RESUMO
OBJECTIVE: To establish a stabilized and reliable detection system of CNVs of HMGR, SQS1, beta-AS gene of Glycyrrhiza uralensis. METHOD: Real time PCR was used to detect the CNVs of HMGR, SQS1, beta-AS gene of G. uralensis. RESULT: In the quantitative detection experiments of HMGR, SQS1, beta-AS gene of G. uralensis, the change of value of C(t) was 25.82-25.88, 29.01-29. 08, 15.52-15.56, 19.06-19.08 respectively, the alue of SD was 0.033, 0.032, 0.024, 0.011 respectively, and the value of CV was 0.12%, 0.22%, 0.16%, 0.06% respectively. CONCLUSION: The repeatability of detection system of Real time PCR was stabilized and reliable, and the method could be used to detect the CNVs of HMGR, SQS1, beta-AS gene of G. uralensis.
Assuntos
Variações do Número de Cópias de DNA , Farnesil-Difosfato Farnesiltransferase/genética , Glycyrrhiza uralensis/genética , Hidroximetilglutaril-CoA Redutases/genética , Tipagem Molecular/métodos , DNA de Plantas , Glycyrrhiza uralensis/enzimologia , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de DNARESUMO
This study is to reveal the correlation between CNVs of HMGR, SQS1, beta-AS gene and genuineness of liquorice. Real-time PCR was used to detect the copy number of HMGR, SQS1, beta-AS gene of liquorice. According to the results, the range of the copy number variation of HMGR gene was between 1 and 3, the copy number of SQS1 gene was 1 or 2, and the copy number of beta-AS gene was only 1. On the basis of the copy number of HMGR, SQS1 and beta-AS gene, there were five groups, type A (2 + 1 + 1), type B (1 + 1 + 1), type C (3 + 2 + 1), type D (2 + 2 + 1) and type E (3 + 1 + 1). There were two types, type A and type B, in Hangjinqi of Inner Mongolia, and the ratio of A to B was 1:1.3. There were also two types, type A and type B, in Chifeng of Inner Mongolia, and the ratio of A to B was 3:1. There were four types, type A, type B, type C and type D, in Yanchi of Ningxia province, and the ratio of A to B was 1:5.1. There were three types, type A, type B and type E, in Minqin of Gansu province, and the ratio of A to B was 2:1. So CNVs mainly existed in the liquorice from Ningxia and Gansu provinces. While the genetic background of liquorice from Hangjinqi of Inner Mongolia was stabilized. The results of the experiment proved that the correlation between CNVs and origins was one of the reasons of genuineness of liquorice.
Assuntos
Variações do Número de Cópias de DNA , Farnesil-Difosfato Farnesiltransferase/genética , Glycyrrhiza uralensis/genética , Hidroximetilglutaril-CoA Redutases/genética , Transferases Intramoleculares/genética , China , DNA de Plantas/genética , Glycyrrhiza uralensis/enzimologia , Reação em Cadeia da Polimerase em Tempo RealRESUMO
OBJECTIVE: To classify Pueraria lobata originated from different geographical regions based on ITS,psbK-psbI and trnH-psbA information. METHOD: Twenty-four samples of P. lobata were collected from northeast China, north China, central China and northwest China. DNA extraction, PCR, sequence and genotypes/haplotypes analysis were performed . RESULT: ITS1, 5.8S and ITS2 varied only 1 bp respectively, psbK-psbI 2 bps; trnH-psbA varied 1 bp and 10 bp deletion. CONCLUSION: Based on the variation of ITS,psbK-psbI and trnH-psbA, 4 genotypes and 2 haplotypes were identified, respectively.
