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Psammosilene tunicoides is a unique perennial medicinal plant species native to the Southwestern regions of China. Its wild population is rare and endangered due to over-excessive collection and extended growth (4-5 years). This research shows that H + -ATPase activity was a key factor for oxalate-inducing programmed cell death (PCD) of P. tunicoides suspension cells. Oxalic acid (OA) is an effective abiotic elicitor that enhances a plant cell's resistance to environmental stress. However, the role of OA in this process remains to be mechanistically unveiled. The present study evaluated the role of OA-induced cell death using an inverted fluorescence microscope after staining with Evans blue, FDA, PI, and Rd123. OA-stimulated changes in K + and Ca 2+ trans-membrane flows using a patch-clamp method, together with OA modulation of H + -ATPase activity, were further examined. OA treatment increased cell death rate in a dosage-and duration-dependent manner. OA significantly decreased the mitochondria activity and damaged its electron transport chain. The OA treatment also decreased intracellular pH, while the FC increased the pH value. Simultaneously, NH 4 Cl caused intracellular acidification. The OA treatment independently resulted in 90% and the FC led to 25% cell death rates. Consistently, the combined treatments caused a 31% cell death rate. Furthermore, treatment with EGTA caused a similar change in intracellular pH value to the La 3+ and OA application. Combined results suggest that OA-caused cell death could be attributed to intracellular acidification and the involvement of OA in the influx of extracellular Ca 2+ , thereby leading to membrane depolarization. Here we explore the resistance mechanism of P . tunicoides cells against various stresses endowed by OA treatment.

Ethnopharmacological Relevance: Gentiana kurroo is a multipurpose critically endangered medicinal herb prescribed as medicine in Ayurveda in India and exhibits various pharmacological properties including anti-cancer activity. The species is rich repository of pharmacologically active secondary metabolites together with secoiridoidal glycosides., Aim of the Study: The study aimed to investigate the chemical diversity in different populations/cytotypes prevailing in G. kurroo to identify elite genetic stocks in terms of optimum accumulation/biosynthesis of desired metabolites and having higher in-vitro cytotoxicity potential in relation to chemotypic diversity., Material and Methods: The wild plants of the species were collected from different ranges of altitudes from the Kashmir Himalayas. For cytological evaluation, the standard meiotic analysis was performed. The standard LC-MS/MS technique was employed for phytochemical analysis based on different marker compounds viz. sweroside, swertiamarin, and gentiopicroside. Different tissues such as root-stock, aerial parts, and flowers were used for chemo-profiling. Further, the methanolic extracts of diploid and tetraploid cytotypes were assessed for cytotoxic activity by using MTT assay against four different human cancer cell lines., Results: The quantification of major bioactive compounds based on tissue- and location-specific comparison, as well as in-vitro cytotoxic potential among extant cytotypes, was evaluated. The comprehensive cytomorphological studies of the populations from NW Himalayas revealed the occurrence of different chromosomal races viz. n = 13, 26. The tetraploid cytotype was hitherto unreported. The tissue-specific chemo-profiling revealed relative dominance of different phytoconstituents in root-stock. There was a noticeable increase in the quantity of the analyzed compounds in relation to increasing ploidy status along the increasing altitudes. The MTT assay of methanolic extracts of diploid and tetraploid cytotypes displayed significant cytotoxicity potential in tetraploids. The root-stock extracts of tetraploids were highly active extracts with IC 50 value ranges from 5.65 to 8.53 μg/mL against HCT-116 colon cancer., Conclusion: The chemical evaluation of major bioactive compounds in diverse cytotypes from different plant parts along different altitudes presented an appreciable variability in sweroside, swertiamarin, and gentiopicroside contents. Additionally, the concentrations of these phytoconstituents varied for cytotoxicity potential among different screened cytotypes. This quantitative difference of active bio-constituents was in correspondence with the growth inhibition percentage of different tested cancer cell lines. Thus, the present investigation strongly alludes towards a prognostic approach for the identification of elite cytotypes/chemotypes with significant pharmacological potential., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Efforts to develop the necessary biotechnologies in Greater Celandine ( Chelidonium majus L.), a leading plant resource for the development of plant-derived medicines, have been hampered by the lack of knowledge about transcriptome and metabolome regulations of its medicinal components. Therefore, this study aimed to examine the effect of abiotic elicitors, methyl jasmonate (MJ) and salicylic acid (SA), at different time courses (12, 24, 48, and 72 h), on expression and metabolome of key benzophenanthridine alkaloids (BPAs) in an optimized in vitro culture. Gene expression analysis indicated the upregulation of CFS (cheilanthifoline synthase) to 2.62, 4.85, and 7.28 times higher than the control at 12, 24, and 48 h respectively, under MJ elicitation. Besides, MJ upregulated the expression of TNMT (tetrahydroprotoberberine N -methyltransferase) to 2.79, 4.75, and 7.21 times at 12, 24, and 48 h respectively, compared to the control. Investigation of BPAs revealed a significant enhancement in the chelidonine content (9.86 µg/mg) after 72 h of MJ elicitation. Additionally, sanguinarine content increased to its highest level (3.42 µg/mg) after 24 h of MJ elicitation; however, no significant enhancement was detected in its content in shorter elicitation time courses. Generally, higher gene expression and BPAs' level was observed through longer elicitation courses (48 and 72 h). Our findings take part in improving the understanding of transcription and metabolic regulation of BPAs in cultured Greater Celandine cells.

