Development and Application of Mitochondrial and Chloroplast Microsatellite Markers for Codonopsis Plants
10.13422/j.cnki.syfjx.20211419
- VernacularTitle:党参线粒体和叶绿体微卫星标记的开发及应用
- Author:
Dan ZHANG
1
;
Chen-hui DU
1
;
Xiang-ping PEI
1
;
Xiao-li LIU
1
;
Hai-xian ZHAN
1
Author Information
1. School of Chinese Medicine and Food Engineering，Shanxi University of Traditional Chinese Medicine，Jinzhong 030619，China
- Publication Type:Research Article
- Keywords:
Codonopsis;
chloroplast;
mitochondria;
microsatellite;
development and application
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2021;27(23):153-162
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To develop the specific molecular markers of Codonopsis plants and better identify their germplasm resources considering the significant difference in active ingredients of Codonopsis Radix from various origins and producing areas. Method:Such bioinformatics software as Primer 5.0， NTSYS-pc 2.10e， and PopGene 32 were used for searching the simple sequence repeat （SSR） markers of C. minima chloroplast genome， C. tsinlingensis chloroplast， and C. lanceolata mitochondrial sequences， and 120 pairs of SSR primers were designed by Primer 5.0. Then 16 pairs of cpSSR primers and 10 pairs of mtSSR primers with good screening effect and high polymorphism were selected for analyzing the interspecific versatility of 20 samples. Result:The results showed that 66 cpSSR primer sites and 26 mtSSR sites were identified from the genome sequences， with 86.20% of single nucleotide， 6.9% of dinucleotide， and 6.9% of trinucleotide for C. minima chloroplast， 83.78% of single nucleotide，13.51% of dinucleotide， and 2.71% of trinucleotide for C. tsinlingensis chloroplast， and 46.15% of single nucleotide and 53.85% of dinucleotide for C. lanceolata mitochondria. As demonstrated by polymerase chain reaction （PCR） identification results， the developed 26 pairs of SSR primers had good applicability in the genus Codonopsis. The analysis by NTSYS-pc 2.10e revealed that the genetic similarity coefficients of 20 samples were within the range of 0.38-1.00， and they were divided into two subgroups at a threshold of 0.69. Four pairs of polymorphic primers were screened out in the diversity analysis of 20 samples using PopGene 32. The number of observed alleles （Na） was 12， and the effective number of alleles （Ne） ranged from 1.362 9 to 2.605 9. The percentage of polymorphic loci （PPL） at each site was 100%， and the average values of genetic parameters Ho， He， and I at each site were 0.555 8， 0.444 2， and 0.753 2， respectively， indicating high polymorphism at each site. The screened four pairs of primers were utilized for DNA fingerprinting of the 20 samples， and it was found that the DNA fingerprints enabled the identification of these 20 samples. Conclusion:This study has provided a molecular basis for the study of the genetic relationship between plants in species Codonopsis and the intraspecific genetic differentiation.
