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本文从线粒体基因片段入手,利用PCR技术扩增出17种枫斗类石斛39个个体的线粒体基因co I、nad 5部分序列及nad 1-intron 2序列,nr DNA ITS及叶绿体基因序列rbc L、mat K、psb A-trn H,同时筛选出合适的基因片段来研究多片段联合在枫斗类石斛植物鉴定方面的应用。结果表明:在7个基因片段中,nr DNA ITS、nad 1-intron 2和psb A-trn H的序列变异度较高,可用于枫斗类石斛的鉴定。但是,仅利用单一序列无法对细茎石斛及霍山石斛进行鉴别。而nr DNA ITS+nad 1-intron 2新型联合片段构建的UPGMA树与其他联合片段相比,更能有效的对枫斗类石斛基原植物进行鉴别并可区分细茎石斛及霍山石斛。此外,借助nr DNA ITS+nad 1-intron 2新型联合片段构建的UPGMA树对3个待检种进行了成功鉴别,分别为霍山石斛、细茎石斛和铁皮石斛。本研究首次将nr DNA ITS与线粒体基因片段联合用于枫斗类石斛的鉴定,为石斛药材的鉴别提供了可靠的科学依据,同时也将对规范枫斗类石斛市场提供理论指导。
The mitochondrial gene co I, nad 5 partial sequences and nad 1-intron 2 sequences, nr DNA ITS and chloroplast gene sequence rbc L were amplified from 39 individuals of 17 species of Dendrobium fimbriatus by PCR. mat K, psb A-trn H, at the same time screening out the appropriate gene fragments to study the multi-fragment combination in Maple bucket Dendrobium plant identification applications. The results showed that the sequence variations of nr DNA ITS, nad 1-intron 2 and psb A-trn H were high in seven gene fragments, which could be used for the identification of Dendrobium species. However, the use of only a single sequence can not distinguish between fine stem Dendrobium and Dendrobium huoshanense. However, compared with other joint fragments, the UPGMA tree constructed by the novel joint fragment of nrDNA ITS + nad 1-intron 2 is more effective in discriminating Dendrobium officinale and Dendrobium huoshanense. In addition, the UPGMA tree constructed with the novel nr DNA ITS + nad 1-intron 2 fragment successfully identified three species to be tested, namely Dendrobium huoshanense, Dendrobium nobile and Dendrobium candidum. This study, for the first time, used nR DNA ITS together with mitochondrial gene fragments for the identification of Dendrobium hunt for Dendrobium. It provided a reliable scientific basis for the identification of Dendrobium officinale and also provided theoretical guidance for the regulation of the Dendrobium candidum Dendrobium market.