Medicinal plants produce valuable secondary metabolites with anticancer, analgesic, anticholinergic or other activities, but low metabolite levels and limited available tissue restrict metabolite yields. Transformed root cultures, also called hairy roots, provide a feasible approach for producing valuable secondary metabolites. Various strategies have been used to enhance secondary metabolite production in hairy roots, including increasing substrate availability, regulating key biosynthetic genes, multigene engineering, combining genetic engineering and elicitation, using transcription factors (TFs), and introducing new genes. In this review, we focus on recent developments in hairy roots from medicinal plants, techniques to boost production of desired secondary metabolites, and the development of new technologies to study these metabolites. We also discuss recent trends, emerging applications, and future perspectives., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

This paper studies modern methods of producing and using callus, suspension cells and root cultures of medicinal plants in vitro. A new solution for natural product production is the use of an alternative source of renewable, environmentally friendly raw materials: callus, suspension and root cultures of higher plants in vitro. The possibility of using hairy root cultures as producers of various biologically active substances is studied. It is proven that the application of the genetic engineering achievements that combine in vitro tissue culture and molecular biology methods was groundbreaking in terms of the intensification of the extraction process of compounds significant for the medical industry. It is established that of all the callus processing methods, suspension and root cultures in vitro, the Agrobacterium method is the most widely used in practice. The use of agrobacteria has advantages over the biolistic method since it increases the proportion of stable transformation events, can deliver large DNA segments and does not require special ballistic devices. As a result of the research, the most effective strains of agrobacteria are identified.

The in vitro tissue culture of medicinal plants is considered as a potential source for plant-derived bioactive secondary metabolites. The in vitro tissue culture of American ginseng has wide commercial applications in pharmaceutical, nutraceutical, food, and cosmetic fields with regard to the production of bioactive compounds such as ginsenosides and polysaccharides. This review highlights the recent progress made on different types of tissue culture practices with American ginseng, including callus culture, somatic embryo culture, cell suspension culture, hairy root culture, and adventitious root culture. The tissue culture conditions for inducing ginseng callus, somatic embryos, cell suspension, hairy roots, and adventitious roots were analyzed. In addition, the optimized conditions for increasing the production of ginsenosides and polysaccharides were discussed. This review provides references for the use of modern biotechnology to improve the production of bioactive compounds from American ginseng, as well as references for the development and sustainable utilization of American ginseng resources., (© 2020 Wiley-VHCA AG, Zurich, Switzerland.)

Catharanthus roseus produces a wide spectrum of monoterpene indole alkaloids (MIAs). MIA biosynthesis requires a tightly coordinated pathway involving more than 30 enzymatic steps that are spatio-temporally and environmentally regulated so that some MIAs specifically accumulate in restricted plant parts. The first regulatory layer involves a complex network of transcription factors from the basic Helix Loop Helix (bHLH) or AP2 families. In the present manuscript, we investigated whether an additional epigenetic layer could control the organ-, developmental- and environmental-specificity of MIA accumulation. We used Whole-Genome Bisulfite Sequencing (WGBS) together with RNA-seq to identify differentially methylated and expressed genes among nine samples reflecting different plant organs and experimental conditions. Tissue specific gene expression was associated with specific methylation signatures depending on cytosine contexts and gene parts. Some genes encoding key enzymatic steps from the MIA pathway were found to be simultaneously differentially expressed and methylated in agreement with the corresponding MIA accumulation. In addition, we found that transcription factors were strikingly concerned by DNA methylation variations. Altogether, our integrative analysis supports an epigenetic regulation of specialized metabolisms in plants and more likely targeting transcription factors which in turn may control the expression of enzyme-encoding genes.

Adulteration in traded medicinal plants is a significant issue nowadays and use of these adulterated medicinal plants can impose harmful impact to end user. However, this problem can be overcome by ensuring the identification of traded medicinal plants which are used in making different herbal medicines. In this regard, palynological markers are considered to be an important taxonomic tool in the identification of original medicinal plant from its adulterant. Hence this study attempted to provide particular reliable palynological markers for distinguishing selected medicinal plants from their adulterants, that is, Cinnamomum verum versus Canella winterana, Cinnamomum tamala versus Cinnamomum obtusifolium, Gymnema sylvestre versus Gymnema lactiferum, Artemisia maritima versus Artemisia absinthium, Achillea millefolium versus Adhatoda vasaka, Sphaeranthus indicus versus Sphaeranthus africanus, Averrhoa carambola versus Butea monosperma, and Morus nigra versus Morus alba. Results demonstrated great variations in multiple palynological characters between original medicinal plant and its adulterant such as in pollen size, shape, colpi length, exine, intine thickness, and fertility. In equatorial view, circular to spheroidal shape of pollen was found in A. millefolium while oblate shape was observed in A. vasaka. Similarly B. monosperma pollen was 34 μm, whereas pollen of its adulterant A. carambola was 21 μm. Moreover, colpi length of A. maritima was 11.8 μm, whereas 4.5 μm in A. absinthium. Hence it can be concluded that palynological characters are commendably helpful in identification of genuine medicinal plant from its adulterant., (© 2019 Wiley Periodicals, Inc.)

Dendrobium is an important medicinal material in China. It has the effect of nourishing the stomach, nourishing yin and clearing heat. In China, there are many types of Dendrobium, and different Dendrobium species have different efficacy. The present study is aimed at distinguishing three major Dendrobium species from morphological and microscopic identification in Ta-pieh mountains area. In this article, the roots, stems and leaves of Dendrobium huoshanense, Dendrobium officinale, and Dendrobium moniliforme are used as materials to compare the differences of tissues of these three Dendrobium species by morphological indexes and microscopic identification of different Dendrobium. The stem morphology of these three Dendrobium species was significantly different except for stem internodes number and the middle part of the stem diameter by measuring the stems of Dendrobium. To ensure the safe use of Dendrobium, we built a fast and convenient method combining normal and fluorescence microscopy was applied in the present study to distinguish D. huoshanense, D. officinale, and D. moniliforme. The microscopic results show that different types of Dendrobium exhibit different states that can be distinguished under normal light normal and fluorescence microscopy. This comparative study of morphology and microscopy contributes to the development of identification and quality evaluation of Dendrobium., (© 2019 Wiley Periodicals, Inc.)

