辽宁省槭属植物种质资源遗传多样性分析及指纹图谱构建

摘要

为研究辽宁省槭属植物种质资源遗传多样性水平并构建分子指纹图谱,为分子水平上鉴定槭属种质提供技术支撑,也为辽宁省槭属植物种质资源开发、保存利用及新品种选育奠定基础,本研究利用SRAP分子标记技术,对辽宁省内的11份槭属植物主栽品种和19份不同种源红花槭种质为材料,利用110对SRAP 引物进行PCR扩增,产物用8%非变性聚丙烯酰胺凝胶电泳分离,并统计多态性引物的总扩增条带数、多态性条带数和多态性比率。利用NTSYS-pc2.1和Excel软件统计SRAP-PCR条带数据,按照相似系数法和非加权组平均法进行聚类分析,计算遗传相似性系数,绘制树状聚类图,并构建30份槭属种质资源指纹图谱。从110对SRAP 引物中共筛选出36对多态性好且重复性高的引物,共扩增得到708条带,其中多态性谱带为542条,多态性比率为76.55%；30份槭属植物材料的遗传相似系数在0.235~0.954之间,其中19份不同种源红花槭的遗传相似系数在0.700~0.954之间,平均为0.849；通过聚类分析将30份材料划分为7个群,在11份本地材料间,假色槭与挪威槭之间遗传相似系数最小(0.235),国王枫与挪威槭相似系数最大(0.703)。SRAP 引物ME6/EM3可有效区分所有材料,并构建出30份槭属材料种质资源特异性分子指纹图谱。结果表明：供试槭属植物材料间具有较丰富的遗传多样性水平,基于36对SRAP引物组合所构建的指纹图谱具有唯一性和高效性,SRAP标记适合用于槭属植物种质遗传多样性分析及品种鉴定。%In order to provide technique support for their identification at molecular level and their exploitation, conservation, utilization and innovation, the genetic diversity of Acer L. germplasms was studied, in addition the DNA fingerprinting were established in Liaoning Province. Eleven materials of maple trees in Liaoning area and 19 different provenances of Acer rubrum were studied by using 110 pair SRAP primers. The PCR amplification products were separated by 8% nondenaturing PAGE electrophoresis, and total bands, polymorphic bands and the percentage. The UPGMA clustering analysis was performed by software NTSYS-pc2.1 and Excel, also attained the genetic similarity coefficient value, and mapped UPGMA tree. Meanwhile, DNA fingerprinting of 30 materials of Acer L. germplasms were constructed, 36 pair polymorphic primers were selected from 110 pair of SRAP primers. The results showed that a total of 708 bands were amplified by these primers, which including 542 polymorphic bands, the average percentage of polymorphic bands was 76.55%. The genetic similarity coefficient of 30 Acer L. germplasms ranged from 0.235 to 0.954, of which the 19 Acer rubrum materials from 0.700 to 0.954, averaging 0.849. The cluster analysis showed that these materials could be divided into seven main groups by using the UPGMA. The clustering among the 11 local materials showed that there was the smallest genetic similarity coefficient between Acer pseudo-sieboldianum(Pax.) Komarov and Acer platanoides, whereas the largest genetic similarity coefficient between Acer platanoides ˊCrimson kingˊ and Acer platanoides. In addition, primer ME6/EM3 could be used to distinguish the 30 Acer L. materials. The molecular fingerprint checking index of Acer L. was constructed according to the special SRAP bands. The research results showed that there was a high level of genetic diversity in Acer L. germplasms, and it was unique and efficient that the molecular identity was established by SRAP primer combinations. The SRAP markers could apply to genetic diversity analysis and cultivar identification of Acer L.