1.Application of synthetic biology to sustainable utilization of Chinese materia medica resources.
Luqi HUANG ; Wei GAO ; Yongjin ZHOU
Acta Pharmaceutica Sinica 2014;49(1):37-43
Bioactive natural products are the material bases of Chinese materia medica resources. With successful applications of synthetic biology strategies to the researches and productions of taxol, artemisinin and tanshinone, etc, the potential ability of synthetic biology in the sustainable utilization of Chinese materia medica resources has been attracted by many researchers. This paper reviews the development of synthetic biology, the opportunities of sustainable utilization of Chinese materia medica resources, and the progress of synthetic biology applied to the researches of bioactive natural products. Furthermore, this paper also analyzes how to apply synthetic biology to sustainable utilization of Chinese materia medica resources and what the crucial factors are. Production of bioactive natural products with synthetic biology strategies will become a significant approach for the sustainable utilization of Chinese materia medica resources.
2.Climate suitable rank distribution of artemisinin content of Artemisia annua in China.
Xiaobo ZHANG ; Lanping GUO ; Luqi HUANG
Acta Pharmaceutica Sinica 2011;46(4):472-8
At the urgent request of Artemisia annua (ART) planting, the paper gets artemisinin content (ARTC) of ART in China from literatures. The paper analyses the relationships between ARTC and ecological factors by statistical analytical methods. The paper also analyses the climate suitable rank distribution of ARTC in China by ArcGIS. The results display that first, ARTC is significantly different in China, that ART from the south regions ARTC is higher. Greatest north parts of China have not suitable climate for the growing of ART and the ARTC is lower than 0.2%, when ART grows above the 34th degree of northern latitude. ARTC is higher and ART grows well, when ART grows under the 34 degrees N and grows at the areas between 100 degrees E and 120 degrees E. Second, subtropical zone is the best suitable climate zone for the growing of ART. ART grows well and ARTC is higher than 0.5%, when ART grows in the subtropical zone. Third, temperature, sunshine duration and rainfall are the main ecological factors that affect the growth of ART and the accumulation of ARTC. That the year temperature between 13.9 degrees C and 22 degrees C, sunshine duration between 853 h and 1507 h, rainfall between 814 mm and 1518 mm, is the best climate for the accumulation of ARTC. Temperature between 13 degrees C and 29 degrees C, rainfall between 600 mm and 1300 mm is the best climate for the growth of ART. Fourth, in northwest of Guangxi, eastern of Sichuan, Guizhou and Yunnan provinces, south Chongqing and west Hunan Province, there are suitable climate for the growth of Artemisia and for the accumulating of ARTC. There are also some suitable climate areas for the growing of artemisia in the south of Hubei, Anhui and Jiangsu provinces.
3.Attributes of Chinese Geoherbs and Its Study Strategy
Luqi HUANG ; Lanping GUO ; Guodong HUA
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(02):-
Objective To discovery the attributes of Chinese geoherbs and find it’s study strategy. Method Literature analysis and theory model structure. Results Both natural science and literae humaniores are attributes of Geoherbs. Conclusions Natural science combined with literae humaniores is the necessary way to study Geoberbs. The conception of Geographical Indications was favorable to discovery the literae humaniores properties. The theory about continuous variation was the key to found the scientific properties of Chinese Geoherbs.
4.Content Determination of Chlorogenic Acid in Shuanghua Eardrop by HPLC
Futian WANG ; Xi YANG ; Luqi HUANG
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(01):-
Objective To establish the method of content determination of chlorogenic acid in Shuanghua eardrop. Methods High performance liquid chromatography (HPLC) was adopted with Kromasil C18 column (5 ?m,4.6 mm?250 mm) and acetonitrile-0.4% phosph acid solution (10∶85) as the mobile phase. The detective wavelength was 328 nm,the flow rate was 1.0 mL/min,column temperature was room temperature. Results Average recovery was 99.8%,relative standard error was 1.51% (n =5). Conclusion The linearity relationship of chlorogenic acid was good at the range of 0.102 4~0.061 44 ?g.
