Luo Yu-ming, Zhang Wei-ming, Ding Xiao-yu, Shen Jie, Bao Shu-lin, Chu Bi-hai, Mao Shan-guo
Jiangsu Key Laboratory for Biodiversity and Technology, College of Life Sciences, Nanjing Normal University, Nanjing 210097, China.
Yao Xue Xue Bao. 2006 Sep;41(9):840-5.
To authenticate all the varieties of Perilla (single-species genus), to analyze sequences of rDNA ITS regions and single nucleotide polymorphism (SNP) within them and based on these, to design allele-specific diagnostic PCR primers.
The rDNA ITS regions of the perilla varieties were sequenced and analyzed by Clustal X 1.8, MEGA 3.0. Allele-specific diagnostic PCR primers that can authenticate all the perilla varieties were designed based on SNPs loci.
The length of rDNA ITS sequences of perilla varieties ranged from 612 to 615 bp in size, including ITS1 (230 -232 bp), 5.8S (179 bp) and ITS2 (203 -204 bp). The GC content is about 61.5% - 61.9%. There is not only SNPs in non-coding region ITS1 and ITS2 (ncSNP), but also three coding SNPs (cSNP) loci in the conservative region of 5.8S. All the SNPs have only two allele loci polymorphism. The cSNP in 5.8S is related to the morphology variation among the varieties. Allele-specific diagnostic PCR primers have been designed according to SNPs loci to authenticate accurately all the seeds and leaves of Perilla varieties.
SNPs in rDNA ITS region can be used as an effective molecular markers to authenticate all the varieties of Perilla.
对紫苏属（单种属）所有品种进行鉴定，分析其核糖体DNA内转录间隔区（rDNA ITS）序列及其中的单核苷酸多态性（SNP），并据此设计等位基因特异性诊断PCR引物。
对紫苏品种的rDNA ITS区域进行测序，并采用Clustal X 1.8和MEGA 3.0软件进行分析。基于SNP位点设计能够鉴定所有紫苏品种的等位基因特异性诊断PCR引物。
紫苏品种rDNA ITS序列长度在612至615 bp之间，包括ITS1（230 - 232 bp）、5.8S（179 bp）和ITS2（203 - 204 bp）。GC含量约为61.5% - 61.9%。非编码区ITS1和ITS2存在单核苷酸多态性（ncSNP），5.8S保守区也存在3个编码单核苷酸多态性（cSNP）位点。所有SNP均只有两个等位基因位点多态性。5.8S中的cSNP与品种间形态变异相关。已根据SNP位点设计等位基因特异性诊断PCR引物，可准确鉴定紫苏品种的所有种子和叶片。
rDNA ITS区域的SNP可作为鉴定紫苏所有品种的有效分子标记。
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