Full-length transcriptome sequence and identification of genes involved in phenylethanol glycoside biosynthesis in Rehmannia glutinosa / 药学学报

italic>Rehmannia glutinosa belongs to the Scrophulariaceae family with important medicinal value. In order to effectively explore the transcriptome information of R. glutinosa and identify the genes encoding enzymes involved in phenylethanol glycoside (PhGs) biosynthesis, the leaves, stems and tuberous roots of R. glutinosa were used for transcriptome sequencing using Pacific Biosiences RS II platform. A total of 27 773 transcripts were generated with an average length of 2 380 bp, and 27 236 coding sequences (CDS) were predicted. Using BLAST software, non-redundant transcript sequences were annotated with NR, NT, GO, COG, KEGG, SwissProt and Interpro databases and a total of 27 399 annotated genes were obtained. Among them, the number of genes related to Sesamum indicum in the NR database was the highest (81.44%), which is consistent with their evolutionary relationship. Enzymes likely involved in the biosynthesis of isoacteoside, echinacoside, cistanosides A, cistanosides F, 2′-acetylacteoside and leonoside F were identified, and 143 genes were identified in R. glutinosa full-length transcriptome. The expression levels of 19 genes correlated with acteoside content in twelve tissues of R. glutinosa, and most showed higher expression levels in leaf tissues and floral organs. This study provides more reliable transcriptome data for screening R. glutinosa for functional genes and provides a foundation for the study of the molecular mechanisms of PhGs biosynthesis.

Identification and biological characteristics of a pathogen causing leaf blight of Rehmannia glutinosa / 中国中药杂志

Leaf blight outbroke in Rehmannia glutinosa plantation in Wenxian county, Henan province in 2019. R. glutinosa plants with diseased leaves were collected from the plantation, and three strains were isolated from the diseased leaf samples. Pathogenicity test, morphological observation, and phylogenetic analysis of ITS, EF1-α, and Tub suggested that they were respectively Fusarium proliferatum, F. oxysporum, and F.acuminatum. Among them, F. acuminatum, as a pathogen of R. glutinosa leaf disease, had never been reported. To clarify the biological characteristics of F. acuminatum, this study tested the influence of light, pH, temperature, medium, carbon source, and nitrogen source on the mycelial growth rate of the pathogen during a 5-day culture period, and explored the lethal temperature. The results showed that the mycelia grew well under the photoperiod of 12 h light/12 h darkness, at 5-40 ℃(optimal temperature: 25 ℃), at pH 4-11(optimal pH: 7.0), on a variety of media(optimal medium: oatmeal agar), and in the presence of diverse carbon and nitrogen sources(optimal carbon source: soluble starch; optimal nitrogen source: sodium nitrate). The lethal temperature was verified to be 51 ℃(10 min). The conclusion is expected to lay a scientific basis for diagnosis and control of R. glutinosa leaf diseases caused by F. acuminatum.

Analysis of Volatile Organic Compounds in Rhizosphere Soil of Rehmannia glutinosa Under Intercropping Zea mays Based on GC-MS Metabonomics / 中国实验方剂学杂志

Objective:To explore the composition characteristics of rhizosphere soil under <italic>Rehmannia glutinosa-Zea mays</italic> intercropping model，and screen out special signal substances in rhizosphere soil of <italic>R. glutinosa</italic> under intercropping <italic>Z. mays</italic>， so as to provide the basis for the study of allelopathic substances in continuous cropping obstacle of <italic>R. glutinosa</italic>. Method:In this experiment，rhizosphere soils of <italic>R. glutinosa</italic> under <italic>Z. mays </italic>intercropping and <italic>R. glutinosa </italic>single cropping models in July，August，September and October were taken as the research objects， and the volatile organic compounds in ethyl acetate fraction were analyzed by gas chromatography-mass spectrometry （GC-MS）. Principal component analysis （PCA）， hierachical cluster analysis （HCA） and orthogonal partial least squares discriminant analysis （OPLS-DA） analysis were performed on the data by SIMCA 14.1 to screen out potential differences in volatile organic compounds between the two models. Result:The types of volatile organic compounds in intercropping and single cropping models were mainly hydrocarbons， alcohols， esters， ketones， amides， acids and other substances. Specifically， the average relative contents of hydrocarbons，esters and amides in intercropping model were 58.46%，32.15% and 5.42% respectively，while the relative contents of hydrocarbons，esters and amides in single cropping model were 37.27%，36.11% and 21.13%. The results of PCA and HCA showed that the characteristics of volatile organic compounds in the ethyl acetate fraction of rhizosphere soil under intercropping and single cropping models could be clearly divided into two categories，the screening results of potential differential components based on OPLS-DA analysis indicated that various components， such as dibutyl phthalate，（<italic>Z</italic>）-9-oleamide，<italic>β</italic>-caryophyllene，dioctyl iso-phthalate， phthalate （2-propylamyl） diester， <italic>n</italic>-hexadecane，octodecane， <italic>n</italic>-heneicosane， were screened from rhizosphere soil under the two models. Conclusion:The <italic>R. glutinosa-Z. mays</italic> intercropping model has certain effects on the volatile organic compounds in the rhizosphere soil of <italic>R. glutinosa</italic>，and the effect of the selected components on the growth and quality characteristics of <italic>R. glutinosa</italic> still need to be further studied.