Euphorbia nivulia Buch.-Ham. (Euphorbiaceae) is commonly known as Indian Spurge Tree in English, and "Saj Thor" or "Jhanami booti" in local language. The plant is used traditionally in the treatment of various diseases like inflammation, fever, worm infection, asthma, cough, wounds and diabetes. In current study fresh as well as dried aerial parts of the plant and cut sections were examined, both macroscopically and microscopically. The study also deals with fluorescence analysis and phytochemical characteristics and other WHO recommended methods for standardization. WHO guidelines on quality control for medicinal plants materials were used for pharmacognostical evaluation of E. nivulia, phytochemical screening helps in determining the predominant classes of active constituents responsible for the activity. The present work will be helpful in identification of the fresh and dried samples of aerial parts pharmacognostically and anatomically. These studies will serve as a reference for correct identification and may be helpful in checking any type of adulteration. These observations will also help in differentiating this species from closely related species of the same genus and family.

The isolation of vacuoles is an essential step to unravel the important and complex functions of this organelle in plant physiology. Here, we describe a method for the isolation of vacuoles from Catharanthus roseus leaves involving a simple procedure for the isolation of protoplasts, and the application of a controlled osmotic/thermal shock to the naked cells, leading to the release of intact vacuoles, which are subsequently purified by density gradient centrifugation. The purity of the isolated intact vacuoles is assayed by microscopy, western blotting, and measurement of vacuolar (V)-H + -ATPase hydrolytic activity. Finally, membrane functionality and integrity is evaluated by measuring the generation of a transtonoplast pH gradient by the V-H + -ATPase and the V-H + -pyrophosphatase, also producing further information on vacuole purity.

One new β-carboline alkaloid 7-methoxy-(9H-β-carbolin-1-il)-(E)-1-propenoic acid (1) together with 9-methoxycanthin-6-one (2) and 9-hydroxycanthin-6-one (3) were isolated from the hairy-root cultures of Eurycoma longifolia. The effects of these compounds on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW264.7 cells were investigated. Compound 1 strongly inhibited the production of NO while 2 and 3 having weak or inactive effect. Consistently, compound 1 decreased the expression of cyclooxygenase-2 and inducible nitric oxide synthase.

Biofertilization with plant growth-promoting rhizobacteria is a potential alternative to plant productivity. Here, in vitro propagation of Handroanthus ochraceus (yellow lapacho), a forest crop with high economic and environmental value, was developed using the Azospirillum brasilense strains Cd and Az39 during rhizogenesis. Epicotiles of in vitro plantlets were multiplied in Woody Plant Medium (WPM). For rooting, elongated shoots were transferred to auxin-free Murashige-Skoog medium with Gamborg's vitamins and WPM, both at half salt concentration (½MSG and ½WPM), and inoculated with Cd or Az39 at the base of each shoot. Anatomical studies were performed using leaves cleared and stained with safranin for optical microscopy and leaves and roots metalized with gold-palladium for scanning electron microscopy (SEM). In ½WPM auxin-free medium, A. brasilense Cd inoculation produced 55% of rooting, increased root fresh and dry weight (45% and 77%, respectively), and led to lower stomata size and density with similar proportion of open and closed stomata. Both strains selectively increased the size or density of glandular trichomes in ½MSG. Moreover, bacteria were detected on the root surface by SEM. In conclusion, the difference in H. ochraceus response to A. brasilense inoculation depends on the strain and the plant culture media. Cd strain enhanced rooting in auxin-free ½WPM and produced plantlets with features similar to those expected in ex vitro plants. This work presents an innovative in vitro approach using beneficial plant-microorganism interaction as an ecologically compatible strategy in plant biotechnology., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The development of plant tissue (including organ and cell) cultures for the production of secondary metabolites has been underway for more than three decades. Plant cell cultures with the production of high-value secondary metabolites are promising potential alternative sources for the production of pharmaceutical agents of industrial importance. Medicinal plant cell suspension cultures (MPCSC), which are characterized with the feature of fermentation with plant cell totipotency, could be a promising alternative "chemical factory". However, low productivity becomes an inevitable obstacle limiting further commercialization of MPCSC and the application to large-scale production is still limited to a few processes. This review generalizes and analyzes the recent progress of this bioproduction platform for the provision of medicinal chemicals and outlines a range of trials taken or underway to increase product yields from MPCSC. The scale-up of MPCSC, which could lead to an unlimited supply of pharmaceuticals, including strategies to overcome and solution of the associated challenges, is discussed.

Gymnema sylvestre R. Br. is a popular herbal medicine. It has been used in ayurvedic system of medicine for thousands of years. It is popularly called as "Gur-mar" for its distinctive property of temporarily destroying the taste of sweetness and is used in the treatment of diabetes. The leaves of gymnema possess antidiabetic, antimicrobial, anti-hypercholesterolemic, anti-sweetener, anti-inflammatory, and hepatoprotective properties and have traditional uses in the treatment of asthma, eye complaints, and snake bite. The leaves contain triterpene saponins such as gymnemic acid which is an active ingredient of Gymnema. Since the cultivation of G. sylvestre is a very slow process and the content of gymnemic acid depends on the environmental factors, cell suspension culture is sought as an alternative means for the production of Gymnema biomass and to enhance the gymnemic acid content. In this chapter, the methods employed for the induction of callus and subsequent establishment of cell suspension cultures for the production of biomass and analysis of gymnemic acid using high performance liquid chromatography are described.

