1.Construction of genetic linkage map of Bupleurum chinense DC. using ISSR and SSR markers.
Qingqing ZHAN ; Chun SUI ; Jianhe WEI ; Shengci FAN ; Jie ZHANG
Acta Pharmaceutica Sinica 2010;45(4):517-23
Abstract: Molecular genetic map is a fundamental organizational tool for genomic research. However, a genetic linkage map for Bupleurum chinense DC. has not been developed. In this study, with the theory of pseudo-testcross, 96 F1 plants from an intraspecific cross of B. chinense were used as mapping populations. Twenty eight ISSR (inter-simple sequence repeat) primers and 44 SSR (simple sequence repeat) primers were used to detect the polymorphisms between the parental plants, and of them, 28 ISSRs and 14 SSRs were selected to analyze the F1 populations. The map consisted of 13 linkage groups which included 80 (72 ISSRs and 8 SSRs) loci, and covered 2 633.9 cM with an average density of 33.4 cM. All 13 linkage groups consisted of 2-31 loci ranging in length from 15.4-1295.7 cM. This map will provide a basis for studies on gene mapping, map-based cloning and maker-assisted selection of important traits in B. chinense.
2.NMR-based analysis of water soluble extracts of different Astragali Radix.
Dong TIAN ; Zhenyu LI ; Shengci FAN ; Jinping JIA ; Xuemei QIN
Acta Pharmaceutica Sinica 2014;49(1):89-94
Water soluble extract (WSE) is an important index for the quality evaluation of Astragali Radix (AR). In this study, the WSE of the wild AR from Shanxi province (SX) and the cultivated AR from Gansu Province (GS) were compared. The WSEs of two types of AR were determined according to the appendix of Chinese pharmacopoeia. Then the WSEs were subjected to NMR analysis, and the obtained data were analyzed using HCA, PCA, OPLS-DA, microarray analysis, and Spearman rank analysis. In addition, the Pearson correlation of differential metabolites were also calculated. The results showed that the WSE content of GS-AR (37.80%) was higher than that of SX-AR (32.13%). The main constituent of WSE was sucrose, and other 18 compounds, including amino acids, organic acids, were also detected. Multivariate analysis revealed that SX-AR contained more choline, succinic acid, citric acid, glutamate, taurine and aspartate, while GS samples contained more sucrose, arginine and fumaric acid. In addition, the Pearson correlations between different metabolites of the two types of AR also showed apparent differences. The results suggested that the WSE of two types of AR differs not only in the content, but also in the chemical compositions. Thus, the cultivation way is important to the quality ofAR. This study supplied a new method for the comparison of extract of herbal drugs.
3.Between Hengshanhuangqi and Chuanhuangqi based on metabolomics and ITS2 sequences.
Pan HE ; Zhenyu LI ; Shengci FAN ; Fusheng ZHANG ; Xuemei QIN ; Guojun DU
Acta Pharmaceutica Sinica 2013;48(10):1595-601
To compare the differences between Hengshanhuangqi (HH) and Chuanhuangqi (CH) at molecular level, 1H NMR based plant metabolomics approach was used to reveal the chemical difference between HH and CH. Then, the contents of astragaloside IV and calycosin-7-O-beta-D-glucoside, the marker compounds specified in China Pharmacopoeia, were determined. In addition, the ITS2 fragments of HH and CH were sequenced. Twenty-three metabolites were identified in the 1H NMR spectrum, and the principal component analysis showed CH and HH could be separated clearly. HH contained more aspartic acid, GABA, citric acid, astragaloside IV and calycosin-7-O-beta-D-glucoside, while CH contained more threonine, alanine, acetic acid, choline, arginine, fructose and sucrose. And the astragaloside IV is almost undetectable in CH. In addition, the ITS2 fragment sequences of HH and CH were different at eight bases. Thus, the HH and CH showed significant differences chemically and genetically.