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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.

The present study was conducted to investigate the rule of sex steroid hormones dynamic in the periodic development of Whitmania pigra gonad. The dynamic of sex steroid hormones in different age of Wh. pigra were detected by enzymolysis, extraction and UPLC-MS/MS. The results showed that the concentrations of estrone, estriol, testosterone and progesterone in the Wh. pigra showed M-type curve and peaked in 6-month-old Wh. pigra. The concentrations of above-mentioned four steroid hormones were the lowest in 9-month-old and then increased slightly. The another peaks of testosterone and progesterone were found in 2-month-old and the another peaks of estrone and estriol were obtained in 8-month-old. The concentrations of testosterone and progesterone are slightly higher than estriol during 1-month-old to 4-month-old, and thereafter strone and estriol showed higher concentration than testosterone and progesterone. In summary, the concentrations of four sex steroid hormones in Wh. pigra increased gradually with the maturation of gonads and decreased rapidly after the discharge period, which indicated that the ratio between estrogen, androgen and progesterone may be greatly related to the specificity of gonad development.

To explore the karyotype difference and to provide basis for genetic resources conservation and breeding of Tulipa edulis.Karyological analysis on nine Tulipa edulis populations was carried out with conventional pressed slice method.Nine Tulipa edulis populations investigated were diploid. Seven populations of Tulipa edulis were diploid with x = 12,and the chromosome numbers was 2n = 2x = 24. While the others were diploid with x = 24,and the chromosome numbers was 2n = 2x =48. Chromosome types of Tulipa edulis consisted of m,sm and st. The ratio of the longest chromosome to the shortest one ranged from1. 88 to 2. 34. Asymmetry coefficient ranged from 62. 99% to 71. 97%. Karyotype inclucled 2A,3A and 3B,in which 3B type was the most type and 2A type was observed for the first time in Tulipa edulis.The karyological type varies with population of Tulipa edulis.

In this paper, on the contrast of healthy leech, the bacterial diversities were analyzed by 16S rDNA sequence analysis of the bacteria of muscle and intestinal tract of Whitmania pigra, the environment water and sediment of cultivating the diseased Wh. pigra in high temperature by high-throughput sequencing to determine the possible pathogenic bacteria of bacterial diseases of Leech in high temperature. The results showed that the original sequence reached over 83 000, and the effective sequences accounted for more than 87%. The GC contents ranged from 52% to 54% and the bacterial diversities were abundant. Bacterial relative abundance analysis showed that the bacterial content of Proteobacteria, Bacteroidetes, and Firmicutes was the most abundant in all treatments. Compared with healthy leech muscles and intestines, the muscle and intestinal tract of pathogenic leech relative abundance of Bacteroides, Pseudomonas, and Desulfovibrio was significantly increased, and it was abundant in water and sediment of diseased leeches, Lead to the possibility that the pathogenic bacteria of this bacterial disease may be Bacteroides, Pseudomonas, Desulfovibrio.

The present study explored the effects of Cu²⁺enrichment and release on the growth and internal quality of Whitmania pigra, and the regularity of enrichment and release in different tissues of Wh. pigra. In the range of safe concentration(SC), a certain concentration of Cu²⁺ was added to the Wh. pigra for 50 days, and stopped adding for another 50 days. The growth index of Wh. pigra, Cu²⁺ content in different tissues, and the internal quality were determined. The results showed that the average body weight of Wh. pigra in 0.014 mg·L⁻¹ of Cu²⁺ group was significantly higher than that in control group during the experimental period (

The present study was conducted to explore the effect of light intensity on growth, bioactivity compounds accumulation and anti-oxidative activity of Sedum sarmentosum. The growth, yield, contents of total flavonoids, total phenolic, quercetin, kaempferol and isorhamnetin, and antioxidant activities were assessed in S. sarmentosum under five light intensities, namely 100% full sunlight (G1), 77% full sunlight (G2), 60% full sunlight (G3), 38% full sunlight (G4), and 16% full sunlight (G5). The results showed that light intensity significantly affected the growth and the chemical compounds accumulation. With the decrease of light intensity, the maximum branch length and the average internode distance increased. G2 treatment greatly promoted the numbers of leaf layers and branches, and G3 treatment remarkably improved the yield. The highest total flavonoids and phenolic contents were obtained in G3 treatment. Meanwhile, the highest quercetin and isorhamnetin contents were obtained in G1 treatment. The difference of kaempferol content was not significant. In addition, based on DPPH, FTC and FRAP methods, the antioxidant activities of the aqueous extracts under G1 treatment were superior to the others. The results indicated that more than 60% full sunlight was the optimum light intensity condition to achieve high yield and quality of S. sarmentosum.

