Phytochemical and pharmacognostic studies of Buddleja asiatica leaves

Buddleja asiatica commonly known as “Booi” is a large deciduous shrub belongs to family Scrophulariaceae, traditionally used as antipyretic, analgesic, hypotensive, antimicrobial, anti-inflammatory, and antifungal agent recorded from essential oil obtained from leaves. The literature revealed that the plant has been widely used for many other purposes. The said plant has been analyzed through pharmacognostic techniques such as morphological, microscopic, and physio-chemical evaluations to keep the originality of the plant and to avoid adulteration. Morphologically, the plant is evergreen shrub, while organoleptic studies showed that the leaf has glabrous upper surface and tomentose lower surface, with slightly bitter taste and aromatic odor. Anatomically, the leaf showed typical dicot histological differentiation with hypostomatic nature having highest frequency (90%) of actinocytic stomata. The average stomatal number and stomatal index were 336 ± 39.5 and 30.01 ± 2.34, respectively. The palisade cell ratio, vein termination, and vein islet number were 9.2 ± 0.72, 10.2 ± 3.1, and 10.3 ± 3.3, respectively. Various tissue fragments have been observed during powdered drug analysis of the leaf. Preliminary phytochemical screening confirmed the presence of proteins, phenols, alkaloids, saponins, flavonoids, tannins, and glycosides. Fluorescence analysis in ordinary day light and under UV light along with extractive values was also analyzed. The above-mentioned studies that have been reported, for the first time, for the said plant may be significant to establish the pharmacognostic and phytochemical standards of the said species.     

Morphological,microscopic, and physicochemical studies of Diospyros montana

Adulteration is the root cause of producing not only a chemically and pharmacologically inferior but also in some instances hazardous or poisonous drug. Despite availability of several techniques, microscopy and physicochemical analyses are the most practical approaches for crude drug authentication. Hence, the present study aimed to evaluate morphological, microscopic, and physicochemical properties of root, bark, leaf, and fruit of Diospyros montana Morphological properties were determined by sensory organs, whereas microscopic features of cross‐sections and powders were determined by light and scanning electron microscopy. The proximate and fluorescence analyses were performed using the standard guidelines. The physical examination of fresh, shade‐dried, and powdered material showed no significant change in color. The identifying cellular structures included cuboidal cork, pitted tracheids, scalariform, reticulate and spiral xylary vessels, and rosettes, raphide, and cuboidal calcium oxalate crystals. The stomatal number, stomatal index, vein‐islet and vein‐termination number, and palisade ratio in the leaf were found to be 293.91 ± 32.68 mm^?2, 64.18 ± 3.42%, 22.00 ± 3.81 mm^?2 and 38.40 ± 5.81 mm^?2, and 3.85 ± 0.60, respectively. Total ash, acid insoluble ash, water soluble ash and sulfated ash of leaf (9.00 ± 0.50%, 1.67 ± 0.23%, 2.00 ± 0.22% and 14.50 ± 0.99%, respectively), foaming index of bark and root (111.11 ± 2.11), and swelling index of fruit (19.00 ± 3.45) were higher than the other parts. The powder of different parts showed characteristic colors in the daylight and UV light upon treatment with various regents. The plant was found to be rich in saponins, fibers, and flavonoids. The results of the present study may serve as identifiers of different parts of Diospyros montana.     

Leaf and stem micromorphology of Byrsonima sericea DC. by light and scanning electron microscopy

Micromorphological studies were carried out using multiple microscopic techniques on the leaves and stem bark of Byrsonima sericea DC. (Malpighiaceae), a species popularly known as “murici” and used medicinally, in order to identify both qualitative and quantitative features of leaf and stem anatomy and histochemistry as differential parameters to support both the quality control of its ethnodrugs and the taxonomy of the genus. The study was conducted using traditional techniques of plant anatomy, histochemical tests, and the stomatal index (SI). Byrsonima sericea has hypostomatic leaves, anomocytic stomata, and its epidermal walls are anticlinal and straight on the adaxial and curved on the abaxial faces. T‐shaped trichomes were observed mainly on the abaxial surface. The leaf epidermis showed waxes syntopism on both surfaces, with the occurrence of different crystalloid forms on a single phylloplane. The mesophyll is dorsiventral, with 3‐4 collateral vascular bundles. Phenolic compounds, starch, and proteins were identified in the petiole and stem. The SI was 14.5 ± 0.53% (p < .05), but did not showed significant variations. A set of characters were found to be distinctive for the studied species, however, constituting parameters that could be used to separate B. sericea from other species of the genus.     