Gymnosporia arenicola Jordaan (Celastraceae) is a shrub or small tree, which naturally occurs in coastal sand dunes of Southern Mozambique and South Africa. Its dried leaf is often used in traditional medicine for the treatment of infectious and inflammatory diseases. Hereby, we present results of studies carried out according to the pharmacopoeia standards for the identification of herbal drugs, in the whole, fragmented, and powdered plant material. These results were complemented with scanning electron microscopy and histochemical techniques. The leaf microscopic analysis revealed a typical dorsiventral mesophyll with a corresponding spongy parenchyma-palisade parenchyma ratio of 0.60, anomocytic and paracytic stomata, papillate cells with a diameter of 4.00 ± 0.40 µm, multicellular uniseriate nonglandular trichomes with a length of 27.00 ± 4.10 µm and cristalliferous idioblasts containing calcium oxalate cluster crystals with a diameter of 23.04 ± 5.84 µm. The present findings demonstrate that the G. arenicola leaf has both nonglandular trichomes and hypoderm, features not previously described in the corresponding botanical section (Gymnosporia sect. Buxifoliae Jordaan). The establishment of these new botanical markers for the identification of G. arenicola leaf is essential for quality, safety and efficacy reasons., (© 2015 Wiley Periodicals, Inc.)

In the present study, a protocol was developed for the in vitro propagation of a rare medicinal plant, Hedysarum theinum (tea sweetvetch), from axillary buds, and identification of the regenerants was performed with the use of ISSR markers. It was demonstrated that Gamborg and Eveleigh medium supplemented with 5 μM 6-benzylaminopurine was the best for H. theinum for initial multiplication. On the other hand, half-strength Murashige and Skoog (MS) basal medium supplemented with 7 μM α-naphthaleneacetic acid proved to be the best for explant rooting. Molecular genetic analysis of the H. theinum mother plants and the obtained regenerants was performed with six ISSR markers. Depending on the primer, four to ten amplified fragments with sizes ranging from 250 to 3000 bp were identified. Our results confirmed the genetic stability of regenerants obtained in five passages and their identity to the mother plant.

Eryngium planum L. has been reported as a medicinal plant used in traditional medicine in Europe. The tissue cultures may be an alternative source of the biomass rich in desired bioactive compounds. The purpose of this study was to investigate the influence of the biotechnological techniques on the selected phenolic acids accumulation in the agitated shoot cultures of E. planum. Qualitative and quantitative analyses of those compounds in 50% aqueous - methanolic extracts from the biomass were conducted by applying the HPLC method. Methyl jasmonate (MeJA), yeast extract (YE) and sucrose (Suc) stimulated accumulation of the phenolic acids: rosmarinic (RA), chlorogenic (CGA) and caffeic (CA) in in vitro shoot cultures. Cultivation of shoots in liquid MS media supplemented with 1.0 mg L(-1) 6-benzyladenine and 0.1 mg L(-1) indole-3-acetic acid in the presence of 100 µM MeJA for 48h was an optimum condition of elicitation and resulted in approximately 4.5-fold increased content of RA + CGA + CA in plant material compared to the control (19.795 mg g(-1) DW, 4.36 mg g(-1) DW, respectively). The results provide the first evidence that the selected phenolic acids can be synthesized in elicited shoot cultures of flat sea holly in higher amount than in untreated shoots.

Medicinal bark refers to structures outside the vascular cambium of stems, branches and roots of gymnospermous and dicotyledonous plants that are used as medicinal materials; bark is an important type of Chinese herbal medicine. However, identification of the species from which the bark comes can be very difficult, especially when the bark is dried and sliced. In our previous studies, we have found that fluorescence microscopy is a powerful tool for the identification of easily confused Chinese herbal medicines, powdered Chinese herbal medicines and decoction dregs. To establish the fluorescent characteristics by which medicinal barks can be identified, for ensuring their safe and effective use, a systematic microscopic investigation by normal light and fluorescence microscope was carried out on transverse section samples of 11 medicinal barks commonly used in China. Specifically, the fluorescent characteristics of mechanical tissues, including stone cells and fibres as well as secretory tissues, have been observed. The microscopic features of medicinal bark are here systematically and comparatively described and illustrated. Under the fluorescence microscope, various tissues emitted fluorescence of different colours, and we found that both the colours and the intensity can be used to distinguish and identify these barks., (© 2014 The Authors Journal of Microscopy © 2014 Royal Microscopical Society.)

Dried leaves of Satureja khuzestanica Jamzad are a popular herbal medicine and dental anesthetic among the nomadic inhabitants of southwestern Iran. The present study establishes criteria for identification of S. khuzestanica dried whole, fragmented and powdered leaves for specification as a herbal substance using macroscopic and microscopic characterization. Quantitative microscopy techniques were also considered. Macroscopically leaves exhibit a grayish-green color, are broadly ovate in shape with an acute apex, attenuate base, and ciliate margin and have a surface covered by an indumentum of glandular and non-glandular trichomes. Microscopically leaves have an isobilateral amphistomatic structure containing peltate glandular trichomes consisting of a multiseriate stalk (five cells) and an enlarged secretory head composed of 12 cells, capitate glandular trichomes of variable morphology together with two types of non-glandular trichomes. Oval-shaped hygromorphic diacytic stomata with an adaxial stomatal index of 13.54, collateral vascular bundles consisting of xylem, and three layers of sclerenchymatous tissue close to phloem together with cluster, prismatic and raphide calcium oxalate crystals were also identified as useful pharmacognostic parameters for identification of S. khuzestanica dried leaves.