Effects of density on growth and gene transcription characteristics of Rehmannia glutinosa / 中国中药杂志

The present study analyzed the effects of planting density on the development, quality, and gene transcription characte-ristics of Rehmannia glutinosa using 85-5 and J9 as materials with three planting densities of 5 000, 25 000, and 50 000 plants/Mu(1 Mu≈667 m~2). The agronomic characteristics of leaves and tuberous roots, the content of catalpol and acteoside, and the changes of gene expression were determined. The results showed that the leaf size, the diameter of tuberous root, leaf biomass, tuberous root number, and tuberous root biomass per plant at low density were significantly higher than those of medium and high densities. The content of catalpol and acteoside in leaves was higher at high density. The content of catalpol in tuberous roots was higher at low density, and the change trend was similar to that in leaves, while the content of acteoside in tuberous roots was higher at high density. Transcriptome analysis found that about 1/2 of the expansin genes could change regularly in response to density treatment, which was rela-ted to the development of tuberous roots. The change trend of the gene expression of multiple catalytic enzymes involved in the biosynthesis of catalpol and acteoside was consistent with that of their content, which was presumedly involved in the accumulation and regulation of density-responsive medicinal components. Based on the analysis of the development, medicinal components, and gene expression characteristics of R. glutinosa at different densities, this study is expected to provide an important basis for regulating the quality and yield of medicinal materials of R. glutinosa by managing the planting density.

Identification and expression analysis of NRT1 family genes in Rehmannia glutinosa / 中国中药杂志

NRT1 family proteins play an important roles for absorbing and transporting of nitrate in different plants. In order to identify the NRT1 family genes of Rehmannia glutinosa, this study used 11 NRT1 homologous proteins of Arabidopsis as probe sequences and aligned with the transcriptome data of R. glutinosa by using NCBI BLASTN software. Resulting there were 18 NRT1 proteins were identified in R. glutinosa. On basis of this, a series of the molecular characteristics of R. glutinosa NRT1 proteins including the conserved domains, the transmembrane structure, the subcellular location and phylogenetic features were in detail analyzed. At same time, it were systematically analyzed that the temporal and spatial expression patterns and characteristics of R. glutinosa NRT1 family genes in response to different stress factors. The results indicated that 18 R. glutinosa NRT1 family genes with the length of coding region from 1 260 bp to 1 806 bp, encoded proteins ranging from 419 to 601 amino acids, and all of they owned the domains of typical peptide transporter with 7 to 12 transmembrane domains. These R. glutinosa NRT1 family proteins mostly were found to locate on cellular plasma membrane, and belonged to the hydrophobic proteins. Furthermore, the evolutionary analysis found that the 18 R. glutinosa NRT1 protein family could be divided into two subfamilies, of which 14 NRT1 family genes might occur the positive selection, and 4 genes occur the passivation selection during the evolution process of R. glutinosa. In addition the expression analysis showed that 18 R. glutinosa NRT1 family genes have the distinct expression patterns in different tissues of R. glutinosa, and their expression levels were also obvious difference in response to various stress. These findings infield that 18 R. glutinosa NRT1 family proteins might have obviously different functional roles in nitrate transport of R. glutinosa. In conclusion, this study lays a solid theoretical foundation for clarifying the absorption and transport molecular mechanism of N element during R. glutinosa growth and development, and at same time for deeply studying the molecular function of R. glutinosa NRT1 proteins in absorption and transport of nitrate.

