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Chrysanthemum indicum L. is widely used in China for significant medicinal value. However, its pharmacological activities were unstable as a result of unreliable flowering time. To study the regulation of flowering time in C. indicum, we cloned the two FT-like genes from C. indicum, which were localized in the cytoplasm and nucleus of tobacco epidermal cells. Phylogenetic relationship analysis showed they were highly conserved and the CiFTL1 gene was strongly induced by short-days. Dual-luciferase assays found that CiCO could specifically bind to the promoter of CiFTL1, but could not directly interact with CiFTL2. Moreover, a CO-responsive element (CORE), localizing between -630 and -430 nucleotides from the ATG, was essential for the CiFT1 gene expression in the flowering process. Therefore, the TGTG(N2-N3)ATG was crucial for CiCO recruitment rather than the CCAAT box, which suggested the CO/FT regulatory module might be conserved in C. indicum. The research will provide sufficient information for revealing regulatory mechanisms of flowering time in C. indicum.

The inflorescence of Chrysanthemum morifolium cv. ‘Hangju’ has been widely used in China due to its antioxidant and anti-inflammatory properties. The biosynthesis and regulation of flavonoids, a group of bioactive components, in C. morifolium are poorly understood. Transcriptome sequencing is an effective method for obtaining transcript information. Therefore, single-molecule real-time (SMRT) sequencing was performed to obtain the full-length genes involved in flavonoid biosynthesis and regulation in C. morifolium. High-quality RNA was extracted from the inflorescence of C. morifolium at different flowering stages and used to construct two libraries (0–5 kb and 4.5–10 kb) for sequencing. Finally, 125,532 non-redundant isoforms with a mean length of 2009 bp were obtained. Of these, 2,083 transcripts were annotated to pathways related to flavonoid biosynthesis, and 56 isoforms were annotated as CHS, CHI, F3H, F3’H, FNS II, FLS, DFR and ANS genes. Based on gene expression levels at different stages, we predicted the major genes involved in flavonoid biosynthesis. By phylogenetic analysis, we found two candidate MYB transcription factors (CmMYBF1 and CmMYBF2) activating flavonol biosynthesis. Based on the full-length transcriptomic data and further quantitative analysis, the major genes involved in flavonoid biosynthesis and regulation in C. morifolium were predicted in our study. The results provide a valuable theoretical basis for the introduction and cultivation of C. morifolium cv. ‘Hangju’.

The genus Isatis is widely distributed throughout the world. In this work, thirty-two Isatis indigotica Fortune germplasms, collected from different regions and geographical locations in China, were analyzed the ploidy levels by flow cytometry. I. indigotica Fort. and Isatis tinctoria Linnaeus distinguished with root tip chromosome compression staining and cell flow cytometry. Microscopic observation showed that the chromosome numbers of I. indigotica Fort. and I. tinctoria L. were 2n = 14 and 2n = 28, respectively. In order to establish a flow cytometric nuclear experiment system suitable for I. indigotica Fort., the leaves of I. indigotica Fort. were used to prepare nuclear suspension with LB01, OTTO, Tris·MgCl2, patent of Luochang and Galbraith extraction buffer. It was found that the histogram generated by LB01, OTTO, Tris·MgCl2 and Patent of Luochang dissociation solution has poor peak shapes and large CV values, and Galbraith dissociation solution extraction buffer was suitable for extracting nuclei from most germplasms. Flow cytometry proved to be a simple, rapid, and highly accurate method for identifying ploidy levels Isatis species.

The novel antimicrobial gene Hirudomacin (Hmc), with a 249-bp cDNA, encodes a mature protein of 61 amino acids and a 22-amino acid signal peptide. Hmc exhibits the highest similarity, at 90.1%, with macin family members found in the salivary gland of the leech Hirudo nipponica Whitman. A mature Hmc protein concentration of 219 μg/mL was detected using the Bradford method. The mature Hmc protein is 6862.82 Da and contains 8 cysteine residues. Antimicrobial assays showed a minimum bactericidal concentration and 50% lethal dose of 1.56 μg/mL and 0.78 μg/mL, respectively, for Staphylococcus aureus and 0.39 μg/mL and 0.195 μg/mL, respectively, for Bacillus subtilis. Transmission electron microscopy revealed membrane integrity disruption in S. aureus and B. subtilis, which resulted in bacterial lysis. The level of Hmc mRNA in the salivary gland during three blood meal stages indicated a remarkable trend of increase (P .05), and western blotting demonstrated that among the three blood meal stages, expression of the mature Hmc protein was highest in both the salivary gland and intestine at the fed stage (P .05). Immunofluorescence further showed the mature Hmc protein to be localized outside the cell nucleus, with the signal intensity in the salivary gland peaking at the fed stage (P .05). In conclusion, the mature Hmc protein exhibits broad-spectrum antimicrobial effects against gram-positive and gram-negative bacteria, and a blood meal upregulates Hmc gene and protein expression in H. nipponica.

Background Monochasma savatieri is a medicinal root hemiparasitic herb that extracts water and nutrients from the host plant via a haustorium. M. savatieri exhibits an enhanced growth after the establishment of parasite-host associations, but little is known about the molecular mechanism responsible. In this study, endogenous hormones, RNA sequencing and small RNA sequencing analysis were performed on M. savatieri before and after establishment of parasite-host associations. Results When grown with the host, decreased contents of jasmonic acid (JA) and indole-3-acetic acid (IAA) and increased abscisic acid (ABA) content were observed in M. savatieri with the established parasitic relationship. When grown with the host, 46,424 differentially expressed genes (DEGs) and 162 differentially expressed miRNAs (DEmiRs) were identified in the comparison between M. savatieri with the established parasitic relationship and without the established parasitic relationship. Analysis of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) showed that these DEGs and targets of DEmiRs mostly participated in plant hormone signal transduction, starch and sucrose metabolism, carbohydrate metabolism, cell growth and death, and transport and catabolism. Furthermore, correlation analysis of mRNA and miRNA revealed that 10 miRNA-target pairs from novel_mir65, novel_mir40, novel_mir80, miR397-5p_1, novel_mir36, novel_mir25 and novel_mir17 may have important roles in regulating the parasitic development of M. savatieri. Conclusions Our study not only expands the understanding of enhanced growth in M. savatieri after the establishment of parasite-host associations, but also first provides abundant resources for future molecular and genetic studies in M. savatieri.

BackgroundMonochasma savatieriis an endangered hemiparasitic medicinal plant with a variety of antioxidant, antimicrobial and anti-inflammatory properties. Despite the urgent need to understand the parasitic biology ofM. savatieri, parasite-host associations have long been neglected in studies ofM. savatieri.MethodsWe conducted a pot cultivation experiment to analyze changes in the growth traits, physiological performance and anatomical structures ofM. savatierigrown with the potential hostGardenia jasminoidesE., before and after the establishment of the parasite-host association.ResultsPrior to the establishment of the parasite-host association, the presence of the host had no significant effect on the maximum root length, leaf indexes or total dry weight ofM. savatieriseedlings, but had significant positive effect on seedling height, number of roots or number of haustoria. When it was continuously grown without a host,M. savatierigrowth was rather slow. The establishment of the parasite-host association enhanced the growth ofM. savatieri, and higher levels of photosynthetic pigments, increased antioxidant enzyme activity and lower malondialdehyde accumulation were observed inM. savatieriwith an established parasite-host association. Furthermore, an analysis of the anatomical structures ofM. savatierishowed that the establishment of the parasite-host association enabled better development of the seedling vegetative organs than that in seedlings without parasite-host associations.ConclusionsOur study demonstrates the physiological and anatomical changes that occurred inM. savatieriafter connection with a host and suggests that the enhanced growth and development ofM. savatieriwere highly dependent on the parasite-host association.

