Silva Rafael de Carvalho, Carmo Lilian Silveira Travassos, Luis Zanderluce Gomes, Silva Luciano Paulino, Scherwinski-Pereira Jonny Everson, Mehta Angela
PPGBIOTEC, Departamento de Biologia, Universidade Federal do Amazonas, CEP 69077-000, Manaus, AM, Brazil.
Embrapa Recursos Genéticos e Biotecnologia, Av. W5 Norte Final, CEP 70770-917, Brasília, DF, Brazil.
J Proteomics. 2014 Jun 2;104:112-27. doi: 10.1016/j.jprot.2014.03.013. Epub 2014 Mar 25.
In the present study we have identified and characterized the proteins expressed during different developmental stages of Elaeis guineensis calli obtained from zygotic embryos. We were interested in the possible proteomic changes that would occur during the acquisition of somatic embryogenesis and therefore samples were collected from zygotic embryos (E1), swollen explants 14days (E2) in induction medium, primary callus (E3), and pro-embryogenic callus (E4). The samples were grinded in liquid nitrogen, followed by total protein extraction using phenol and extraction buffer. Proteins were analyzed by two-dimensional electrophoresis (2-DE) and the differentially expressed protein spots were analyzed by MALDI-TOF mass spectrometry (MS and MS/MS). Interestingly, we have identified proteins, which can be used as potential candidates for future studies aiming at the development of biomarkers for embryogenesis acquisition and for the different stages leading to pro-embryogenic callus formation such as type IIIa membrane protein cp-wap13, fructokinase and PR proteins. The results obtained shed some light on the biochemical events involved in the process of somatic embryogenesis of E. guineensis obtained from zygotic embryos. The use of stage-specific protein markers can help monitor cell differentiation and contribute to improve the protocols for successfully cloning the species.
Understanding the fate and dynamics of cells and tissues during callus formation is essential to understand totipotency and the mechanisms involved during acquisition of somatic embryogenesis (SE). In this study we have investigated the early stages of somatic embryogenesis induction in oil palm and have identified potential markers as well as proteins potentially involved in embryogenic competence acquisition. The use of these proteins can help improve tissue culture protocols in order to increase regeneration rates. This article is part of a Special Issue entitled: Environmental and structural proteomics.
在本研究中，我们鉴定并表征了从合子胚获得的油棕愈伤组织不同发育阶段所表达的蛋白质。我们对体细胞胚胎发生过程中可能发生的蛋白质组变化感兴趣，因此从合子胚（E1）、在诱导培养基中培养14天的肿胀外植体（E2）、初级愈伤组织（E3）和胚性愈伤组织（E4）中采集样本。样本在液氮中研磨，随后使用苯酚和提取缓冲液进行总蛋白提取。通过二维电泳（2-DE）分析蛋白质，并通过基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS和MS/MS）分析差异表达的蛋白点。有趣的是，我们鉴定出了一些蛋白质，它们可作为未来研究中的潜在候选物，旨在开发用于胚胎发生获得以及导致胚性愈伤组织形成的不同阶段的生物标志物，如IIIa型膜蛋白cp-wap13、果糖激酶和病程相关蛋白（PR蛋白）。所获得的结果揭示了从合子胚获得的油棕体细胞胚胎发生过程中涉及的生化事件。使用阶段特异性蛋白质标志物有助于监测细胞分化，并有助于改进成功克隆该物种的方案。
了解愈伤组织形成过程中细胞和组织的命运及动态对于理解全能性以及体细胞胚胎发生（SE）过程中涉及的机制至关重要。在本研究中，我们研究了油棕体细胞胚胎发生诱导的早期阶段，并鉴定出了潜在标志物以及可能参与胚胎发生能力获得的蛋白质。使用这些蛋白质有助于改进组织培养方案以提高再生率。本文是名为“环境与结构蛋白质组学”的特刊的一部分。
