日本落叶松胚性愈伤组织蛋白组分双向电泳分析

Analysis of Total Proteins in the Embryogenic Tissues of Japanese Larch (Larix leptolepis) by two-Dimensional Electrophoresis

对日本落叶松(Larixleptolepis)胚性愈伤组织进行了组成特点和蛋白质组分双向电泳的研究.研究表明,胚性愈伤组织是由不同发育阶段的早期胚组成的细胞团,早期胚主要由长的胚柄细胞及其顶端的胚头细胞组成,而后者又具不同的分裂相.以三氯乙酸/丙酮法提取胚性组织的总蛋白,通过双向凝胶电泳获得了完整的全蛋白质图谱.运用图像分析软件(I mage Master2D)对电泳图谱分析表明:在凝胶上展现出475个蛋白质组分,并进一步确定每个蛋白质相应的分子质量、等电点和相对含量.从本研究所获得的具高分辨率日本落叶松胚性愈伤组织蛋白质图谱,将为今后与愈伤组织的分化能力相关的蛋白质的检测、分离和基因克隆,以及胚性能力相关蛋白标准化的制定等提供参考.

The embryogenic tissues of Larix leptolepis have been investigated by two-dimensional electrophoresis in the current study. It shows that the embryogenic tissues are composed of a group of small meristematic cells at one end （embryoproper cells） , which are at different phages of cell division and many long cells at the other end （embryo-suspensor cells）. The total proteins of the embryogenic tissues are extracted by trichloroacetic acid/acetone （TCA） method,and then separated by isoelectric focusing as the first dimension and SDS-PAGE as the second dimension. The protein spots are visualized by staining with Coomassie Brilliant Blue. After analysis with ImageMaster 2D, 475 different proteins resolve on the gel, their isoelectric point （pI）,molecular weight （MW） and relative volume of each protein in the embryogenic tissues are also discovered. The present study has built a solid foundation for identification,characterization,gene cloning and standardization of relation proteins in the embryogenic tissues of Larix leptolepis.