The present paper deals with cytological studies on 31 populations covering 17 species belonging to 10 genera of Apiaceae from Western Himalayas. The chromosome numbers in the two species as Chaerophyllum capnoides (n = 11) and Heracleum brunonis (n = 11), along with additional cytotypes for Pimpinella acuminata (n = 9) and Sium latijugum (n = 12) have been reported for the first time on world-wide basis. The genus Pleurospermum, although cytologically worked out earlier from outside India, its species densiflorum (n = 11) makes first representation of the genus from India. Besides, the chromosome number in Chaerophyllum aromaticum (n = 11) have been worked out for the first time from India. The course of meiosis varies from normal to abnormal in different populations of Chaerophyllum villosum, Pimpinella achilleifolia and Sium latijugum while abnormal meiotic course has been observed in all the studied populations of Chaerophyllum acuminatum, C. aromaticum, C. capnoides, Pimpinella acuminata, P. diversifolia, Pleurospermum densiflorum and Vicatia coniifolia. Such taxa are marked with meiotic abnormalities in the form of cytomixis, chromatin stickiness, formation of laggards and bridges resulting into abnormal microsporogenesis. The occurrence of structural heterozygosity has been recorded in the Chaerophyllum acuminatum and C. aromaticum. The effect of these abnormalities is clearly seen on the pollen size and fertility.

Based on the research of plant taxonomy and botanical investigation, microscopic characteristics of the root, stem, leaf transverse section and powder of Atropa belladonna were studied for identification of the herb. The research detailed and made clear to the description identification and microscopic characteristics of officinal parts of the herbs. The work provided reference for the identification of A. belladonna herbs and pieces of work in the future, as well as a theoretical basis for the further research, development, medicinal use and the upgrading of quality standards.

Objective: To observe microscopic characteristics of rhizome of Alpinia kwangsiensis, Alpinia platychilus, Alpinia blepharocalyx, and to provide basis for their identification., Methods: Microscopic identification of root transverse section by paraffin tissue section and free-hand section and powder were carried out to distinguish them., Results: The microscopic identification can be made by cell structure of epidermis and endodermis, the existence of nonglandular hair, the lignification degree of vascular bundle fiber, the number and existence style of vascular bundle fiber, the number and size of tube, and so on. The powder identification can be made mainly according to the existence of nonglandular hair and spiral vessel, the shape of starch grain, and so on., Conclusion: The microscopic characteristics of rhizome can provide basis for the identification of three Alpinia species.

Objective: To compare macroscopic and microscopic characteristics of the fruits of Amomum tsao-ko and Amomum paratsao-ko., Methods: Transverse section and powder of pericarp and seeds of these two species were identified., Results: The fruits of Amomum tsao-ko and Amomum paratsao-ko had similar microscopic characteristics. They could be distinguished from each other by the peri-carp color, furrows and ribs, and the shape of seeds., Conclusion: Above characteristics can be used as basis of pharmacognostical study of Amomum tsao-ko and Amomum paratsao-ko.

Objective: To study the feasibility and significance of the method to identify medicinal plants through the observation and statistics of 9 species of Sabia medical plants from Guizhou province., Methods: Leaf epidermis characteristics were observed, measured by optical microscope and analyzed by SPSS 17.0 software., Results: All of these plants had some differences in indumentum, cell morphology and size, and had significant difference in the length and circumference of lower epidermis cells., Conclusion: The method combining microscopic observation with statistics can be used as the classification and identification basis of medicinal plants and materials of Sabia genus.

Objective: To provide macroscopic and microscopic identification basis for Ligularia przewalskii., Methods: Macroscopic and microscopic identification of roots, stems and leaves of Ligularia przewalskii were carried out with the methods of paraffin section, leaves epidermal section and powder transdermal section., Results: The microscopic characteristics included: Open collateral vascular bundles in stem were not in the same size and arranged in two rings; Lots of fiber bundles scattered in the column parts; There were two vascular bundles in principal vein of leaf; Anticlinal wall of upper epidermis cells was thickened like moniliform, lower epidermis were like waves with irregular; The type of stoma was anomocytic; Calcium oxalate acicular crystal could be seen in the powder., Conclusion: These features can provide references for identification of Ligularia przewalskii.

Objective: To provide the identification basis for Mongolian medicine Potentilla glabra., Methods: Macroscopic and microscopic identification were applied to observe macroscopic, histological, superficial, and powder characteristics of its stems and leaves., Results: The following characteristics were observed: The cork layer sandwiched with sclerenchyma ring which was main composed of fibers, accompanied by stone cells and large cell layer; Catheter with fiber existed in xylem; Beaded thickened anticlinal wall in leaf epidermal cell; Stomatal infinitive; Small clusters of calcium oxalate crystal and different type of leaf transverse section., Conclusion: These characteristics can provide evidences for the identification and quality control of Potentilla glabra.

Objective: To establish microscopic identification standards for four species from Uncaria genus., Methods: The microscopic characteristics of transvers section and powder of these four species rattan were observed., Results: As for these four species from Uncaria genus, the microscopic identification of rattan transvers section could be made by the existence of cork cambium and pericycle in the cortex, and the location of phloem fiber and stone cells, while the powder identification could be made according to the existence of scalariform vessel and calcium oxalate crystal, and the quantity of stone cells., Conclusion: The microscopic characteristics of rattan can provide information for identification of these four species from Uncaria genus.

