Identification of Corydalis yanhusuo,C. turtschaninovii,C. decumbens by allele-specific PCR.
10.19540/j.cnki.cjcmm.20190527.108
- Author:
Qing-Yun CHAN
1
;
Lu JIANG
1
;
Ming-En CHENG
1
;
Shan-Shan CHU
1
;
Da-Qing YU
1
;
Jin XIE
2
;
Liang-Ping ZHA
3
;
Hua-Sheng PENG
3
Author Information
1. School of Pharmacy,Anhui University of Chinese Medicine Hefei 230012,China.
2. School of Pharmaceutical Sciences,Anhui Medical University Hefei 230032,China.
3. School of Pharmacy,Anhui University of Chinese Medicine Hefei 230012,China Institute of Conservation and Development of Traditional Chinese Medicine Resources,Anhui Academy of Chinese Medicine Hefei 230012,China.
- Publication Type:Journal Article
- Keywords:
Corydalis decumbens;
Corydalis turtschaninovii;
Corydalis yanhusuo;
molecular identification;
specific polymerase chain reaction
- MeSH:
Alleles;
Corydalis;
classification;
genetics;
Genes, Plant;
Genetic Markers;
Polymerase Chain Reaction
- From:
China Journal of Chinese Materia Medica
2019;44(15):3261-3267
- CountryChina
- Language:Chinese
-
Abstract:
To establish a DNA molecular markers method for identification of Corydalis yanhusuo,C. turtschaninovii and C. decumbens,the mat K,trn G and psb A-trn H sequences of 56 samples from 14 species of C. yanhusuo,C. turtschaninovii,C. decumbens and their related species were obtained by sequencing. The SNP loci were obtained by Bio Edit 7. 2. 2 software. The primers for AS-PCR identification were designed based on the mutation sites,and the conditions of PCR were optimized to identify C. yanhusuo,C. turtschaninovii,and C. decumbens according to the specific bands. The results showed that the amount of template( 0. 6-1 200 ng)and annealing temperature( 42-60 ℃) had little influence on the amplification results,and the number of cycles had much influence on the amplification results. When the number of cycles was 20,the specific bands of 297 bp( mat K),353 bp( trn G) and 544 bp( mat K) were amplified from C. yanhusuo,C. turtschaninovii and C. decumbens,respectively. The method established in this study had a minimum detection limit of 6 ng for C. yanhusuo,60 ng for C. decumbens and less than 0. 6 ng for C. turtschaninovii. Thus,the allelespecific PCR method established in the research can specifically identify C. yanhusuo,C. turtschaninovii,and C. decumbens.
