Laboratoire des Biotechnologies Végétales Appliquées à l'Amélioration des Cultures, Faculté des Sciences de Sfax, Sfax, Tunisia.
J Proteomics. 2009 Nov 2;73(1):161-77. doi: 10.1016/j.jprot.2009.07.003. Epub 2009 Jul 18.
Two-dimensional gel electrophoresis coupled to mass spectrometry has been used to compare the proteome of date palm (Phoenix dactylifera L. cv. Deglet Nour) zygotic and somatic embryos. Proteins were trichloroacetic acid-acetone-phenol extracted, quantified, and resolved by 2-DE in the 5 to 8 pH range. Total protein content and number of resolved spots were higher in zygotic (110+/-14.5mg/g DW; 349 spots) than in somatic (70.96+/-4.8mg/g DW; 210 spots) embryos. The 2-DE map of both systems showed qualitative (263) and quantitative (72) differences. Statistical analysis of spot intensity was performed by PCA, obtaining two accurate groupings of the samples and determining the most discriminating spots. Samples were also clustered using Euclidean distance with average linkage algorithm of the Genesis software package. Sixty-three variable spots were subjected to mass spectrometry analysis, resulting in 23 identifications. Identified proteins were classified in the following functional categories; glycolysis (8 proteins), citrate cycle (1), ATP synthesis (1), carbohydrate biosynthesis (2), amino acids metabolism (1), stress related (4), storage (3), and with no function assigned for three of them. Most of the somatic embryo specific proteins identified belonged to glycolysis pathways, whereas those of the zygotic embryo to storage and stress-related proteins. Differences are discussed in terms of metabolism and biology of both types of embryos.
双向电泳结合质谱技术被用于比较椰枣(Phoenix dactylifera L. cv. Deglet Nour)合子胚和体细胞胚的蛋白质组。蛋白质用三氯乙酸-丙酮-酚提取、定量,并用 2-DE 在 5 到 8 pH 范围进行分离。合子胚的总蛋白含量和分辨斑点数高于体细胞胚(110+/-14.5mg/g DW;349 个斑点)(70.96+/-4.8mg/g DW;210 个斑点)。两个系统的 2-DE 图谱显示出定性(263)和定量(72)差异。通过 PCA 对斑点强度进行了统计分析,获得了样品的两个准确分组,并确定了最具区分性的斑点。还使用欧几里得距离和 Genesis 软件包的平均链接算法对样品进行聚类。对 63 个可变斑点进行质谱分析,得到 23 个鉴定结果。鉴定出的蛋白质被分为以下功能类别:糖酵解(8 种蛋白质)、柠檬酸循环(1 种)、ATP 合成(1 种)、碳水化合物生物合成(2 种)、氨基酸代谢(1 种)、应激相关(4 种)、储存(3 种),其中 3 种没有功能分配。鉴定出的大多数体细胞胚特异性蛋白质属于糖酵解途径,而合子胚的蛋白质则属于储存和应激相关蛋白质。根据两种胚胎的代谢和生物学讨论了差异。