As a medicinal and edible plant, Chrysanthemum morifolium cv. "Hangju" has economic value and is widely used as a Chinese herbal medicine and tea. However, the metabolomic difference between "Hangju" flowers with white and yellow petals is unclear. In this study, Chrysanthemum variety ZXYJ with white petals and its bud mutation strain JJYH with yellow petals were used as research materials. A preliminary study found that the relative content of total flavonoids in JJYH was 45.4% higher than that in ZXYJ. Combined transcriptomic and metabolomic analysis revealed that the phenylpropanoid biosynthetic pathway and the phenylalanine metabolic pathway had the highest degree of differentially expressed genes and significantly changed metabolites enrichment, respectively. The redirection of metabolic flux may contribute to increased accumulation of total flavonoid levels in JJYH, which may begin as early as phenylalanine metabolism. The results of this research provide a scientific basis for the development of products related to different colored medicinal chrysanthemums and references for the breeding of new varieties and the biosynthesis of phenylpropanoids.

Amana edulis (Miq.) Honda is an important medicinal plant with a variety of anti-cancer properties, and it is of great importance to improve its reproductive rate through micropropagation technology to meet increasing demand. In the present study, in order to establish a rapid in vitro bulblet propagation protocol for A. edulis, an L16 (45) orthogonal design was used to investigate the effects of sucrose, 6-benzyladenine (6-BA), α-naphthaleneacetic acid (NAA), and macro-elements on bulblet induction. The results show that the sucrose concentration was the crucial factor for A. edulis bulblet initiation, followed by 6-BA and macro-elements, while NAA had the weakest effect. The optimal medium for in vitro bulblet induction was Murashige and Skoog medium supplemented with 0.1 mg·L−1 6-BA, 0.01 mg·L−1 NAA, and 100 g·L−1 or 80 g·L−1 sucrose (pH 5.8), in which A. edulis shoot clusters (without roots) were cultured at 5(±2)°C for 35 d or 60 d and then incubated at 23(±2)°C for 90 d. The entire cultivati...

Chrysanthemum morifolium. cv &ldquo, Hangju&rdquo, is an important medicinal material with many functions in China. Flavonoids as the main secondary metabolites are a major class of medicinal components in &ldquo, and its composition and content can change significantly after flooding. This study mimicked the flooding stress of &ldquo, during flower bud differentiation and detected its metabolites in different growth stages. From widely targeted metabolomics data, 661 metabolites were detected, of which 46 differential metabolites exist simultaneously in the different growth stages of &ldquo, The top three types of the 46 differential metabolites were flavone C-glycosides, flavonol and flavone. Our results demonstrated that the accumulation of flavonoids in different growth stages of &ldquo, was different, however, quercetin, eriodictyol and most of the flavone C-glycosides were significantly enhanced in the two stages after flooding stress. The expression of key enzyme genes in the flavonoid synthesis pathway were determined using RT-qPCR, which verified the consistency of the expression levels of CHI, F3H, DFR and ANS with the content of the corresponding flavonoids. A regulatory network of flavonoid biosynthesis was established to illustrate that flooding stress can change the accumulation of flavonoids by affecting the expression of the corresponding key enzymes in the flavonoid synthesis pathway.

Background: Prunella vulgaris L. has been an important medicinal plant for the treatment of thyroid gland malfunction and mastitis in China for over 2000 years. There is an urgent need to select effective wavelengths for greenhouse cultivation of P. vulgaris as light is a very important factor in P. vulgaris growth. Here, we described the effects of natural light (control) and UV solar exclusion on the morphological and physiological traits, secondary metabolites contents and antioxidant activities of P. vulgaris. Results: The results showed that UV solar exclusion resulted in remarkable alterations to morphological and biomass traits; significantly reduced the chlorophyll a, chlorophyll b and total chlorophyll contents; significantly enhanced the ratio of chlorophyll a to b; and significantly increased the carotenoid and anthocyanin contents in P. vulgaris. UV solar exclusion significantly increased the catalase (CAT) and peroxidase (POD) activities, increased superoxide dismutase (SOD) and ascorbate peroxidase (APX) activities and slightly decreased the glutathione (GSH) content. UV solar exclusion significantly increased the soluble sugar and H2O2 contents and increased the soluble protein content but significantly decreased the proline content and slightly decreased the MDA content. The secondary metabolite contents (total phenolics, rosmarinic acid, caffeic acid, hyperoside, ursolic acid and oleanolic acid) and in vitro antioxidative properties (DPPH· and ABTS·+scavenging activities) were significantly increased in P. vulgaris spicas under UV solar exclusion. Additionally, the total polysaccharide and total flavonoids contents were slightly increased by UV solar exclusion. The salviaflaside content was significantly reduced by UV solar exclusion. Conclusion: Our study demonstrated that P. vulgaris activates several antioxidant defence systems against oxidative damage caused by UV solar exclusion.

We have examined the influence of four phosphorus (P) levels on growth performance, photosynthetic efficiency and bioactive phytochemical production of Prunella vulgaris L. in hydroponic culture. Results that 0.20 mM P was enhanced the dry weight, shoot height, spica and root weights, spica length and number, total chlorophyll and carotenoid contents, net photosynthetic rate, transpiration rate, and stomatal conductance were determined after three months' treatment. A supply of surplus P resulted in a higher concentration of foliar P and was negatively correlated with biomass. Both P-deficient (0 mM) and high P (10.00 mM) resulted in increased concentrations of ursolic acid and oleanolic acid, with the exception of flavonoids. An increase in water extract from P. vulgaris spicas was noted with the application of increasing P concentrations. Our results indicated that the application of 0.20 mM P improves the biomass production and the yield of bioactive constituents of P. vulgaris.

