To specifically identify Platycodon grandiflorum and its miscible products, a method was established by optimizing DNA extraction method,designing specific primers based on the ITS(internal transcribed spacer)sequence, and carrying out specific PCR amplification. The authenticity of P.grandiflorum was quickly determined by the amplification success rate. The results showed that the modified CTAB(cetyltrimethylammonium bromide)method gave relatively pure DNA and amplification was 100% successful, specific primers U1/D1 and specific PCR were designed based on the specific sites of P. grandiflorum 293 and 538. In the identification, only P.grandiflorum can be amplified to obtain a 264 bp specific band, and other medicinal materials were negative. The ITS sequence and the designed specific primers can be applied to identification of the P.grandiflorum and its common miscible products, which should provide scientific evidence for origin identification of P. grandiflorum.
ZHAO Xinyue
,
LIU Rui
,
FENG Hong
,
MAO Wenwen
,
CAO Fei
,
ZHANG Lanlan
. Molecular identification of Platycodon grandiflorum and its common miscible using DNA barcording[J]. Journal of Hebei University(Natural Science Edition), 2021
, 41(1)
: 60
-67
.
DOI: 10.3969/j.issn.1000-1565.2021.01.009
[1] 国家药典委员会.中华人民共和国药典一部[S].北京:中国医药科技出版社,2015:277.
[2] 朱立芬,王冰.桔梗皂苷D防御口腔黏膜上皮细胞感染白色念珠菌的作用[J].中国病理生理杂志,2017,33(1):161-165. DOI:10.3969/j.issn.1000-4718.2017.01.027.
[3] 陆文总,杨亚丽,贾光锋,等.桔梗多糖对U-14宫颈癌抗肿瘤作用的研究[J].西北药学杂志,2013,28(1):43-45. DOI:10.3969/j.issn.1004-2407.2013.01.018.
[4] 左军,尹柏坤,胡晓阳.桔梗化学成分及现代药理研究进展[J].辽宁中医药大学学报,2019,21(1):113-116. DOI:10.13194/j.issn.1673-842x.2019.01.031.
[5] 高山,马环宇.桔梗及其误用品的鉴别[J].中国中医药现代远程教育,2010,8(10):91. DOI:10.3969/j.issn.1672-2779.2010.10.080.
[6] 陈优芬.桔梗及其伪品的鉴别[J].中国药业,2010,19(10):74. DOI:10.3969/j.issn.1006-4931.2010.10.058.
[7] 王千枝.桔梗的真伪鉴别[J].科学技术创新,2011(3):44. DOI:10.3969/j.issn.1673-1328.2011.03.042.
[8] 万秋娥,刘汉平,张鹤鸣,等.激光拉曼光谱法无损鉴别人参及其伪品[J].光谱学与光谱分析,2012, 32(4):989-992. DOI:10.3964/j.issn.1000-0593(2012)04-0989-04.
[9] 陈士林,郭宝林,张贵君,等.中药鉴定学新技术新方法研究进展[J].中国中药杂志,2012,37(8): 1043-1055. DOI:10.4268/cjcmm20120801.
[10] CHEN S L,YAO H,HAN J P,et al. Validation of the ITS2 region as a novel DNA barcode for identifying medicinal plant species[J]. PLoS ONE,2010,5:8613. DOI:10.1371/journal.pone.0008613.
[11] 罗焜,陈士林,陈科力,等.基于芸香科的植物通用DNA条形码研究[J].中国科学:生命科学,2010,40(4):342-351. DOI:10.1007/s11427-010-0089-1.
[12] 朱英杰,陈士林,姚辉,等.重楼属药用植物DNA条形码鉴定研究[J].药学学报,2010,45(3):376-382. DOI:10.16438/j.0513-4870.2010.03.004.
[13] GAO T,YAO H,SONG J Y, et al. Evaluating the feasibility of using candidate DNA barcodes in discriminating species of the large Asteraceae family[J].BMC Evol Biol,2010,10:324. DOI:10.1186/1471-2148-10-324.
[14] 赵月梅,程敏,吴珍,等.基于ITS序列对桔梗及其伪品的鉴别[J].陕西农业科学,2018,64(3):76-78. DOI:10.3969/j.issn.0488-5368.2018.03.024.
[15] 王丹依,陈京,徐攀,等.ITS2序列作为DNA条形码鉴定紫堇属植物的有效性研究[J].浙江中医药大学学报,2017,41(2):97-102,111. DOI:10.16466/j.issn1005-5509.2017.02.001.
[16] 戴善光桔梗与其混淆品的鉴别[J].广西中医学院学报,2003,6(2):42-43. DOI:10.3969/j.issn.1008-7486.2003.02.027.
[17] 纪宝玉,裴莉昕,陈随清,等.山茱萸药材鲜品与干品总DNA提取方法比较[J].中国实验方剂学杂志, 2015,21(10):70-72. DOI:10.13422/j.cnki.syfjx.2015100070. (