基于SNP位点快速鉴定药用、斑点野生稻的dCAPS标记体系的建立

The Establishment of dCAPS Markers System in Oryza punctata and Oryza officinalis on the Basis of the SNP

为防止野生稻重要基因资源流失,便于口岸准确快速鉴定,探讨了基于SNP位点建立dCAPS分子标记体系。采用生物信息学软件寻找了药用野生稻和斑点野生稻上7个基因碱基序列上适合设计错配引物的SNP位点,设计合成适当的错配引物进行PCR扩增,选择相应的限制性内切酶进行酶切。在这2种野生稻DNA的matk基因上分别找到1个合适的SNP位点,设计了3对错配引物,PCR后3种相应的限切酶都可以特异性地切开PCR产物,将药用野生稻或斑点野生稻与栽培稻和其它野生稻区分开来。本结果证明了dCAPS分子标记可以在口岸查验上应用。

The dCAPS molecular markers system is established on the basis of the SNP to facilitate identification of rice quickly and prevent the loss of important genetic resources in wild rice. Nucleotide sequences of seven genes in O. punctata and O. officinalis were analyzed by bio-informatics software to search for SNP loci that are suitable for designing appropriate mismatch primer（s）, which allow relevant enzymes to cut on after PCR. In matK gene of two wild rice spieces DNA, one appropriate SNP loci were found respectively, on the basis of which three mismatch primer pairs were designed and distinguished O. punctata or O. officinalis from other Oryza spp after restriction enzyme digestion of PCR products. This result supported that dCAPS molecular markers can be used in port inspection.