We aimed to assess the effect of Radix Isatidis dietary supplementation on resistance to Proteus mirabilis infection in Poecilobdella manillensis . Six groups of leeches housed in tanks were randomly divided into five treatment groups (fed a basal diet supplemented with 2.5, 5.0, 7.5, 10.0, or 12.5 g/kg of Radix Isatidis ) and one control group (fed a basal diet). Compared with the control group, the specific growth rate (SGR) and weight gain (WG) were significantly decreased in the treatment groups supplemented with Radix Isatidis . However, supplementation with 2.5 to 10.0 g/kg of Radix Isatidis did not produce an effect on mortality. Mortality was recorded after the leeches were challenged with P. mirabilis , and changes were observed in the serum cortisol, glucose, alanine aminotransferase (ALT), 3,5,30-triiodothyronine (T3), thyroxin (T4), alkaline phosphatase (ALP), total protein (TP), globin, and lysozyme contents. In the ingluvies, changes were observed in the total anti-oxidative capacity, superoxide dismutase (SOD) and catalase (CAT) activity, malondialdehyde (MDA) content and heat shock protein 70 (HSP70) gene expression over a period of 7 d. Treatment with 5.0 to 10.0 g/kg of Radix Isatidis increased the following parameters: total anti-oxidative capacity of the ingluvies prior to infection and at 1, 2 and 7 d after infection; serum ALP activity and serum globin concentrations prior to infection and at 1 and 2 d after infection; T4 concentrations prior to infection; serum lysozyme activity at 1 d after infection; SOD and CAT activity in the ingluvies prior to infection and 7 d after infection; T3 concentrations at 1 and 2 d after infection; and the relative level of HSP70 mRNA expression in the ingluvies at 2 d after infection. However, treatment with 5.0 to 10.0 g/kg of Radix Isatidis increased the following parameters: ALT contents prior to infection and 1, 2 and 7 d after infection; serum cortisol concentrations and serum glucose at 1, 2 and 7 d after infection; MDA contents of the ingluvies at 1 and 2 d after infection; and T4 concentrations at 2 and 7 d after infection. Compared with the control group, significantly lower mortality was observed in the groups supplemented with 5.0 to 10.0 g/kg of Radix Isatidis . These results suggest that supplementation with 5.0 to 10.0 g/kg of Radix Isatidis in the basal diet could enhance resistance to P. mirabilis infection in P. manillensis. Statement of relevance We believe that this manuscript is appropriate for publication in Aquaculture because it provides a natural method of controlling disease in P. manillensis , thereby avoiding the use of antibiotics and the development of antibiotic resistance. Furthermore, the results of our study provide new information on the physiological and molecular mechanisms that underlie the protective effect of Radix Isatidis .

The dried roots of Zanthoxylum nitidum (Roxb.) DC have been widely used in Chinese medicine for the treatment of rheumatoid arthritis, toothache, stomach ache, bruises, and snakebites. Alkaloid content, particularly for nitidine chloride (NC), is a crucial indicator of herb quality. Abuse of plant sources of Z. nitidum, such as the usage of NC content not adhering to the Chinese Pharmacopoeia specification, has appeared in herb markets. Thus, a distribution survey was conducted on wild samples from 35 sites grown in 5 provinces, including the evaluation of morphological and phytochemical differences (three alkaloids, one flavonoid, and one lignin). Consequently, the surveyed plants were attributed to four types: three species (type I, II, and III) and one variety (type IV). The NC content of most type I, II, and III plants met quality guidelines, however, all three type IV plants from Guangxi Province exhibited low NC content; the use of plants from that area should be carefully considered. To identify the biosynthetic pathway of NC and other active compounds, nine transcriptomes of type III (root, stem and leaf, three replicates for each tissue) and three of type I (three leaf replicates) were performed to mine candidate unigenes. A set of 505,386 unigenes with an average length of 866 bp were identified. Coexpression analysis of tissue expression patterns were used to narrow down the candidate genes. Consequently, 23 candidate unigenes involved in the alkaloid biosynthesis pathway, 38 unigenes in the lignan biosynthesis pathway, and 16unigenes in the flavonoid biosynthesis pathway were screened. The distribution survey and phytochemical and transcriptome analyses conducted in this study provided valuable insights into resource preservation and the potential of molecular breeding to improve the quality of Z. nitidum products.

The red stigmas of saffron are one of the most expensive spices in the world and serve as a traditional Chinese medicine. More saffron has been cultivated in China, and different drying technologies have been studied. However, a comprehensive and comparative analysis of different drying approaches has not been well studied. In this study, we compared electric oven and vacuum oven drying approaches on saffron. We found saffron was dried quicker under high vacuum drying mode with high temperature and the quicker drying rate provided, the more open microstructural interstices on the saffron surface. Both methods were best fit to Midilli and Kucuk model. Besides, the coloring, aroma and bitterness strength after drying showed the similar results. In sum, our data suggested the optimal drying temperature was 100 °C for 20 min for two evaluated methods, however considering the machine cost, the electric oven drying would be the first choice. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10068-018-0487-x) contains supplementary material, which is available to authorized users.

A new cytoplasmic small heat shock protein gene was cloned from Pinellia ternata (Thunb.) Breit and designated PtHSP18.2 (GenBank number: KF693218). The PtHSP18.2 gene has an open reading frame of 711 bp that encodes 159 amino acids, and a molecular mass of 18.20 kDa. The phylogenetic tree indicated high homology of the PtHSP18.2 protein with that of cytosolic class I small heat shock proteins (CI sHSPs) from other plant species. PtHSP18.2 is expressed in root, leaf, and stem tissues. The expression of PtHSP18.2 was increased under heat and cold stress but was not induced by drought stress. When overexpressed in E. coli, PtHSP18.2 was correlated with the preservation of viability under heat stress. The CI sHSP family is the largest sHSP subfamily, and several different members of this subfamily are present in P. ternata. Two-dimensional electrophoresis (2-DE) western blot analysis with an anti-AtHSP17.7 (a member of CI sHSPs) antibody revealed five 18 kDa heat shock-induced P. ternata proteins. In combination, our findings suggest that PtHSP18.2 is potentially significant in the response to thermal stress.

Coreopsis tinctoria Nutt. from Mt. Kunlun (located in Tibet, Xinjiang) is a new Chrysanthemum tea material, and its volatile components are reported here for the first time. C. tinctoria produces a greater quantity and better quality of volatile components compared with two other Chrysanthemum tea materials (Chrysanthemum morifolium cv. ‘Hangju’ and Chrysanthemum morifolium cv. ‘Gongju’). The volatile oil content was 0.42%, with eighty-one components. The major components included six monoterpenes (limonene, 11.308%; α-phellandrene, 3.884%; p-acetyltoluene, 2.761%; carvone, 2.666%; 1R-α-pinene, 2.656%; and α-campholenal, 2.483%), three sesquiterpenes (α-bergamotene, 7.314%; tricyclo[6.3.0.0(1,5)]undec-2-ene-4-one,2,3,5,9-tetramethyl-, 3.454%; and naphthalene, 1,2-dihydro-1,1,6-trimethyl-, 3.024%), and a fatty acid component (n-hexadecanoic acid, 2.713%). Antioxidant activity study was revealed that the volatile oil of C. tinctoria had the better activity than the two compared samples. With next further study, C. tinctoria could be a more promising medicinal raw material than other popular chrysanthemum tea materials.

