目的 设计特异性聚合酶链式反应(PCR)引物,建立川麦冬PCR鉴定方法。方法 回收随机扩增多态性DNA标记(RAPD)扩增筛选到的川麦冬分子标记CM503基因,将其连接进入T-载体克隆并测序。根据测序结果设计一对特异性引物CM1/CM2,以麦冬基因组DNA为模板进行特异性PCR扩增。结果 PCR鉴定结果为在退火温度68 ℃时,川麦冬在297 bp处出现特异性扩增条带,其他麦冬品种则未出现条带。结论 实验室建立的川麦冬PCR鉴定方法具有操作简单、准确、灵敏、重复性好等优点,应用前景理想。
Abstract
OBJECTIVE To design specific PCR primers and establish the PCR identification method of Ophiopogon japonicas from Sichuan.METHODS The gene footprint of Ophiopogon japonicas from Sichuan named CM503 was screened from random amplified polymorphic DNAC(RAPD) amplification. Reclaimed CM503 gene was inserted into T-vector to be cloned and sequenced. One pair of specific primers CM1/CM2 were designed according to the CM503 sequence and applied in specific PCR using the genomic DNA of Ophiopogon japonicas from Sichuan as template.RESULTS A specific band around 297 bp was detected in Ophiopogon japonicus from Sichuan at 68 ℃, while nothing appeared for the other varieties.CONCLUSION The method is convenient, reproducible, and precise, with broad application prospects.
关键词
川麦冬 /
随机扩增多态性DNA标记 /
中药 /
分子鉴定 /
PCR鉴定 /
分子鉴定标记
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Key words
Sichuan Ophiopogon japonicus /
random amplified polymorphic DNAC /
traditional Chinese medicine /
molecular identification /
PCR identification /
molecular identification marker
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中图分类号：
R284
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参考文献
[1] Ch. P(2015)Vol Ⅰ(中国药典2015年版.一部)[S]. 2015:155-156.
[2] ZHANG L T. Quality evaluation on resources of Radix Ophiopogonis[D]. Nanjing:Nanjing Agricultural University,2009.
[3] LIU F,XU Z Y,YAN W,et al. The determination and analysis of effective content in geo-authentic crude drugs in sichuan province-Ophiopogon japonicus[J]. J China West Nor Univ (Nat Sci)(西华师范大学学报:自然科学版),2011,32(4):362-365.
[4] JIN H,WANGH D,HE L,et al. Comparison on histology of root tubers and fibrous roots of Ophiopogon japonicus and ophiopogonins content[J]. Chin Tradit Herb Drugs(中草药),2014,45(7):1002-1005.
[5] LIU C L,GUAN C Y,LI X. Studies on the authenticity of plant RAPD marker[J]. Biotechnol Bull(生物技术通报),1999,(2): 31-34.
[6] GAO J. A study on genetic diversity of wild Sichuan Province-Ophiopogon japonicus population[D]. Chendu:Sichuan Agricultural University,2009.
[7] LI L Y,CHEN D X,ZHONG G Y, et al. Analysis on genetic relationship of plants in Curculigo Gaertn. from China by RAPD [J]. Chin Tradit Drugs(中草药),2011,42(5): 980-984.
[8] LI M,ZHAO X. RAPD analysis of three species of Fritillaria in southern China[J]. J Zhejiang Univ Technol (浙江工业大学学报),2012,40(6):634-638.
[9] LI M, HUANG L M, ZHAO X. Specific PCR identification of Fritillaria thunbergii[J]. Chin Tradit Drugs(中草药), 2014, 45(12): 1754-1757.
[10] LI M, HUANG L M, CHEN Q, et al. The screening of Ohpiopogon japonicus molecular marker by RAPD analysis[J]. J Zhejiang Univ Technol (浙江工业大学学报),2014,42(5):504-508.
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