The issues of disordering production and non-standard pesticide application are common in the production of Chinese herbal medicines. Aimed to above problems, research groups built the pollution-free and precision cultivation system of medicinal plants. This system mainly included the precise site selection of medicinal plants based on the GIS technology, modern omics-assisted breeding, metagenomics guiding the soil complex improvement, and the precise field management based on rational application of fertilizer and comprehensive control of disease. At present, the production and distribution of medicinal plants were performed in the many poor counties of the whole nation. The breeding platform of resistant varieties was built, and certificates of new and well-bred varieties were received, in the base of genetic backgrounds of the original species of medicinal plants. The disease incidences were declined after application of these resistant varieties. Additionally, chemical pesticide consumption of medicinal plants (such as Panax ginseng, P. notoginseng, Salvia miltiorrhiza, P. quinquefolium, Schisandra chinensis, Platycodon grandiflorum and P. grandiflorus etc.) reduced by 20%-80% based on the genetic testing technologies of plant diseases and insect pests and safety evaluation of pollution-free pesticides. The application of pollution-free and precision cultivation system of Chinese herbal medicines achieve significantly social, economic and ecological benefits.

To explore the effect and mechanism of Astragali Radix on growth and immunity of Whitmania pigra, 0, 0.01%, 0.03%, 0.05%, 0.07%, 0.09% of Astragali Radix were added to the daily feeding of Wh. pigra. After 60 days of feeding, the growth performance, activities of digestive enzyme and anti-reverse enzyme, inner quality, the expression levels of GH, IGF-1 and digestive enzyme-related genes were measured. Meanwhile, the effects of heat stress on the living conditions of Wh. pigra were observed and counted, and the expression levels of HSP70 and immune related genes were measured. The results showed that the final weight, weight gain rate, specific growth rate, activities of digestive enzyme and anti-reverse enzyme, the expression levels of GH, IGF-1 and digestive enzyme-related genes in the Astragali Radix group were higher than those in the control group, and with the increase of Astragali Radix concentration, the above-mentioned indexes increased initially and then decreased, and significantly higher in the 0.05% of Astragali Radix group than in the other groups (

Through the determination of the growth, physiological and active components of Glechoma longituba, the effect of soil bulk density on growth and quality of G. longituba was explored. Through pot experiment, six different bulk density treatments were set for 0.8, 0.9, 1.0, 1.1, 1.2, 1.3 g·cm⁻³ , the growth index, physiological index and the content of active components were determined. The results showed that the growth indexes and biomass of G. longituba firstly increased and then decreased with the increase of soil bulk density. The dry weight of G. longituba achieved the maximum 5.70 g in the treatment of 1.0 g·cm⁻³ . The photosynthetic pigments, soluble sugars and free amino acids firstly increased and then decreased with the increase of soil bulk density. The suitable soil bulk density was beneficial to the accumulation of photosynthetic pigment, soluble protein and free amino acid synthesis. The content of malondialdehyde increased gradually with the increase of soil bulk density. The contents of ethanol-soluble extractives of G. longituba were greater than 25%, meeting its quality criterionin Chinese Pharmacopoeia 2015, achieved the maximum of 40.66% in the treatment of 1.0 g·cm⁻³ . The total flavonoids content of G. longituba increased gradually with the increase of soil bulk density, triterpenic acid and phenolic acids decreased gradually with the increase of soil bulk density. Soil bulk density has a significant effect on the growth and quality of G. longituba, and it is conducive to the growth and its medicinal quality at the soil bulk density of 1.0 g·cm⁻³.

The current study was conducted to explore the relationship between property of traditional Chinese medicines (TCM) and the light conditions of their original plant in habitats and advance a "light-cold and heat property" hypothesis under the guidance of theory of TCM. In the present study, we selected 127 common Chinese herbs. Association rules mining method was used to find out the association relationship between the cold-heat properties of different TCM and the habitats of their original plant, while using statistical methods to analyze the correlation between light condition and cold-heat property of TCM based on the description of the habitats in the Flora of China and investigation. The results showed that the habitats associated to TCM with cold property are in the order of grasslandcurbhillsidewastelandgullyfarmlandriverside. The habitats that associated to TCM with heat property are: undergrowthshrubgullyjungle. For 68 TCMs with cold property, 81.2% of their original plants are light-demanding, 8.7% are shade-demanding, and 8.7% of them showed no obvious light preference. For 59 TCMs with heat property, 62.7% of their original plants are shade-demanding, 20.3% are shade-demanding, and 17.0% of them showed no obvious light preference. Meanwhile, Chi-square test showed that there was a significant difference between the cold-heat property and light preference (