Tulipa edulis (Liliaceae) is the botanical origin of the traditional Chinese medicine (TCM) “Guangcigu”. Due to overexploitation that induced a decline in natural sources, many dried bulbs from other species of Tulipa have been used, adulterating the medicine in recent years. This practice may cause a series of inconsistent therapeutic effects and quality control problems in the herbal medicine industry. Hence, three DNA regions ( mat K, psb A- trn H and rbc L) were evaluated as barcodes for identifying T. edulis and its adulterants. All candidate DNA barcodes were successfully amplified from leaf samples. Based on the sequence divergences, rbc L and psb A- trn H can assign T. edulis and its adulterants to the correct genus, while mat K can accurately differentiate T. edulis and its adulterants. Thus, at the DNA level, the mat K intergenic region is a more suitable, accurate and applicable identification of T. edulis and its adulterants than rbc L and psb A- trn H.

Context: Changium smyrnioides Wolff (Apiaceae) is an endangered medicinal plant with numerous pharmacological uses. Objective: To investigate the effect of light intensity levels on the growth and accumulation of secondary metabolites of C. smyrnioides, cultivated seedlings were subjected to different relative light intensities via sun-shading. Materials and methods: Changium smyrnioides seedlings were subjected to five irradiance treatments (100, 60.54, 44.84, 31.39, and 10.56% sunlight) in glasshouse for 9 months. Enzymatic and non-enzymatic antioxidants with spectrophotometric method, photosynthetic parameters with Li-6400XT, dry matter accumulation and active component contents in the root with spectrophotometric and HPLC method were analyzed. Results: With an increase in relative light intensity levels, activities of enzymatic and non-enzymatic antioxidants, and malondialdehyde (MDA) contents were increased overall, while net photosynthetic rate (Pn) and dry matter accumulation patter first increased and then declined. The highest net photosynthetic rate (30.68 μmol/m2·s) and dry root weight (5.07 g) were achieved under 60.54% sunlight. Lower relative light intensity levels stimulated the accumulation levels of bioactive compounds in the roots so that the highest contents of mannitol (1.35%) and choline (405.58 μg/g) were recorded under 31.39% sunlight, and the highest polysaccharide content (10.80%) were achieved under 44.84% sunlight. With a decrease in the relative light intensity levels, the water-soluble component content increased first and then decreased. Discussion and conclusion: The results revealed that 31.39–60.54% sunlight serve as appropriate relative light intensity conditions for cultivated C. smyrnioides.

Soil is the natural medium for plant growth, and its chemical property is one of the most important factors for metabolic process of phytochemicals. Therefore, revealing the soil chemical property-crop effects will strongly improve the cultivation technology for higher yield and quality. Glechoma (G.) longituba has been long treated as a dedicated edible and medicinal herb. Here, the relationships between soil chemical properties and bioactive compounds of G. longituba were analyzed, and the effects of the most two related soil indicators on the plant were investigated and evaluated. The soil from thirty G. longituba provenances with different chemical properties were selected to test their effects on G. longituba. Based twelve soil indicators, the elements of phosphorus (P) and iron (Fe) were screened out as the most primary influential factors attributing to the high fluctuations of the contents of bioactive compounds. Through a factorial design of P and Fe by five different levels, significant influences of P, Fe, and their interaction on the bioactive compounds and biomass of G. longituba were validated. All the contents of the bioactive compounds in G. longituba expressed the same trend, which increased firstly and then decreased with the growing additions of P or Fe, but were slightly different in the peak levels. In all, the optimum extra addition was the combination of 0.9 g kg−1 P and 0–0.24 g kg−1 Fe. These findings could theoretically elucidate the connections between soil chemicals and bioactive compounds, and provide targeted cultivated strategies for improving the biological yield and medicinal quality of G. longituba.

Transcriptome sequencing data (6.5 Gb) of the salivary glands of the haematophagous leechHirudo nipponiawas obtained by using the BGIseq-500 platform. After identification and analysis, one transcript (Unigene5370) was annotated to hirudin HV3 fromHirudo medicinaliswith ane-value of 1e-29 and was named hirudin-HN. This transcript was a new thrombin inhibitor gene belonging to the proteinase inhibitor I14 (hirudin) family. Hirudin-HN, with a 270-bp cDNA, encodes an 89-aa protein containing a 20-aa signal peptide. The mature hirudin-HN protein contains the typical structural characteristics of hirudin, e.g., three conserved disulfide bonds and the PKP and DFxxIP motifs. Proteins (HirandM-Hir) were obtained via prokaryotic expression, and the mature hirudin-HN protein was shown to have anticoagulant activity and thrombin affinity by using the chromogenic substrate S2238 and surface plasmon resonance (SPR) interaction analysis, respectively. The N-terminal structure of the mature hirudin-HN protein was shown to be important for anticoagulant activity by comparing the activity and thrombin affinity ofHirandM-Hir. The abundances of Hirudin-HN mRNA and protein were higher in the salivary glands of starving animals than in those of feeding or fed leeches. These results provided a foundation for further study on the structure-function relationship of hirudin-HN with thrombin.

The objective of this study was to obtain an overview of the genetic relationships among and within Whitmania pigra and Hirudo nipponica using the ISSR and SRAP markers that were derived from related accessions, 11 W. pigra and 4 H. nipponica, from 8 provinces in China. All populations were uniquely fingerprinted by two markers. Mean genetic similarities were estimated at 0.76 and 0.79 using the ISSRs and SRAPs. Two main clusters that separated the W. pigra and H. nipponica populations were produced using UPGMA analysis. The Mantel test resulted in a good fit (SRAP = 0.80609) to the fit (ISSR = 0.76006) of the cophenetic values. Comparing the two marker systems, the ISSR and SRAP similarity indices were correlated (r = 0.8564). Finally, an appropriate strategy for conserving the Hirudo germplasm is proposed.

An assessment of the genetic diversity and variation of Pinellia ternata collected from 43 populations in China was undertaken using SRAP + TRAP markers. A total of 13 SRAP primers in addition to 3 TRAP primer combinations yielded 292 bands in total of which 286 were polymorphic (98.0%), with an average of 16 for each. The PIC value ranged from 0.88 to 0.95, with a mean polymorphic information content (PIC) of 0.92 over all the primers. A cluster analysis was conducted based on the unweighted pair group method with arithmetic average (UPGMA), and a principle coordinate analysis (PCA) was performed relying on Nei's genetic distance, both of which showed similar outcomes in that the results of clustering did not correlate to the geography locations but showed an association with morphology. An analysis of molecular variance (AMOVA) was performed to detect the source of variation; the results indicated that the main variation existed within populations (67.2%), whereas there was still 32.8% differentiation existing among the populations.

► The best harvest time of Chrysanthemum morifolium is when 50% tubular and 70% ray florets open. ► Dynamic accumulation of four bioactive components showed the trend of “low-high-low”. ► Genotype Wanyang (Tongxiang) exhibited the highest bioactive components contents. ► Genotype, origin and harvest time affect significantly the quality of C. morifolium.

This study assessed the molecular genetic diversity and relationships among 25 wild Prunella vulgaris L. populations and one cultivated population in China. The results showed that all the 26 P. vulgaris populations possessed high levels of genetic diversity. However, polymorphism was relatively lower in the cultivated population. Seventeen SRAP primer combinations generated 153 amplification bands, of which 134 were polymorphic. However, 195 polymorphic bands were detected out of 219 bands that were amplified using 20 ISSR primers. ISSR markers showed a slightly higher proportion of polymorphic loci (PPL = 89.19%) than did the SRAP markers (PPL = 87.93%). The results also showed that the 26 populations could be clustered into three groups by cluster analysis, which matched the geographic distribution of this species. The data indicated that ISSR and SRAP are both reliable and effective tools for analyzing genetic diversity in P. vulgaris .

