Li Tongxiang, Wang Jinke, Lu Zuhong
Chien-Shiung Wu Laboratory, Southeast University, Nanjing 210096, P.R. China.
J Biochem Biophys Methods. 2005 Feb 28;62(2):111-23. doi: 10.1016/j.jbbm.2004.10.006. Epub 2004 Nov 21.
About 63 species of Dendrobium are identified in China, making the identification of the origin of a particular Dendrobium species on the consumer market very difficult. We report evaluation of multiple species-specific probes screened from genomic DNA for closely related Dendrobium species identification, based on DNA array hybridization. Fourteen species-specific probes were screened from five closely related Dendrobium species, D. aurantiacum Kerr, D. officinale Kimura et Migo, D. nobile Lindl., D. chrysotoxum Lindl. and D. fimbriatum Hook., based on the SSH-Array technology we developed. Various commercial Dendrobium samples and unrelated samples were definitely identified. The specificity and accuracy of the multiple species-specific probes for species identification was assessed by identifying various commercial Dendrobium samples (Herba Dendrobii). Hybridization patterns of these multiple probes on digested genomic DNAs of Dendrobium species indicated that there are distinct polymorphic sequence fragment in the higher eukaryotes. This is the first report on detection and utilization of multiple species-specific probes of Dendrobium in whole genomic DNA, and this could be useful tools not only for a new technical platform for the closely related species identification but also for epidemiological studies on higher eukaryotes.
中国已鉴定出约63种石斛属植物，这使得在消费市场上鉴定某一特定石斛属植物的来源变得非常困难。我们报告了基于DNA芯片杂交技术，对从基因组DNA中筛选出的多种物种特异性探针进行评估，以用于近缘石斛属物种的鉴定。基于我们开发的SSH-Array技术，从五种近缘石斛属植物——橘黄石斛、铁皮石斛、金钗石斛、鼓槌石斛和流苏石斛中筛选出了14种物种特异性探针。各种市售石斛样品和无关样品都得到了明确鉴定。通过鉴定各种市售石斛样品（石斛），评估了多种物种特异性探针用于物种鉴定的特异性和准确性。这些多种探针在石斛属植物消化后的基因组DNA上的杂交模式表明，高等真核生物中存在明显的多态性序列片段。这是关于在全基因组DNA中检测和利用多种石斛属物种特异性探针的首次报道，这不仅可以作为近缘物种鉴定的新技术平台，也可用于高等真核生物的流行病学研究。