We cloned flavonol synthase gene (named as CmFLS) by RACE from Chrysanthemum morifolium cv. 'Hangju' based on transcriptome database. Sequencing results showed that 1 235 bp sequence was acquired with the largest open reading frame (ORF) of 1 008 bp, which encoded 335 amino acids. The predicted CmFLS encoded protein had an isoelectric point (pI) of 5.41. The phylogenetic tree analysis indicated that CmFLS was highly homologous to other FLSs, which identified from the species of Compositae. The recombinant fusion protein, with a molecular mass of 43 kDa, was successfully expressed by prokaryotic expression system. Meanwhile, Ni-NTA resin was used to purify the recombinant fusion protein, and the Ni-Native Buffer containing 250 mmol·L⁻¹ imidazole was most favorable for elution. The purified recombinant fusion protein was subjected to in vitro catalytic reaction, and then the products were extracted and analyzed by HPLC. The results showed that the recombinant fusion protein CmFLS was able to catalyze the production of quercetin by dihydroquercetin under specific buffer and reaction conditions, which indicated that the functional protein encoded by CmFLS had dioxygenase activity in the biosynthetic pathway of flavonoids biosynthesis in Ch. morifolium cv. 'Hangju'. The above results laid the foundation for further studying on CmFLS, and provided new ideas for the regulation of flavonoids metabolism from the molecular level and the catalytic synthesis of flavonols in vitro.

The present study cloned wpGuamerin gene from a non-bloodsucking leech (Whitmania pigra), and the bioinformatics analysis of the sequence was performed. Additionally, the effects of feeding duration on the expression profile of the wp Guamerin gene were explored. The results showed that its sequence consisted of 295 nucleotides encoding a peptide of 83 amino acids(Genbank: KX768545), and its relative molecular weight is 9 145.95 Da. wp Guamerin does not encode proteins with a signal peptide, belonging to the hydrophilic protein. Its secondary structure is mainly composed of α-helix, extending chain, folding and random curl. Its similarity with other blood-sucking leeches ranges from 29% to 65%. The results revealed that wpGuamerin mRNA was detected higher expression in muscle than in salivary glands of Wh. pigra, and did not expressed in ingluvies and intestine. Its expression in muscle and salivary glands showed a single peak curve after feeding and the peak was observed in the 1st and 3rd after feeding, respectively. In summary, wp Guamerin in Wh. pigra is a small molecule polypeptide protein and is different from the Guamerin in blood-sucking leeches. wpGuamerin does not express in the digestive tract of Wh. pigra, and mainly express in muscle. Feeding behavior would stimulate the expression of wpGuamerin gene in muscle and salivary glands, but not in digestive tract.

To study the suitable arousal modes of hibernating Whitmania pigra, the biological characteristics, activity of amylase, lipase, and protease as well as morphologic structure of digestive tract were investigated by direct observational method and 3, 5-dinitrosalicylic acid colorimetry, p-nitrophenyl palmitate ester（p-NPP）colorimetry, folin-phenol and histological methods. The results revealed that Wh. pigra activity was increased with increases of the water temperature and prolonging treating duration. Except for the feeding groups of direct arousal mode at 24 h and 48 h, none of the other Wh. pigra groups died. Compared with that of normal groups, the digestive tract structure of hibernating Wh. pigra was looser and the mucosal folds of craw were more sparse. No obvious recovery was observed for the structure of the digestive tract before 48 h of direct arousal mode or the 5th day of 15 °C gradient arousal mode. The mucosal folds of craw increased and the muscularis were incrassated after 72 h of direct arousal mode or the 8th day of 20 °C gradient arousal mode, that indicated the tissue structure was approximately restored to the normal state. The digestive enzyme activities were increased with prolonging treating duration. And the feeding groups recovered faster than that of the no feeding groups. Additionally, the enzyme activities of feeding groups were significantly higher than that of no feeding groups (

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.

At present, there were few studies about the effects of cultivation measures on the quality and pharmacological activity of medicinal plants. To explore the hepetoprotective activity of Sedum sarmentosum aqueous extracts after different water treatments, S. sarmentosum were planted under five water treatments for 60 days, namely 15%-20% FC (field capacity, S1), 35%-40% FC (S2), 55%-60% FC (S3), 75%-80% FC(S4), and 95%-100% FC (S5) and CCl₄ drug-induced liver injury model in vitro was used. Cell viability, cell cycle, and cell apoptosis were individually detected by MTT, PI single staining, and Annexin-V FITC/PI double staining assays. Additionally, ALT, AST and antioxidant index in supernatant were determined by colorimetry. The results showed that, compared with the model group, S. sarmentosum aqueous extract could significantly improve the HepG2 cell viability. Among the five S. sarmentosum groups, the cell viability of S4 (75%-80% FC) treatment was the highest, and the cell apoptosis was the least. Meanwhile, the level of ALT, AST, and MDA in S4 was the lowest, but it achieved the highest level of SOD and GSH. Taken together, different water treatments had great influence on hepatoprotective effect of S. sarmentosum, and the soil moisture of the 75%-80% FC is beneficial to the hepetoprotective activity of S. sarmentosum.