To optimize the purification process of pigments from Coreopsis tinctoria with macroporous resins by establishing second regression model with response surface methodology. The experiment showed that XDA-7 resin had the best purification effect for pigments from C. tinctoria. The optimal absorption conditions for pigments from C. tinctoria were determined as follows： concentration of pigments solution 2.7 g•L⁻¹, flow rate 6 mL•min⁻¹, pH 6. Under these conditions, the absorption rate of pigments was up to 94.16%. Optimal desorption conditions were as follows： ethanol concentration 64%, flow rate 5 mL•min⁻¹, elution dosage 4 BV. Under these conditions, pigment desorption rate was as high as 98.72%.

Genetic variation and structure in 36 wild populations of Chrysanthemum indicum L. were assessed using chemical and molecular analyses in this paper. Using 24 selected ISSR primers 238 bands were generated, of which 205 bands were polymorphic (86.13%), and with 28 selected SRAP primer combinations 290 amplified bands were observed with 253 polymorphic bands (87.22%). The statistical results indicated that there was a high genetic differentiation among populations (GST 0.421 and 0.436). The UPGMA clustering results using ISSR, SRAP or ISSR + SRAP combination showed the genetic distance had no significant correlation with geographic distance. However, cluster results based on the contents of quercetin, luteolin, apigenin and acacetin did not keep in good accordance with molecular analysis, indicating that environmental and genetic factors may play an essential role in determining the bioactive compound levels of C. indicum.

Essential oils present in the dried flowers, leaves and stems of Prunella vulgaris were investigated for their chemical diversity using capillary GC and GC-MS. A total of 68 compounds were isolated from these three components: 28 from the flowers (representing 83.4 % of the essential oils present), 19 (80.1 %) from the leaves and 21 (79.5 %) from the stem. The most abundant components in the flower oil included hexadecanoic acid (70.0 %), (Z)-9-octadecenoic acid hexyl ester (3.8 %) and 2,2,3-trimethyl-decane (2.7 %). Aromadendrene (55.4 %), cucumber alcohol (8.5 %) and phytol (5.1 %) were the main components in the leaf. In the stem, hexadecanoic acid (38.4 %), trans-traumatic acid (22.4 %) and phytol (6.6 %) were the major components.

The diversity and genetic relationship among 29 populations of Chrysanthemum morifolium , one of Chrysanthemum indicum and one of Chrysanthemum nankingense from China were analyzed using morphological traits and molecular markers. Twenty morphological traits were scored as well as 182 ISSR marker-fragments, as amplified by 22 primers [the percentage of polymorphic bands (PPB): 81.87%], and 243 SRAP marker-fragments as generated by 26 primer pairs (PPB: 75.72%). Mantel’s test indicated significant correlation ( r = 0.624) of morphological trait and SRAP. By contrast, the morphological trait showed low correlation with ISSR ( r = 0.246). Cluster analysis showed groupings of the accessions according to all four methods correlated well with their geographic region of origin, and most populations from the south of China were classified into one cluster and most populations from the north of China were classified into another cluster. Finally, an appropriate strategy for conserving the C. morifolium germplasm was proposed.

Using chloroplast DNA (cpDNA) trnL-F sequences, we studied the phylogeographic pattern and demographic history of Chrysanthemum indicum with 81 individuals from 27 populations representing the partial geographic range in China. Fifteen haplotypes were identified based on nucleotide variation. Relatively high levels of haplotype diversity (Hd = 0.783) and low levels of nucleotide diversity (Pi = 0.00521) were detected in C. indicum. An analysis of molecular variance (AMOVA) indicated that most of the genetic variation (59.9%, P

Thesium chinense is a root hemiparasite and the whole weed serves as a traditional medicine in China. The resource of T. chinense is sharply declining, due to unsustainable exploitation and environmental destruction. At the same time, the obligate hemiparasitic nature and the relationship of the T. chinense-host association are not well understood. In order to protect and cultivate T. chinense, we aimed to find a superior host for T. chinense by studying the effect of different hosts (eight host species and without host) on the growth, chlorophyll and nutrient content of pot-cultured T. chinense. The results showed that the presence of a host species significantly promoted the growth of T. chinense but considerable differences existed among different host species. Imperata cylindrica, Prunella vulgaris, Chrysanthermum indicum were more efficient in promoting the growth of T. chinense than the other hosts. Thesium chinense chlorophyll (chl) concentration and chl a:chl b ratio increased significantly when grown with a host. Asp and Glu were the most common amino acids in T. chinense. Dry weight of T. chinense per plant was significantly positively correlated to the concentration of Pro (r=0.870, p

The effects of UV-B radiation on the content of bioactive components and the antioxidant activity of Prunella vulgaris L. spica during development were studied. The experimental design involved two levels of UV-B radiation intensity (0 and 120 μW cm−2 nm−1). The results showed that the contents of total flavonoids, rosmarinic acid, caffeic acid and hyperoside, as well as the antioxidant capacities (DPPH● and ABTS•+ scavenging activities), in the spicas significantly decreased during spica development. The content of salviaflaside in the spicas significantly increased during development. The highest contents of total flavonoids, rosmarinic acid, and caffeic acid and the highest antioxidant activities were found in spicas in the full-flowering stage, while the highest content of hyperoside was found in spicas in the bud stage. In addition, the highest content of salviaflaside was found in spicas in the mature-fruiting stage. UV-B radiation significantly promoted the synthesis of secondary metabolites, increased the contents of the main bioactive components in the three developmental stages of isolated dried spicas, and significantly increased the DPPH● and ABTS•+ scavenging activities of P. vulgaris spicas in the mature-fruiting stage. Moreover, the total flavonoids content was positively correlated with the DPPH● and ABTS•+ scavenging activities, and the correlation with the DPPH● scavenging activity was very strong. This result shows that the highest contents of the main bioactive components in the spicas were not all found in the same developmental stages of P. vulgaris. Our research revealed that the best stage for harvesting P. vulgaris spica was between the bud stage and the full-flowering stage since harvesting at this point provides a higher content of bioactive components and a higher antioxidant capacity, which is relevant for medicinal applications.

