| 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. |
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