Botanical features for identification of Gymnosporia arenicola dried leaf

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. Microsc. Res. Tech. 78:1001–1009, 2015. © 2015 Wiley Periodicals, Inc.     

Microscopic investigations and pharmacognostic techniques for the standardization of the fruits of Rosa laxa Retz

Rosa laxa Retz., a shrub belonging to the family Rosaceae, is widely distributed in the northern foothills of the Tianshan Mountains in Xinjiang, China. The fruits of R. laxa (FRL) has antibacterial, hypolipidemic, and antioxidant effects. In this study, FRL was subjected to pharmacognostic identification of its source, morphology, microscopic characteristics, and physicochemical properties. The microscope showed that the cross-sectional features of FRL were obvious, and the FRL powder contained vessel, parenchyma cells, exocarp cells, pollen grains, and cluster crystals. Scanning electron microscopy (SEM) analysis results show that numerous villi and many small particles (particle size of 5–50 μm) were observed in the FRL powder, and there are many gullies on the surface of the particles. In addition, the secondary metabolites of FRL were characterized via ultraviolet–visible (UV–Vis) spectroscopy, Fourier transform infrared (FTIR) spectroscopy, and thin-layer chromatography (TLC). Results showed that FRL contains various secondary metabolites, including flavonoids, phenolic acids, glycosides, and tannins. Water as the extraction solvent had the highest extraction rate and the contented of total flavonoids was 2.88 mg/g, and the contented of total polyphenols was 54.89 mg/g. Moreover, TLC identification revealed that it contains catechin and tiliroside. These parameters of FRL, which are reported herein, are important to the development of the pharmacognostic standards, as well as in the identification and quality control of FRL.     

Authentication of herbal drug Tukhm‐e‐balango (Lallemantia royleana Benth.) using microscopic,pharmacognostic, and phytochemical characterization

The study is aimed to provide a comprehensive account on authentication of herbal drug named as Tukhm‐e‐balango (Lallemantia royleana Benth.) from the seeds of Ocimum basilicum by using microscopic, pharmacognostic, and phytochemical characterization. The crude medicinal plants and their parts are often adulterated or substituted in market due to improper identification by the consumers while among herbal plant sellers, taxonomic confusion is caused due to morphological similarities of the plant parts and lack of a standard identification system.In microscopy, both herbarium and fresh specimens were studied using qualitative and quantitative morphological characteristics of leaves, seeds, and pollen. For pharmacognosy, solubility, fluorescence, and physicochemical characterizers were analyzed whereas a total phenolic and flavonoids contents was determined in addition to DPPH radical scavenging activity. In current study, microscopic, pharmacognostic, and phytochemical characterization clearly differentiated L. royleana from O. basilicum. The major problem in herbal drug industry is caused due to confusion and controversy of certain synonyms used for more than one or two drugs. Sometimes, under the same common or local name, entirely different taxa are being sold in herbal markets. It is concluded that correct and proper identification of medicinal plants is very crucial to ensure the safety and efficacy of herbal medicines, as many medicinal plants are intentionally or unintentionally adulterated with similar species or varieties. In herbal market, the seeds of L. royleana are adulterated with seeds of O. basilicum due to their similar morphology.     

