Hewezi Tarek, Petitprez Michel, Gentzbittel Laurent
Laboratoire de Biotechnologies et Amèlioration des Plantes, Ecole Nationale Supérieure Agronomique de Toulouse, Avenue de l'Agrobiopôle, BP 107, Auzeville Tolosane, Castanet Tolosan, 31326 France.
Planta. 2006 Apr;223(5):948-64. doi: 10.1007/s00425-005-0151-6. Epub 2005 Nov 24.
The early stage of embryo development is a critical step in plant production. To identify genes with potential roles in the early sunflower seed development, a cDNA microarray approach was employed. We developed a thematic cDNA microarray containing clones representing high sequence similarities with known or predicted Arabidopsis genes implicated in different metabolic and signal transduction pathways. This 800-element cDNA array was used to compare the expression patterns in leaves and immature embryos (2 mm and 6 mm). Statistical analysis, using two-step ANOVA, revealed that 143 cDNA clones can be considered as differentially expressed. Of these, 62 clones were found to be up-regulated in leaves, 81 in embryos whereas only seven clones displayed increased level of mRNA in the 6 mm embryos when compared with 2 mm embryos. The differentially expressed clones are distributed among many metabolic and signal transduction pathways. For example, genes related to fatty acid metabolism and amino acid biosynthesis exhibited preferential expression patterns in immature embryos. Also, clones potentially encoding enzymes involved in the metabolism of ascorbate and aldarate, pyruvate, propanoate and inositol, and citrate cycle were found to be up-regulated in embryos. In contrast, cDNA clones putatively involved in energy metabolism were more abundant in leaves than embryos. Clones encoding potential signal transduction components including receptors, protein kinases, protein phosphatases, and transcription factors were also identified, with preferential expression profiles in immature embryos. The expression patterns derived from this study provide initial characterization of metabolic pathways and signalling transduction networks occurring in the early stage of sunflower seed development.
胚胎发育早期是植物生产中的关键步骤。为了鉴定在向日葵种子早期发育中具有潜在作用的基因,采用了cDNA微阵列方法。我们开发了一个主题cDNA微阵列,其中包含与参与不同代谢和信号转导途径的已知或预测拟南芥基因具有高度序列相似性的克隆。这个包含800个元件的cDNA阵列用于比较叶片和未成熟胚(2毫米和6毫米)中的表达模式。使用两步方差分析的统计分析表明,143个cDNA克隆可被视为差异表达。其中,62个克隆在叶片中上调,81个在胚中上调,而与2毫米胚相比,只有7个克隆在6毫米胚中mRNA水平升高。差异表达的克隆分布在许多代谢和信号转导途径中。例如,与脂肪酸代谢和氨基酸生物合成相关的基因在未成熟胚中表现出优先表达模式。此外,发现可能编码参与抗坏血酸和醛糖代谢、丙酮酸、丙酸和肌醇代谢以及柠檬酸循环的酶的克隆在胚中上调。相反,推测参与能量代谢的cDNA克隆在叶片中比在胚中更丰富。还鉴定了编码包括受体、蛋白激酶、蛋白磷酸酶和转录因子在内的潜在信号转导成分的克隆,它们在未成熟胚中具有优先表达谱。本研究得出的表达模式为向日葵种子发育早期发生的代谢途径和信号转导网络提供了初步特征。