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.

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.

To examine the histological changes of diabetic rats' skin and the effects on the percutaneous absorption of hydrocortisone (HC, a glucocorticoid), male Wistar rats were randomly divided into five groups: control group, diabetes one-week group (W1), two-week group (W2), three-week group (W3), and four-week group (W4), while each group contained 6 rats. Diabetes mellitus (DM) rat model was prepared with the method of streptozocin (STZ, 40 mg x kg(-1)) intraperitoneal injection. Abdominal skin was cut to carry out an in-vitro penetration experiment on an improved Franz diffusion cells, and phosphate buffer (PBS, pH 7.4) was used as receptor solution. The solution was analyzed with HPLC, and then the penetrating rate can be calculated. Meanwhile, rats' abdominal skins of different DM periods were HE stained and made into tissue slices to find if any histological changes occurred. The penetrating rate of control, W1, W2, W3, and W4 groups were 2.39 +/- 1.25, 3.22 +/- 1.72, 3.02 +/- 1.89, 3.63 +/- 2.02 and 5.00 +/- 3.36 microg x h(-1) x cm(-2), respectively. There was significant difference between the control and the W4 group (P < 0.05), but no significant differences were found between any other two groups (P > 0.05). The tissue slices showed that compared to the normal rats' skin, little change was observed in one-week DM rats' skin, but the skin of one-month DM rats' skin was observed thinner, and it became much thinner than that of rats with two-month diabetes, especially the epidermis. After making a rat into diabetic, the rats' skin goes through a pathological change, and this change is closely interrelated with the increase of the permeation of HC. Therefore, it is necessary to adjust the dose while some drug was applied on the skin in case of diabetes mellitus.

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.

Patent technology is regarded as technological trends under the market economy condition. The case showed the information form patent literature can be widely used in technology or economy. In this study, we analyzed the patent technology status quo and development trend for Chinese herbal medicines based on China patent database. The patent technology status quo is divided from the technology of biotechnology, quality control, cultivation and herb processing on Chinese herbal medicines. Furthermore, some recommendations of technology development and advices on patent protection for Chinese herbal medicines were suggested.
China ; Drugs, Chinese Herbal ; chemistry ; Herbal Medicine ; legislation & jurisprudence ; trends ; Humans ; Patents as Topic ; Plants, Medicinal ; chemistry ; genetics ; Quality Control

By reverse transcription-polymerase chain reaction (RT-PCR), an open reading frame of pathogenesis-related protein 1 (PR1) was isolated from Panax notoginseng and named as PnPR1. Molecular and bioinformatic analyses of PnPR1 revealed that an open reading frame of 501 bp was predicted to encode a 166-amino acid protein with a deduced molecular mass of 18.1 kD. Homology analysis showed that the deduced amino acid sequence of PR1 protein of Panax notoginseng had a high similarity with other higher plants had the same conservative structure domain of cysteine-rich secretory protein (CAP). The recombinant expressed plasmid pET28a(+)-PnPR1 was expressed in Escherichia coli BL21. The expression conditions were optimized by induction at different times, different temperatures, different IPTG concentrations and different giving times. The optimum expression condition was 0.4 mmol.L-1 IPTG at 28 degrees C for 20 h. The successful expression of PnPR1 provides some basis for protein purification and preparation of the monoclonal antibody.

