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.

Diterpenes, an important class of natural compounds, are widely distributed in nature. As the valuable diterpenoids continue to be found, diterpene synthase in the course of diterpene synthesis get as much attention as possible. The multiformity of end-product-diterpenoids were also due to the diversity of diterpene synthase. This paper focuses on the advances in recent biosynthesis pathway of diterpene and types, cloning, catalytic mechanism, synthetic biology application.

Objective:To observe the clinical efficacy of pressing-kneading manipulation combined with herb-insulated moxibustion at Shuidao(ST28)for postpartum urinary retention after labor analgesia and its effect on bladder urination function. Methods:A total of 154 patients with postpartum urinary retention after labor analgesia were randomly divided into a Western medication group and a herb-insulated moxibustion group,with 77 cases in each group.In the Western medication group,neostigmine sulfate was injected into Zusanli(ST36).In the herb-insulated moxibustion group,after pressing-kneading manipulation at Shuidao(ST28),herb-insulated moxibustion was applied to Shuidao(ST28)with self-made Tong Quan San.Both groups were treated once,and the clinical efficacy was evaluated 5 h after treatment.The first urination time,first urination volume,average urinary flow rate,bladder residual urine volume,hospitalization days,and costs were recorded. Results:The total effective rate and markedly effective rate of the herb-insulated moxibustion group were higher than those of the Western medication group(P<0.05),the time to the first urination and residual urine volume in the bladder of the herb-insulated moxibustion group were shorter or smaller than those of the Western medication group(P<0.01),the first urination volume and average urine flow rate of the herb-insulated moxibustion group were larger than those of the Western medication group(P<0.01).There were no significant differences in the hospitalization days and costs between the two groups(P>0.05). Conclusion:Pressing-kneading manipulation combined with herb-insulated moxibustion at Shuidao(ST28)can effectively treat postpartum urinary retention after labor analgesia and improve bladder urination function.

This study reported the obtainment of the full-length cDNA of Salvia miltiorrhiza hairy roots (Abbr: SmHDR, GenBank number: JX233817), via extracting Salvia miltiorrhiza hairy roots total RNA, designing specific primers according to the transcriptome data and using the RACE strategy, and then analyzed it with bioinformatics approaches. On this basis, using the real-time PCR to detect SmHDR gene expression after Ag+ induction, and testing tanshinones contents of corresponding samples by UPLC. SmHDR has 1 647 nucleotides, and an open reading frame (ORF) encoding a protein of 463 amino acid residues. The deduced protein has isoelectric point (pI) of 5.72 and a calculated molecular weight about 51.88 kD. In the secondary structure, the percentage of alpha helix, beta turn and random coil were 35.64%, 20.30% and 44.06%, respectively. Sequence alignment and phylogenetic analysis demonstrated that SmHDR had relative close relationship to the HDR of Picrorhiza kurrooa, similar to HDR from other species of plants. Real time PCR results indicated that elicitor of Ag+ stimulated the increase of mRNA expression of SmHDR. At the same time, results of ultra performance liquid chromatography (UPLC), used to examine the accumulation of diterpenoid tanshinones in hairy roots, showed that the contents of diterpenoid tanshinones in hairy roots of Salvia miltiorrhiza were increased dramatically at 12 h after treated with Ag+, and then decreased significantly. This result showed a positive correlation between the levels of mRNA expression and tanshinones accumulation in Salvia miltiorrhiza stimulated by Ag+. The content of tanshinones was gradually raised, and it had an obvious increase at 120 h. The bioinformatics analysis and gene expression indicated that SmHDR might be involved in tanshinones biosynthesis, which laid the foundation for further study of secondary metabolic regulation mechanism of tanshinones.

This paper introduced a new method of "ingredient difference phonetypical cloning" for functional gene clone of medicinal plants, which might solve the difficulties in isolating genes encoding enzymes for the biosynthesis of secondary metabolites by usual ways. Concepts, mechanisms and methods were systematically introduced and possibility was proved by experiments. The method showed the extra superiority of for the isolation of the genes belonged to unknown metabolic pathway and little information about its sequences. The method provides a new way to isolate functional gene cloning from Chinese herbs and a fundament for the further study on medicinal plant genetic engineering.
Cloning, Molecular ; Drugs, Chinese Herbal ; metabolism ; Genes, Plant ; genetics ; Genetic Engineering ; methods ; Phenotype ; Plants, Medicinal ; genetics ; metabolism ; Reproducibility of Results ; Time Factors

OBJECTIVETo clone and sequence the open reading frame and genomic sequence of squalene synthase (SQS) from Glycyrrhiza uralensis.

METHODThe primers were designed according to cDNA sequence of SQS from G. glabra reported by Hiroaki HAYASHI, SQS cDNA was cloned with total RNA extracted from roots of G. uralensis. Specific fragments were amplified by RT-PCR and then were cloned and sequenced. SQS DNA was cloned with total DNA extracted from roots of G. uralensis. Specific fragments were amplified by PCR and then were cloned and sequenced.

RESULTGuSQS1 (GenBank accession number: GQ266154) was 1 242 bp in length encoding proteins with 412 amino acid. NCBI Blast x search results showed GuSQS1 had the highest amino acid similarity to the corresponding proteins from G. uralensis. The identities of GuSQS1 with the two proteins were 98. 55% and 88. 62%. SQS (GenBank accession number: GQ180932) gene with 4 484 bp containing 13 exons and 12 introns was then amplified by PCR with genomic DNA extracted from roots of G. uralensis.

