Zhao Jian, Li Hui, Fu Shuangbin, Chen Bo, Sun Wenting, Zhang Junqi, Zhang Jinfeng
National Engineering Laboratory for Tree Breeding, Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, The Tree and Ornamental Plant Breeding and Biotechnology Laboratory of State Forestry Administration, College of Biological Science and Biotechnology, Beijing Forestry University, Beijing 100083, China.
PLoS One. 2015 Mar 17;10(3):e0119987. doi: 10.1371/journal.pone.0119987. eCollection 2015.
Prince Rupprecht's larch (Larix principis-rupprechtii Mayr) is a native high-value forest tree species in North China whose clonal propagation through somatic embryogenesis (SE) has the potential to rapidly capture the benefits of breeding or genetic engineering programs and to improve raw material uniformity and quality. To date, research has focused on clarifying the molecular mechanism of SE, but proteomic studies are still in the early stages. In this study, isobaric tags for relative and absolute quantitation (iTRAQ) analysis was performed on three developmental stages of SE in L. principis-rupprechtii in an attempt to identify a wide range of proteins that are regulated differentially during this process. Proteins were extracted and analyzed from the pro-embryogenic mass (PEM), globular embryo (GE), and cotyledon embryo (CE) stages of embryo development. We detected 503 proteins in total and identified 96 proteins expressed differentially during different developmental stages. The identified proteins were analyzed further to provide information about their expression patterns and functions during SE. Four clusters of proteins based on shared expression profiles were generated. Functional analysis showed that proteins involved in primary metabolism, phosphorylation, and oxidation reduction were upregulated during somatic embryo development. This work provides novel insights into the process of larch embryo development in vitro and a basis for further study of the biological process and opportunities for practical application of this knowledge.
华北落叶松(Larix principis-rupprechtii Mayr)是中国北方本土的高价值林木树种,通过体细胞胚胎发生(SE)进行克隆繁殖,有潜力迅速获取育种或基因工程计划的益处,并提高原材料的一致性和质量。迄今为止,研究主要集中在阐明体细胞胚胎发生的分子机制,但蛋白质组学研究仍处于早期阶段。在本研究中,对华北落叶松体细胞胚胎发生的三个发育阶段进行了相对和绝对定量的等压标签(iTRAQ)分析,试图鉴定在此过程中差异调节的多种蛋白质。从胚胎发育的原胚性细胞团(PEM)、球形胚(GE)和子叶胚(CE)阶段提取并分析蛋白质。我们总共检测到503种蛋白质,并鉴定出96种在不同发育阶段差异表达的蛋白质。对鉴定出的蛋白质进行进一步分析,以提供其在体细胞胚胎发生过程中的表达模式和功能信息。基于共享表达谱生成了四类蛋白质。功能分析表明,参与初级代谢、磷酸化和氧化还原的蛋白质在体细胞胚胎发育过程中上调。这项工作为落叶松胚胎体外发育过程提供了新的见解,并为进一步研究这一生物学过程以及该知识的实际应用机会奠定了基础。