Thesium chinense (Santalaceae) is an obligate root hemiparasite and the whole weed serves as a traditional medicine in China. This paper examined chlorophyll (Chl) concentration, transpiration (E), photosynthesis (Pn), stomatal conductance (g), respiration nighttime (Rd), water use efficiency (WUE), and mineral elements of T. chinense grown with different hosts or without host. The results showed that Chl concentrations, Pn, Pnmax, and WUE of T. chinense grown with host were significantly higher, approximately 1.5–3 folds, than that grown without host. E and g value of T. chinense greatly exceeded that of their hosts during the daytime—they increased about 1.5–3.5 folds (except of Gnaphlium affine) and 1.2–3.0 folds, respectively, while maintaining little higher or equal to their host during night but very much lower than daytime. The WUE value of T. chinense was generally much less than that of their hosts, except Eremochloa ophiuroides. T. chinense grown with hosts had higher concentration of K, P, Cu and K/Ca ratio, and lower Na concentration compared with that without host plants. There were also differences among different host species backgrounds on all examined parameters of T. chinense. We pointed out that Imperata cylindrica, Prunella vulgaris, and Gnaphlium affine were more suitable hosts than E. ophiuroides. As an additional result T. chinense can mostly be independent as for C supply.

Seven dark-septate endophytic (DSE) fungi have been isolated from the roots of Epimedium wushanense T. S. Ying (Berberidaceae), an important medicinal plant with various pharmacological activities.The current study explores the effects of seven DSE fungi on the growth and accumulation of bioactive compounds in E. wushanense.Each 1-month-old E. wushanense seedling was inoculated with one of the seven DSE fungi and was grown under greenhouse conditions for 90 d. The molecular identification of the fungi was based on the ITS1-5.8S-ITS2 nuclear ribosomal gene cluster.The results showed that the influence of DSE fungi inoculation varied between strains. Inoculation with DSE8 not only significantly enhanced plant height, root length, leaf area, leaf number, and shoot and root biomass but also improved the total flavonoid and icariin content, with an increase ranging from 20.24% to 237.97%. Three of the seven DSE fungi caused the inoculated plants to die, and the remaining three DSE strains showed neutral or negative effects on plant growth and the accumulation of bioactive compounds. According to the ITS sequence, DSE8 is congeneric to the genus Leptodontidium.The findings indicate that application of DSE8 may be valuable to facilitate the cultivation of E. wushanense with a higher biomass and improved medicinal quality.

Prunella vulgaris L (Labiatae) is commonly used as a traditional medicinal herb in some Asian and Europe countries. To date, few studies have been conducted to determine the influence of [Formula: see text] - N/[Formula: see text] - N ratio on growth, physiological development, and bioactive phytochemical accumulation in hydroponically grown P. vulgaris.The current study was conducted to evaluate the effect of five [Formula: see text] - N/[Formula: see text] - N ratios on growth, nitrogen metabolism, photosynthetic efficiency, and bioactive phytochemical production in P. vulgaris.Hydroponically cultivated P. vulgaris were fertilized with five [Formula: see text] - N/[Formula: see text] - N ratios in a greenhouse for 85 d. Dried weight of root, stem, leaf and spica, leaf area, photosynthetic efficiency, activities of nitrate reductase (NR), glutamine synthetase (GS), and the concentrations of N, soluble protein, and free amino acids in the leaves, as well as the contents of rosmarinic acid (RA), ursolic acid (UA), and oleanolic acid (OA) in the spicas were measured.Both [Formula: see text] - N and [Formula: see text] - N as the sole source of nitrogen had inhibitory effects on P. vulgaris growth. P. vulgaris fertilized with the 25/75 ([Formula: see text] - N/NO3 - N) ratio had the highest leaf area, photosynthetic rate, and chlorophyll content. The 25/75 ([Formula: see text]/[Formula: see text]) ratio increased the spica biomass by 1828%, nitrate-reductase (NR) activity by 98%, and soluble protein concentration by 29.45% compared with the 100/0 ([Formula: see text]/[Formula: see text]) treatment. Additionally, 25 [Formula: see text] - N/75 NO3 - N resulted in the highest contents of RA and total flavonoids as well as relatively high contents of UA and OA; therefore, this ratio had the highest yield of RA, UA, OA, and total flavonoids in spicas.The use of 25 [Formula: see text] - N/75 [Formula: see text] - N is recommended to improve biomass production and medicinal quality of P. vulgaris.

Pinellia ternata is an important traditional Chinese medicinal plant. The growth of P. ternata is sensitive to high temperatures. To gain a better understanding of heat stress responses in P. ternata, we performed a comparative proteomic analysis. P. ternata seedlings were subjected to a temperature of 38 °C and samples were collected 24 h after treatment. Increased relative ion leakage and lipid peroxidation suggested that oxidative stress was frequently generated in rice leaves exposed to high temperature. Two-dimensional electrophoresis (2-DE) was used to analyze heat-responsive proteins. More than 600 protein spots were reproducibly detected on each gel; of these spots, 20 were up-regulated, and 7 were down-regulated. A total of 24 proteins and protein species were successfully identified by MALDI-TOF/TOF MS. These proteins and protein species were found to be primarily small heat shock proteins (58%) as well as proteins involved in RNA processing (17%), photosynthesis (13%), chlorophyll biosynthetic processes (4%), protein degradation (4%) and defense (4%). Using 2-DE Western blot analysis, we confirmed the identities of the cytosolic class II small heat shock protein (sHSPs-CII) identified by MS. The expression levels of four different proteins [cytosolic class I small heat shock protein (sHSPs-CI), sHSPs-CII, mitochondrial small heat shock protein (sHSPs-MIT), glycine-rich RNA-binding protein (GRP)] were analyzed at the transcriptional level by quantitative real-time PCR. The mRNA levels of three sHSPs correlated with the corresponding protein levels. However, GRP was down-regulated at the beginning of heat stress but then increased substantially to reach a peak after 24 h of heat stress. Our study provides valuable new insight into the responses of P. ternata to heat stress.

Prunella vulgaris L. is an important medicinal plant with a variety of pharmacological activities, but limited information is available about its response to potassium chloride (KCl) supplementation. P. vulgaris seedlings were cultured in media with four different KCl levels (0, 1.00, 6.00 and 40.00 mM). Characteristics relating to the growth, foliar potassium, water and chlorophyll content, photosynthesis, transpiration, nitrogen metabolism, bioactive constituent concentrations and yield were determined after three months. The appropriate KCl concentration was 6.00 mM to result in the highest values for dry weight, shoot height, spica and root weight, spica length and number in P. vulgaris. The optimum KCl concentration resulted in a maximum net photosynthetic rate (Pn) that could be associated with the highest chlorophyll content and fully open stomata conductance. A supply of surplus KCl resulted in a higher concentration of foliar potassium and negatively correlated with the biomass. Plants that were treated with the appropriate KCl level showed a greater capacity for nitrate assimilation. The Pn was significantly and positively correlated with nitrate reductase (NR) and glutamine synthetase (GS) activities and was positively correlated with leaf-soluble protein and free amino acid (FAA) contents. Both KCl starvation (0 mM) and high KCl (40.00 mM) led to water loss through a high transpiration rate and low water absorption, respectively, and resulted in increased concentrations of ursolic acid (UA), oleanolic acid (OA) and flavonoids, with the exception of rosmarinic acid (RA). Moreover, the optimum concentration of KCl significantly increased the yields of RA, UA, OA and flavonoids. Our findings suggested that significantly higher plant biomass; chlorophyll content; Pn; stronger nitrogen anabolism; lower RA, UA, OA and flavonoid accumulation; and greater RA, UA, OA and flavonoid yields in P. vulgaris could be expected in the presence of the appropriate KCl concentration (6.00 mM).

