大豆种子发育过程中差异表达蛋白的蛋白质组分析(英文)

Proteomic Analysis of Differentially Expressed Proteins in Developmental Soybean [Glycine max(L.)Merr.]Seed

采用蛋白质组学技术研究了大豆N2899种子发育过程中蛋白质的差异表达。运用PDQuest软件比较分析不同发育时期(15,20,30,40,50DAF和成熟种子)大豆种子蛋白的双向电泳图谱,在考染的2-D胶上共检测到337个蛋白点。有些蛋白质在整个发育过程中都出现,而另外一些只出现在发育早期或成熟的种子中。利用基质辅助-激光解吸/电离-飞行时间-质谱(MALDI-TOF-MS)技术,分析了不同发育时期30个差异表达蛋白,并用Pro-found(http://www.prowl.rockefeller.edu)工具,对质谱产生的肽质量指纹(PMF)数据进行NCBInr数据库检索,结果鉴定了18个蛋白质。比较发现,这些蛋白主要参与种子的成熟(如伴豆球蛋白)、逆境胁迫反应(如抗坏血酸过氧化酶)、细胞分裂(如Skp1)和蛋白运输(如钙网蛋白)等。研究表明,种子发育过程十分复杂,所鉴定的蛋白质,可为从分子水平上研究大豆种子发育进程中蛋白的积累和调控奠定基础。

A proteomic approach was applied in the study on the changes in proteins that occur during seed development in soybean [ Glycine max（ L. ）Merr. ] cv N2899. Protein expression at different developmental stages（ 15,20,30,40,50 DAF and mature seed）were compared using PDQuest software（Bio-Rad） after two-dimensional gel electrophoresis. About 337 low-salt extractable protein spots were resolved in gels. It was found that some proteins were present throughout the whole developmental stages, while others were associated with early seed filling or seed maturation. Among total spots 18 of 30 highly differentially expressed proteins were identified by MALDI- TOF- MS, querying peptide mass fingerprinting data in NCBInr database by Profound（ http ：//www. prowl, rockefeller, edu）. It was found that these identified proteins are mainly involved in seed maturation（ e. g. conglycinin ）, stress tolerance （e. g. ascorbate peroxidase）, cell division （e. g. Skp1 ） , and proteins transportation （e. g. calreticulin）. These results indicate that the seed development process is complex, and the identified genes would provide a basis for molecular elucidation of the temporal effects on protein accumulation in soybean seed.