是南非木材生产效率最高的树种之一。影响实木产品质量和产量的最严重问题是原木端部开裂的发生。因此,选择具有优良木材品质的巨桉种植材料是南非林业产业的首要任务。我们结合表达谱分析,利用基于微阵列的DNA扩增片段长度多态性(AFLP)分析技术,对7棵不同巨桉树木的木材形成组织进行指纹图谱构建和基因表达谱分析。
通过将来自7棵个体巨桉树木的768个cDNA-AFLP克隆和810个cDNA文库克隆排列在硅烷化载玻片上,构建了一个包含1578个探针的cDNA微阵列。结果显示,32%的斑点片段呈现出明显的表达模式(倍数变化至少为1.4或-1.4,p值为0.01),这些片段可被归类为代表共表达基因的簇。对微阵列上cDNA-AFLP片段的二元分布进行评估表明,可以区分个体基因型。
开发了一种简单但通用的方法,用于对具有不同开裂特性和木质素含量的巨桉树木的木材形成组织进行基因分型和表达谱分析。对基因表达谱和芯片上cDNA-AFLP片段的二元分布进行评估表明,所开发的原型芯片可用于转录谱分析,并有助于在商业育种计划中鉴定具有优良木材品质性状的桉树。