Internal Phloem Research Articles

Chemical analysis of different parts from agarwood columns by artificially agarwood-inducing method based on GC–MS and UPLC-TOF-MS

Agarwood is resin-containing wood produced by plants that have been injured. It is widely used in herbal medicine, incense, decorative items, and so on. In this study, we conducted resin area statistical analysis, determined starch particle and reducing sugar contents, and performed multivariate statistical analysis of chemical composition by GC–MS and UPLC-Q-TOF-MS to explore the different components in sections cut from an agarwood column, designated as A1–A4. The results showed that after stimulation by Agar-Bit inducer, the internal phloem parenchyma cells of the column started to form agarwood, and then starch granules were converted into soluble reducing sugars and agarwood resin. Section A1 showed rapid loss of starch granules, resulting in higher contents of reducing sugars and resin. The resin areas of agarwood in the respective sections were different, gradually decreasing on going from A1 to A4. Total numbers of metabolites of 87 and 63 were identified by GC–MS and UPLC-Q-TOF-MS, respectively. Of these, 10 and 16 metabolites with significant differences (variable importance projection >1) were selected through multivariate statistical analysis. These metabolites included chromones, sesquiterpenes, alkanes, and fatty acids. Among them, 6-methoxy-2-(2-phenylethyl)chromone and 6,7-dimethoxy-2-(2-phenylethyl)chromone were significant markers detected by both GC–MS and UPLC-Q-TOF-MS, which may be essential substances responsible for differences in the agarwood-forming capacities of the cut sections. In conclusion, there has been limited research on the different agarwood-forming capacities of agarwood columns. Here, we explored the differences in various sections of agarwood through chemical analysis to provide a more comprehensive and in-depth understanding of its constitution.

The spruce bark volatiles and internal phloem chemical profiles after the forest gap formation: the annual course

Abstract Our study explores the impact of sudden gap formation on the bark volatile and internal chemical profiles of Norway spruce trees during the initial dry year of research plot 2018 following gap formation. We investigated the annual variation in two main physiological traits of Norway spruce trees at the forest edge (FE) and in the forest interior (FI): bark monoterpene (MT) emission spectra and internal phloem MT composition. Given that gap formation increases the solar radiation dose and temperature for trees at the forest edge, we hypothesized that the concentrations of airborne terpenes released from the tree bark and internal phloem terpenes will increase as a consequence of induced tree defenses. Our findings demonstrate significant increases in both airborne terpene concentrations and internal terpene composition in trees at the forest edge compared to the control trees in the forest interior. This study provides novel insights into the annual dynamics of bark monoterpenes following forest edge establishment and underscores the physiological changes experienced by trees in response to the gap formation.

Anatomical observation and transcriptome analysis of branch-twisted mutations in Chinese jujube

BackgroundPlant organs grow in a certain direction and organ twisted growth, a rare and distinctive trait, is associated with internal structure changes and special genes. The twisted branch mutant of Chinese jujube jujube, an important fruit tree native to China and introduced to nearly 50 countries, provides new typical materials for exploration of plant twisted growth.ResultsIn this study, the cytological characteristics and related genes of twisted branches in Chinese jujube were revealed by microscopy observation and transcriptome analysis. The unique coexistence of primary and secondary structures appeared in the twisted parts of branches, and special structures such as collateral bundle, cortical bundles, and internal phloem were formed. Ninety differentially expressed genes of ‘Dongzao’ and its twisted mutant were observed, in which ZjTBL43, ZjFLA11, ZjFLA12 and ZjIQD1 were selected as candidate genes. ZjTBL43 was homologous to AtTBL43 in Arabidopsis, which was involved in the synthesis and deposition of cellular secondary wall cellulose. The attbl43 mutant showed significant inflorescence stem bending growth. The transgenic lines of attbl43 with overexpression of ZjTBL43 were phenotypically normal.The branch twisted growth may be caused by mutations in ZjTBL43 in Chinese jujube. AtIQD10, AtFLA11 and AtFLA12 were homologous to ZjIQD1, ZjFLA11 and ZjFLA12, respectively. However, the phenotype of their function defect mutants was normal.ConclusionIn summary, these findings will provide new insights into the plant organ twisted growth and a reference for investigation of controlling mechanisms of plant growth direction.

Anastomosing laticifer in the primary and secondary structures of Calotropis procera (Aiton) W.T.Aiton (Apocynaceae) stems.

