Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal.
Physiol Plant. 2012 Oct;146(2):236-49. doi: 10.1111/j.1399-3054.2012.01633.x. Epub 2012 May 10.
Medicago truncatula is a model legume, whose genome is currently being sequenced. Somatic embryogenesis (SE) is a genotype-dependent character and not yet fully understood. In this study, a proteomic approach was used to compare the induction and expression phases of SE of both the highly embryogenic line M9-10a of M. truncatula cv. Jemalong and its non-embryogenic predecessor line, M9. The statistical analysis between the lines revealed 136 proteins with significant differential expression (P < 0.05). Of these, 5 had a presence/absence pattern in M9 vs M9-10a and 22 showed an at least twofold difference in terms of spot volume, were considered of particular relevance to the SE process and therefore chosen for identification. Spots were excised in gel digested with trypsin and proteins were identified using matrix-assisted laser desorption ionization-time of flight/time of flight. Identified proteins indicated a higher adaptability of the embryogenic line toward the stress imposed by the inducing culture conditions. Also, some proteins were shown to have a dual pattern of expression: peroxidase, pyrophosphatase and aspartate aminotransferase. These proteins showed higher expression during the induction phases of the M9 line, whereas in the embryogenic line had higher expression at stages coinciding with embryo formation.
蒺藜苜蓿是一种模式豆科植物,其基因组目前正在测序中。体细胞胚胎发生(SE)是一种依赖于基因型的特征,尚未完全理解。在这项研究中,采用蛋白质组学方法比较了高度胚胎发生的 M. truncatula cv。Jemalong 的 M9-10a 品系和其非胚胎发生的前体品系 M9 的 SE 诱导和表达阶段。对这两个品系之间的统计分析显示,有 136 种蛋白质具有显著差异表达(P<0.05)。其中,5 种在 M9 与 M9-10a 之间存在/不存在模式,22 种在斑点体积方面表现出至少两倍的差异,被认为与 SE 过程特别相关,因此被选择用于鉴定。用胰蛋白酶消化凝胶后切除斑点,并使用基质辅助激光解吸电离飞行时间/飞行时间进行蛋白质鉴定。鉴定出的蛋白质表明,胚胎发生品系对诱导培养条件施加的应激具有更高的适应性。此外,一些蛋白质表现出双重表达模式:过氧化物酶、焦磷酸酶和天冬氨酸氨基转移酶。这些蛋白质在 M9 系的诱导阶段表达较高,而在胚胎发生系中,与胚胎形成阶段同时表达较高。