5.Investigation of Resources and Varieties of Coptis Crude Drug Original Plant on Chinese Market
Xin LIU ; He HUANG ; Luqi HUANG ; Hezhen WU ; Yanfang YANG
China Pharmacist 2014;(10):1691-1695
Objective::To understand the resources distribution and varieties of crude drug Coptis chinensis Franch. on Chinese market to provide scientific basis for the resource conservation and variety collection and studies of the Chinese medicine. Methods:On the basis of literature survey, field investigation, mandatory survey and interview survey were conducted. Results:Due to the chan-ges in environment for many years and the predatory development in Coptis resources at the end of 20th century, the ecological environ-ment of wild Coptis resources changed significantly with destructed resources and decreased reserves, especially wild Yalian and Yun-lian were on the verge of extinction. The mainstream product on Chinese market was Weilian followed by Yunlian mainly used in small-scale medical clinic and medicine market in the distribution area, while Yalian commodity medicine could not found. Conclusion::The supply and demand on Coptis market should be stabilized to ensure herbalist interests. The production technology of Coptis need further improvement, cultivated area and suitable variety should be confirmed to guarantee the quality of Coptis medicinal materials. State should pay attention to the changes in Coptis resources and strengthen protection for wild Coptis resources to ensure the sustainable development of Coptis resources. Relevant government departments should guide the production of Coptis reasonably.
6.Cloning and polymorphism analysis of SmERF in Salvia miltiorrhiza.
Guanghong CUI ; Hua FENG ; Wenyuan LI ; Wanyi WANG ; Luqi HUANG
Acta Pharmaceutica Sinica 2010;45(9):1188-93
The transcription factor of ethylene responsive factor binding protein (ERF) is belonged to AP2/ERF superfamily, which is known to be unique in plants. AP2/ERF proteins have important functions in the transcriptional regulation of a variety of biological processes related to growth and development, as well as various responses to environmental stimuli. An ERF gene from Salvia miltiorrhiza is cloned and divided into ERF gene family group VII of Arabidopsis and Rice. It contains a MCGGAI (I/L) motif referred to as CMVII-1 and a single intron in the 5'-flanking region of the AP2/ERF domain. Sequence analysis reveals that the region of second extron has abundant polymorphism sites. There are 21 single nucleotide polymorphism sites (SNPs) in the 264 bp region, among them, 14 SNPs are synonymous substitutions and 7 SNPs are non-synonymous substitutions. Though analysis of 181 samples from Shandong, Shaanxi and Sichuan Provinces, it reveals that each production area has its own special genotypes, 5 SNPs show significant difference. Cluster based on UPGMA method reveals that different populations from specific province have clustered together. It shows that SmERF gene will be a candidate molecular marker for the identification of Salvia miltiorrhiza from different areas.
7.Molecular cloning and SNP analysis of a acetyl-CoA C-acetyltransferase gene (SmAACT) from Salvia miltiorrhiza
Guanghong CUI ; Xueyong WANG ; Hua FENG ; Jingxue ZHAO ; Luqi HUANG
Acta Pharmaceutica Sinica 2010;45(6):785-90
Acetyl-CoA C-acetyltransferase (AACT) is the first enzyme in the terpene synthesis pathway, catalyzed two units of acetyl-CoA to acetoacetyl-CoA. In order to study the tanshinone biosynthesis in Salvia miltiorrhiza, a novel AACT gene, SmAACT, was cloned using cDNA microarray and RACE strategy. The full length cDNA of SmAACT is 1 623 bp (accession No. EF635969), which contained a 1 200 bp open reading frame (ORF) encoding a 399 amino acid protein. Nine introns were found in the genomic sequence. SmAACT was upregulated by YE and Ag+ elicitors both with cDNA microarray and quantitative RT-PCR analyses along with the accumulation of tanshinones. Sequence homology comparison and phylogenetic analysis all suggested that SmAACT belonged to the class of acetyl-CoA C-acetyltransferase. The transcription level of SmAACT was relatively higher in root than that in stem and leaf tissues. SNP analysis revealed that SmAACT was highly variable in the region of 6 to 9 introns with 33 SNPs in the 600 bp region, there are 5 SNPs in the cDNA region while they are all synonymous cSNPs. Some special genotypes were found in Salvia miltiorrhiza from different areas. SmAACT will be an useful gene for further analyze the mechanism of gene regulation among the tanshinones biosynthesis.
