摘要
[目的]通过对4种蛋白质提取方法对比分析,筛选出适于早春植物叶片蛋白质的提取方法。对3种蔷薇科植物不同时期叶片中蛋白质含量及组分进行测定分析,揭示叶片中蛋白质含量和组分的变化规律,为后期开展相应的蛋白组学研究提供理论基础。[方法]4种蛋白质提取方法:酚提取法,改良酚提取法,改良Tris-HCl法和TCA-丙酮法。通过考马斯亮蓝(G-250)染色法和SDS-PAGE法对3种早春植物叶片的蛋白质含量和组分进行测定。[结果]与其他3种蛋白质提取方法相比,利用TCA-丙酮法提取叶片中蛋白质,后期所得蛋白图谱效果最好,蛋白质条带多且清晰。4月初至6月初5个时期叶片中蛋白质含量整体呈"先增长后下降"的变化趋势,不同生长时期华北珍珠梅叶片蛋白质含量均明显低于日本晚樱和西府海棠。SDS-PAGE电泳表明:3种早春植物叶片蛋白质组分丰富,变化多样。花期为4-5月的日本晚樱和西府海棠叶片蛋白质表达的种类相对较多,条带也较清晰;花期为6-7月的华北珍珠梅叶片蛋白质条带和发生明显变化的蛋白质种类都比较少。[结论]TCA-丙酮法是针对3种蔷薇科植物最优蛋白质提取方法。早春蔷薇科植物花期与叶片蛋白质含量相关,开花前蛋白质含量持续增长,在花期达到最高;且花期较早的日本晚樱和西府海棠比花期晚的华北珍珠梅叶片蛋白质含量和蛋白组分高。
[Objective]The suitable method of leaf protein extraction for early spring plant was screened out from four different ways.The variation of protein content and composition at different leaf stagesin three rosaceous species were determined and analyzed..The results provided the theoretical basis for the corresponding proteomics research in the later stage.[Methods]Four kinds of protein extraction methods,known as phenolic extraction,modified phenolic method,modified Tris-HCl method,and TCA-acetone extraction method were employed.The protein content and composition of three early spring plant leaves were determined by（G-250）staining and SDS-PAGE.[Results]Study indicated that TCA-acetone extraction method produced better protein profile and higher quality of gel electrophoresis pattern compared to other extraction methods.During April to early June,the protein content in the leaves at five different growth stages showed ＂first increase,then decrease＂trend＂,and in different growth stages Sorbaria kirilowii leaf protein content was significantly lower than Cerasus serrulata,lannesiana,and Malus micromalus.SDS-PAGE electrophoresis showed that three early spring plant leaves were rich in various protein components.From April to May,the protein expression of Cerasus serrulata lannesiana and Malus micromalus leaves were relatively higher and the bands were clearer.The protein bands of Sorbaria kirilowii leaves and the protein types that were changed during flowering stage from June to July were relatively fewer than other species.[Conclusion]The TCA-acetone extraction method was the optimal protein extraction method for three kinds of rosaceous plants.In early spring,protein content of rosaceous plants was closely affected by the flowering time,and the protein content before flowering was constantly in-creased and reached the peak at full-blooming stage.The protein content of early flowering species,such as Cerasus serrulata lannesiana and Malus micromalus,was greater than that of late flowering species,such as
作者
陈虹
李涛
白晋华
郭红彦
Chen Hong;Li Tao;Bai Jinhua;Guo Hongyan(College of Forestry,Shanxi Agricultural University,Taigu 030801,China)
出处
《山西农业大学学报（自然科学版）》
CAS
北大核心
2018年第6期29-34,共6页
Journal of Shanxi Agricultural University(Natural Science Edition)