Effect of strawberry vein banding virus and strawberry mottle virus co-infection on the growth and development of strawberry

Strawberry mottle virus (SMoV) and strawberry vein banding virus (SVBV) cause diseases on strawberry plants, but the effect of coinfection of SMoV and SVBV on the growth, development, and defense system of strawberry (Fragaria × ananassa Duchesne) remains unknown. We investigated the effect of SMoV and SVBV co-infection on strawberry cultivar ‘Benihope’. The results showed that stem diameter, leaf size, leaf number, relative chlorophyll content, total chlorophyll content, photosynthetic parameters, and stomatal aperture of SMoV and SVBV co-infected strawberry (VIS) plants were in a weaker level than uninfected control plants, indicating that viruses inhibited the growth and photosynthesis of strawberry plants. Furthermore, the initiation of flowering and fruiting stages of VIS plants were delayed by about three weeks compared with the controls, and the fruiting period was shortened, demonstrating that the reproduction of VIS plants was inhibited. Fruit quality was damaged in VIS plants due to a significant increase in fruit firmness and titratable acidity and decrease in total soluble solid content than control fruits. More dead cells and H₂O₂ accumulated along the veins of VIS leaves, and the content of abscisic acid and catalase activity significantly increased, whereas anthocyanin content was lower than that of control plants. The results demonstrate that SVBV and SMoV coinfection inhibits the growth and development of ‘Benihope’ strawberry plants, and the plants respond to viruses by regulating stomatal aperture, the accumulation of ABA and antioxidants. To our knowledge, this study contributes information to understand how both viruses impair the strawberry growth and development for the first time.

     

Microscopic and phytochemical techniques as a tool for authentication of herbal drug chiraita: Swertia cordata (G. Don) C.B. Clarke

Swertia cordata (G. Don) C.B. Clarke is one of the potential medicinal plants extensively used in eastern traditional medicine such as Unani, Ayurveda, Siddha, and in traditional Tibetan and Chinese medicine. S. paniculata is the common adulterant of S. cordata at herbal shops and markets but S. paniculata is also used in number of herbal formulations. The present study was conducted to use microscopic, pharmacognostic, and phytochemical techniques as a tool for the authentication of herbal drug chiraita (S. cordata). In herbal markets, mixing, adulteration, and use of spurious materials as substitute have become a major concern for herbal practitioners, local user, and industry for reasons of safety and efficacy. Therefore, authentication of medicinal plants is of utmost importance at each level of drug research. In the present study, anatomical features of two species showed a great diversity, as irregular epidermal cells and nonglandular, unicellular trichomes were found in S. cordata while in S. paniculata epidermal cells were hexagonal in shape and trichomes were A‐shaped. Antioxidant activity of two species showed a great variation where IC₅₀ value recorded for S. cordata was 208 μg/mL, while for S. paniculata IC₅₀ was 624 μg/mL. The study can serve as an important source of information to achieve the authenticity and to evaluate the quality and purity of the plant material in accordance to WHO guidelines. As this species is greatly exploited, so conservation is highly recommended.     

Time‐lapse microscopy using smartphone with augmented reality markers

A prototype system that replaces the conventional time‐lapse imaging in microscopic inspection for use with smartphones is presented. Existing time‐lapse imaging requires a video data feed between a microscope and a computer that varies depending on the type of image grabber. Even with proper hardware setups, a series of tedious and repetitive tasks is still required to relocate to the region‐of‐interest (ROI) of the specimens. In order to simplify the system and improve the efficiency of time‐lapse imaging tasks, a smartphone‐based platform utilizing microscopic augmented reality (μ‐AR) markers is proposed. To evaluate the feasibility and efficiency of the proposed system, a user test was designed and performed, measuring the elapse time for a trial of the task starting from the execution of the application software to the completion of restoring and imaging of an ROI saved in advance. The results of the user test showed that the average elapse time was 65.3 ± 15.2 s with 6.86 ± 3.61 μm of position error and 0.08 ± 0.40 degrees of angle error. This indicates that the time‐lapse imaging task was accomplished rapidly with a high level of accuracy. Thus, simplification of both the system and the task was achieved via our proposed system. Microsc. Res. Tech. 77:243–249, 2014. © 2014 Wiley Periodicals, Inc.     

