Cloning and prokaryotic expression analysis of HDS from Salvia miltiorrhiza bge.f.alba / 药学学报

According to the designed specific primers of gene fragment based on the Salvia miltiorrhiza transcriptome data, with the method of reverse transcription polymerase chain reaction (RT-PCR), this study cloned full-length cDNA sequence of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase gene from Salvia miltiorrhiza bge.f.alba, this sequence is named as SmHDS and its GenBank registration number is KJ746807. SmHDS, 2 529 bp long, contains an ORF of 2 229 bp, encodes 742 amino acids, including 5' UTR 170 bp and 3' UTR 130 bp. Using bioinformatics software, having made a homology analysis of the obtained sequence, we can have a conclusion that SmHDS have a close genetic relationship with HDS of Salvia miltiorrhiza. Analysis result of prokaryotic expression revealed that in Escherichia coli, SmHDS expressed target proteins which in size are comparable with the protein predicted. Meanwhile, the 4 factors which can influence the protein expression were optimized, the 4 factors are inducing temperature, inducing time, IPTG concentrations and density of inducing host bacterium (A600). The optimal expression conditions of SmHDS were 30 degrees C until the A600 is 0.6, and add IPTG to a final concentration of 0.2 mmol x L(-1), and the induction time of 20 h. It provides theoretical basis for the further study of the function of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase in the biosynthesis of tanshinone compounds.

Construction of medicinal plant core collection based on molecular phylogeography / 中国中药杂志

The paper introduces the current situation and the characteristics of the medicinal plant core collection. It expounds the significance and research methods for the medicinal plant core collection based on molecular phylogeography. Guided by molecular phylogeography, the essay explores the feasibility and methods of medicinal plant core collection for the medicinal plants with rich wild resources and without wild resources. It further forecasts the application of medicinal plant core collection methods on the basis of molecular phylogegraphy.

Locality identification of Chinese medicinal plant Scutellaria baicalensis (Lamiaceae) population-level DNA barcoding / 中国中药杂志

Scutellaria baicalensis is an important traditional Chinese medicine and Scutellaria flavonoids have received worldwide attention in recent years. It is the basis of controlling quality of S. baicalensis to develop a reliable genetic marker system used to identify locality of origin. Because of the characteristics of maternal inherited and high-rate of evolution, the cpDNA intergenic spacer can effectively elucidate the degree of genetic variation in different areas of the same species (populations), which can be used as the population-level DNA barcoding to locality identify. In this study, we have used the molecular phylogeography analysis for the three cpDNA intergenic spacers atpB-rbcL, trnL-trnF and psbA-trnH of 17 wild populations from different localities, which reveals the 20 haplotypes, including 13 polymorphic sites and constitutes a shallow gene tree. The authers have divided the haplotypes of S. baicalensis into three grades of population-level DNA barcoding according to the frequence and geographic distribution: 3 highest-frequency haplotypes as area-population-level DNA barcoding, 3 haplotypes were mainly shared by 2-3 adjacent populations as region-population-level DNA barcoding, and there are also 8 unique-population haplotypes as unique-population-level DNA barcoding. The result of this study reveals that population-level DNA barcoding is a reliable genetic marker used to locality identify of S. baicalensis.

Application of molecular pharmacognosy in research of Mongolian medicine / 中国中药杂志

Molecular pharmacognosy has developed as a new borderline discipline. Using the method and technology of molecular pharmacognosy, a wide range of challenging problems were resolved, such as the identification of Mongolian medicinal raw materials, etiology of endangerment and protection of endangered Mongolian medicinal plants and animals, biosynthesis and bioregulation of active components in Mongolian medicinal plants, and characteristics and the molecular bases of Dao-di Herbs. So molecular pharmacognosy will provide the new methods and insights for modernization of Mongolian medicine.

Population genetic study of Magnolia officinalis and Magnolia officinalis var. biloba / 中国中药杂志

<p><b>OBJECTIVE</b>population genetic study of Magnolia officinalis and M. officinalis var. biloba was performed in this paper. The objectives of this study are to provide baseline data of molecular pharmaceutics for quality control of Cortex Magnoliae Officinalis.</p><p><b>METHOD</b>A total of 15 populations of M. officinalis and M. officinalis var. biloba were collected. PCR amplification and sequencing were performed with two chloroplast intergenic spacers psbA-trnH and trnL-trnF. Chloroplast haplotype frequencies were calculated, genetic diversity and genetic structure were estimated by using the program HAPLONST, and a haplotype network depicting the mutational relationships among distinct haplotypes was drawn following the principle of parsimony by TCS version 1.13.</p><p><b>RESULT</b>The differentiation of haplotype frequencies between M. officinalis and M. officiunalis var. biloba was significant although there was no private haplotypes of themselves.</p><p><b>CONCLUSION</b>There was certain genetic divergence between M. officinalis and M. offcinalis var. biloba although the monophyletic clade of themselves was not still formed.</p>

Isolation and identification of endophytic fungi from different swollen root of Rehmannia glutinosa / 中国中药杂志

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.

Prospect of application of molecular phylogeography in study of geoherbs / 中国中药杂志

This paper firstly introduces the concept, method and current research of molecular phylogeography and then discusses its application in the study of geoherbs. The relativity between three genetic differentiation patterns of plant inferred by molecular phylogeography (i.e. allopatric fragmentation, restricted gene flow with isolation by distance and range expansion) and the formation of genuine character is analysed. Molecular authentication of geoherbs based on molecular phylogeography has the advantage of former molecular identification at technology and knowing genetic differentiation of geoherbs. Using molecular phylogeography for study on changing history of geoherbs habitat is also explicated. The problem of germplasm degeneration in cultural geoherbs could be effectively resolved by molecular phylogeography method. The application of molecular phylogeography in these subjects opens up prospects for study on geoherbs by using the principle and method of molecular phylogeography.