8.High resolution melting and its application in identity study of traditional Chinese medicine.
Kang CHEN ; Chao JIANG ; Yuan YUAN ; Luqi HUANG
Acta Pharmaceutica Sinica 2015;50(12):1581-8
High resolution melting (HRM), based on melting curve analysis, requires not only saturating dyes that fluoresce in the presence of double-stranded DNA, but also higher resolution detection equipment. The melting curve is a novel method for sequence matching, genotyping and mutation scanning. The technology is simple, accurate, rapid, closed-tube, low-cost, and high-throughput, which make it gain more and more applications. This review article presents the basic principles, key factors and both the advantage and limitations of HRM. The potential application is discussed in the study of molecular identity of traditional Chinese medicine.
9.Development of microsatellites and genetic diversity analysis of Scutellaria baicalensis Georgi using genomic-SSR markers.
Linjie QI ; Ping LONG ; Chao JIANG ; Yuan YUAN ; Luqi HUANG
Acta Pharmaceutica Sinica 2015;50(4):500-5
A total of 12 775 SSRs were identified from Scutellaria baicalensis Georgi genomic database, accounting for 2.56% of the total genomic sequences. The result showed that S. baicalensis SSRs were based on 68.32% dinucleotide and 18.63% trinucleotide repeats; CT/GA and TTC/GAA were predominant in the dinucleotide motifs and the trinucleotide motifs respectively. Nine primers were selected to produce highly reproducible SSR bands and were used in studying the genetic diversity of S. baicalensis, 50 individuals from ten populations. 68 SSR polymorphic loci were detected, these loci were polymorphic and displayed 4 to 12 alleles per locus with a mean number of 7; the effect number of alleles was 3. Expected heterozygosities were 0.6 and were far more greater than the average in dicotyledonous plants. PIC (polymorphism information content) was 0.72, Shannon's information index was 1.32, these all proved that S. baicalensis had a high genetic diversity in general. Genetic differentiation among population Gst was 0.131, genetic variation among population accounted for 13.1% and genetic variation within population accounted for 86.9%. The cluster analysis showed that 10 populations S. Baicalensis were classified into 2 groups, but it was not associated with geographical distribution.
10.Bioinformatics analysis of DNA demethylase genes in Lonicera japonica Thunb.
Linjie QI ; Yuan YUAN ; Chong WU ; Luqi HUANG ; Ping CHEN
Acta Pharmaceutica Sinica 2015;50(3):367-71
The DNA demethylase genes are widespread in plants. Four DNA demethylase genes (LJDME1, LJDME2, LJDME3 and LJDME4) were obtained from transcriptome dataset of Lonicera japonica Thunb by using bioinformatics methods and the proteins' physicochemical properties they encoded were predicted. The phylogenetic tree showed that the four DNA demethylase genes and Arabidopsis thaliana DME had a close relationship. The result of gene expression model showed that four DNA demethylase genes were different between species. The expression levels of LJDME1 and LJDME2 were even more higher in Lonicera japonica var. chinensis than those in L. japonica. LJDME] and LJDME2 maybe regulate the active compounds of L. japonica. This study aims to lay a foundation for further understanding of the function of DNA demethylase genes in L. japonica.