The effects of four kinds of different plant populations on the soil physical and chemical properties and soil enzyme activities of Salvia miltiorrhiza were analyzed by setting up four kinds of mixed planting groups, such as S. miltiorrhiza and Cassia obtusifolia, Capsicum annuum, Perilla frutescens and Zea mays. And through the root isolation treatment, we preliminarily explored the formation mechanism of the four kinds of matching plants of soil physical and chemical properties and soil enzyme activities of S. miltiorrhiza, and chose the matching plants suitable for the establishment of the compound population with S. miltiorrhiza,and provided the basis for constructing high efficiency compound planting pattern of S. miltiorrhiza. The results showed that there were significant differences in soil physical and chemical properties and soil enzyme activitiesof Salvia miltiorrhiza in different compound population mixed.The combination of C. obtusifolia, P. frutescens, Z. mays and S. miltiorrhiza mixed planting were all reduced the soil cation exchange capacity and soil enzyme activities, and increased the soil electric conductivity. The compound planting of P. frutescens and S. miltiorrhiza increased the soil cation exchange capacity and soil enzyme activities, and reduced the soil electric conductivity. Therefore,The compound planting population of P. frutescens and S. miltiorrhiza is conducive to improve soil physical and chemical properties and increase soil microbial activity, which is the best collocation species of S. miltiorrhiza.

ICP-MS was used to determine the contents of five heavy metals such as lead (Pb), cadmium (Cd), mercury (Hg), arsenic (As) and copper (Cu) in Chrysanthemum indicum and soil samples. Pearson correlation and path analysis were used to analyze the effects of latitude and longitude and altitude on the content of heavy metal elements in Ch. indicum. To explore the relationship between geographical location of heavy metal content in Ch. indicum and the content of heavy metal elements in soil. The results showed that the Ch. indicum had high enrichment of Cd and had a synergistic effect on the absorption of five heavy metal elements. Elevation directly affected the accumulation of Pb and Cd in Ch. indicum and was a positive effect. The direct positive effect of latitude acts on the accumulation of Hg and Cu in Ch. indicum. The effect of longitude on the Cd content was the negative effect of longitude. The geographical location had different effects on the accumulation of heavy metal elements in Ch. indicum. There was a certain interaction between direct and indirect effects. Ch. indicum had different absorption and enrichment characteristics for different heavy metal species. When controlling the heavy metal content and medicinal material quality of Ch. indicum, we must fully consider the influence of geographical location on the quality of medicinal materials, select suitable geographic regions to carry out acquisitions, and choose a good ecological environment. Regional establishment of Chinese herbal medicine planting bases, or adopting certain agricultural measures to reduce the absorption of heavy metal elements in Ch. indicum herbs, while conducting research on chemical composition, strengthening the research on the effective forms of heavy metals, and paying attention to the safety of medicinal herbs.

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 is an important medicinal plant with a variety of anti-cancer properties. There is an urgent need to reduce water consumption in T. edulis under greenhouse cultivation. A greenhouse experiment was performed to study the effect of well-watered (80% of field capacity, Control), persistent drought (50% of field capacity, D) and alternate wetting and drying (50% and 80% of field capacity, AWD) regimes on the growth traits, photosynthetic pigment contents, chlorophyll fluorescence parameters, photosynthetic characteristics and yield of T. edulis . The results showed that compared with the well-watered control, persistent drought inhibited plant growth and yields dramatically, presumably due to the significantly lower photosynthetic rate caused by a decrease in photosynthetic pigments and a reduced chlorophyll fluorescence efficiency (Fv/Fm, Fv/F0). In contrast, the AWD plants showed a capacity for photosynthesis comparable to the well-watered control, which could be associated with similar photosynthetic pigment levels and chlorophyll fluorescence parameters (Fv/Fm, Fv/F0). AWD reduced the irrigation water input by 25.09% while leading to comparable yields and increased water productivity by 21.21% compared with the well-watered control. The results suggested that an alternate wetting and drying (50% and 80% of field capacity) regime can be an effective means of saving water, promoting bulb production, and improving water production for T. edulis .