Thermosensitive liposomal doxorubicin plus radiofrequency ablation increased tumor destruction and improved survival in patients with medium and large hepatocellular carcinoma: A randomized, double-blinded, dummy-controlled clinical trial in a single center

Background: Lyso-thermosensitive liposomal doxorubicin (LTLD, ThermoDox) consists of doxorubicin encapsulated contained within a heat-sensitive liposome. Aims and Objectives: We sought to evaluate whether the use of combined radiofrequency ablation (RFA) and LTLD would result in larger coagulation volume and longer overall survival (OS) compared with the use of RFA alone in patients with 3–7 cm unresectable hepatocellular carcinoma (HCC). Materials and Methods: Between 2010 and 2012, 22 HCC patients were randomly assigned to one of two treatments in our center: (1) ultrasound-guided percutaneous RFA plus intravenous (IV) infusion of LTLD (combination, n = 11) or (2) RFA plus IV dummy (RFA, n = 11). Four patients withdrew from the study, and the remaining 18 patients entered the final analysis. There were 14 male and 4 female patients with an average age of 61.1 ± 9.3 years (range: 40–73 years). The average tumor size was 4.2 ± 1.0 cm (range: 3.1–6.1 cm). One-month enhanced computed tomography was used to evaluate the ablation efficacy and coagulation volume after RFA. Regular follow-up after RFA was performed to assess toxicity, local response rates, and OS rates. Results: A major complication (empyema) occurred in one case in the combination group. Combination treatment region did not induce any additional toxicity beyond doxorubicin. The primary ablation success rate was 93.3% (14/15 tumors) in the combination group and 77.8% (7/9 tumors) in the RFA group (P = 0.308). The difference in coagulation volume between pre- and post-RFA in the combination group was significantly larger than that of the RFA group (105.7 ± 73.8 cm 3 vs. 37.3 ± 8.5 cm 3, P = 0.013). The follow-up period ranged from 11 to 80 months (average: 49.1 ± 24.8 months). The local progression rate was 6.7% (1/15 tumors) in the combination group and 22.2% (2/9 tumors) in the RFA group. The mean OS for the combination group was 68.5 ± 7.2 months, which was significantly greater compared with the RFA group (46.0 ± 10.6 months, P = 0.045). Conclusions: RFA with heat target delivery chemotherapy facilitated better tumor coagulation necrosis without additional toxicity. This combined treatment may improve the clinical efficacy of RFA or free doxorubicin and prolong survival in patients with medium to large HCC

Cloning and expression analysis of LncRNA-RgAGT2 responding to consecutive monoculture in Rehmannia glutinosa / 中国中药杂志

The consecutive monoculture obstacle is a major problem in the field of Rehmannia glutinosa( R. glutinosa),has severely declined the yield and quality of R. glutinosa. Here,using hi TAIL-PCR and RACE techniques,we have cloned the full-length transcript( 1 573 bp) of Unigene 29334＿All screened by DGE as a consecutive monoculture obstacle response gene of R. glutinosa. Based on ORF Finder prediction,all ORFs detected in the full-length transcript were less than 300 nt,which suggested that the above transcript was confirmed to be a long non-coding RNA( LncRNA). With alignment in R. glutinosa transcriptome,this LncRNA was partially homologous to alanine glyoxylate transaminase 2 gene( Rg AGT2),which was named LncRNA-RgATG2. To further explore the function of LncRNA-RgAGT2,we have examined expression patterns of LncRNA-RgAGT2 and Rg AGT2 at five critical development stages( seedling,elongation,pre-expanding,mid-expanding,late-expanding) in the first and second year replanting of R. glutinosa,respectively. The results indicated that LncRNA-RgAGT2,as a potential regulator,is possible to play a vital role in Rg AGT2 expression regulation. Meanwhile,LncRNA-RgAGT2 has presented significant variation in all development stages of R. glutinosa,which could be used as a " diagnostic label" to assess consecutive monoculture obstacle. This study,for the first time,showed that LncRNA was responsible for the response and regulation of consecutive monoculture obstacle,which would be a powerful supplement to reveal the molecular mechanisms of consecutive monoculture obstacle of R. glutinosa.