Electron microscopic observations of stomata,epicuticular waxes,and papillae in Chamaecyparis obtusa: Reconsidering the traditional concept of Y‐shaped white stomatal bands

The foliar morphological characters of hinoki (Chamaecyparis obtusa) were revisited using optical and scanning electron microscopy. In C. obtusa, typical Y‐shaped white stomatal bands were evident on the abaxial leaf surfaces. Two facial leaves and two lateral leaves were observed at the same node. Waxy papillae and oval stomata were arranged in two or three rows with protuberant rims on the abaxial leaf surfaces. Higher magnifications revealed the deposition of epicuticular waxes (tubules) on the Y‐shaped white stomatal bands. Given the absence of stomatal bands after dewaxing with organic solvents, the white stomatal bands in C. obtusa were related to the epicuticular waxes rather than the presence of aggregated stomata alone. In contrast to C. obtusa, a single median leaf and two lateral leaves were observed at the same node of oriental arborvitae (Platycladus koraiensis). Neither stomatal bands nor papillae were observed on P. koraiensis leaves. The stomatal density and epicuticular waxes in the stomatal regions of C. obtusa were higher than those of P. koraiensis. This study suggests that the traditional concept of Y‐shaped white stomatal bands in C. obtusa should be revised to describe the arrangement of the aggregated waxy stomata that occur in rows.     

Leaf epidermal characteristics of Cissampelos L. (Menispermaceae) species from Northeastern Brazil

The morphological similarities among the species of Cissampelos are remarkable and the difficult to distinguish them as well. This article presents a comparative anatomical study of the leaves of common Northeastern Brazilian species of Cissampelos, carried out using light and scanning electron microscopy. The leaf epidermal was studied to obtain data on epidermal characteristics and to evaluate their taxonomic significance. As results, some micromorphological characters on the leaf epidermal like the cuticular waxes, the presence of papillae in epidermis and nonglandular trichomes, the anticlinal walls epidermal cells, the distribution, density and type of trichomes, and also the type and distribution of epicuticular wax proved to be the most useful characteristics to distinguish the species in taxonomic studies. Microsc. Res. Tech., 2011. © 2010 Wiley‐Liss, Inc.     

Morphology and nanoindentation properties of mouthparts in Cyrtotrachelus longimanus (Coleoptera: curculionidae)

Microstructure and nanoindentation properties of the mouthparts for the skillful driller Cyrtotrachelus longimanus are presented. The composition and morphological examinations are made using light, fluorescent, scanning electron microscopy, and Energy Disperse Spectroscopy, respectively. Nanoindentation was carried out to measure the elastic modulus and the hardness of mouthparts. The hardness and modulus for “dry” samples is 0.202 ± 0.065 and 8.604 ± 0.838 GPa, respectively, and the values of “fresh” ones is 0.126 ± 0.0196 and 6.951 ± 0.065 GPa, respectively. These results are critical for analyze the drilling mechanism of the weevil and provide a biological template to inspire the biomimetic design.     

Light microscopy of Pakistani Berberis leaf cuticles and its taxonomic implications

Taxonomy of the genus Berberis is quite complex, due to overlapping morphological characters, making it very difficult to differentiate the species within the genus. In order to resolve this taxonomic complexity, the foliar anatomy of 10 Berberis L. species was carried out, for the first time from Pakistan, using light microscopy (LM). Significant variation in terms of epidermal cells shape, size, cell wall pattern, and stomata type was observed. B. baluchistanica has the largest epidermal cells, Adaxial: length = 45–(53.9 ± 3.6)–62.5 μm; and width = 22.5–(26.3 ± 1.3)–30 μm; Abaxial: length = 37.5–(43.25 ± 2.5)–50 μm; and width = 20–(22.6 ± 0.8)–25. The highest number of stomata was observed in B. glaucocarpa as 62 on the abaxial surface while the lowest number of stomata was recorded in B. baluchistanica as 8 on the adaxial surface. Of 10 investigated species, 6 possess anomocytic type stomata, while 2 species that is, B. aitchisonii and B. parkeriana have both anomocytic and anisocytic stomata while B. baluchistanica and B. calliobotrys have only paracytic type stomata. The highest number of cells per unit area was present on the adaxial surface of B. calliobotrys ranging from 245–(252.4)–260 followed by B. parkeriana with 209–(227.8)–250 on the abaxial surface. Stomatal index (SI) also varied considerably and was the lowest (2.6) percentage in B. baluchistanica and highest (31.9) percentage in B. kunawurensis. A taxonomic key based on micro‐morphological characters is provided for species identification.     