OBJECTIVE:To provide reference for the classification of the specification grade of Dioscorea opposite.METHODS:Using the Delphi method,17 experts,who were associated with the study of the distribution and clinical application of D.opposite,were selected to conduct two rounds of consultation on 18 samples of medicinal herbs.The importance of the sensory evaluation indexes of D.opposite was screened and the overall satisfaction was evaluated.The specification grade of D.oppositewas determined preliminarily.RESULTS:Totally 17 questionnaires were issued for each round of consultation,and 17 were recovered with recovery rate of 100％.According to statistical analysis,the average value of expert's authority coefficient was 0.77 +0.07,and average recurrence rate of individual review was (91.18 + 7.64)％.The average recurrence rates of group review in two rounds of consultation were (66.67 + 13.50)％ and (65.97 + 14.01)％.The influence of working life,business background and educational background on recurrence rates of group review for interviewed experts was comprehensive.The full score ratio of appearance and cross-sectional characteristics importance was more than 80％,and the average value was more than 0.8.The full score ratio and average value of iron yam were higher.CONCLUSIONS:It is accurate and scientific to evaluate the specification grade of Chinese medicinal herbs with sensory experience.The most important evaluation indicators of D.opposite were the appearance,shape and cross-sectional characteristics.And it is divided into iron yam and non-iron yam preliminarily.The iron yam grade in descending order:Henan sandy iron yam,Shandong sandy iron yam,Henan loquat iron yam.The non-iron yam grade in descending order:Henan Huaiqingfu D.opposite,Hebei Xiaobaizui D.opposite,Hebei Ma D.opposite and Shanxi Chang D.opposite.

To find the identification method of seeds from different locations, we analyzed the SDS-PAGE electrophoretogram of Magnolia officinalis seeds from 5 different locations of Enshi district, Hubei province, the genuine producing area of M. officinalis, and compared band numbers, band intensity and protein molecular weight of different locations. The result showed that there were significant differences among seeds from different locations. 9 bands with the molecular weight about 80.34, 63.07, 56.48, 51.32, 40.64, 25.18, 22.63, 21.70, 18.14 kDa were involved in all seeds electrophoretograms and could be as standard bands of M. officinalis seeds. Seeds from Badong county, Jianshi county and Enshi city had their own special bands which were very different from other locations. So this electrophresis method can be applied to identify different locations of M. officinalis seeds.
Electrophoresis, Polyacrylamide Gel ; Magnolia ; chemistry ; Molecular Weight ; Plant Proteins ; analysis ; Seeds ; chemistry

OBJECTIVETo investigate the effects of light on baicalin,baicalein accumulation and related genes expression in suspension cultures of Scutellaria baicalensis.

METHODThe content of baicalin, baicalein in suspension cultures of S. baicalensis was determined by HPLC. The related genes expression was analyzed by semi-quantitative PCR.

RESULTActive ingredients in suspension cultures of S. baicalensis were promoted significantly by light, PAL gene transcription under the conditions of light was higher than that under the dark conditions, the UBGAT gene expression levels was significantly related to the baicalein (r = -0.995).

CONCLUSIONActive ingredients and related genes (PAL, UBGAT) in suspension cultures of S. baicalensis were promoted significantly by light.

Antioxidants ; pharmacology ; Cell Culture Techniques ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; pharmacology ; Flavanones ; analysis ; genetics ; radiation effects ; Flavonoids ; analysis ; genetics ; pharmacology ; radiation effects ; Free Radical Scavengers ; pharmacology ; Light ; Medicine, Chinese Traditional ; methods ; Phytotherapy ; methods ; Plant Extracts ; chemistry ; Rehmannia ; chemistry ; Scutellaria baicalensis ; chemistry ; growth & development ; Suspensions ; chemistry

The swollen root of Rehmannia glutinosa is used as one kind of important Chinese traditional medicine. The root of R. glutinosa usually swelled in rotational cropping but not in continuous cropping. The rhizosphere microorganisms of R. glutinosa under different farming condition were thought related to that. In this study, the endophytic fungi in the root of R. glutinosa growing in various soil conditions were isolated for the study of the relationship between the microorganisms and the root enlargement of their host plants. The dominant endophytes, Verticillium spp., Fusarium oxysporum, F. redolens and Ceratobasidium spp. were identified by morphological observation and 18S rDNA and ITS sequence analysis. The preliminary investigation showed that the excessive growth of Verticillium and Fusarium genus fungi is unfavorable for the R. glutinosa root swelling, but Ceratobasidium fungi has no effects on the root enlargement.
DNA, Fungal ; genetics ; DNA, Ribosomal ; genetics ; Fungi ; classification ; genetics ; isolation & purification ; Molecular Sequence Data ; Plant Roots ; microbiology ; Rehmannia ; microbiology