【作者】 王利；

【导师】 邢世岩； 束怀瑞；

【作者基本信息】 山东农业大学 ， 园艺， 2006， 博士

【摘要】 以来自国内外的90个银杏种质为材料,采用CTAB法提取幼嫩银杏叶片总DNA,从64对EcoRI/MseI引物(其中Msel引物为荧光标记物)中筛选出7-8对清晰、多态性高的引物,对90个银杏材料的遗传多样性进行了AFLP分析。采用GeneScan3.1软件对图像进行处理,构建0、1数学矩阵。利用POPGENE32 version1.32软件计算遗传多样性指数,采用NTSYS.pc2.11f计算软件,得到Dice相似性系数矩阵,并用UPGMA法进行聚类分析,研究结果如下:(1)利用8对引物首次对来自美国、荷兰、日本、法国和我国的21个银杏观赏品种的遗传多样性进行了研究,共产生1117条谱带,229个特异位点(包括单态带单态带175条和缺失带54条),多态带983条,多态带的比例为88%;每对引物鉴别效率为100%。品种之间的相似系数为0.49-0.84。当相似系数为0.73时,供试观赏品种可分为四类。聚类结果表明:品种并没有全部按来源地聚类,各来源地的品种之间存在着广泛的交流。来源地相同的品种并不单独聚成一类,中国和法国的品种分别属于其中的三类。根据对观赏品种的特异位点、相似系数、聚类结果等进行综合分析表明,‘塔形银杏’、‘垂乳银杏’、‘筒叶银杏’、‘大耳银杏’、‘斑叶银杏’、‘展冠银杏’、‘垂枝银杏’、‘沂源叶籽银杏1’这8个品种是银杏观赏品种中的重要特异种质。14个国外银杏观赏种质平均多态带的比例高于7个国内种质。(2)首次利用7对引物对对来自国内的29个银杏雄株种质的遗传多样性进行了研究,7对引物共产生1173条谱带,187个特异位点和1145条多态带,多态带的比例平均为97.6%。每对引物鉴别效率达到100%。所有种质之间相似系数在0.38-0.82之间,所试种质遗传变异大。当相似系数为0.51时,全部种质可以分为2大类,山东的大部分种质聚为一类。‘天目山6’、‘宁国2’、‘郯城5’可能是比较原始的种质。29个国内雄株的遗传多样性略高于21个银杏观赏种质。(3)利用7对引物对来自国内外的49个银杏雌株种质的遗传多样性进行了研究,共产生1219条谱带,131个特异位点(包括单态带120条和缺失带11条),多态带1200条,多态性为98.44%。每对引物的鉴别效率为100%。各雌株种质之间相似系数在0.39-0.83之间。在SC=0.51、0.64处,供试材料分为2类和7类,所试山东银杏雌株种质占总类型的100%,山东银杏雌株种质存在着较宽的遗传基础。49个国内雌株的遗更多还原

【Abstract】 Ninety germplasms of Ginkgo biloba from foreign and domestic countries were used as tested materials, genomic DNA of them were extracted using the cetyltrimethyl ammonium bromide (CTAB) method. Seven-eight primer combinations that produced clearly and highly polymorphic bands were screened from 64 EcoRI/MseI primer combinations (MseI fluorescent labeled). The genetic diversity of 90 germplasms of G. biloba from the U. S., Holland, Japan, France and China was analyzed. Band patterns were dealt with Genescan3.1 software (Apllied Biosystems, Foster city, CA. USA), the binary data matrix was constructed. The Dice similarity coefficient (SC) matrix was obtained by using NTSYS.pc2.11f software (Apllied Biosystems, Setauket, NY. USA), and cluster analysis based on SC matrix was performed with unweighted pair-group methods of arithmetic averages (UPGMA). The results were obtained as following:Genetic diversity of ornamental cultivars of G. biloba: The genetic diversity of twenty-one G. biloba ornamental cultivars from the U S, Holland, Japan, France and China was analyzed with 8 primer combinations. These primer combinations produced a total of 1119 bands, 229 specific loci (including 54 absent bands, 175 monomorphic bands), 983 polymorphic bands, the percentage of polymorphic bands(PPB) was 88%, and the identification percentage (IP) per primer combination was 100%. Genetic similarity coefficient (SC) among all cultivars varied from 0.49 to 0.84, and all cultivars were divided into four clusters when SC was set at 0.73. The cultivars from the same origin did not fall into the same group. The cultivars from France and China were classified into three groups, respectively. According to the comprehensive analysis based on specific loci, similarity coefficient, and clustering results, 8 cultivars‘Fastigiata’,‘Tit’,‘Tubifolia’,‘Daeryinxing’,‘Variegata’,‘Horizontalis,‘Pendula’, and‘Yiyuanyeziyinxing’were considered to be important germplasms of ornamental cultivars of Ginkgo biloba. The average PPB (35.86%) of 14 foreign G. biloba ornamental cultivars was higher than that (31.51%) of 7 domestic G. biloba ornamental cultivars.更多还原