An in-depth understanding of the development and distribution of laticifer (latex secretory structure) will be important for the production of both rubber and medicines and will support studies on plant adaptations to their environments. We characterize here and describe the ontogenesis of the laticifer sytem in Calotropis procera (Apocynaceae), an invasive subshrub species in arid landscapes. Anatomical and histochemical evaluations of the primary and secondary structures of the stem were carried out on a monthly basis during a full year, with ultrastructural evaluations of laticifer on the stem apex during the rainy season. In the primary structure, laticifer differentiate early from procambium and ground meristem cells of the cortex and medulla and become concentrated adjacent to the external and internal phloem of the bicollateral bundles. In the secondary structure, laticifer differentiates from fusiform derivative cells of the phloem close to the sieve-tube elements. The laticifer is of the articulated, anastomosing, branched type, and it originates from precursor cells that loose the transversal and longitudinal walls by dissolution. Latex is a mixture of terpenes, alkaloids, flavonoids, mucilage, and proteins. The apical meristem and vascular cambium where the laticifer system begins its development are active throughout the year, including during the dry season. The vascular cambium produces phloem with laticifer precursor cells during the rainy season, with high temperatures and long days. The ability of C. procera to grow under water deficit conditions and produce laticifer throughout the year contribute to its wide distribution in arid environments.

Actual aspects of anatomical research of medicinal plant material of Vinca minor L

The article presents the results of a microscopic study of the medicinal plant material Vinca minor L. (Apocynaceae), used in the pharmaceutical industry to obtain indole alkaloids with antitumor activity. When carrying out anatomical analysis of the leaves and stems of V. minor, the main anatomical and diagnostic signs was established, this can be used in the identification and assessment of the authenticity of medicinal plants. Analysis of the anatomical structure showed that the leaves of V. minor are dorsoventral; hypostatic sheet plastic. The stomatal apparatus of plants is paracytic. The main vein of the leaf blade is represented by a bicollateral conducting bundle; lateral leaf veins had closed collateral bundles. The cross-sectional shape of the stem is grooved with four ribs. The stem of V. minor was characterized by a non-bundle type of anatomical structure. The peculiarity of the stem structure was the presence of internal phloem. The characteristic marker features of the stem and leaf of the plant include the presence of straight, non-segmented, non-anastomosing lactic acidae with an excretory substance of a greenish-yellow color. The information obtained can serve as a basis for the development of the section “Microscopy” when writing pharmacopoeial articles.

Formation of medullary phloem in Argyreia nervosa (Burm. f.) Bojer

Histologically, family Convolvulaceae is characterised by the presence of successive cambia, medullary (internal/intraxylary) and interxylary phloem in majority of the species, whereas some of the members are devoid of successive cambia and medullary bundles. The present study on Argyreia nervosa (Burm. f.) Bojer showed the presence of medullary bundles while internal phloem was absent during the primary growth. As the secondary growth progressed, successive cambia initiated from the pericyclic parenchyma. Development of medullary bundles began along with the regular protoxylem and protophloem while formation of intraxylary phloem was observed only after the initiation of secondary growth. Medullary/intraxylary sieve elements began to develop from the marginal pith cells. In thick stems, small segments of internal cambium initiated between the protoxylem and internal phloem. This internal cambium was functionally unidirectional and produced internal phloem centripetally. Developmental particulars are described in details along with its significance.

Inter-organ transport of secologanin allows assembly of monoterpenoid indole alkaloids in a Catharanthus roseus mutant

The medicinal value of the monoterpenoid indole alkaloids (MIAs) such as 3′,4′-anhydrovinblastine, as well as their chemical complexity have stimulated extensive efforts to understand the biochemical and molecular pathways involved in their biosynthesis in plants such as Catharanthus roseus, Rawvolfia serpentina and others. Ethyl methane sulphonate (EMS) mutagenesis has been used successfully together with simple MIA thin layer chromatography screening to identify C. roseus mutants with altered MIA profiles. This study describes the isolation of very low iridoid and MIA containing C. roseus mutant (M2-1582) that accumulates MIAs when the plant is provided with secologanin by feeding mutant roots or by grafting the mutant scion onto wild type roots. The observed low iridoid and MIA content was correlated with lowered expression of BIS1/BIS2 transcription factors and several genes involved in secologanin biosynthesis that are expressed in internal phloem parenchyma cells of leaves. When exogenous secologanin was applied to the roots of the mutant plant, secologanin levels rose more than 13-fold, while two major MIAs catharanthine and vindoline rose more than 8- and 4- fold, respectively. Grafting the mutant on WT stocks led to 27-, 11- and 27-fold increases in secologanin, catharanthine and vindoline, respectively in leaves of the scion one week after graft initiation. Other minor MIAs (serpentine, anhydrovinblastine, vindolidine, deacetylvindoline, tabersonine and 16-methoxytabersonine) that were not detected in the mutant, became detectable in leaves of the scion. These results provide strong evidence for a secologanin transport mechanism that mobilizes this iridoid between different plant organs in C. roseus and that secologanin transport to the mutant across the graft union permits the formation of MIAs in leaves of the mutant.