Objective: To study the macroscopic and microscopic characteristics of Zhuang medicine Tetrastigma planicaule., Methods: Macroscopic and microscopic identification were studied., Results: The microscopic characteristics were significant. The root had more than 10 layers of cork cells arranging in line. The starch grain and calcium oxalate cluster crystals distributed in the phloem parenchymas, and mucilage cells scattered in the cortex of the stem. There were 8 vascular bundles in the vein of leaves., Conclusion: This study provides a scientific basis for the identification and utilizing of Tetrastigma planicaule.

Objective: To identify the macroscopic and microscopic characteristics of Chloranthus henryi and provide a basis for establishing its quality standard., Methods: The plants were identified by original plant, macroscopic and microscopic identification., Results: The radical vascular bundle of Chloranthus henryi was primary xylem tetrarch. The secretory canals dispersed in cortex. The vascular bundle in rhizome was amphivasal bundle, stone cell scattered in cortex, and there were solitary crystals in the core parenchyma cell. Several vascular bundles in caudex were connected like a circel by interfascicular fibers. Meanwhile, two vascular bundles lay in main vein, like a converse Chinese letter eight., Conclusion: This paper reports the microscopic characteristics of Chloranthus henryi. It provides a basis for the quality standard of Chloranthus henryi.

Objective: To establish a method of microscopic identification of Tibetan medicinal herb " Songdi" (Saxifraga umbellulata var. pectinata)., Method: The different characteristics and microscopic identification of 4 species of Songdi were compared, including the main variety (Saxifraga umbellulata var. pectinata ) and approximate varieties (S. unguiculata, S. przewalskii and S. tanguTi,) were studied and compared., Result: The botanical anatomy characteristics of Saxifraga umbellulata var. pectinata (including roots, stems, leaves, flowers, stalks and fruits) have been measured. The methods for powder identifications and transverse section of stems root and leaves of four species of Songdi were established., Conclusion: Four species of Songdi can be identified by the growth pattern of basal leaves, type of hair, type of inflorescence, number of flowers, proportion of tissue structures in stem, form of pollen grains.

PNS (Panax notoginseng saponins) is the main medical bioactive component in Panax notoginseng. The medical value of PNS cannot be extended because of its low production. With the deep study of saponins biosynthetic pathway, the control of PNS biosynthesis through metabolic engineering has gradually become possible. In this study, the Squalene synthase (SS) over-expression vector was established. By the way of agrobacterium-mediated method, the vector was transfered and integrated into the Panax notoginseng genome. The result of the PCR detection and the saponin content detection shows that over-expression SS is able to produce high level of Panax notoginseng saponins, and confirms the regulatory function of SS in the biosynthesis of ginsenosides in Panax notoginseng. It provides a theoretical basis and technical basis for the construction of PNS homologous or heterologous efficient expression system in the future.

China consumes and exports traditional Chinese medicinal resources the most in the world. However, we cannot anchor our hope on field production of traditional Chinese medicinal materials and their active ingredients, due to limited land resources. Therefore, the development of biotechnology is of great importance for China to solve the problem of traditional Chinese medicinal resources. Plant cell culture is an important approach for the sustainable development of precious medicinal resources. This essary summarizes the optimization of conditions for medicinal plant cell culture, the regulation of secondary metabolic pathways and cell bioreactor culture, and realizes that the authentic commercial production of more medicinal plants requires efforts from all aspects.

Premise of the Study: Microsatellite markers were isolated and characterized in a medicinal plant, Phyllanthus emblica, to study population genetics for designing an effective in situ and ex situ conservation of genetic resources of the species. •, Methods and Results: Six microsatellite markers were developed using an enrichment and magnetic separation protocol. They were characterized in two natural populations of P. emblica. Out of the six microsatellites, five showed polymorphism, with the number of alleles ranging from four to seven. Observed and expected heterozygosities ranged from 0.360 to 0.760 and 0.499 to 0.806, respectively. •, Conclusions: The five polymorphic microsatellite markers will be useful for studying the genetic structure, reproductive biology, and for identification of clones and provenances of this important medicinal plant.

The column chromatography on silica gel, Sephadex LH-20 and semi-preparative HPLC were used to separate and purify the compounds from the EtOAc extract of medium and MeOH extract of cell cultures of Morus alba. Eight compounds were isolated. Based on physico-chemical properties and spectroscopic data, their structures were identified as isobavachalcone (1), genistein (2), norartocarpetin (3), albanin A (4), guangsangon E (5), mulberrofuran F (6), chalcomoracin (7), kuwanon J (8). Compounds 3-6 were isolated from the cell cultures of M. alba for the first time.

Polygonum L. s. str., belonging to Polygonaceae family, is a big genus with abundant medicinal plants. More than 10 plants are specified in Chinese Pharmacopoeia and many local medicinal standards and over 50 species are used as folk medicines. Owing to the similar morphologies and very small flowers and fruits, they are uneasily identified and often confused with each other and misused clinically. In order to provide a basis for identification of Polygonum s. str. plants, a histological study on stems and leaves of 30 species from Polygonum was undertaken by a routine/polarized light microscopy for the first time. The results showed that: (1) the transverse sections of stems of Polygonum are relatively similar, sclerenchyma such as xylem and fibres with strong polarization effects; (2) the surface views of leaves of Polygonum are distinguishable on distributions and types of stomata, with or without attachments (such as glandular hairs/scales or non-glandular hairs) and the polariscopic features of epidermal cell walls, stomata and cell contents. Observed under polarized light, it was found for the first time that stomata on leaf surface of some plants have a Maltese-cross effect with the arms of the cross intersecting at the stomatal opening. As a result, a key combining the microscopic and polariscopic characteristics of the stems as well as leaves was provided for identifying the 30 medicinal plants of Polygonum. The polarized light microscopic method was proven to be one of the quick, simple and effective techniques for the identification of medicinal plants and botanic crude materials.

