水稻种子胚人工老化过程中蛋白质差异表达分析

Analysis of Differential Expression of Seed Embryo Protein in the Artificial Aging Process in Rice

采用2-DE蛋白质双向电泳技术,对不同老化程度水稻种子的胚蛋白质进行了比较研究,采用17 cm、pH 3~10 IPG胶条,上样量1 mg蛋白,检测到约320个蛋白质点,对其中11个重复性比较好且差异表达在2倍以上的蛋白质点进行胶内酶切后,采用基质辅助激光解析分离飞行时间质谱进行肽指纹图谱分析,然后进行数据库搜索.其中2个蛋白质点分别与含有VHS,ENTH和ANTH结构域蛋白及异戊烯转移酶蛋白相匹配.含有VHS,ENTH和ANTH结构域蛋白能介导蛋白的内吞作用、增加膜的通透性且具有生长抑制活性功能.异戊烯转移酶蛋白的REF和SRPP共有保守域存在于部分与逆境相关的蛋白中,其中的SRPP广泛存在于动、植物中,与植物的抗逆境反应具有一定关系.结果表明：含有VHS,ENTH和ANTH结构域蛋白质的上调表达与异戊烯转移酶蛋白的下调表达可能与水稻种子的快速老化有关,但它们在老化过程中所起的作用,尚需进一步研究.

The seeds of rice were exposed to artificial accelerated aging.By 2-DE technology of protein bidirectional electrophoresis,significant viability and vigor changes were found in the seeds during the extension of the accelerated aging time.The proteins of rice seeds which were in different aging levels were separated first by isoelectric focusing with immobilized gel （pH 3-10 and 17 cm） gradients and then by SDS-PAGE.The coomassie brilliant blue-stained protein spots were analyzed using PDQUEST software,and about 320 detectable spots on each 2D-ge1 with the protein of 1 mg were found.With direct MALDI-TOF mass spectrometry analysis and protein database searching,only 2 out of 11 protein spots with high repeatability and double expression difference were identified with significant data.They were identified as a member of the VHS,ENTH and ANTH domain superfamily and the prenyltransferase domain-containing protein,respectively.As the VHS,ENTH and ANTH domain superfamily is an evolutionarily conserved protein module found primarily in proteins that participate in mediated endocytosis and the prenyltransferase domain-containing protein is widespread in nature and is related to the anti-adversity reaction of plants,the above results of this research suggested that the VHS,ENTH and ANTH domain superfamily and the prenyltransferase domain-containing protein may be related to the artificial accelerated aging of rice.However,their exact roles in ageing remain unclear and further study is needed.