山西中医药大学学报2025，Vol.26Issue(3)：265-270,6.DOI:10.19763/j.cnki.2096-7403.2025.03.06
基于ISSR-PCR鉴别青葙子及其混伪品
Identification of Celosiae Semen and its adulterants based on ISSR-PCR
摘要
Abstract
Objective:To identify Celosiae Semen,Cockscomb Semen and Green Amaranth by using ISSR-PCR mo-lecular marker technique.Methods:The CTAB method was used to extract DNA from the samples,and the concentration and quality of the sample DNA were detected by a nucleic acid protein analyzer.The quality of the sample DNA was verified by amplification according to ITS2 universal primers.According to the literature,30 ISSR primers were selected for PCR amplification and electrophoresis.The annealing temperature,primer amount,template amount and cycle times were inves-tigated to determine the best reaction system and conditions for ISSR-PCR amplification.Results:The ISSR-3 primer was selected as the best primer.There was a specific band around 400bp and 750 bp in the Celosiae Semen,and the 400bp band was brighter.There were specific bands between 100-250 bp,about 250 bp and 1000-2000 bp of Cockscomb characteristic band,and the 250bp band was more bright.No bands were produced between 250 and 500 bp in Green Amaranth.By opti-mizing the PCR amplification system and conditions,the optimal annealing temperature,primer amount,template amount,and cycle times were 50.8℃,2 μl,1 μl,and 35 times,respectively.The ISSR-3 primers with different bands were se-lected and could be used to identify Celosiae Semen,Cockscomb Semen and Green Amaranth.Conclusion:ISSR-3 primer can accurately identify Celosiae Semen and its adulterants under the optimized PCR reaction system and conditions.关键词
青葙子/ISSR-PCR/鉴别/混伪品Key words
Celosiae Semen/ISSR-PCR/identification/adulterants分类
医药卫生引用本文复制引用
王孟虎,孙一帆,马进进,刘冉冉,程鹤,范啸天,孟祥松,左亚锋,汤建,栗进才..基于ISSR-PCR鉴别青葙子及其混伪品[J].山西中医药大学学报,2025,26(3):265-270,6.基金项目
安徽高校协同创新项目(GXXT-2023-073) （GXXT-2023-073）
亳州学院项目(bzzc2023030,KJ2019ZD80,202312926021,YQYB2023064) （bzzc2023030,KJ2019ZD80,202312926021,YQYB2023064）
亳州职业技术学院项目(2023AH053216、BYK2303) （2023AH053216、BYK2303）
