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微卫星锚定片段长度多态性(MFLP)是一种新型分子标记技术,本研究依据该技术原理,通过将M13通用荧光接头连接在设计的SSR引物上对扩增产物进行荧光标记,建立了适合于烟草的荧光MFLP标记体系;在此基础上,利用该技术从144对选择性扩增引物组合中筛选出15对适宜的引物组合,对32份来自于不同国家和地区的烟草种质材料的遗传多态性进行了分析。试验结果显示,这些引物组合扩增的产物在32个烟草种质中的多态性范围为2～13个,平均4.3个;利用种质材料间遗传多态性开展的遗传相似性分析表明,32份烟草种质间的相似系数在0.40到0.83之间。运用UPGMA聚类分析和主坐标分析法,有效地将这些烟草种质材料的来源和类型进行了分类。结果表明,荧光MFLP标记技术能有效地发现供试烟草材料的DNA多态性,表明该技术适用于烟草遗传特性分析的研究和应用。
The microsatellite anchor fragment length polymorphism (MFLP) is a novel molecular marker technology. According to the principle of the technology, the M13 universal fluorescent linker is connected to the designed SSR primer to fluorescently amplify the amplified product, and a Based on this, 15 pairs of suitable primer combinations were selected from 144 pairs of selective amplification primer combinations, 32 pairs of tobacco germplasm materials from different countries and regions The genetic polymorphisms were analyzed. The results showed that the polymorphism range of 32 tobacco germplasm ranged from 2 to 13 with an average of 4.3. The genetic similarity analysis based on the genetic polymorphism of the germplasm showed that, The similarity coefficient of 32 tobacco germplasms ranged from 0.40 to 0.83. The sources and types of these tobacco germplasm materials are effectively classified using UPGMA cluster analysis and principal-co-ordinate analysis. The results showed that the fluorescence MFLP labeling technology can effectively find the DNA polymorphism of tested tobacco materials, indicating that the technology is suitable for the study and application of genetic analysis of tobacco.