Transcriptomic and functional analysis of cucumber (Cucumis sativus L.) fruit phloem during early development.

The phloem of the Cucurbitaceae has long been a subject of interest due to its complex nature and the economic importance of the family. As in a limited number of other families, cucurbit phloem is bicollateral, i.e. with sieve tubes on both sides of the xylem. To date little is known about the specialized functions of the internal phloem (IP) and external phloem (EP). Here, a combination of microscopy, fluorescent dye transport analysis, micro-computed tomography, laser capture microdissection and RNA-sequencing (RNA-Seq) were used to study the functions of IP and EP in the vascular bundles (VBs) of cucumber fruit. There is one type of VB in the peduncle, but four in the fruit: peripheral (PeVB), main (MVB), carpel (CVB) and placental (PlVB). The VBs are bicollateral, except for the CVB and PlVB. Phloem mobile tracers and 14 C applied to leaves are transported primarily in the EP, and to a lesser extent in the IP. RNA-Seq data indicate preferential gene transcription in the IP related to differentiation/development, hormone transport, RNA or protein modification/processing/transport, and nitrogen compound metabolism and transport. The EP preferentially expresses genes for stimulus/stress, defense, ion transport and secondary metabolite biosynthesis. The MVB phloem is preferentially involved in photoassimilate transport, unloading and long-distance signaling, while the PeVB plays a more substantial role in morphogenesis and/or development and defense response. CVB and PlVB transcripts are biased toward development of reproductive organs. These findings provide an integrated view of the differentiated structure and function of the vascular tissue in cucumber fruit.

A Contribution on the Microcharacters of Barleria prionitis L. Useful in Pharmcognosy

Barleria prionitis (Acanthaceae) is an important medicinal plant distributed throughout the various regions of India and useful in urinary and paralytic affection, stomach disorders, catarrh, cough and anasaraca and lacerated sole. In the present investigation an attempt was made to study the anatomy of root, stem, lamina, midrib, petiole, stomata and trichomes as well as histochemical localization of various substances viz., calcium oxalate, lignin, suberin, lipids in the organs of Barleria prionitis. The present study revealed that the plant distinguishing microcharacters viz., bicollateral vascular bundles, internal phloem, uni - biseriate xylem rays, inverted omega shape vascular bundle, diacytic stomata and eglandular as well as glandular trichomes which are useful in solving taxonomic problems and pharmacognosy. They have significance in identification of crude drugs from this taxon.

Foliar micromorphology and anatomy of Ugni molinae Turcz. (Myrtaceae), with particular reference to schizogenous secretory cavities

Ugni molinae Turcz. is one of the most studied species of South American Myrtaceae due to its edible fruits and foliar medicinal compounds. However, there is no anatomical study of the leaves or secretory cavities. This paper seeks to describe the leaf micromorphology and anatomy of the species using standard protocols for light and scanning electron microscopy. Secretory cavities were anatomically characterized in young and mature leaves. Histochemical staining of the cavities was performed. The leaves of U. molinae are hypostomatic, have a wavy surface and possess scattered hairs. Leaf anatomical features include dorsiventral mesophyll, two to three layers of palisade parenchyma with abundant chloroplasts, calcium oxalate crystals and internal phloem in vascular bundles. Schizogenous secretory cavities are present on the abaxial surface and are mainly located on the margins of the leaves. Histochemical tests of these cavities suggest the presence of lipophilic substances. This is the first study of secretory cavities in Chilean Myrtaceae. In general, micromorphological and anatomical characters are similar to other species of the family. The present findings could provide valuable anatomical information for future research in South American Myrtaceae.

How to catch the patch? A dendrometer study of the radial increment through successive cambia in the mangrove Avicennia.