Phenotypic Character and HPLC Fingerprints of Radial Striations from Different Germplasms Rehmanniae Radix / 中国实验方剂学杂志

Objective: To discuss the phenotypic character and the HPLC fingerprints of radial striations from different germplasms Rehmanniae Radix. Method: The changes in the shape and column diameter of the radial striations of Rehmanniae Radix were observed and measured in the whole growth period. Besides,the HPLC fingerprints of the root,radial and un-radial striations were established to sign the chemical quality and analyzed by principal component analysis(PCA)and systematic cluster analysis. Result: There were significantly differences and regularities in the shape and proportion of the radial striations of different germplasms Rehmanniae Radix. The fingerprints showed the consistency between different types of chemical ingredients,and the differences in chemical quality characteristics mainly lay in the content of chemical compositions and theirs relative ratio. The results of PCA indicated that active ingredients, such as acteoside,catalpol,rehmaionoside D,rehmaionoside A and leonuride, were involved in the quality expression of different parts from various germplasms of Rehmanniae Radix,but each ingredient had a distinctive contribution rate to the differential quality expression between different parts from various germplasms of Rehmanniae Radix. However,the other components involved in the differential quality expression had different contribution rates in different germplasms.The systematic cluster analysis indicated that great differences in the chemical quality between the radial striations and un-radial striations of Beijing-1,Qinhuai,Qinhuai Zheng and 1706 germplasms,but with small differences in 85-5 and Baixuan germplasms. Conclusion: There are differences in phenotypic character of the radial striations and HPLC fingerprints between different germplasms Rehmanniae Radix.

Progress on germplasm enhancement and breeding of Rehmannia glutinosa / 中国中药杂志

The history of Rehmannia glutinosa breeding has already beyond 100 years. There are rich cultivated varieties and wild germplasm resources in R. glutinosa. However, there also exist a lot of problems, such as, the pedigree of the existing varieties is not clear, the genetic basis is narrow, backward method of germplasm enhancement and breeding. Breeding of new varieties has been unable to meet the demand of R. glutinosa production in the new era. This paper summarizes the species of Rehmannia and their distribution, the diversity of plant morphology and the quality of R. glutinosa germplasm resources, as well as the progress of R. glutinosa breeding in recent 100 years. For ensuring the orderly, effective and safe production of R. glutinosa, the authors suggest to establish the wild resources protection area and germplasm resources garden, deeply study the genetic base of quality, strengthen application of new breeding method such as mutation breeding, haploid breeding and gene editing.

Qingkailing Injection () for Treatment of Children Pneumonia Induced by Respiratory Syncytial Virus: A Meta-Analysis of Randomized Controlled Trials / 中国结合医学杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of Qingkailing Injection (, QKL) for treatment of children pneumonia caused by respiratory syncytial virus (RSV).</p><p><b>METHODS</b>Randomized clinical trials (RCTs) comparing QKL with ribavirin injection in the treatment of children pneumonia induced by RSV were searched in PubMed, Science Direct, Cochrane Library, Chinese VIP database, CNKI and Wanfang databases from their inception to March 2014. Meta-analyses were performed using RevMan 5.2 software. The methodological quality of the selected RCTs was evaluated by the Modified Jadad Score. The primary outcome measures were effective rate and the secondary outcomes were relief time of fever and cough.</p><p><b>RESULTS</b>Seven RCTs with 992 cases published from 2008 to 2013 were identified. The meta-analysis results indicated that QKL was more effective in cure rate [risk ratios (RR)=1.32, 95% CI (1.17, 1.50), P<0.01], total effective rate [RR=1.07, 95% CI (1.02, 1.13), P=0.009] and less fever clearance time [mean difference=-0.73, 95% CI (-1.22,-0.23), P=0.004], compared with ribavirin injection in the treatment of RSV-induced children pneumonia. No dead case was reported in all trials. There were 3 trials mentioned adverse events, 2 reported no obvious adverse event occurred while 1 reported adverse events described as skin hypersensitivity, elevation of ALT, a mild abnormal of hepatic and renal function in both QKL and ribavirin group.</p><p><b>CONCLUSIONS</b>QKL was an effective and relatively safe option for the treatment of RSV-induced children pneumonia. These therapeutic effects were promising but need to be interpreted with caution due to variations in the treatment and methodological weakness in the studies.</p>