The objective of this study was to obtain an overview of the genetic relationships within Perinereis aibuhitensis using Inter-Simple Sequence Repeat (ISSR) and Sequence-Related Amplified Polymorphism (SRAP) markers that were derived from related populations residing in the Chinese coasts. The percentage of polymorphic bands, Nei's gene diversity and Shannon's information index revealed a high level of genetic diversity at the species level. The analysis of molecular variance revealed that 81.22% (ISSR) and 76.29% (SRAP) of variability were partitioned among individuals within populations, which indicated the coherent trend by Nei's genetic differentiation (Gst) (0.2568/0.2876). The gene flow number (Nm) was 1.4470/1.2385, which indicated that there was limited gene exchange between populations. The phylogenetic tree of the ten P. aibuhitensis populations was separated into four major clusters using the neighbor-joining (NJ) method. These results provide a simple and useful basis for P. aibuhitensis germplasm research and aquaculture breeding.

Isatis indigotica Fort. (Cruciferae) is a biennial medicinal plant. In order to protect the decreasing natural genetic resources of I. indigotica , three candidate DNA barcodes (ITS2, trnL-F and rbc L) were employed to establish an accurate and effective identification system for I. indigotica . The results demonstrated that all three candidate DNA barcodes have performed very well in I. indigotica . The interspecific genetic distances were obviously greater than the intraspecific distance among I. indigotica as indicated by ITS2, trnL-F and rbc L. Sequence alignment analysis of I. indigotica genotypes revealed that four SNPs (54, 108, 146 and 181 bp) located in ITS2, three (2, 30, 709 bp) in trnL-F and one (531 bp) in rbc L, respectively. UPGMA phylogenetic tree constructed from trnL-F and rbc L could allote I. indigotica to the correct corresponding genus, whereas rbc L could not distinguish I. indigotica from its adulterants. Meanwhile, UPGMA tree of ITS2 could accurately identify I. indigotica from its adulterants according to the corresponding species. Consequently, it can be concluded that ITS2 is a more suitable and accurate DNA barcode for identifying I. Indigotica and its adulterants than trnL-F and rbc L.

The contents of 22 kinds of mineral elements in different parts of Changium smyrnioides and in the rhizosphere soil of 10 different populations were determined by microwave digestion-inductively coupled plasma atomic emission spectrometry (ICP-AES). The characteristics of mineral elements in the plants and the soil of main distribution area of Ch. smyrnioides was analyzed and the mechanism of the quality formation of Ch. smyrnioides was explored to provide the basis for the quality evaluation and cultivation regulation of Ch. smyrnioides.The results showed that the quality of soil environment was better in the main distribution area of Ch. smyrnioides, the content of trace elements was higher, the contents of P and K were lower and the contents of mineral elements in the soil of each distribution area was significantly different. The three elements of Se, P and K are significantly accumulated in the root of Ch. smyrnioides. There were significant differences in the total contents of mineral elements in the roots of Ch. smyrnioides in different producing areas. The contents of mineral elements in different parts of Ch. smyrnioides were significantly different. Ch.smyrnioides of the main distribution area of Ch. smyrnioides belonged to the safety level, the distribution of mineral elements in the plants can be used as an indicator of the quality of medicinal herbs.

The effects of four kinds of different plant populations on the morphology, the dry matter accumulation, active ingredient content and antioxidant activty in vitro of Salvia miltiorrhiza were analyzed by setting up four kinds of mixed planting groups, such as S. miltiorrhiza and Cassia obtusifolia, Capsicum annuum, Perilla frutescens and Zea mays. And through the root isolation treatment, we preliminarily explored the formation mechanism of the four kinds of matching plants of the yield and quality of S. miltiorrhiza, and chose the matching plants suitable for the establishment of the compound population with S. miltiorrhiza,and provided the basis for constructing high efficiency compound planting pattern of S. miltiorrhiza. The results showed that there were significant differences in plant morphology, dry matter accumulation of root, active ingredient content and antioxidant activty in vitro of S. miltiorrhiza in different compound population mixed. The growth and yield of S. miltiorrhiza were unfavorable to the combination planting of Cassia obtusifolia, Z. mays and Salvia miltiorrhiza.The compound planting of P. frutescens and S. miltiorrhiza significantly promoted the growth of S. miltiorrhiza, but significantly reduced the quality of S. miltiorrhiza.The yield and quality of S. miltiorrhiza were significantly improved by the combination of C. annuum and S. miltiorrhiza. Therefore, among the four plants of C. obtusifolia, C. annuum, P. frutescens, and Z. mays, the P. frutescens of Solanaceae is the best matching species for the construction of compound planting group with S. miltiorrhiza.