Objective To compare botanical characteristics of cultivated Chrysanthemum morifolium cv. 'Hangju' of different origins in order to provide the basis for introduction and cultivation of Ch. morifolium cv. 'Hangju'. Method The characteristics of plants, leaves and capitulum of Ch. morifolium cv. 'Hangju' were measured, and the obtained data were analyzed and compared. Result The range of plant height was 60.87-99.47 cm, number of branches 2.76-5.20, leave length 4.90-8.40 cm, leave width 3.25-5.38 cm, aspect ratio of leave 1.35-1.83, number of leave split 1.92-3.08. Numbers of capitulum were 21.92-53.12, diameter of capitulum 3.41-5.48 cm, lays of ray florets 3.28-7.16, number of ray florets 55.32-114.60, ray florets length 1.58-2.37 cm, ray florets width 0.50-0.69 cm, aspect ratio of ray florets 2.90-3.99, diameter of tubular flower 1.10-1.58 cm. Conclusion The botanical characteristics of cultivated Ch. morifolium cv. 'Hangju' were distinguished from different origins. With the cultivation environment change, the botanical characteristics of the cultivars are changed.

OBJECTIVE To study the influence of host species on growth and development and active component content of Thesium chinense. METHOD Plant morphology and active component content of T. chinense grown with different hosts were measured. The hosts were evaluated by using index-sum method. RESULT Hosts significantly promoted the growth of T. chinense by increasing height, per plant weight, stem diameter, leaf area, the number of seed and haustorium,and decreased RW/SW ratio compared to without host treatment. Considerable differences existed among the effect of different host species treatments. Synthetical evaluation score of Gnaphlium affine was the highest (37), followed by Imperata cylindrical and Prunella vulgaris (36). It is suggested that they were superior hosts for T. chinense. But the scores of Triticum aestivum (25) and Eremochloa ophiuroides (17) were lower, so they were unsuited hosts for T. chinense. CONCLUSION Hosts significantly promoted the growth of T. chinense. Considerable differences existed among the growth and development of T. chinense grown with different hosts, as well as active component content. Gnaphlium affine, Imperata cylindrical and Prunella vulgaris were superior hosts for T. chinense. While Triticum aestivum and Eremochloa ophiuroides were unsuited hosts for T. chinense.

OBJECTIVE To study the in vitro embryo culture of Epimedium wushanense and provide scientific basis for large scale production of tissue culture. METHOD Cullus and buds were induced from embryo of E. wushanense on a MS medium supplemented with different 2,4-D,6-BA, NAA, IBA. RESULT The optimal compositions of medium that induced callus and buds from embryo were the MS medium supplemented with 2,4-D 2 mg x L(-1), IBA 2 mg x L(-1) and NAA 0.5 mg x L(-1) and the MS medium supplemented with IBA 2 mg x L(-1) and 6-BA 0.5 mg x L(-1), respectively. The optimum medium for callus differentiation was MS + 6-BA 1 mg x L(-1) + NAA 0.5 mg x L(-1) + IBA 1 mg x L(-1), and MS +6-BA 1.0 mg x L(-1) + NAA 0.5 mg x L(-1) for shoots proliferation. CONCLUSION Using embryo as explants, the method of induction and culture of E. wushanense was established by the callus and buds, and the embryo of E. wushanense can be quickly propagated.

OBJECTIVE To achieve the rapid identification for seedlings of medicinal Chrysanthemum morifolium, the discriminant equation was established and the software for rapid identification was designed. METHOD Leaf structure of medicinal Chrysanthemum of 12 cultivars was analyzed to establish the discriminant equation based on variance analysis and discriminant analysis. On this basis, the identification program and software (based on the python language) were designed. RESULT Through the analysis of variance and multiple comparisons for the 11 leaf parameter index data of 12 different cultivars, it was found that that the leaf parameters were significant different from each other and reached significant levels. The discriminant equation and the rapid identification software were set up based on the analysis of various indicators. CONCLUSION The rapid identification system of seedlings of medicinal Chrysanthemum could be achieved through the establishment of discriminant equation combined with computer technology.

OBJECTIVE To provide the basis for standardization cultivation of medicinal Chrysanthemum morifolium, the relationship between the seedling grading and plant growth, yield and quality were studied. METHOD The morphological index of the seedlings was measured and the method of principal component and correlation analysis were used to determine the grading index, and step-wise cluster analysis was applied for clustering analysis. Pot experiments were used to measure the indicators of plant growth and development, yield and quality. RESULT The height and ground diameter were determined for the quality indicators of the seedlings grading, and the standard quality grading of seedlings of Ch. morifolium was initially set up. The ground diameter of the class I and II were larger than that of the class III, and the number of branches of class I and II was more than that of the class III, on the contrary, the plant height of the class III was higher than that of the class I and II. The shape and appearance of the plant had no effect on the intrinsic quality. Flower center diameter and tubular floret number of the class I and II were significantly larger than those of the class III, so as the yield. The seedling grading had no obvious effect on the internal quality of medicinal material. CONCLUSION Seedlings of the class I and class II were suitable for transplanting.

Prunella vulgaris L. (Labiatae) is a perennial plant common in China and Europe and is rich in rosmarinic acid (RA), ursolic acid (UA), and oleanolic acid (OA). The dried spica of P. vulgaris has been used as traditional medicine in China for over a hundred years. To our best knowledge, no study has been conducted to determine the influence of harvesting time on concentrations of bioactive compounds of P. vulgaris.In the current study, changes in the bioactive compounds present in spicas were investigated at five harvest times over 2 months.Plant material were collected at five fixed dates: 5th May, 20th May, 7th June, 15th June, and 25th June and assayed for chemical contents by high-performance liquid chromatography (HPLC).Among the different harvest times, the highest levels of RA (56.81 mg·g⁻¹), UA (2.77 mg·g⁻¹), and OA (0.91 mg·g⁻¹) were found on 5th May, whereas the lowest levels of RA (1.66 mg·g⁻¹), UA (2.27 mg·g⁻¹), and OA (0.43 mg·g⁻¹) were observed on 25th June.As each medicinal product has its own content requirement for different bioactive components, the optimum harvest time might be determined according to the accumulation dynamics of target compound in dried spicas of P. vulgaris. These results may be useful for determining the optimal harvest time when bioactive components are at the maximum level, which is in early May.

Prunella vulgaris L. (Labiatae) contains a variety of structurally diverse natural products, primarily rosmarinic acid (RA), ursolic acid (UA) and oleanolic acid (OA), which possess a wide array of biological properties. In the present study, P. vulgaris was harvested at three developmental stages (vegetative, full-flowering and mature-fruiting stages), dissected into stem and leaf tissues and assayed for chemical contents using high performance liquid chromatography. Significant changes in the concentrations of the major secondary metabolites (RA, UA and OA) were observed at the different development stages. The highest concentrations of RA, UA and OA were found at the full-flowering stage (15.83 mg/g dry weight (DW) RA, 1.77 mg/g DW UA and 0.65 mg/g DW OA). Among the different aerial parts of the plant, the concentrations of RA, UA and OA were higher in the leaves than in the stems at the different developmental stages. These results suggest that the full-flowering stage is characterized by the highest concentrations of bioactive compounds. Therefore, this stage may be the optimum point for harvesting P. vulgaris plants. In additional, the leaves of P. vulgaris demonstrated higher RA, UA and OA concentrations than the stems, suggesting higher utilization potential.