Study on herbal textual evolution and flavonoids and their pharmacological of Spatholobi Caulis / 中国中药杂志

Spatholobi Caulis, the vine stem of Spatholobus suberectus and widely used in China and Southeast Asian nations, has the effects on nourishing the blood and promoting blood flow, regulating menstruation and relieving pain, and invigorating the nerves. Through consulting the herbal textual and local chronicles, we summarized the original textual research and medicinal evolution on Spatholobi Caulis to analyze the changes of varieties in different historical periods. Further, the major active ingredient in Spatholobi Caulis was discussed. According to the literature to date, 60 flavonoids compounds have been isolated and could be divided into isoflavones, dihydroflavones, flavanols, dihydroflavonols, procyaninides, chalcones, pterocarpans, isoflavanols, isoflavanones and aurone according to their molecular structures. These indicative ingredients in Spatholobi Caulis showed pharmacological activities on regulation of the blood system, anti-tumor, anti-oxidation, anti-virus, anti-bacteria and inhibition of melanin deposition. This review will provide reference and basis for the sustainable use of resources and industry development on Spatholobi Caulis.

Identification and analysis of the calcium signaling system genes responding to continuous cropping injury in Rehmannia glutinosa L / 药学学报

Calcium signaling plays a critical role in response to various abiotic and biotic stresses in plants. Preliminarily evidence showed that calcium signaling perceived and transduced the harmful signaling generated from continuous cropping stress in R. glutinosa. To investigate the roles of calcium signaling in continuous cropping injury formation, the key genes involved in calcium signaling transduction were identified in R. glutinosa transcriptome through bioinformatic methods. Furthermore, the calcium ion concentration in both normal and continuous cropping R. glutinosa root cells were measured by potassium pyroantimonate precipitation and calcium fluorescence method. As a result, a set of 84 calcium signaling-related genes, including 5 CaMs, 12 CBLs, 21 CDPKs, 21 CIPKs, 16 CMLs, and 9 CRKs were captured in R. glutinosa transcriptome. The analysis of expression profile in continuous cropping compared to normal growth R. glutinosa indicated that continuous cropping stress significantly increased the expression of calcium signaling-related genes in continuous cropping R. glutinosa. At the same time, the abundance levels of 12 calcium signaling-related genes quantified by qPCR further validated the high expression of calcium signaling-related genes presented in continous cropping R. glutinosa. In addition, the continuous cropping condition significantly promoted the accumulation of intracellular calcium ions in R. glutinosa based on two methods of potassium pyroantimonate precipitation and calcium fluorescence. This study verified the possible roles of calcium signaling in the formation of continuous cropping injury on molecular and cellular level, which lays a solid foundation for illuminating formation mechanism of continuous cropping injury on molecular level.

Comparison of chemical quality characteristics between radial striations and non-radial striations in tuberous root of Rehmannia glutinosa / 中国中药杂志

An HPLC method was established to determine the contents of catalpol, acteoside, rehmaionoside A, rehmaionoside D, leonuride in three part of Rehmanni glutinosa in Beijing No.1 variety R. glutinosa during the growth period, This method, in combination with its HPLC fingerprint was used to evaluate its overall quality characteristics.The results showed that：① the content of main components of R. glutinosa varied in different growth stages ;② there was a great difference of the content of main components between theradial striations and the non-radial striations; ③ the two sections almost have the same content distribution of catalpol, acteoside and rehmaionoside D; ④the content of rehmaionoside A in non-radial striations was higher than that in radial striations,while the content of leonuride in radial striations was higher than that in non-radial striations.; ⑤the HPLC fingerprint of radial striations, non-radial striations and whole root tuber were basically identical, except for the big difference in the content of chemical components. The result of clustering displayed that the radial striations, non-radial striations, and whole root were divided into two groups. In conclusion, there was a significant difference in the quality characteristics of radial striations and non-radial striations of R. glutinosa. This research provides a reference for quality evaluation and geoherbalism of R. glutinosa.