To study the effects of ingestion on antithrombin activities in different tissues of Whitmania pigra, the salivary glands, ingluvies, intestine and muscle of adult leeches were weighted on the 1st d, 3rd d, 5th d, 7th d and 11th d after feeding, respectively, and meanwhile antithrombin activity was measured by antithrombin titration method. The results showed that the antithrombin activity of salivary glands in different stages was significantly higher than that in other tissues (

The reproductive system and gonad development and germ cell occurrence of Whitmania pigra have been studied by using tissue section electron microscope techniques. W. pigra has completely independent male and female reproduction system, which lasts 11 months. The development of spermary started before the development of ovary. When egg cell is only a primordial germ cell, sperm has an initially complete form. Meanwhile, sperm cells and egg cells orderly development and synchronously mature. According to the development of sperm cells and egg cells, the development of cycle of the spermary could be divided into 6 stages: proliferating stage (1-3 months of age), growing stage (4-5 months of age), resting stage (6-8 months of age), maturing stage (9 months of age), spawning stage (10 months of age) and degradation stage (11 months of age). The development of cycle of the ovary could be divided into 6 stages: forming stage (1-2 months of age), proliferating stage (3-4 months of age), growing stage (5-8 months of age), maturing stage (9 months of age), spawning stage (10 months of age) and resting stage (11 months of age). W. pigra is a synchronous hermaphrodite animal, the development of cycle of the spermary and ovary each has six stages, sperm cells and egg cells orderly development and synchronously mature.

In this study, four light color treatments (red blue, white, natural light) were tested to determine effects of light spectrum on the growth of larval Whitmania pigra (30 days old) at the same intensity. The experiment lasted for 50 days. The specific growth rate (SGR) under blue light condition was significantly higher during 30 days to 40 days. At the end of the experiment, specific growth rate (SGR) and weight gain rate under red light condition were higher (P0.05) than those under other groups. Meanwhile, the enzymes activities of digestive enzymes (protease, amylase and lipase) were higher under red light condition. However, the enzymes activities of antioxidant enzymes (SOD, CAT, ALP) were lower under white light condition compared with the other treatments（P＜0.05）. In addition, no significant changes were observed for water content, total ash, acid insoluble ash, pH and antithrombin activity in the all treatments. After 50 days, the digestive enzymes and growth rate were higher under red light condition but antioxidant enzymes were lower under white light condition. Furthermore, light spectrum has no significant effect on the internal quality of Wh. pigra.

The study is aimed at determine the content of anthocyanins and expressions of relative genes and activity of relative enzymes. The effects of flood stress on anthocyanins synthesis with relative genes and enzymes of Chrysanthemum morifolium cv. 'Hangju' were analyzed. The expression of CHS and the content of CHS presented the trend of first rising and after downward with the increase of flowering degree. The content of anthocyanins, the expression of DFR and the activity of DFR presented the trend of first downward and after rising with the increase of flowering degree. There was a positive correlation among anthocyanins,DFR gene and DFR. However there was no significant correlation among anthocyanins,CHS gene and CHS. Flood stress has significant effects on anthocyanins synthesis with relative genes and enzymes of Ch. morifolium cv. 'Hangju',but don't change the patterns of genes expression and anthocyanins and enzymes accumulation. DFR and DFR are the key gene and key enzyme of anthocyanins synthesis.

Marsdenia tenacissima is a well-known anti-cancer medicinal plant used in traditional Chinese medicine due to bioactive constituents of polyoxypregnane glycosides, such as tenacissosides, marsdenosides and tenacigenosides. Genomic information regarding this plant is very limited, and rare information is available about the biosynthesis of polyoxypregnane glycosides. To facilitate the basic understanding about the polyoxypregnane glycoside biosynthetic pathways, de novo assembling was performed to generate a total of 73,336 contigs and 65,796 unigenes, which represent the first transcriptome of this species. These included 27 unigenes that were involved in steroid biosynthesis and could be related to pregnane backbone biosynthesis. The expression patterns of six unigenes involved in polyoxypregnane biosynthesis were analyzed in leaf and stem tissues by quantitative real time PCR (qRT-PCR) to explore their putative function. Furthermore, a total of 15,295 simple sequence repeats (SSRs) were identified from 11,911 unigenes, of which di-nucleotide motifs were the most abundant.

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.

