变性高效液相色谱技术在大豆品种鉴定中的应用

Application of Denaturing High-Performance Liquid Chromatography for Soybean Variety Identification

单核苷酸多态性(SNPs)和插入-删除标记(In Del)在基因组中广泛分布,日益成为主要作物的重要遗传标记。利用变性高效液相色谱(DHPLC)技术检测大豆品种的SNP和In Del的多态性,可为大豆种质的品种鉴定提供一种新型的快速检测方法。基于公共数据库中的大豆基因信息,筛选了3条DNA序列作为目标分析片段,以大豆种质粤夏119作为参比基因型,利用DHPLC技术平台对10份大豆品种进行了鉴定。结果表明:混合参比DNA后的大豆样本产生DHPLC谱图均具有多态性,谱型与测序基因型一一对应。综合3条目标片段的DHPLC特征谱图可以区分所有大豆品种。试验结果验证了DHPLC应用于大豆品种鉴定的可行性和可靠性。由于DHPLC分辨率高、快速、操作简单、安全等优点,在大豆的品种鉴定中具有良好的应用前景。

Single nucleotide polymorphisms （SNP） and insertions/deletions （InDel） can be found within every genomic se- quence, which have been used as genetic markers for staple crops. Denaturing high-performance liquid chromatography （ DH- PLC） could be used to test the SNP and InDel, which would be applied for the rapid identification of diverse soybean varie- ties. In order to test the feasibility and reliability of DHPLC for discriminating between the diverse soybeans, the DHPLC anal- ysis on 10 soybean varieties were conducted. In this study, based on the information in soybean genome database, three DNA fragments were screened. The reference homozygous DNA from Yuexia 119 was added to PCR product of each soybean variety for the DHPLC analysis. The results showed the 3 screened DNA fragments were polymorphic across mixed samples of soybean variety and Yuexia 119 with distinct DHPLC elution profiles produced. Sequence analysis confirmed that the distinct DHPLC patterns corresponded to different DNA sequences. The set of distinct characteristic profiles in 3 DNA fragments differentiated in all the soybean varieties. The study demonstrates the power and usefulness of this technique for identifying soybean varie- ties. Due to the advantages of high resolution, high through-put, simplicity and safety, DHPLC is a promising technology for identification of soybean.