Proteome analysis of male gametophyte development in rice anthers
Proteome analysis of male gametophyte development in rice anthers
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DOI：
10.1002/pmic.200300424
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发表时间：
2003-05-01
期刊：
影响因子：
3.4
通讯作者：
Rolfe, BG
中科院分区：
文献类型：
--
作者：
Kerim, T;Imin, N;Rolfe, BG
我们用蛋白质组学方法研究了严格控制生长条件下水稻花药发育过程中蛋白质合成的变化规律。通过细胞学分析和外部生长测量(如花药长度、耳廓距离和开花前天数)来确定花粉发育阶段。这使得收集代表六个不同花粉发育阶段的同步花药材料成为可能。从花药样品中提取蛋白质，用双向凝胶电泳法进行分离，得到蛋白质组图谱。比较了不同发育阶段的花药蛋白质组图谱，并用基质辅助激光解吸电离飞行时间质谱仪分析了150个在发育过程中不断变化的蛋白质点，产生了多肽质量指纹图谱(PMF)数据。使用这些PMF数据进行数据库搜索，发现了所分析的40个蛋白质点的身份。这40种蛋白质代表了33种独特的基因产物。对PMF分析不能鉴定的4个蛋白质点进行N末端微测序分析。鉴定出多种电荷异构体，包括空泡酸性转化酶、果糖激酶、β-膨胀素和Profilin。这些蛋白质与糖代谢、细胞伸长和细胞扩张密切相关，所有这些都是花粉萌发所必需的细胞活动。同一蛋白质的多种异构体的存在表明，在花粉发育过程中，这些蛋白质发生了某种翻译后修饰。
We used proteomic analysis to investigate the changing patterns of protein synthesis during pollen development in anthers from rice plants grown under strictly controlled growth conditions. Cytological analysis and external growth measurements such as anther length, auricle distances and days before flowering were used to determine pollen developmental stages. This allowed the collection of synchronous anther materials representing six discrete pollen developmental stages. Proteins were extracted from the anther samples and separated by two-dimensional gel electrophoresis to produce proteome maps. The anther proteome maps of different developmental stages were compared and 150 protein spots, which were changed consistently during development, were analysed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry to produce peptide mass fingerprint (PMF) data. Database searches using these PMF data revealed the identities of 40 of the protein spots analyzed. These 40 proteins represent 33 unique gene products. Four protein spots that could not be identified by PMF analysis were analysed by N-terminal microsequencing. Multiple charge-isoforms of vacuolar acid invertase, fructokinase, beta-expansin and profilin were identified. These proteins are closely associated with sugar metabolism, cell elongation and cell expansion, all of which are cell activities that are essential to pollen germination. The existence of multiple isoforms of the same proteins suggests that during the process of pollen development some kind of post-translational modification of these proteins occurs.