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李雄英, 罗育, 吴耀生, 蒋军富, 赵瑞强, 蓝秀万. RAPD技术在药用植物绞股蓝鉴别中的初步研究[J]. 植物科学学报, 2008, 26(3): 251-254.
引用本文: 李雄英, 罗育, 吴耀生, 蒋军富, 赵瑞强, 蓝秀万. RAPD技术在药用植物绞股蓝鉴别中的初步研究[J]. 植物科学学报, 2008, 26(3): 251-254.
LI Xiong-Ying, LUO Yu, WU Yao-Sheng, JIANG Jun-Fu, ZHAO Rui-Qiang, LAN Xiu-Wan. Initial Research on Authentication of Medicinal Plant Gynostemma pentaphyllum by RAPD Method[J]. Plant Science Journal, 2008, 26(3): 251-254.
Citation: LI Xiong-Ying, LUO Yu, WU Yao-Sheng, JIANG Jun-Fu, ZHAO Rui-Qiang, LAN Xiu-Wan. Initial Research on Authentication of Medicinal Plant Gynostemma pentaphyllum by RAPD Method[J]. Plant Science Journal, 2008, 26(3): 251-254.

RAPD技术在药用植物绞股蓝鉴别中的初步研究

Initial Research on Authentication of Medicinal Plant Gynostemma pentaphyllum by RAPD Method

  • 摘要: 运用RAPD技术对绞股蓝(Gynostemma pentaphyllum)及其伪品进行DNA指纹图谱的鉴别研究。采用改进的CTAB法提取七叶绞股蓝、五叶绞股蓝、乌蔹莓(Cayratia japonica)3种植物的总DNA,主要以七叶绞股蓝DNA为模板,采用随机引物WGS001进行PCR扩增,对反应体系包括模板、Mg2+Taq酶、牛血清白蛋白(BSA)、退火温度进行优化。优化的反应体系总体积25μL,含MgCl22 mmol/L、dNTP 0.2 mmol/L、引物0.4μmol/L、模板60 ng、Taq酶1 U、BSA2μg/μL,退火温度58℃。用10条含20个碱基的随机引物对以上3种植物总DNA作PCR扩增,进行引物筛选。筛选得到的两条随机引物(WGS001、WGS004)可扩增得到识别这些物种基因组DNA的多态性片段。这些片段可以有效地鉴别绞股蓝和乌蔹莓。

     

    Abstract: The random amplified polymorphic DNA(RAPD) analysis was applied to identify the resources of medicinal plant Gynostemma pentaphyllum and its spurious breed with DNA fingerprints.The modified CTAB method was used to extract the genomic DNA of several plants,including seven leaves Jiaogulan,five leaves Jiaogulan,Cayratia japonica.With the DNA extracted from seven leaves Jiaogulan as template,the random primer WGS001 for PCR has been done meanwhile RAPD reaction system has been optimized so that it could be determined for the suitable annealing temperature and concentrations of template,Mg2+,primer,Taq DNA polymerase,BSA.In the optimized system of 25 μL,MgCl2 2 mmol/L,primer 0.4 μmol/L,template 60 ng,Taq DNA polymerase 1 U,BSA 2 μg/μL and annealing temperature 58℃ were used.Based on the optimized system,ten primers of 20 bp were applied to do random amplification with total DNAs of the above mentioned three plants,respectively.The polymorphic fragments of DNA fingerprints of the above mentioned three plants were obtained with two screened random primers(WGS001,WGS004).These polymorphic fragments can be used to distinguish G.pentaphyllum with C.japonica obviously.

     

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