A consideration on screening of allelochemicals by using "knock-out/in" of targeting ingredients based on identification model of medicinal effect / 中国中药杂志

This paper clarified in detail the definition, characteristics of allelopathy and its association with consecutive monoculture problem.Most of studies have indicated that it is critical to parse the formation mechanisms of consecutive monoculture problem that identification of allelochemicals and verification of its function. Here, we proposed a new approach to separate and identify the allelochemical group precisely and effectively, in which the "knock-out/in" methods of targeting ingredients in the model of medicinal effect identification and quality control were applied. This method will contribute to deep understanding plant allelopathy, and provide theoretical basis and technical support for alleviating consecutive monoculture problems simultaneously.

Research progress of relationship between continuous cropping obstacles and microRNAs in Rehmannia glutinosa / 中国中药杂志

The efficacy of Rehmannia glutinosa which as a large quantity of traditional Chinese medicine is significant. However, the land must be given up after one season of R. glutinosa cultivation or replanted after a period of 8-10 years because of the severe continuous cropping obstacles. MicroRNAs is a class of endogenous non-coding small RNAs, which participate in regulation of physiological activities by target mRNA cleavage or translational repression in plants. In recent years,studies on the role of miRNAs in plants have made significant progresses,especially in medicinal plants．MiRNAs from some different medicinal plant species have been identified with regulatory effects．When plants are exposed to environmental stress, miRNAs act on stress-related genes and initiate stress-resistance mechanisms in the body against adverse factors. R. glutinosa is also a kind of environmental stress. It is conducive to deciphering the molecular mechanism of continuous cropping obstacles for us by researching miRNAs. This article reviews the production of miRNAs, mechanism, research approaches and characteristics of resisting the environmental stresses in plants, the development trends and future prospect of R. glutinosa miRNAs research.

Advances on molecular mechanisms of Rehmannia glutinosa consecutive monoculture problem formation in multi-omics era / 中国中药杂志

Although consecutive monoculture problems have been studied for many years, no effective treatments are currently available. The complexity of systems triggered the formation of consecutive monoculture problems was one major cause. This paper elaborated the physiological and ecological mechanisms of consecutive monoculture problem formation based on the interaction relationship among multiple factors presented in the rhizosphere soil of consecutive monoculture plants. At same time, in this paper the multiple interactions among cultivated medicinal plants, autotoxic allelochemicals and rhizosphere microbial were proposed to be most important causes that derived the formation of consecutive monoculture problem. The paper also highlighted the advantage of 'omics' technologies integrating plant functional genomics and metabolomics as well as microbial macro-omics in understanding the multiple factor interaction under a particular ecological environment. Additionally, taking R. glutinosa as an example, the paper reviewed the molecular mechanism for the formation of R. glutinosa consecutive monoculture problem from the perspective of the accumulation of allelopathic autotoxins, the rhizosphere microecology catastrophe and theresponding of consecutive monoculture plants. Simultaneously, the roles of mutilple 'omics' technologies in comprehending these formation mechanism were described in detail. This paper provides finally a new insight to solve systematically the mechanism of consecutive monoculture problem formation on molecular level.

Cloning and expression analysis of a tyrosine decarboxylase gene from Rehmannia glutinosa / 中国中药杂志

Tyrosine decarboxylase (TyrDC) is an important enzyme in the secondary metabolism of several plant species, and was hypothesized to play a key role in the biosynthesis of phenylethanoid glycosides. Based on the transcriptome data, we cloned the full-length cDNA (GenBank accession NO. KU640395) of RgTyDC gene from Rehmannia glutinosa, and then performed bioinformatic analysis of the sequence. Further, we detected the expression pattern in different organs and hair roots treated with four elicitors by qRT-PCR. The results showed that the full length of RgTyDC cDNA was 1 530 bp encoding 509 amino acids. The molecular weight of the putative RgTyDC protein was about 56.6 kDa and the theoretical isoelectric point was 6.25. The RgTyDC indicated the highest homology with Sesamum indicum SiTyDC and Erythranthe guttata EgTyDC, both of them were reached 88%. RgTyDC highly expressed in R. glutinosa leaf, especially in senescing leaf, and rarely expressed in tuberous root. After the treatment of SA and MeJA, the relative expression level of RgTyDC mRNA was substantially increased. The results provide a foundation for exploring the molecular function of RgTyDC involved in phenylethanoid glycosides biosynthesis.

