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根据细叶石斛及其它37种枫斗类和黄草类石斛的rDNAITS序列,我们设计了位点特异性PCR鉴别引物XY-JB01S和XY-JB01X,对细叶石斛进行了成功的DNA分子鉴别。在进行位点特异性鉴别PCR之前,首先运用扩增ITS区的通用引物P1、P2对模板DNA进行扩增,以验证模板的可靠性和扩增的合适浓度。当退火温度上升为64℃,只有细叶石斛的模板DNA能被扩增出来,而其它的37种石斛属植物均为阴性。该鉴别反应重复性好,已在鉴别细叶石斛中发挥重要作用。与DNA测序鉴别方法相比,位点特异性PCR具有简单、省时、高效、准确等优点。
Based on the rDNA ITS sequences of Dendrobium nobile and other 37 species of Dendrobium species and Dendrobium candidum, we designed site-specific PCR primers XY-JB01S and XY-JB01X to identify DNA molecules successfully. Prior to performing site-specific PCR, the template DNA was first amplified using universal primers P1 and P2 in the amplified ITS region to verify the reliability of the template and the appropriate concentration for amplification. When the annealing temperature rose to 64°C, only template DNA of Dendrobium could be amplified, while the other 37 Dendrobium species were negative. The reproducibility of the identification reaction is good and has played an important role in identifying the Dendrobium nobile. Compared with DNA sequencing identification methods, site-specific PCR has the advantages of simplicity, time saving, high efficiency, and accuracy.