Chrysanthemum morifolium Ramat. is not only a famous floricultural crop but also a plant of pharmacological importance. An experiment was carried out to evaluate the effects of different colored polyethylene shade on the growth and accumulation of chlorogenic acid, total flavonoids, quercetin, apigenin and acacetin in C. morifolium capitulum. The results showed that colored polyethylene shade decreased the flower number and biomass yield while increasing the flower diameter and plant height, and the magnitude of effect was in the order of blue>red>yellow. The chlorogenic acid content was observed to be highest under yellow and blue polyethylene cover at different stage of flower development. Blue shaded polyethylene led to lower contents of total flavonoid, quercetin, apigenin, and acacetin. The contents of total flavonoids and quercetin under different colored shaded polyethylene peaked at different stage throughout the study period. The contents of acacetin and apigenin in chrysanthemum capitulum from four shaded polyethylene initially increased and then decreased throughout the study period. The results suggested that different color of polyethylene could induce changes in the microclimate that could act through natural regulatory system within chrysanthemum and affect growth and bioactive compounds accumulation. Key words: Chrysanthemum morifolium Ramat. cv. ‘Chuju’, colored polyethylene shade, growth, bioactive compounds.

Objective To study the biological and growth characteristics of Thesium chinense. Method The wild population was observed in certain sites, pot and indoor experiments were applied for cultivated plant. Result and conclusion T. chinense is an obligate root hemiparasite and perennial herb, stem and leaf were green, there were haustoriums on its root, can only achieve its whole life process when parasitized to host. T. chinense grows mostly in the humid wasteland and herbaceous community where water is generally abundant. T. chinense likes light, warm climate and acidic to neuter soil. The seeds of T. chinense have dormancy characteristics. The life history of T. chinense can divide into 5 stages: seedling, ramify, florescence, fructification and withering periods.

Glechoma longituba (Nakai) Kupr. (Labiatae) is an important medicinal plant with various pharmacological activities. Current study was carried out to establish the variation pattern in the contents of ethanol-soluble extractive, total flavonoids, ursolic acid (UA) and oleanolic acid (OA) in G. longituba in relation to harvesting time and geographical origin. G. longitubawas harvested at one month interval during ontogenetic development phases and twenty-nine populations were collected from different distribution areas in China. The results showed that the contents of ethanol-soluble extractive, total flavonoids, UA and OA in G. longituba were related to harvesting time and distribution origin. The optimum harvesting time should be in mid-April in terms of the contents of ethanol-soluble extractive and total flavonoids, and be in mid-August or in mid-March in terms of the contents of UA and OA. Populations of G. longituba showed remarkable differences in chemical composition depending on the provenance of plants. And the differences among 29 populations are likely to be genetically controlled since all populations were grown under uniform conditions. Our results on seasonal and environmental factors will be useful for commercial producers of G. longituba in determining the optimum harvesting time and the most appropriate plants for germplasm evaluation and breeding. Key words: Glechoma longituba (Nakai) Kupr., harvesting time, inter-populational variability, total flavonoids, ursolic acid, oleanolic acid.

OBJECTIVE The accumulation of dry matter, nitrogen, phosphorus and potassium in Prunella vulgaris and their relationships under field condition were studied, in order to provide a basis for the rational fertilization. METHOD Through the dynamic sampling of P. vulgaris in different growing stages, and the dry weight of plant, the amount of dry matter and the content of N, P and K within each organ of the plant were measured. RESULT The dry weight of P. vulgaris linearly increased with the increase of nitrogen, phosphorus and potassium accumulation, there existed a significant correlation at whole growth period. The maximum stage accumulation rate of dry matter from mid-April to the beginning of May, reached 63.71%. The amount of dry matter was mainly distribution in leaves from late March to mid-April. The distribution ratio in stem was the highest from beginning of May to late May. Until mid-June, the distribution ratio of dry matter in spicas was higher than that in other organs. The absorption amounts of N, P and K were lower before mid-April, and the nutrient element was concentrated in plant leaves. From late April to beginning of May, the N, P and K accumulation reached maximum of total absorptive capacity, which accounted for 72.35%, 59.82%, 87.41%, respectively, and the mineral elements were concentrated distribution of plant stems. When transferred to reproductive stage, both accumulation amount and distribution rate of nutrient elements in spicas increased rapidly, which reached the maximum until mid-June. From elongation stages to quaring period was the period of maximal accumulation of dry matter and showed the maximal efficiency of N, P and K nutrition. CONCLUSION The time from elongation stages to quaring period is the most crucial period in the growth development stage of P. vulgaris, and should be paid more attention to the supply of mineral nutrition and moisture of plant to promote spicas formation and dry matter accumulation in this stage. The average N, P2O5 and K2O absorption amount per 100 kg dry matter of spicas were 1.62, 0.36, 2.88 kg respectively, and the ratio of N-P2O5,-K2O was 1 : 0.22: 1.77.

Chrysanthemum indicum L. is a significant medicinal plant, which flowers (capitula) are usually used as Chinese medicine. The phytochemical test analyzed the regularity of biosynthesis and degradation of flavonoids and phenolic acids in flowers which grown under colored polyethylene and harvested from five flowering stages. Dry weight of one thousand buds or flower heads from different maturation stages and total number of flowers from individual plant were evaluated as biomass yield. The results indicated that the total number of buds and flower heads of individual plant grown under red polyethylene film was substantially higher than other treatments (P

Objective The contents of mineral elements in soil and each organ from cultivated and wild Prunella vulgaris were determined in order to provide a theoretical basis for exploitation and protection of wild resources and GAP study. Method The contents of mineral elements (P, K, Ca, Mg, Na, Fe, Cu, Zn, Mn, Al) in soil and each organ from cultivated and wild P. vulgaris were determined by ICP-AES, and the results were analyzed by correlation analysis. Result The contents of mineral elements in soil and different organs of P. vulgaris were significant different between cultivated and wild species; the contents of P and Cu ranked in the order of spica > leaf > stem, the order of Mg, Ca, Zn, Fe, Mn, Al from high to low was leaf > spica > stem, the concentrations of K, Na were mainly distributed in leaf and stem, less in spica. A distinctly positive correlation between soil and spica for Fe content, and the relationship between soil and leaf, stem were significant positive correlation, the concentrations of Zn in spica, leaf, stem were positively related to soil, K contents in soil and stem were significant positive correlation. The different organs of cultivated and wild P. vulgaris had a strong tendency to accumulate P. Conclusion There was no direct relationship between the concentrations of mineral elements in P. vulgaris, and their corresponding soil P. vulgaris adjusts the concentration of mineral elements mainly by active absorption.