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目的应用ITS2序列对93份岗梅药材及其近缘物种进行鉴定研究。方法对药材、基原植物样本进行基因组DNA提取,PCR扩增ITS2序列,用MEGA5.0进行序列分析,采用相似性搜索法、最近距离法、构建NJ树对序列鉴定能力进行评估。结果岗梅药材ITS2序列长度为241 bp,平均GC含量为63.4%,种内K2P遗传距离为0~0.030,岗梅药材及其近缘种的种间最小K2P距离为0.043。岗梅药材和基原物种在NJ树单独聚为一支,能有效地与其他物种分开,相似性搜索法结果表明ITS2序列能够准确鉴定岗梅药材及其近缘种。结论 ITS2序列能够准确鉴定岗梅药材及其近缘物种,可为保障岗梅药材临床用药安全提供新的技术手段。
Objective To identify 93 germplasm resources of Gangmei and its related species using ITS2 sequence. Methods Genomic DNA was extracted from the samples of medicinal plants and primordial plants. ITS2 sequences were amplified by PCR. Sequence analysis was carried out by using MEGA 5.0. Similarity search method and nearest distance method were used to construct NJ tree to evaluate sequence identification ability. Results The length of ITS2 sequence was 241 bp with the average GC content of 63.4%. The genetic distance of K2P in species was 0 ~ 0.030. The minimum K2P distance of interspecific linkage was 0.043 in Gangmur and its related species. Gangmian medicinal materials and base species are clustered together in NJ tree, which can be effectively separated from other species. The similarity search results show that ITS2 sequence can accurately identify Gangmur and its related species. Conclusion ITS2 sequence can accurately identify Gaomi medicinal materials and their related species, which can provide new technical means to ensure the clinical drug safety of Gangmei medicinal materials.