Multi-element analysis of Marsdenia tenacissima samples was used to develop a reliable method of tracing the geographical origins. 27 elements in 128 samples from four provinces of China were analyzed by inductively coupled plasma-atomic emission spectroscopy. First, we used ANOVA and PCA for a preliminary analysis. Then, machine-learning tools for identification and classification were applied to verify the feasibility of using data mining tools to find the region where M. tenacissima samples originated. The study clearly indicated that the support vector machine, the radial basis function neural network, and the random forest chemometric tools had the potential to identify the origins of M. tenacissima . The results revealed that the SVM-grid model was superior to all the other mathematical methods with an average discrimination rate of 98.875% for the training set and 100% for the test set. The order of successful identification rates is as follows: SVM-grid > SVM-ga > RF > RBF-NN > SVM-pso. Moreover, there have been few relevant studies about the application of these machine-learning tools combined with multi-element analysis for tracing the geographical source, so this paper can serve as a reference to identify the origin and perform quality assurance in the field of medicinal plants.

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 isolate microsatellite loci to analyze the genetic diversity of Whitmania pigra . Four new microsatellite markers of W. pigra were developed from an enriched library and ten from a modified SAMPL assay. A total of 127 alleles were detected, with an average of 9.1 alleles per locus. The expected heterozygosity (He) of each microsatellite locus varied from 0.451 to 0.857, with an average of 0.688. The polymorphism information content (PIC) of each microsatellite locus ranged from 0.361 to 0.838, with an average of 0.640. Analysis of molecular variance showed that the main variation component existed within the populations (81.64%) rather than among the populations (18.36%). Phylogenetic tree for 15 populations of Hirudo using the NJ method by MEGA 5.1 software were divided into two major clusters. These microsatellite markers will contribute to research on the individual identification, genetic diversity, population structure, genome mapping and conservation biology of Hirudo .

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 .

The biological characteristics, oxygen consumption, oxygen consumption rate, and activities of amylase, lipase and protease of Whitmania pigra at different temperature were studied by using direct observational method, the still water method and 3, 5-dinitrosalicylic acid colorimetry, right-nitrophenyl palmitate ester（ρ-NPP）colorimetry and folin-phenol method.The results revealed that with decreasing water temperature, the daily activity and the daily feeding ration were decreased. As the temperature was lowered to 4 ℃, the head and tail of Wh.pigra curved, showing a crescent-shape without feeding and daily activity. Oxygen consumption, oxygen consumption rate and digestive enzyme activities reduced along with temperature drops. The downward trend slowed below 10 ℃, began to stabilize below 4 ℃ and doesn't change with the decrease of temperature since then. During the 40 days treatment at 4 ℃, the changes of amylase were not significant, the lipase and protease activity decreased at the 20th day, and the lipase showed an slightly increase after the decrease and finally remained at a low level.In conclusions, the pivotal temperature of hibernation of Wh.pigra is 4 ℃ and the crescent shape can be considered as a symbol of hibernation.

The oxygen consumption, oxygen consumption rate and suffocation point of different quality Whitmania pigra and Bellamya purificata were determined by hydrostatic breathing room method. The effects of feeding modes on growth of W.pigra were determined by biomass. The results showed that the oxygen consumption correlated positively with the weight of W.pigra and B. purificata（P

MicroRNAs (miRNAs) are a class of endogenous, non-coding small RNAs that play an important role in transcriptional and post-transcriptional gene regulation. However, the sequence information and functions of miRNAs are still unexplored in Tulipa edulis. In this study, high-throughput sequencing was used to identify small RNAs in stolon formation stages (stage 1, 2, and 3) in T. edulis. A total of 12,890,912, 12,182,122, and 12,061,434 clean reads were obtained from stage 1, 2, and 3, respectively. Among the reads, 88 conserved miRNAs and 70 novel miRNAs were identified. Target prediction of 122 miRNAs resulted in 531 potential target genes. Nr, Swiss-Prot, GO, COG, and KEGG annotations revealed that these target genes participate in many biologic and metabolic processes. Moreover, qRT-PCR was performed to analyze the expression levels of the miRNAs and target genes in stolon formation. The results revealed that miRNAs play a key role in T. edulis stolon formation.

Twelve populations of Inula lineariifolia were used as materials to measure morphological characteristics, photosynthetic parameters and chemical constituents. It aims to provide a theoretical basis for germplasm resources evaluation. The results showed that I. lineariifolia had relatively rich morphological diversity, there were significant differences of morphological characteristics, photosynthetic parameters and chemical constituents among populations. There was positive correlation on morphological characteristics and P(n). Twelve populations were divided into three-type. The three populations of Xuyi, Mingguang and Fengyang were of narrower-longer leaf, bigger biomass,better photosynthetic and higher chemical constituents. Then they were classified for a similar group. It proved that the three populations were more suitable for cultivation and promotion.