Successive vascular cambia are involved in the secondary growth of at least 200 woody species from >30 plant families. In the mangrove Avicennia these successive cambia are organized in patches, creating stems with non-concentric xylem tissue surrounded by internal phloem tissue. Little is known about radial growth and tree stem dynamics in trees with this type of anatomy. This study aims to (1) clarify the process of secondary growth of Avicennia trees by studying its patchiness; and (2) study the radial increment of Avicennia stems, both temporary and permanent, in relation to local climatic and environmental conditions. A test is made of the hypothesis that patchy radial growth and stem dynamics enable Avicennia trees to better survive conditions of extreme physiological drought. Methods Stem variations were monitored by automatic point dendrometers at four different positions around and along the stem of two Avicennia marina trees in the mangrove forest of Gazi Bay (Kenya) during 1 year. Patchiness was found in the radial growth and shrinkage and swelling patterns of Avicennia stems. It was, however, potentially rather than systematically present, i.e. stems reacted either concentrically or patchily to environment triggers, and it was fresh water availability and not tidal inundation that affected radial increment. It is concluded that the ability to develop successive cambia in a patchy way enables Avicennia trees to adapt to changes in the prevailing environmental conditions, enhancing its survival in the highly dynamic mangrove environment. Limited water could be used in a more directive way, investing all the attainable resources in only some locations of the tree stem so that at least at these locations there is enough water to, for example, overcome vessel embolisms or create new cells. As these locations change with time, the overall functioning of the tree can be maintained.

Structure And Development Of Internal Phloem In Solanum Pseudocapsicum (Solanaceae)

The development of internal phloem in the Jerusalem cherry, Solanum pseudocapsicum L. (Solanaceae), was studied in young and mature stems. The early presence of primary internal phloem is succeeded by the development of secondary internal phloem from an internal cambium situated between the protoxylem and primary internal phloem. In the second and third visible internodes of the young stem, procambial derivatives begin to differentiate as discrete strands of internal protophloem in a perimedullary position prior to the differentiation of protoxylem and external protophloem. In 6–8 mm diameter stems, sieve elements of the internal phloem become non-conducting, begin to collapse, and undergo obliteration. In 15–20 mm diameter stems internal cambium is initiated from the parenchyma cells situated between the protoxylem and primary internal phloem. The development of internal phloem and an internal cambium in S. pseudocapsicum is compared with that in other taxa. There seems to be a gradual variation in the origin of an internal cambium from either remnants of the procambium or dedifferentiation of peripheral pith cells across dicotyledons with an internal cambium.

Morphoanatomy of Schultesia pachyphylla (Gentianaceae): a discordant pattern in the genus

Schultesia Mart. comprises 21 closely related taxa. The revision of the genus showed that Schultesia pachyphylla Griseb. and Xestaea lisianthoides Griseb. are different from other species. The aim of this study was to describe the morphoanatomy of S. pachyphylla, comparing it with other Schultesia species and X. lisianthoides, and to provide data on their geographical distribution and habitat. Morphological and anatomical data were subjected to multivariate analysis. Schultesia pachyphylla is endemic to the state of Bahia, Brazil, and is separated from the other species by having 5-merous flowers, 5-winged calyx, wings between the calyx lobes, loss of interveinal tissue on sepals during fruit maturation, dark red colored immature calyces, capsules opening from base to apex, 3-anaporate pollen grains, and epidermal papillae evident in the field. Schultesia pachyphylla has amphistomatic leaves with anisocytic stomata, uniseriate epidermis, dorsiventral mesophyll, and bicollateral vascular bundles. The stem has small wings, cortical and medullary parenchyma, and internal phloem. Foliar colleters and nectaries on leaves and stem were observed. We believe that morphological and anatomical data could support the creation of a monotypic genus or the relocation of this species into another genus. Phylogenetic analyses are in progress to define the species’ positioning.

A 7-Deoxyloganetic Acid Glucosyltransferase Contributes a Key Step in Secologanin Biosynthesis in Madagascar Periwinkle