The lignan nordihydroguaiaretic acid (NDGA) and its derivatives existing in Larrea divaricata species show a wide range of pharmacological activities which makes this genus an interesting target to consider the plant in vitro cultivation systems as a feasible alternative source for their production. These compounds are potentially useful in treating diseases related to heart condition, asthma, arteriosclerosis, viral and bacterial infections, inflammation and cancer. In the present study, calli, cell suspension cultures, and in vitro and wild plants of L. divaricata were investigated for their potential to synthesize phenolic compounds. Calli, both with and without organogenesis, produced NDGA and quercetin, as did plantlet and wild plants. NDGA was also produced by the cell suspension cultures, together with p-coumaric acid, ferulic acid and sinapyl alcohol. The capacity of undifferentiated tissues to form phenolic compounds is very limited, but when the calli underwent organogenesis, developing mainly adventitious shoots, the phenolic compound production increased significantly. Plantlets regenerated from adventitious shoots of L. divaricata calli did not show the same phenolic pattern as wild plants, with levels of NDGA and quercetin being 3.6- and 5.9-fold lower, respectively., (Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.)

Spilanthes acmella Murr. has a plethora of highly valuable biologically active compounds and has been listed as one of the important medicinal plants of the world. However, no perceptible biotechnological advances have been made for this genus to exploit or enhance its utility. To nullify the effect of seasonal variations, the present report is the first attempt to establish in vitro cell suspension cultures and to evaluate the production of spilanthol from them, a key component of the plant responsible for most of its pharmaceutical activities. The study examined the biomass growth in relation to the consumption of major nutrients and sucrose, agitation speed and dynamic change in pH. Results indicated that the consumption of phosphate resulted in the onset of decline phase in cultures. Spilanthol production was observed to be growth associated and maximum production occurred on the 15th day. Among the carbon sources, the highest production of spilanthol as 91.4 µg g(-1) DW was recorded in the medium supplemented with sucrose, followed by glucose which produced 56.8 µg g(-1) DW of spilanthol. Spilanthol could not be detected in fructose containing medium. Maximum viable cultures were obtained at a rotation speed of cells at 120 rpm. This study signifies the utility of Spilanthes suspension cultures for biosynthesis and constant production of spilanthol, throughout the year. The results of present study are useful for further scale-up process.

Objective: To provide reference for identification and development of Basella rubra., Methods: Microscopic identification and UV spectrometry methods were used., Results: Obvious characteristics were found in the tissue structures of the roots, stems and leaves . And Basella rubra showed some UV spectrometry characteristics., Conclusion: The results can be used as the reference for analyzing the quality of the crude drug.

Two new neolignans (1, 2) and six sesquiterpenes (3- 8) were isolated from the cell cultures of Stellera chamaejasme. Their structures and absolute configurations were elucidated by extensive spectroscopic and computational methods. Compound 4 exhibited significant protective effects against CCl (4)-induced hepatotoxicity in HepG2 cells, reducing aspartate aminotransferase release by 29.49 % at 10 µM. These compounds have not been isolated from plant material, which implies that in vitro plant cell cultures may offer alternative and effective sources of bioactive natural compounds., (© Georg Thieme Verlag KG Stuttgart · New York.)

Nain-e Havandi (Andrographis paniculata Nees.) (AP) is an annual herbaceous plant belonging to the family Acanthacea. Only a few species of Andrographis genus out of 28 are medicinally concerned of which AP is the most important. Knowledge about the arrival of AP to Iran is extremely lacking but most probably it has been imported from India. However, evidence implies the familiarity of Iran's folkloric medicine with this plant, but it has been disappeared from contemporary medicine for unknown reasons. Presence of active ingredients from diterpenoids group such as andrographolide, neoandrographolide and 14-deoxy-11,12-didehydroandrographolide has given incredible unique medicinal properties to the plant. Traditionally, Nain-e Havandi has been used in the role of a non-farm plant as a remedy for skin problems, flu, respiratory disease, and snakebite in East and Southeast Asia for centuries. Recently, it has been utilized as a treatment for HIV, hepatitis, diabetes, cancer and kidney disorders. Intensive cultivation of the herb started only in the past decade in countries such as China, India, Thailand, Indonesia, West Indies, Mauritius and to some extent, in Malaysia. Availability of different ecological zones in Iran complies with reestablishment of AP in tropical and temperate regions of the country. This is killing two birds with one stone, supporting the conservational and economic aspects.

Objective: To compare and identify Celastrus aculeatus and Kadsura heteroclita with pharmacognosy methods for analyzing the quality of the crude drug., Methods: Pharmacognosy study on Celastrus aculeatus and Kadsura heteroclite was carried out through plant identification, crude drug identification and microscopic characteristics identification. The characteristics of Celastrus aculeatus and Kadsura heteroclite were compared., Results: There were significant differences between Celastrus aculeatus and Kadsura heteroclite in appearance of cork, attachments on internal surface of cork,shape of leave edge, number of lateral vein, type of stoma and vessel, and the crystals, the stone cells and the fibers in the same part of both drugs., Conclusion: The pharmacognosy characteristics of both crude drugs can be used for identification and quality control on Celastrus aculeatus and Kadsura heteroclite.

