| 龙眼体细胞胚发生早期的蛋白质组学 |
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| 引用本文： | 赖呈纯,赖钟雄,方智振,何园,姜顺日. 龙眼体细胞胚发生早期的蛋白质组学[J]. 中国农业科学, 2012, 45(9): 1775-1790. DOI: 10.3864/j.issn.0578-1752.2012.09.013 |
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| 作者姓名： | 赖呈纯 赖钟雄 方智振 何园 姜顺日 |
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| 作者单位： | 1.福建农林大学园艺植物生物工程研究所，福州 3500022.福建省农业科学院农业工程技术研究所，福州 350003 |
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| 基金项目： | 国家自然科学基金项目(31078717,39870544);福建省亚热带果树及特种经济作物种质资源共享平台(2008N2001)项目 |
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| 摘 要： | 【目的】通过龙眼（Dimocarpus longan Lour.）体细胞胚发生早期5个阶段的蛋白质组学研究，为植物胚胎发育相关蛋白基因分离和植物体细胞胚发生相关机制等的深入研究奠定基础。【方法】利用已建立的龙眼体细胞胚发生再生系统，经同步化培养获得龙眼体细胞胚发生早期5个阶段胚性培养物，并采用双向电泳和质谱技术对龙眼体细胞胚发生早期的蛋白质组变化进行分析。【结果】龙眼体细胞胚发生早期5个阶段的蛋白质2D表达谱可检测到1 203-1 798个蛋白点，其在龙眼体细胞胚发生早期各阶段大部分具有阶段差异表达特性，有小部分是阶段特异表达，根据其蛋白数目、表达丰度、分子量和等电点等变化，确定了龙眼体细胞胚发生早期的ECII到CpECGE阶段是体细胞胚发生早期的关键性阶段。成功鉴定45个差异蛋白，成功率为37%，其中以能量、碳水化合物代谢相关蛋白和氧化胁迫反应相关蛋白占多数，分别为22%和27%；通过其功能分析可以推断出，龙眼体细胞胚发生早期，能量和糖代谢是体细胞胚发生的基础，而氧化胁迫反应是体细胞胚发生的先决条件，并通过参与细胞骨架的稳定、氮代谢、信号转导、基因调控、蛋白质的翻译加工修饰和定位等功能的蛋白，构成一个庞大的龙眼体细胞胚发生蛋白质调控网络系统，保证体细胞胚的正常发育。【结论】对龙眼体细胞胚发生早期过程的蛋白质表达变化有了较全面的了解，蛋白质表达数量呈先减低后升高再减低的趋势，龙眼体细胞胚发生早期能量和糖代谢旺盛，氧化胁迫反应相关蛋白对体细胞胚早期的发生和发育有重要的调控作用。
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| 关 键 词： | 龙眼 体细胞胚发生 蛋白质组学 双向电泳 质谱 |
| 收稿时间： | 2011-11-23 |
Proteomic Analysis of Early Somatic Embryogenesis in Longan(Dimocarpus longan Lour.) |
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| LAI Cheng-chun , LAI Zhong-xiong , FANG Zhi-zhen , HE Yuan , JIANG Shun-ri. Proteomic Analysis of Early Somatic Embryogenesis in Longan(Dimocarpus longan Lour.)[J]. Scientia Agricultura Sinica, 2012, 45(9): 1775-1790. DOI: 10.3864/j.issn.0578-1752.2012.09.013 |
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| Authors: | LAI Cheng-chun LAI Zhong-xiong FANG Zhi-zhen HE Yuan JIANG Shun-ri |
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| Affiliation: | 1(1Institute of Horticultural Biotechnology,Fujian Agriculture and Forestry University,Fuzhou 350002;2Institute of Agricultural Engineering and Technology,Fujian Academy of Agricultural Sciences,Fuzhou 350003) |
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| Abstract: | 【Objective】The differential expressions of proteome patterns at 5 early developmental stages were analyzed during longan somatic embryogenesis.The results would contribute to the further research for isolation of plant embryo development associated protein genes and studies of plant somatic embryogenesis mechanisms.【Method】Five embryogenic cultures at early developmental stages from the established longan somatic embryogenesis and regeneration system were obtained after synchronized culture.And their changes of proteomic patterns were investigated by the immobilized pH gradients two-dimensional electrophoresis technology and matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF/TOF).【Result】From 1 203 to 1 798 protein spots could reproducibly be resolved in two-dimensional gels,most of them displayed obvious differences in the expression abundances at different stages and small part of them were of specific expression.On the basis of variations of protein numbers,expression abundances,molecular weight and isoelectric point,the key stages of early somatic embryogenesis in longan were determined,namely from ECII(Embryogenic callus II) to CpECGE(Compact pro-embryogenic cultures globular embryos).Forty-five differentially expressed proteins were identified and the identification rate was 37%.These findings suggested that energy and carbohydrate metabolism and oxidative stress response have a large majority,22% and 27% respectively.It could be deduced by analyzing the functions of these proteins that the material foundation of longan somatic embryogenesis at early stages was of energy and carbohydrate metabolism,and the precondition of somatic embryogenesis was of oxidative stress response.To ensure the normal development of somatic embryos,these proteins together with the proteins related to cytoskeleton stability,nitrogen metabolism,signal transduction,gene regulation,protein synthesis or modification,and positioning function formed an enormous protein network system of early somatic embryogenesis in longan.【Conclusion】There had more overall understanding to expression changes of proteome patterns during the early longan somatic embryogenesis.The number of expressed proteins had a trend of increase after the first decrease,and then reduced again.During the early longan somatic embryogenesis,energy and carbohydrate metabolism is very vigorously,and the proteins related to oxidative stress response may play an important role in modulating the early somatic embryogenesis and development. |
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| Keywords: | longan(Dimocarpus longan Lour.) somatic embryogenesis proteomics two-dimensional electrophoresis mass spectrometry |
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