相似分析在结构仿生设计中的应用研究

指出了仿生相似分析内容和步骤,并对生物骨架与小型梁肋机翼结构进行了模糊相似评价,表明叶脉承力性能与小型梁肌机翼具有高的相似度;针对机翼结构设计要求,借鉴叶脉结构特征,设计了仿生型机翼结构.有限元分析表明,仿生型机翼具有更高的结构效能.     

AFM studied the effect of celastrol on β1 integrin‐mediated HUVEC adhesion and migration

Integrin‐mediated human umbilical vein endothelial cells (HUVECs) adhesion to the extracellular matrix plays a fundamental role in tumor‐induced angiogenesis. Celastrol, a traditional Chinese medicine plant, has possessed anticancer and suppressed angiogenesis activities. Here, the mechanism underling the antiangiogenesis capacity of celastrol was investigated by exploring the effect of celastrol on β1(CD29) integrin‐mediated cell adhesion and migration. Flow cytometry results showed that the HUVECs highly expressed CD29 and cell adhesion assay indicated that celastrol specifically inhibited the adhesion of HUVECs to fibronectin (FN) without affecting nonspecific adhesion to poly‐L‐lysine (PLL). After cell FN adhesion being inhibited, the cell surface nanoscale structure and adhesion force were detected by atomic force microscope (AFM). High‐resolution imaging revealed that cell morphology and ultrastructure changed a lot after being treated with celastrol. The membrane average roughness (Ra) and the major forces were decreased from 31.34 ± 4.56 nm, 519.60 ± 82.86 pN of 0 μg/ml celastrol to 18.47 ± 6.53 nm, 417.79 ± 53.35 pN of 4.0 μg/ml celastrol, 10.54 ± 2.85 nm, 258.95 ± 38.98 pN of 8.0 μg/ml celastrol, respectively. Accompanying with the decrease of adhesion force, the actin cytoskeleton in the cells was obviously disturbed by the celastrol. All of these changes influenced the migration of HUVECs from the wound‐healing migration assay. Taken together, our results suggest that celastrol can be as an inhibitor of HUVEC adhesion to FN. This work provides a novel approach to inhibition of tumor angiogenesis and tumor growth. SCANNING 35:316‐326, 2013. © 2012 Wiley Periodicals, Inc.     

Palyno-anatomical characters and their systematic significance in the family Apiaceae from Chitral,eastern Hindu Kush,Pakistan

The present study was performed to provide a detailed explanation of leaf epidermal anatomy and pollen micromorphological features of selected species of family Apiaceae from Chitral, eastern Hindu Kush region as the basis of forthcoming studies. In the present article pollen morphology of eight species and foliar epidermal of seven species of family Apiaceae have been examined through microscopic techniques. In results two types of pollen prolate (five species) and perprolate (three species) with three colpi have been recorded. The exine ornamentation was found to be regulate, striate, and cerebroid. Largest pollen was found in Heracleum leucocarpum with the polar diameter of 43.25 μm and equatorial diameter of 21.6 μm. Smallest pollen was observed in Elaeosticta chitralica with the polar diameter of 18.4 μm. The P/E ratio varied from 1.59 to 2.16. Regarding to foliar epidermal anatomy, three types of epidermal cells including rectangular, irregular, and polygonal with variation in anticlinal wall pattern were determined. In the selected species three kinds of stomata comprising anisocytic, anomocytic, and paracytic type were reported in the current research. The size of epidermal cells ranged from 106 × 42.50 μm in Bupleurum falcatum subsp. cernuum and 77.25 × 26.35 μm in Prangos pabularia in adaxial surface. Largest stomatal complex was found in Prangos pabularia both in adaxial 33.55 × 20.05 μm and abaxial 50.25 × 39.40 μm. All the observed quantitative and qualitative features of the species were proved to be useful in the delimitation of species at generic and species level.     