Effects of different drying methods including sun drying, steamed, boiled, constant temperature drying (at 40, 50, 60 °C) on appearance, hardness, rehydration ratio, dry rate, moisture, total ash, extractive and polysaccharides contents were studied to provide the basis of standard processing method for Tulipa edulis bulbus. The results showed that the treatments of sun drying and 40 °C drying showed higher rehydration ratios, but lower dry rate, higher hardness, worse color, longer time and obvious distortion and shrinkage in comparison with other drying methods. The treatments of 60 °C constant temperature drying resulted in shorter drying time, lower water and higher polysaccharides content. Drying time is shorter and appearance quality is better in the treatment of steaming and boiling compared with other treatments, but the content of extractive and polysaccharides decreased significantly. The treatments of 50 °C constant temperature drying led to similar appearance quality of bulb to commercial bulb, and it resulted in lowest hardness and highest dry rate as well as higher rehydration ratio, extractive and polysaccharides content, moderate moisture and total ash contents among these treatments. Based on the results obtained, 50 °C constant temperature drying is the better way for the processing of T. edulis bulbus.

There is time-honored history and culture of medicinal plant cultivation in China. In the present review, the medicinal plant cultivation history in china was summarized, its current situation and question were analyzed, and the prospects of medicinal plant cultivation research were pointed out, with the purpose of accelerating the growth of medicinal plant cultivation research.

Tulipa edulis (Miq.) Baker is an important medicinal plant with a variety of anti-cancer properties. The stolon is one of the main asexual reproductive organs of T. edulis and possesses a unique morphology. To explore the molecular mechanism of stolon formation, we performed an RNA-seq analysis of the transcriptomes of stolons at three developmental stages. In the present study, 15.49 Gb of raw data were generated and assembled into 74,006 unigenes, and a total of 2,811 simple sequence repeats (SSRs) were detected in T. edulis. Among the three libraries of stolons at different developmental stages, there were 5,119 differentially expressed genes (DEGs). A functional annotation analysis based on sequence similarity queries of the GO, COG, KEGG databases showed that these DEGs were mainly involved in many physiological and biochemical processes, such as material and energy metabolism, hormone signaling, cell growth, and transcription regulation. In addition, qRT-PCR (quantitative real-time PCR) analysis revealed that the expression patterns of the DEGs were consistent with the transcriptome data, which further supported a role for the DEGs in stolon formation. This study provides novel resources for future genetic and molecular studies in T. edulis.

The effects of harvest and different processing methods on the anti-thrombin activity of Poecilobdella manillensis were respectively studied. The indicators included processing methods (vacuum freeze drying, fresh homogenate, drying under sunlight, freezing, scalding, baking under different temperatures), different parts (entire body, cephalon, pygidium, exudate) and body weights (≤10, 10-20, 20-30, 30-40, ≥40 g). The anti-thrombin activities of P. manillensis with different processing methods were evaluated by direct anti-thrombin titration. The results indicated that the processing methods significantly affected the anti-thrombin activities of P. manillensis. Among the 11 groups, the anti-thrombin activity of P. manillensis processed with vacuum freeze drying (1 303.56 U•g⁻¹) was significantly highest than the other groups (P

The effect of stereoscopic cultivation on the growth, photosynthetic characteristics and yield of Tulipa edulis was studied to explore the feasibility of stereoscopic cultivation on efficient cultivation of T.edulis. Total leaf area and photosynthetic parameters of T.edulis under stereoscopic cultivation (the upper, middle and the lower layers ) and the control were measured using LI-3100 leaf area meter and LI-6400XT photosynthesis system in the growing peak period of T.edulis.Plant biomass and biomass allocation were also determined.In addition, the bulb regeneration and yield of T.edulis were measured in the harvesting time.The results indicated that in the middle layer of stereoscopic cultivation, leaf biomass proportion was the highest, but total bulb fresh and dry weight and output growth (fresh weight) were the lowest among the treatments.And total bulb fresh weight in the middle of stereoscopic cultivation reduced significantly, by 22.84%, compared with the control.Light intensity in the lower layer of stereoscopic cultivation was moderate, in which T.edulis net photosynthetic rate and water use efficiency were higher than those of the other layers of stereoscopic cultivation, and bulb biomass proportion was the highest in all the treatments.No significant difference was detected in the total bulb fresh weight, dry weight and output growth (fresh weight) between the middle layer of stereoscopic cultivation and the control.In general, there was no significant difference in the growth status of T.edulis between stereoscopic cultivation and the control.Stereoscopic cultivation increased the yield of T.edulis by 161.66% in fresh weight and 141.35% in dry weight compared with the control in the condition of the same land area, respectively.In conclusion, stereoscopic cultivation can improve space utilization, increase the production, and achieve the high density cultivation of T.edulis.

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.