Iridoids form a broad and versatile class of biologically active molecules found in thousands of plant species. In addition to the many hundreds of iridoids occurring in plants, some iridoids, such as secologanin, serve as key building blocks in the biosynthesis of thousands of monoterpene indole alkaloids (MIAs) and many quinoline alkaloids. This study describes the molecular cloning and functional characterization of three iridoid glucosyltransfeases (UDP-sugar glycosyltransferase6 [UGT6], UGT7, and UGT8) from Madagascar periwinkle (Catharanthus roseus) with remarkably different catalytic efficiencies. Biochemical analyses reveal that UGT8 possessed a high catalytic efficiency toward its exclusive iridoid substrate, 7-deoxyloganetic acid, making it better suited for the biosynthesis of iridoids in periwinkle than the other two iridoid glucosyltransfeases. The role of UGT8 in the fourth to last step in secologanin biosynthesis was confirmed by virus-induced gene silencing in periwinkle plants, which reduced expression of this gene and resulted in a large decline in secologanin and MIA accumulation within silenced plants. Localization studies of UGT8 using a carborundum abrasion method for RNA extraction show that its expression occurs preferentially within periwinkle leaves rather than in epidermal cells, and in situ hybridization studies confirm that UGT8 is preferentially expressed in internal phloem associated parenchyma cells of periwinkle species.

Interxylary phloem: Diversity and functions

Interxylary phloem is here defined as strands or bands of phloem embedded within the secondary xylem of a stem or root of a plant that has a single vascular cambium. In this definition, interxylary phloem differs from intraxylary phloem, bicollateral bundles, pith bundles, and successive cambia. The inclusive but variously applied terms included phloem and internal phloem must be rejected. Histological aspects of interxylary phloem are reviewed and original data are presented. Topics covered include duration of interxylary phloem; relationship in abundance between sieve tubes in external phloem and interxylary phloem; distinctions between interxylary and intraxylary phloem; presence of parenchyma, fibers, and crystals in the interxylary phloem strands; development of cambia within interxylary phloem strands; three-dimensionalization and longevity of phloem, systematic distribution of interxylary phloem; physiological significance; and habital correlations. No single physiological phenomenon seems to explain all instances of interxylary phloem occurrence, but rapidity and volume of photosynthate transport seem implicated in most instances.

Anatomical characteristics as taxonomic tools for the species of Curtia and Hockinia (Saccifolieae–Gentianaceae Juss.)

The delimitation of species and even support of the genera Curtia and Hockinia, monotypic genus, are still controversial. To define anatomical characteristics that help resolve taxonomic problems, the stem and leaf anatomy of seven species of Curtia and Hockinia montana are described. Some characteristics are found in both genera: leaves with uniseriate epidermis, dorsiventral mesophyll, cells of the palisade parenchyma with invaginations, basal acrodromous venation with an incomplete marginal ultimate venation, presence of colleters and nectaries, stem with small wings and internal phloem. The types of stomata, the midrib vascular bundle, and the type and distribution of nectaries were used to separate the genera. The presence/absence of crystals, the sinuosity of the anticlinal walls of the epidermal cells, the type and distribution of stomata, and the distribution of nectaries distinguish species of Curtia. Our results suggest that these anatomical characteristics are potentially useful for clarifying the taxonomy of Gentianaceae, and that they represent an additional tool for distinguishing genera and identifying species of Curtia, particularly pointing out the need to revise the synonymy between C. patula and C. tenuifolia. We also highlight the first report of leaf nectaries and colleters in the Saccifolieae and the first record of stem nectaries in the Gentianaceae.

Characterization of the plastidial geraniol synthase from Madagascar periwinkle which initiates the monoterpenoid branch of the alkaloid pathway in internal phloem associated parenchyma

Madagascar periwinkle (Catharanthus roseus [L.] G. Don, Apocynaceae) produces monoterpene indole alkaloids (MIAs), secondary metabolites of high interest due to their therapeutic value. A key step in the biosynthesis is the generation of geraniol from geranyl diphosphate (GPP) in the monoterpenoid branch of the MIA pathway. Here we report on the cloning and functional characterization of C. roseus geraniol synthase (CrGES). The full-length CrGES was over-expressed in Escherichia coli and the purified recombinant protein catalyzed the conversion of GPP into geraniol with a Km value of 58.5μM for GPP. In vivo CrGES activity was evaluated by heterologous expression in a Saccharomyces cerevisiae strain mutated in the farnesyl diphosphate synthase gene. Analysis of culture extracts by gas chromatography-mass spectrometry confirmed the excretion of geraniol into the growth medium. Transient transformation of C. roseus cells with a Yellow Fluorescent Protein-fusion construct revealed that CrGES is localized in plastid stroma and stromules. In aerial plant organs, RNA in situ hybridization showed specific labeling of CrGES transcripts in the internal phloem associated parenchyma as observed for other characterized genes involved in the early steps of MIA biosynthesis. Finally, when cultures of Catharanthus cells were treated with the alkaloid-inducing hormone methyl jasmonate, an increase in CrGES transcript levels was observed. This observation coupled with the tissue-specific expression and the subcellular compartmentalization support the idea that CrGES initiates the monoterpenoid branch of the MIA biosynthetic pathway.

