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以导入红海榄总DNA的耐盐番茄及其野生型番茄为材料,于3~4片真叶期分别用调和海水(约1%盐度)和自来水进行灌溉,处理14 d后,采用BPP法提取番茄叶片的全蛋白质,通过等电聚焦和第二向SDSPAGE电泳得到清晰的番茄叶片全蛋白质双向电泳图谱。利用Image Master 2D platinum 6.0软件对海水处理及对照条件下耐盐番茄及野生型番茄差异表达的蛋白进行分析发现,蛋白质双向电泳图谱大约有800个蛋白点,共检测到123个差异蛋白点,每张图谱的匹配率达90%。海水处理下的耐盐番茄与野生型番茄的叶片全蛋白质图谱中共检测到82个高丰度的差异蛋白点,其中42个蛋白点的表达量为上调,40个蛋白点的表达量为下调。对差异表达的蛋白质点进行了基质辅助激光解吸电离串联飞行时间质谱(MALDI-TOF-TOF/MS)鉴定,得到了相关的差异蛋白质信息,为揭示耐盐番茄的耐盐机理、为克隆相关耐盐基因提供了帮助。
The salt-tolerant tomato and its wild-type tomato, which were introduced into total DNA of Rhizophora stylosa, were used to irrigate seawater (about 1% salinity) and tap water in 3 ~ 4 true leaf stage respectively. After 14 d of treatment, The whole protein of tomato leaves was extracted and the clear two-dimensional gel electrophoresis map of tomato leaves was obtained by isoelectric focusing and second-dimension SDSPAGE electrophoresis. Using Image Master 2D platinum 6.0 software to analyze the proteins differentially expressed in salt-tolerant tomato and wild-type tomato under seawater treatment and control conditions, there are about 800 protein spots in the two-dimensional electrophoresis map of protein, and 123 differential protein spots were detected, each Matching rate of 90% of the map. A total of 82 highly abundant differential protein spots were detected in salt-tolerant tomato and wild tomato leaves under seawater treatment, of which 42 protein spots were up-regulated and 40 spots were down-regulated. The differentially expressed protein spots were identified by MALDI-TOF / MS, and the related differential protein information was obtained. To reveal the salt tolerance mechanism of salt tolerant tomato, The salt gene provided help.