不同检测方法对甜菜SCoT和DAMD扩增产物多态性的影响  

Effect of Different Detection Methods on Polymorphism of Beet SCoT and DAMD Amplification Products

作　　者：梁雪梅 吴则东[1,2] LIANG Xue-mei;WU Ze-dong(College of Advanced Agriculture and Ecological Environment,Heilongjiang University,Harbin 150080,China;Key Laboratory of Sugar Beet Genetic Breeding,Heilongjiang University,Harbin 150080,China)

机构地区：[1]黑龙江大学现代农业与生态环境学院,哈尔滨150080 [2]黑龙江省普通高校甜菜遗传育种重点实验室/黑龙江大学,哈尔滨150080

出　　处：《中国甜菜糖业》2022年第3期1-8,共8页China Beet & Sugar

基　　金：“财政部和农业农村部:国家现代农业产业技术体系资助(CARS-170111)”。

摘　　要：为筛选更优的SCoT和DAMD扩增产物多态性的检测方法,为构建甜菜品种分子身份证,建立一套快速鉴定甜菜品种真实性的体系提供依据。试验以8个不同的甜菜品种对7条DAMD引物和15条SCoT引物进行筛选,选出多态性最好,分辨率最高的核心引物;再利用1%、2%琼脂糖凝胶和6%、8%的非变性聚丙烯酰胺凝胶两种电泳方法对扩增产物的多态性进行检测并对比分析。结果表明,从7条DAMD引物和15条SCoT引物中筛选出6条带型清晰,多态性高的核心引物。这6条核心引物在8%的非变性聚丙烯酰胺凝胶电泳检测产物时,条带难以分离辨认;而在6%的非变性聚丙烯酰胺凝胶中共扩增出131条带,其中130条为多态性条带,总多态性占比为99.2%。同时在用1%的琼脂糖凝胶检测产物的条带图虽能显示一定的多态性,但条带分离较差,出现重叠的情况;2%琼脂糖凝胶电泳中,扩增总条带57条,43条多态性条带,总的多态性占比为75.4%;前者扩增的总条带数比后者扩增出的总条带数多了近2.3倍,而在总多态性占比中,前者是后者的3倍。因此非变性聚丙烯酰胺凝胶电泳比琼脂糖凝胶电泳的检测更适于对DAMD和SCoT扩增产物的检测,用于构建甜菜品种指纹图谱构建,这种需要检测样品数量大,且需获得更准确遗传信息的研究,可以提高检测效率。To screen better detection methods for SCoT-PCR and DAMD-PCR amplification product polymorphism,that provide the basis for constructing a molecular identity card for sugar beet varieties and establishing a system for rapid identification of authenticity of sugar beet varieties.The 7 DAMD and 15 SCoT markers were screened with 8 varieties,and the core primers with the best polymorphism and highest resolution were selected,then the polymorphisms of the amplified products were detected and compared with each other using two electrophoresis methods:1%and 2%agarose gels and 6%and 8%non-denaturing polyacrylamide gels.The results showed that 6 core primers with clear band shape and high polymorphism were selected from 7 DAMD and 15 SCoT primers to identify the authenticity of sugar beet varieties.The 6 core primers were difficult to separate and identify when the products were detected by electrophoresis on 8%non-denaturing polyacrylamide gels,while 131 bands were amplified on 6%non-denaturing polyacrylamide gels,of which 130 were polymorphic,with a total polymorphism of 99.2.Meanwhile,although the band plots of the products detected on 1%agarose gels showed some polymorphism,but the bands were poorly separated and overlapped.In 2%agarose gel electrophoresis,there are 57 amplicon including 43 polymorphic bands,with a total polymorphism percentage of 75.4.The total number of bands amplified by the former was nearly 2.3 times higher than that of the latter,while the former was 3 times higher than the latter in terms of the total polymorphism ratio.Therefore,non-denaturing polyacrylamide gel electrophoresis is more suitable than agarose gel electrophoresis for detecting DAMD and SCoT amplification products for constructing fingerprint profiles of sugar beet varieties,and such studies require testing a large number of samples to obtain more accurate genetic information,which can improve detection efficiency.

关 键 词：甜菜 DAMD SCoT 非变性聚丙烯酰胺凝胶电泳 琼脂糖凝胶电泳 

分 类 号：S566.3[农业科学—作物学]