Determination of protoberberine alkaloids in Coptidis Rhizoma by novel extraction and high-performance liquid chromatography

An alternative, effective, and solvent-free microextraction method has been developed using weighing paper as the adsorbent for five protoberberine alkaloids from the methanolic extract of Coptidis Rhizoma (C. Rhizoma). Several variables influencing extraction efficiency were optimized. 1.0?×?1.0?cm glassine weighing paper was employed for the isolation of the analytes followed by high-performance liquid chromatography. Under the optimum conditions, the limits of detection were between 0.2 and 0.4?ng/mL with enrichment factors from 41–47. The calibration curves were linear across the concentration ranges examined with correlation coefficients exceeding 0.992. All relative standard deviations were less than 10.0%. The method was employed for the determination of the analytes in C. Rhizoma. Berberine, palmatine, coptisine, epiberberine, and jatrorrhizine were present at concentrations between 9.62?±?0.87?mg/g and 0.48?±?0.05?mg/g.     

Design and construction of a new temperature-controlled chamber for light and confocal microscopy under monitored conditions: biological application for plant samples

A new light microscope–temperature‐controlled chamber (LM–TCC) has been constructed. The special feature of the light microscope–temperature‐controlled chamber is the Peltier‐element temperature control of a specimen holder for biological samples, with a volume capacity of 1 mL. This system has marked advantages when compared to other approaches for temperature‐controlled microscopy. It works in a temperature range of −10°C to +95°C with an accuracy of ±0.1°C in the stationary phase. The light microscope–temperature‐controlled chamber allows rapid temperature shift rates. A maximum heating rate of 12.9°C min⁻¹ and a maximum cooling rate of 6.0°C min⁻¹ are achieved with minimized overshoots (≤1.9°C). This machinery operates at low cost and external coolants are not required. Especially with samples absorbing irradiation strongly, temperature control during microscopy is necessary to avoid overheating of samples. For example, leaf segments of Ficaria verna exposed to 4500 μmol photons m⁻² s⁻¹ in a standard microscopic preparation show a temperature increase (δT) of 18.0°C, whereas in the light microscope–temperature‐controlled chamber this is reduced to 4°C. The kinetics of microscope‐light induced δT are described and infrared thermography demonstrates the dissipation of the temperature. Chloroplasts of the cold adapted plant Ranunculus glacialis show the tendency to form stroma‐filled protrusions in relation to the exposure temperature. The relative number of chloroplasts with protrusions is reduced at 5°C when compared to 25°C. This effect is reversible. The new light microscope–temperature‐controlled chamber will be useful in a wide range of biological applications where a rapid change of temperature during microscopic observations is necessary or has to be avoided allowing a simulation of ecologically relevant temperature scenarios.     

Foliar micromorphology and its role in identification of the Apocynaceae taxa

In the present study, we evaluate the importance of foliar epidermal micromorphological characteristics of Apocyanaceae for accurate identification and classification. The species were collected from the University of Peshawar's main campus in the spring season to observe its qualitative and quantitative features. The length and width of guard cells, stomatal pore and subsidiary cells, trichomes, and crypts on both sides of the leaf were examined. Many species were observed to be hypostomatic. Plumeria rubra, Raulfia serpentine, Thevetia peruviana, Trachelospermum lucidum, Alstonia scholaris, and Catharanthus roseus demonstrated hypostomatic leaves. Nearly all the investigated species had anisocytic type of stomata only or in combination with other types of stomata on the upper and lower epidermis. Carissa carandas had anomocytic, anisocytic, and cyclocytic type of stomata on the upper epidermis, and the lower epidermis showed variations in stomatal type, such as anomocytic, stephanocytic, brachyparacytic, and hemiparacytic. Nerium oleander had no specific shape of stomata but showed stomatal crypts in which the stomata were enclosed inside many trichomes. The taxonomic key based on stomatal types, epidermal cells, stomatal index value, and statistical analysis, along with the variations in the epidermal cells, shows the link between the selected plants species, which will provide a baseline for future anatomical studies. This study highlights many undocumented micromorphological characteristics. The anatomical characteristics observed in this study will be helpful for taxonomic identification and species delimitation of the family Apocynaceae.