CONCLUSIONThese findings of cloning and sequencing the open reading frame and genomic sequence of squalene synthase (SQS) from G. uralensis brought some new clues for the further exploration of SmSQS function in sterol and terpenes biosynthesis.

Amino Acid Sequence ; Cloning, Molecular ; methods ; DNA, Complementary ; chemistry ; Farnesyl-Diphosphate Farnesyltransferase ; chemistry ; Glycyrrhiza uralensis ; chemistry ; enzymology ; Molecular Sequence Data ; Open Reading Frames ; Plant Roots ; chemistry ; enzymology ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA ; methods

Flavonoids are an important effective component of traditional Chinese medicine, which are widely distributed in the plant kingdom. The biosynthesis of flavonoid in plants is affected and regulated by various environmental factors. For a necessary environmental factor to plant growth and development, mineral nutrients are paid more and more attention on the regulation to the metabolism of flavonoids in medicinal plants. In this paper, an overview of flavonoids biosynthetic pathway, and the macroelements, microelements and rare earth elements on the metabolism of flavonoids in medicinal plants are presented. And the regulation mechanism of them are also analyzed and discussed.
Flavonoids ; analysis ; metabolism ; Minerals ; analysis ; metabolism ; Nutrition Assessment ; Plants, Medicinal ; chemistry ; metabolism

Secondary metabolites are produced during the growth and development of plants along with the adaptation of outer environment, as a rule they are the main active ingredients in medicinal plants and ensure the quality of crude drugs. Since biogenesis is quite complex, the production and accumulation of secondary metabolites are influenced by various biotic and abiotic factors either from gene or environments, the complexity may affect quality control of crude drugs and utilization of the active ingredients. The thought and approach adopted in systems biology is a powerful tool to explore biology fully, along with the development of modern molecular biology and information biology, omics integration like genomics, transcriptomics, proteomics, and metabolomics will bring new opportunities for the study of secondary metabolites of medicinal plant. It has great significance to apply this holistic and systematic method in researches on biosynthetic pathway, signal transduction, ecological environment and metabolic engineering of the formation of the secondary metabolites of medicinal plants, and in building secondary metabolite biosynthesis gene expression and regulation system model, in order to explain the origin of the active ingredients of medicinal plants, formation mechanism of the Chinese herbs, metabolic engineering effecting active ingredients of medicinal plants, and the rational exploitation and utilization of resources of medicinal plants systematically.
Models, Biological ; Models, Theoretical ; Plants, Medicinal ; metabolism ; Systems Biology ; methods

OBJECTIVETo study the mechanism of secondary metabolites of some phenolic acids in the hairy roots of Salvia miltiorrhiza induced by methyl jasmonate.

METHODThe hairy roots of S. miltiorrhiza were induced with methyl jasmonate (100 micromol x L(-1)) and collected at 0, 12, 24, 36 h after treatment. Real-time quantitative PCR was used for detecting the mRNA expression level of the key enzyme genes on the secondary metabolites pathway of rosmarinic acid, while a LC-MS method was developed to determine the content of rosmarinic acid, caffeic acid and salvianolic acid B.

RESULT AND CONCLUSIONThe concentration of phenolic acids grew up and accumulated quickly in the hairy roots with exogenous signal molecule MJ induced, and it was showed that the content of CA and RA reached the maximum after 24 h and the content of LAB reached the maximum in 36 h by MJ induced. The induction mechanism may be activated with different levels of RA synthesis in PAL, 4CL, C4H genes on the key enzyme phenylalanine pathway and TAT, HPPR genes on tyrosine pathway. The time of gene expression was different, among them, 4CL and PAL genes were more important. In a word, the result can provide some basis data about the mechanism of secondary metabolites of phenolic acids for further research.

Cyclopentanes ; analysis ; metabolism ; Gene Expression Regulation, Plant ; Hydroxybenzoates ; analysis ; metabolism ; Oxylipins ; analysis ; metabolism ; Plant Proteins ; genetics ; metabolism ; Plant Roots ; chemistry ; genetics ; metabolism ; Salvia miltiorrhiza ; chemistry ; enzymology ; genetics ; metabolism

OBJECTIVETo establish the new EST-SSR markers for analyzing the genetic variation of different population of Salvia miltiorrhiza.

METHODIt was dealt with ESTs newly downloaded from Genbank and that of acquired from HMPL lab EGassembler software, and then carried out SSR loci search and SSR type analysis by SSRIT software. After that, it was designed the EST-SSR primer pairs for PCR amplification condition optimization.

RESULTAbundant and high coverage of SSR loci distribution were found in S. miltiorrhiza with having one SSR per 5.8 kb ESTs. Among them, the occurrences of different repeat units were mainly the di- (63.0%) and tri- (35.5%). The CT/AG was the most frequent motif in dinucleotide motif type and the GAA/TCC was the most frequent motif in trinucleotide repeats. Out off 36 primer pairs, 29 primer pairs (80.5%) were successfully amplified in all samples of S. miltiorrhiza while the rest failed to give PCR products at various annealing temperature and Mg2+ concentrations. The selected primer pairs also showed the polymorphism in samples from different S. miltiorrhiza populations.

CONCLUSIONThe newly establishment of EST-SSR markers showed high SSR loci coverage and genetic polymorphisms in S. miltiorrhiza population. It could be used for genetic variation analysis.

Alleles ; Expressed Sequence Tags ; Gene Frequency ; Genetic Markers ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Salvia miltiorrhiza ; genetics