Resource situation investigation and analysis about endangered Chinese herbal medicine Daphnes Cortex / 中国中药杂志

In order to figure out the status and distribution of the wild and cultivated resources of traditional Chinese medicine Daphnes Cortex, its suitable habitat and endangering factors were analyzed to provide the basis for its rational use, protection and cultivation.Our research group tooka resources survey in Shanxi, Gansu, Sichuan and Qinghai provinces, which include 23 counties. Investigation and sampling investigation combined with interview were carried out. The total reserve of resources was estimated through route-quadrat method in combination with the vegetation and soil-type map area method. The results indicated that there was no obvious change between the present distribution ranges of the wild Daphnes Cortex and its historical records, but the density of the population has undergone major changes. The wild reserves resources has declined seriously, even on the verge of exhaustion in some regions. According to the survey results, the current total reserve of the wild Daphnes Cortex in the four provinces was no more than 600 tons. Simultaneously, we only found the cultivated resource in a mountain at an altitude of about 2 800 m in Kang county of Gansu province, which cropping scope was about 33 000 m². The cultivated resource can't provide medicinal products at present, because their growing period is too short to have curative effect. Destructive excavation and the longer growth cycle result in a sharp decline of the wild resources reserves, even to the point of extinction. Artificial cultivation of product will become the main source of medicinal resources in the future. Therefore, we must protect its suitable habitat, formulate rational harvesting policy, strengthen the supervision of government departments, collect and establish the germplasm nursery and seed bank. On the basis, we must carry out studies into seed-selecting and breeding as well as rapid propagation and growth technology at once.

Observation of prime position and driving zones in process of tuberous root expanding and expression analysis of phytohormone relative genes in Rehmannia glutinosa / 中国中药杂志

In order to study the development characteristics of Rehmannia glutinosa tuberous root expansion and reveal the regulation mechanism of the genes related to hormones in this process, R. glutinosa "wen-85" was used as the experimental material in this study. R. glutinosa tuberous roots of different developmental stages were collected to observe phenotype and tissue morphology using resin semi-thin sections method. The genes related to hormone biosynthesis and response were chosen from the transcriptome of R. glutinosa, which was previously constructed by our laboratory, their expression levels at different development stages were measured by real-time quantitative PCR. The results showed that the root development could be divided into six stages: seeding, elongation, pre-expanding, mid-expanding, late-expanding and maturity stage. The anatomic characteristics indicated that the fission of secondary cambium initiated the tuberous root expansion, and the continuous and rapid division of secondary cambium and accessory cambium kept the sustained and rapid expansion of tuberous root. In addition, a large number oleoplasts were observed in root on the semi-thin and ultra-thin section. The quantitative analysis suggested that the genes related to biosynthesis and response of the IAA, CK, ABA,ethylene, JA and EB were up-regulated expressed, meanwhile, GA synthesis and response genes were down-regulated expressed and the genes of GA negative regulation factors were up-regulated expressed. The maximum levels of most genes expression occurred in the elongation and pre-expansion stage, indicating these two stages were the key periods to the formation and development of tuberous roots. Oleoplasts might be the essential cytological basis for the formation and storage of the unique medicinal components in R. glutinosa. The results of the study are helpful for explanation of development and the molecular regulation mechanism of the tuberous root in R. glutinosa.

Index components contents in honeysuckle change trend at different time in a day and different stubbles in a year / 中国中药杂志

The index components contents of different time and different stubbles in honeysuckle were measured by HPLC, and were analysis by using the method of SPSS. Results showed that the content of index ingredients of different time had differences, and firstly decreased, then increased with time. The content of index ingredients of different stubbles had significantly differences, and firstly decreased, then increased with time. The chlorogenic acid contents were 2.059%-3.593%. The luteolosid contents were 0.110%-0.171%. Results indicated that the best picking buds time is before seven o'clock in the morning and evening at before and after seven o'clock, the index component content is higher. Picking buds in spring and at autumn index component content is higher; Picking buds in summer index component content is low. The experiment provides theoretical support for quality control in the whole process of the honeysuckle harvested and comprehensive utilization of honeysuckle.