Objective: To determine cytology chromosome numbers, doubling, type, karyotype formulae of seven kinds of Taraxacum., Method: Seven kinds of dandelion in Northeast China were karyomorphologically studied by conventional pressed slice method., Result: The interphase nuclei and prophase chromosomes of all species were found to be of the complex chromocenter type and the interstitial type respectively. The somatic chromosomes showed polysomaty within or among species. The range of chromosome numbers was from 16 to 32, and 24 was preponderant. It is firstly confirmed that the chromosome numbers of T. ohwianum were 2n = 2x = 16 of and those of T. variegatum 2n = 4x = 32 of, and those of other five kinds of dandelion 2n = 3x = 24. Karyotype was diverse and consisted of metacentric, submetacentric and satellites. The relative lengh of chromosome varies from 3.74 to 27.68, asymmetry index was between 59.68% and 64.02%; The karyotype type was belonged to "1A", "2A"and "2B" extensively, to ensure the T. ohwianum and T. variegatum evolutional karyotype type and genetic inheritance pattern., Conclusion: According to results of the cytology karyotype we suggest that T. antungense and T. urbanum could be merged, the result was consistent with Flora Reipublicae Popularis Sinicae (FRPS). This paper reports systematically cytology karyotype feature of seven kinds of dandelions in Northeast, provides the cytology theoretical basis for further development and use of the resource and genetic breeding research of dandelions.

Objective: To study the morphological character and microscopic identification of Embelia parviflora., Methods: Researches on the morphological character and microscopic identification of roots and stems of Embelia parviflora were carried on respectively., Results: Significant microscopic characteristics of Embelia parviflora were confirmed. Such as single stone cells scattered in the root cortical, lots of stone cell groups and fiber bundles scattered in the column parts and became ring intermittently. Stem phloem was broad, large scale secretory cells scattered near cambium region and stone cell groups in ray parts. Crystal stone cells of thick and thin could be seen in powder., Conclusion: These features can be used as the reference for quality standard of Embelia parviflora.

Objective: To study the microscopic and UV-Vis spectrum characteristic of Sapium sebiferum., Methods: The microscopic identification and UV-Vis spectrum identification were adopted., Results: There were cluster crystals in cortical cells of roots and stems and parenchyma cells of the leaves. A few stone cells were found in cortex of the stem. The vascular bundles arranged in the form of a circle in the main vein of the leaves. It could be observed that many crystals and crystal fibers in the powder. Six kinds of solvent of crude drug had absorption peaks between 200 - 800 nm., Conclusion: These results can provide the scientific evidence for the establishment of standard for quality control and further study.

Objective: To identify Ferula syreitschikowii by identification of pharmacognosy and provide the scientific evidence for its identification and application., Methods: The crude drug identification, morphological, UV spectrum identifications, microscopic identification and thin layer chromatography (TLC) were adopted., Results: There were specialization in morphology, microscopic characteristic, UV spectrum identifications and thin layer chromatography (TLC)., Conclusion: These characteristics can be used for scientific basis for gruffs identification of crude drug and providing reference information for further development.

Objective: To identify Calystegia soldanella and provide foundation for its further study and application., Methods: Scanning electron microscope (SEM), upright microscope and UV were applied in the research., Results: Laticifers were observed in the cortex and phloem of root, sclerenchymatous cells were cyclized in the outer phloem of stem, the stomas present on both adaxial and abaxial epidermis, the stomatal type was paracytic. Observed by scanning electron microscope, stomas distributed on and sank into both adaxial and abaxial epidermis; The surface of guard cells were smooth, and subsidiary cells were smooth or veined, the veins were perpendicular to guard cells on adaxial epidermis, while the veins were irregular on subsidiary cells of abaxial epidermis. And the absorption peaks were significantly in UV scanning spectrum: the ethanol extract had an absorption peak at 324nm; While the chloroform extract's at 241nm, 296nm and 316nm., Conclusion: Bases on the characteristic identification, micro-identification and physicochemical identification, SEM is further used in the identification of micro-morphological characteristics of Calystegia soldanella, and it will raise the accuracy of the identification of Calystegia soldanella.

Context: Light microscopy is in most cases a quick method for the identification and discrimination of medicinally used plant drugs; moreover, this technique is very inexpensive. Reliable descriptions of the anatomy of plants and their adulterations are prerequisites for necessary purity controls., Objective: The anatomy of the subterranean organs of 18 pharmaceutically useful as well as related but inconsiderable Asteraceae species from nine genera (Taraxacum F. H. Wigg., Leontodon L., Scorzoneroides Moench, Hypochaeris L., Crepis L., Aposeris Neck., Cichorium L., Scorzonera L., and Tragopogon L.; tribe Cichorieae, Asteraceae) is described in detail and graphically illustrated. Features characterizing and discriminating the studied taxa are presented and discussed., Materials and Methods: The roots/rhizomes of various species were examined by means of light microscopy., Results: Useful anatomical characters were found for the discrimination between the species, and some of them were examined for the first time., Discussion and Conclusion: Discrimination of most genera and species investigated was possibly based on the anatomy of their underground parts. The identified characters may be effectively used for quality control of commercial drugs and the identification of adulterations.

The medicinal herbs derived from genus Senecio have been commonly used in Chinese medicine and triggered attention in recent decades for that they contain the hepatotoxic pyrrolizidine alkaloids. Therefore the botanical pharmacognostic study to authenticate those herbs based on their macroscopic and microscopic characteristics is important for the assurance of safety when they are applied as raw material for extracts or for finished products. In this paper, 13 taxa (11 species and 2 varieties) of Senecio plants were collected and their macroscopic and microscopic characteristics were observed and described by digital microscopic illustration. The results showed that the distribution of collenchyma in the cortex, the level of development for pericycle, the location of the phloem, and the ratio of pith in transverse sections of the stems, and the morphology of the leaf epidermal cells, the stomatal types and the non-glandular hairs in leaf surface view were found to be the main microscopic characteristics for authentication of different Senecio species. The herbs derived from genus Senecio can be distinguished from each other on the basis of their macroscopic and microscopic characteristics, and those observation can be used for the identification of commercial crude drugs from Senecio plants.