【作者】 任国臣；

【导师】 闫桂琴；

【作者基本信息】 山西师范大学 ， 植物学， 2010， 硕士

【摘要】 翅果油树属胡颓子科胡颓子属,为中国二级珍稀濒危保护植物,主要分布于我国山西省的翼城、平陆和乡宁;已有的研究表明其具有非常重要的科学经济价值和生态价值。本文以翅果油树种子蛋白为研究对象,对种子蛋白提取和电泳条件的相关主要指标进行优化实验,在此基础上对翅果油树三种生态型种子贮藏蛋白的含量进行测定和组分分析实验,以及利用种子总贮藏蛋白为遗传标记对翅果油树翼城、平陆和乡宁3个主要种群的共73粒种子进行遗传多样性分析实验;以期为翅果油树种子蛋白的进一步深入研究和应用研究提供一些基础理论依据,同时也为翅果油树的保护生物学提供一定的理论参考。用考马斯亮蓝G-250染色法和SDS-PAGE电泳技术对三种生态型翅果油树种子贮藏蛋白进行含量测定和电泳图谱分析,结果表明:大宫灯、长果型、小宫灯三种生态型种子中总贮藏蛋白的百分含量分别为33.753±0.746、32.075±1.111和26.633±1.091;翅果油树种子贮藏蛋白含量以谷蛋白含量最高,其次为清蛋白和球蛋白,醇溶蛋白的含量最低;三生态型间的总贮藏蛋白含量存在显著差异;电泳图谱显示三生态型之间清蛋白、球蛋白、醇溶蛋白和谷蛋白的蛋白质亚基条带信息都存在差异,表明翅果油树生态型间种子贮藏蛋白具有多态性。以种子总贮藏蛋白为遗传标记对翅果油树翼城、平陆和乡宁3个主要种群共73粒种子进行遗传多样性分析实验结果表明:(1)平陆种群的多态百分率为44.44%、乡宁和翼城两个种群的多态位点百分率均55.56%;种群平均多态位点百分率为51.85%,总多态位点百分率77.78%。(2)种群间的的遗传分化系数为GST=0.1539,说明翅果油树种群的遗传分化主要存在于种群内。(3)基于遗传分化系数估算的基因流是Nm=2.7492,大于1,表明种群间存着在一定的基因交流。(4) UPGMA聚类结果显示3个种群形成了明显的两支,平陆种群单独成为一支,乡宁和翼城种群聚为一支;遗传距离和地理距离的Mantel相关分析(相关系数r=0.99094>0.8)表明来自相似地理环境的或地理距离较近的种群具有相似的遗传背景。综合上述指标显示:翅果油树种群具有较高的遗传多样性,不存在着遗传上的衰退现象,能够适应多变自然生存环境;其濒危的主要原因可能是人类活动造成生境的破坏和种子萌发率低。针对对翅果油树种子蛋白遗传多样性研究的这一结果,提出了以下保护策略:加强对翅果油树的组织培养快繁技术、种子贮藏保存技术及种子育苗繁殖技术的研究;建立翅果油树自然保护区,加强就地保护;加强翅果油树的经济价值开发研究,使人们充分认识到保护翅果油树的生态价值和经济价值。更多还原

【Abstract】 Elaeagnus mollis diels(E.mollis diels) belongs to Elaeagnus (Elaeagna-ceae). It is considered a rare and endangered plant,and national second-class protected species in China. It only exists in China,and mainly distributes Xiang ning,Yi cheng,Ping lu of Shan xi’s province. And many studies in the past showed that it has the very important economical and ecological value.This study used seed protein of E.mollis diels as the research object. Firstly, some key elements about extraction and electrophor- etic conditions of seed storage proteins were optimized in this thesis. Based on the results of the fist experiments, contents of storage protein of three kinds of ecotypes in E.mollis diels seeds were determined and components of its seed storage proteins were also analysized. The genetic diversity of three populations of E.mollis diels by seed total storage proteins were studied in the third experiment,and three major populations were separately Yi cheng, Ping lu and xiang ning. These research results of above experiments can provide not only some basic theories for further theoretical and applied research about seed proteins but also some theoretical references for Conservation Biology on E. mollis diels.Seed storage proteins of three ecotypes in E. mollis diels were studied on the content determination and electrophoresis analysis by using the staining with coomassie brilliant blue G-250 and a sodium dodecylsulfate electrophoresis performed polyacrylamide gels(SDS- PAGE).The results indicated that the percentage content of total storage proteins in the seeds of three ecotypes,which are named Da-Gong Deng,Chang-Guo Xing and Xiao-Gong Deng,is respectively 33.753±0.746, 32.075±1.111,26.633±1.091.For E. mollis diels ,most of storage proteins are glutelin.The next is respectively albumin and globulin.There is a little alcohol soluble protein. There are significant differences in the content of total storage proteins between three ecotypers. Electrophoresis maps reveral that three ecotypers are differentiated by the information of protein subunits on glutelin, albumin,globulin and alcohol soluble protein ;these also show that seed storage proteins display obvious polymorphisms among three ecotypes.This thesis studied the genetic diversity of three populations of E. mollis diels by using the genetic marker of seed storage proteins, the results revealed: (1) The genetic diversity of total 73 individuals from three populations were investigated. Seved within the total nine loci detected were polymorphic. The percentage of polymorphic loci in three populations(Yi cheng, Ping lu and xiang ning) are respectively 55.6%,44.44% and 55.6%. The mean percentage of polymorphic loci of three populations is 51.85%, and the total percentage of polymorphic loci of three populations is 77.78%. (2)With seed storage protein marker data, coefficient of genetic differentiation(GST) among three populations was 0.1539. This showed that the majority of genetic variation occurred within populations of E. mollis diels. (3) Based on the data of GST the estimated value of gene flow (Nm) was 2.7492, indicating that relatively high level of gene flow existed among populations. (4) The results of UPGMA (Unweighted Pair Group Method with Arithmetic mean) clustering analysis showed that three populations of E. mollis diels were clearly divided into two clusters. Ping lu population clustered into a single class. two populations of Xiang ning and Yi cheng clustered into the other class . The Mantel’test analysis on genetic distance and geographic distance (correlation coefficient r=0.99094>0.8) showed that those population- ns from similar geographical environment condition or closer geographical distance lied in similar genetic background. Acoording to these analysis above, E. mollis diels populations had higher genetic diversity, and there is no genetic deterioration among populations. This indicated that E. mollis diels could adapt to the changing natural habitats; so habitats destruction caused mainly by human activity and lower seed germination rate resulted that this speices was in endanger.Finally, aiming at the research conclusions from seed protein genetic diversity of E.mollis diels in the end of this thesis, some corresponding protection strategyies are suggested that are :(1)strengthening basic theory and biotechnology studies on E.mollis diels such as tissue culture propagation techniques,seed preservat- ion storage and seedling breeding technology (2)establishing E.mollis diels nature reserves in order to strengthen in situ conservation;(3) developing the economic value research of E.mollis diels so as to make people fully understand the ecological value and economic value of protecting E.mollis diels.更多还原