无患子科树木细子龙营养贮藏蛋白质的分离鉴定

doi:10.3969/j.issn.1000-2561.2004.04.016

CORC > 北京大学 > 生命科学学院

	无患子科树木细子龙营养贮藏蛋白质的分离鉴定; Identification and Distribution of Vegetative Storage Proteins in a Sapindaceae Tree Amesiodendron chinense (Merr.) Hu
	王旭初 ; 史敏晶 ; 陈月异 ; 田维敏
刊名	热带作物学报
	2004
关键词	细子龙营养贮藏蛋白质细胞学免疫印迹间接免疫荧光定位 Amesiodendron chinense (Merr.) Hu vegetative storage proteins cytology Western-blotting immuno-fluorescence localization
DOI	10.3969/j.issn.1000-2561.2004.04.016
英文摘要	采用光学显微技术、十二烷基磺酸钠-聚丙稀酰胺凝胶电泳(SDS-PAGE)、免疫印迹和间接免疫荧光定位技术,分离鉴定了无患子科树木细子龙(Amesiodendron chinense)的营养贮藏蛋白质(VSP).在经汞-溴酚蓝染色的石蜡切片中,观察到枝条、树干和根的次生木质部的木薄壁细胞中有深蓝色颗粒状物质,而在上述枝条、树干和根的次生韧皮部的韧皮薄壁细胞和韧皮射线薄壁细胞中则为均一状的淡蓝色物质.这些被染成不同程度蓝色的物质具有蛋白质性质.SDS-PAGE分析表明,在枝条、树干和根的树皮及木质部中,有2种含量高的蛋白质,其分子质量约23 kDa和26 kDa.使用荔枝(一种无患子科树木)的22 kDa VSP的多克隆抗体对这些部位的树皮和木质部可溶性蛋白质进行免疫印迹分析,在免疫印迹图谱上,该抗血清只识别23 kDa和26kDa蛋白质,且23 kDa蛋白质谱带颜色较深,说明它们与荔枝的22 kDa VSP有一定的同源性.间接免疫荧光定位结果表明,只有颗粒状和均一状蛋白质内含物发出异硫氰酸荧光素特异的蓝绿色荧光,说明23 kDa和26 kDa蛋白质是这些颗粒均一状内含物的主要成分.树木抽新梢后,这2种蛋白质含量在一定程度上减少.可见,23 kDa和26 kDa蛋白质符合树木VSP的3条标准,应该是细子龙的VSP.韧皮部和木质部的VSP在形态上的差异可能与蛋白质的种类不同有关.; Vegetative storage proteins (VSPs) in the Sapindaceae tree species Amesiodendron chinense were identified by using optical microscopy, SDS-PAGE, Western-blotting and indirect immunohistochemistry. Under opticaal microscope, deep blue masses in a granular appearance were observed in the secondary xylem parenchyma cells while light blue masses in homogeneous form were present in the secondary phloem parenchyma cells in the cross-section of barks and woods of branches, trunk and roots stained with mercury-bromophenol blue. These blue masses were obviously proteinaceous and the cells with the blue masses were called protein-storing cells (PSCs). Two major proteins, with molecular weights of about 23 kDa and 26 kDa, were isolated by SDS-PAGE. With the polyclonal antibodies of the 22kDa VSPs in Litchi chinensis, a Sapindaceae tree, Western-blotting was carried out. Among the soluble proteins from the bark tissues and xylem ofAmesiodendron chinense, only the 23 kDa and 26 kDa proteins reacted with the antibodies,with much deeper color of the 23 kDa protein band, illustrating that they were immuno-related with the 22 kDa VSPs in Litchi chinensis. Immunofluorescence localization showed that some substances corresponding to the granular and homologeous proteinaceous masses in PSCs emitted FITC (fluorescein isothiocyanate)-specific blue-green fluorescence,demonstrating that the 23 kDa and 26 kDa proteins were the major components of the proteinaceous masses. The two proteins disappeared to some extent during new shoot development. From the combined data, the 23 kDa and 26 kDa proteins should be the VSPs in Amesiodendron chinense. The different forms between the VSPs in bark tissues and that in xylem may be ascribed to the differences in their biochemical properties.; 国家科学基金; 中国科学引文数据库(CSCD); 0; 4; 78-83; 25
语种	中文
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/241733]
专题	生命科学学院
推荐引用方式 GB/T 7714	王旭初,史敏晶,陈月异,等. 无患子科树木细子龙营养贮藏蛋白质的分离鉴定, Identification and Distribution of Vegetative Storage Proteins in a Sapindaceae Tree Amesiodendron chinense (Merr.) Hu[J]. 热带作物学报,2004.
APA	王旭初,史敏晶,陈月异,&田维敏.(2004).无患子科树木细子龙营养贮藏蛋白质的分离鉴定.热带作物学报.
MLA	王旭初,et al."无患子科树木细子龙营养贮藏蛋白质的分离鉴定".热带作物学报 (2004).