Morpho-anatomy of the leaf and stem of Eugenia pyriformis

Eugenia pyriformis Cambess., known as uvaia, is a species of Myrtaceae native to Brazil. Its leaves are used in folk medicine to treat gout because they possess the property of inhibiting xanthine oxidase, an enzyme involved in the conversion of xanthine into uric acid. The objective of this work was to study the leaf and stem morpho-anatomy of E. pyriformis, in order to contribute to what is known about the Brazilian flora, and this medicinal plant and potential vegetal drug. Samples of mature leaves and young stems were fixed and sectioned by freehand, or embedded in glycol methacrylate and sectioned with a microtome, and then stained. In addition, microchemical tests and scanning electron microscopy were performed. The leaf is simple, symmetric, elliptic-lanceolate, with an acute apex and base, and an entire margin. The epidermis is uniseriate and coated with a moderately thick cuticle. The stomata are anomocytic and inserted at the same level as the adjacent cells. Unicellular non-glandular trichomes are abundant on the abaxial surface. The mesophyll is dorsiventral. In transverse section, the midrib is plano-convex and the petiole is circular, and both of these structures have a single bicollateral vascular bundle. In the stem, the vascular cylinder consists of external phloem, xylem and internal phloem, traversed by narrow rays. Phenolic compounds, druses and prismatic crystals of calcium oxalate are also present in the leaf and stem.

A single gene encodes isopentenyl diphosphate isomerase isoforms targeted to plastids, mitochondria and peroxisomes in Catharanthus roseus

Isopentenyl diphosphate isomerases (IDI) catalyze the interconversion of the two isoprenoid universal C5 units, isopentenyl diphosphate and dimethylally diphosphate, to allow the biosynthesis of the large variety of isoprenoids including both primary and specialized metabolites. This isomerisation is usually performed by two distinct IDI isoforms located either in plastids/peroxisomes or mitochondria/peroxisomes as recently established in Arabidopsis thaliana mainly accumulating primary isoprenoids. By contrast, almost nothing is known in plants accumulating specialized isoprenoids. Here we report the cloning and functional validation of an IDI encoding cDNA (CrIDI1) from Catharanthus roseus that produces high amount of monoterpenoid indole alkaloids. The corresponding gene is expressed in all organs including roots, flowers and young leaves where transcripts have been detected in internal phloem parenchyma and epidermis. The CrIDI1 gene also produces long and short transcripts giving rise to corresponding proteins with and without a N-terminal transit peptide (TP), respectively. Expression of green fluorescent protein fusions revealed that the long isoform is targeted to both plastids and mitochondria with an apparent similar efficiency. Deletion/fusion experiments established that the first 18-residues of the N-terminal TP are solely responsible of the mitochondria targeting while the entire 77-residue long TP is needed for an additional plastid localization. The short isoform is targeted to peroxisomes in agreement with the presence of peroxisome targeting sequence at its C-terminal end. This complex plastid/mitochondria/peroxisomes triple targeting occurring in C. roseus producing specialized isoprenoid secondary metabolites is somehow different from the situation observed in A. thaliana mainly producing housekeeping isoprenoid metabolites.

Anatomy and Pharmacognosy ofLeptadenia Reticulata, an important medicinal plant of India

Leptadenia reticulata, an important medicinal plant of India, was subjected to a detailed pharmacognostical analysis to locate distinguishing biomarkers. Multicellular uniseriate smooth trichomes, bi-collateral vascular bundles, anisocytic stomata and absence of any cell inclusions were the features seen in leaf. Stem possessed broad patches of sclereids in the pericycle and irregular patches of intraxylary phloem and internal phloem consisted of radial rows of angular cells with starch grains. Secondary xylem consisting of large vessels and tracheids and prismatic crystals of calcium oxalate in cortex are very distinct. Large sclereids, starch grains and prismatic calcium oxalate crystals in cortex, bast and parenchyma differentiate roots. X-shaped pits and multiseriate medullary rays were seen in stem whereas roots possessed simple pits and mostly uniseriate medullary rays. Other micromorphological characters were stomatal index 11.67±0.68, palisade ratio 6.0±0.39, vein islet number 9.5±0.97, and vein termination number 11–12.