Identification of Astragalus and its adulterants based on ITS2sequence and secondary structure
10.16438/j.0513-4870.2019-0869
- VernacularTitle:基于ITS2序列及二级结构对药用黄芪及混伪品的鉴别研究
- Author:
Ya-ling LIU
1
;
Ya-ping GENG
1
;
Fang WANG
1
;
Xiao-dong XIE
1
;
Peng-fei ZHANG
2
;
Jian-ping LIANG
1
;
De-wang LIU
3
Author Information
1. College of Life Science, Shanxi Agricultural University, Jinzhong 030801, China
2. College of Horticulture, Shanxi Agricultural University, Jinzhong 030801, China
3. College of Pharmacy, Inner Mongolia Medical University, Hohhot 010059, China
- Publication Type:Research Article
- Keywords:
italic>Astragalus;
ITS2 sequence;
secondary structure;
molecular identification
- From:
Acta Pharmaceutica Sinica
2020;55(3):522-529
- CountryChina
- Language:Chinese
-
Abstract:
To effectively identify the Astragalus and its adulterants based on ITS2 sequence and secondary structure, in this study, 32 portions of Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Beg.) Hsiao and Astragalus membranaceus (Fisch.) Bge. collected were conducted ITS2 sequence amplification and bidirectional sequencing, whose results were then spliced by CExpress software remove the 5.8S and 28S sequences at both ends to obtain a complete ITS2 sequence. In addition, 3 ITS2 sequences for each of the adulterants of Astragalus, respectively, Oxytropis coerulea, Caragana sinica, Hedysarum polybotrys, Althaea rosea were downloaded from GenBank. The intra-specific and inter-specific genetic distances were calculated by the software MEGA7 to analyze the difference of each sequence; the Neighbor-joining (NJ) method was used to construct the phylogenetic tree based on ITS2 sequence (primary structure) as well as joint ITS2 sequence and its secondary structure. The results showed that the average ITS2 sequence length of both A. mongolicus and A. membranaceus was 216 bp, and their average GC content was 50.00% and 50.46%, respectively. The similarity of ITS2 sequence length and GC content between the two kind of Astragalus and Oxytropis coerulea was the highest, while the ITS2 sequence length and GC content of Althaea rosea showed great differences with those of Astragalus. The inter-specific distance between Astragalus and Oxytropis coerulea was the smallest, while that between the medicinal Astragalus and Hedysarum polybotrys, Caragana sinica as well as Althaea rosea was great. The phylogenetic trees constructed based on the ITS2 sequence (primary structure) and joint ITS2 sequence and its secondary structure showed that the topological relations of the two phylogenetic trees were basically the same, and both could effectively identify the Astragalus and its adulterants. What’s more, the addition of secondary structure information made end branch of the phylogenetic tree become more in its construction, and the distinguish ability and approval rating were also improved, which further reflected the genetic relationship of Astragalus and its adulterants. This provides some scientific basis for classification and accurate identification of Astragalus and its adulterants.
