The Proteome of Seed Development in the Model Legume Lotus japonicus
The Proteome of Seed Development in the Model Legume Lotus japonicus
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DOI：
10.1104/pp.108.133405
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发表时间：
2009-03-01
期刊：
影响因子：
7.4
通讯作者：
Stougaard, Jens
中科院分区：
文献类型：
--
作者：
Dam, Svend;Laursen, Brian S.;Stougaard, Jens
我们的特点是在模式豆科植物百脉根种子的发展。像大豆（Glycine max）和豌豆（Pisum sativum）一样，莲花会长出直的豆荚，每个豆荚包含大约20颗种子，在40天内成熟。组织切片显示了豆科植物种子发育的三个阶段，以及干燥种子中胚、胚乳和种皮的存在。此外，蛋白质，油，淀粉，植酸和灰分含量测定，这表明成熟莲子的组成更接近大豆比豌豆。在第一次尝试，以确定种子蛋白质组，两个二维聚丙烯酰胺凝胶电泳的方法和基于凝胶的液相色谱-质谱法的方法被使用。球蛋白进行了分析，通过二维聚丙烯酰胺凝胶电泳，和5个豆球蛋白，LLP 1至LLP 5，和两个convicilins，LCP 1和LCP 2，通过基质辅助激光解吸电离四极杆/飞行时间质谱法进行了鉴定。对于两个不同的发展阶段，种子填充和干燥，凝胶为基础的液相色谱-质谱法的方法，并确定了665和181个独特的蛋白质对应的基因登录号的两个阶段，分别。所有的蛋白质组数据，包括实验数据和质谱谱峰，都收集在一个数据库中，该数据库可通过Web界面（http：//www.cbs.dtu.dk/cgi-bin/lotus/db.cgi）提供给科学界。该数据库为莲种子发育的生理、生化和调控奠定了基础。连同一个新的Web界面（http：//bioinfoserver.rsbs.anu.edu.au/utils/Pathing4legumes/）收集所有的蛋白质鉴定的莲花，苜蓿，和大豆种子蛋白质组，该数据库是一个宝贵的资源比较种子蛋白质组学和途径分析的豆类家庭内外。
We have characterized the development of seeds in the model legume Lotus japonicus. Like soybean (Glycine max) and pea (Pisum sativum), Lotus develops straight seed pods and each pod contains approximately 20 seeds that reach maturity within 40 days. Histological sections show the characteristic three developmental phases of legume seeds and the presence of embryo, endosperm, and seed coat in desiccated seeds. Furthermore, protein, oil, starch, phytic acid, and ash contents were determined, and this indicates that the composition of mature Lotus seed is more similar to soybean than to pea. In a first attempt to determine the seed proteome, both a two-dimensional polyacrylamide gel electrophoresis approach and a gel-based liquid chromatography-mass spectrometry approach were used. Globulins were analyzed by two-dimensional polyacrylamide gel electrophoresis, and five legumins, LLP1 to LLP5, and two convicilins, LCP1 and LCP2, were identified by matrix-assisted laser desorption ionization quadrupole/time-of-flight mass spectrometry. For two distinct developmental phases, seed filling and desiccation, a gel-based liquid chromatography-mass spectrometry approach was used, and 665 and 181 unique proteins corresponding to gene accession numbers were identified for the two phases, respectively. All of the proteome data, including the experimental data and mass spectrometry spectra peaks, were collected in a database that is available to the scientific community via a Web interface (http://www.cbs.dtu.dk/cgi-bin/lotus/db.cgi). This database establishes the basis for relating physiology, biochemistry, and regulation of seed development in Lotus. Together with a new Web interface (http://bioinfoserver.rsbs.anu.edu.au/utils/PathExpress4legumes/) collecting all protein identifications for Lotus, Medicago, and soybean seed proteomes, this database is a valuable resource for comparative seed proteomics and pathway analysis within and beyond the legume family.