摘要
目的:建立一种能特异性检测青葙子及其混伪品的分子鉴定方法。方法:针对青葙子SRAP标记筛选并设计特异性引物,各地采集青葙子、鸡冠花子、苋菜子、刺苋子和反枝苋子样品,提取DNA作为模板,建立并优化PCR反应条件,利用特异性引物对不同产地的青葙子及其混伪品进行扩增和电泳检测,根据特异性条带的大小及有无进行真伪及掺伪鉴别,并对其检出限、耐受性和适用性进行考察与验证。结果:退火温度为60℃、循环次数为30次时,青葙子、鸡冠花子分别在206和581 bp处出现单一明亮条带,其他混伪品及空白对照均无此条带;该方法对青葙子药材DNA的最低检测限为1 ng;对于青葙子中掺杂鸡冠花子的检出限是2%。结论:建立的特异性PCR方法可准确鉴定青葙子及其混伪品。
Objective:To establish the specific molecular identification method for the detection of Celosia argentea seeds and its adulteration.Methods:The specific primers were designed and screened by the SRAP markers of Celosia argentea seeds.The seeds of Celosia argentea,Celosia cristata,Amaranthus tricolor,Amaranthus spinosus,Amaranthus retroflexus from different places were collected as samples,and the DNA was extracted as PCR templates.The PCR reaction conditions were established and optimized,the Celosia argentea seeds and its adulterants from different regions were amplified with specific primers and detected by electrophoresis,and the authenticity and adulteration were identified according to the size and presence of specific bands.The detection limit,tolerance and applicability of the method were investigated and verified.Results:When the annealing temperature was 60℃and the cycle number was 30,the single bright bands appeared at 206 and 581 bp of the seeds of Celosia argentea and Celosia cristata,respectively,there was no such band for other adulterants and blank control.The DNA minimum detection limit of this method was 1 ng for Celosia argentea seeds and 2%to detect the adulterated Celosia cristata seeds in Celosia argentea seeds.Conculsion:The allele-specific PCR method established in the research can accurately identify Celosia argentea seeds and its adulterants.
作者
谭新宁
吴文如
来慧丽
梁婉晴
行冰楠
TAN Xin-ning;WU Wen-ru;LAI Hui-li;LIANG Wan-qing;XING Bing-nan(School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Guangdong Food and Drug Vocational College,Guangzhou 510520,China)
出处
《中药材》
CAS
北大核心
2021年第8期1839-1843,共5页
Journal of Chinese Medicinal Materials
基金
国家中医药管理局全国中药资源普查项目(GZY-KJS-2018-004)
全国中医药创新骨干人才培训项目(国中医药人教函[2019]128号)
广州中医药大学省级大学生创新创业训练计划项目(S202010572075)
中央本级重大增减支项目(2060302)
关键词
青葙子
鸡冠花子
聚合酶链式反应
分子鉴定
掺伪
Seeds of Celosia argentea L.
Seeds of Celosia cristata L.
Polymerase chain reaction
Molecular identification
Adulteration
