Comparative foliar anatomical and pollen morphological studies of Acanthaceae using light microscope and scanning electron microscope for effective microteaching in community.

In this study, foliar anatomy and pollen morphology of 10 species of Acanthaceae has been investigated using light and scanning electron microscopy. The study was aimed to highlight the role of microscopy in microteaching at community for proper characterization of plants using palyno-anatomical characters including pollen type, exine sculpturing, shape of epidermal cells, pattern of anticlinal wall, type and size of stomata, and trichome. Most of the species have polygonal cell shapes but some species have irregular, tetragonal, and pentagonal shape of epidermal cells. The largest epidermal cell length on adaxial and abaxial surface were observed in Asystasia gangetica 66.95 and 87.40 µm whereas least was observed on adaxial surface in Justicia adhatoda 36.9 µm and on abaxial surface in Barleria cristata 35.65 µm. In anatomy, species have diacytic type of stomata, whereas stomata of paracytic type observed in two species, while in A. gangetica cyclocytic type of stomata are present. Quantitively on abaxial surface, largest stomata length 29.9 µm and width 24.30 µm was noted in B. cristata. While shortest stomata length was observed in Ruellia prostrata 25.95 µm whereas minimum width of stomata was examined in Barleria acanthoides 2.05 µm. The diversity of trichomes are present in all species except in Ruellia brittoniana. Acanthaceae can be characterized by exhibiting different pollen morphology having five types of pollen shapes, prolate, spheroidal, perprolate, subprolate, and oblate spheroidal. Exine peculiarities showing variations such as reticulate, granulate, coarsely reticulate, lophoreticulate, perforate tectate, and granulate surface were examined.

Anatomical, Phytochemical, and Histochemical Study of Juniperus rigida Needles at Different Altitudes.

Needles of Juniperus rigida are used in Chinese traditional medicine for the treatment of brucellosis, dropsy, skin disease, and rheumatoid arthritis. This is the first study that reports anatomical structures of the J. rigida needles collected at different altitudes. The most common anatomical, phytochemical, and histochemical techniques and methods are used. The results show that anatomical structures and chemical composition change significantly at different altitudes. The main anatomical characters are significant xeromorphic structures (thick epidermis, hypodermis, and cuticle), a stomatal band, a developed vascular bundle, and a marginal resin duct. The xeromorphic structures become more pronounced with increasing altitude. The phytochemical and histochemical results demonstrate that the content of the main chemical compounds (phenols and terpenoids) basically increases at a higher elevation. Histochemical analysis localizes the phenols in epidermal cells, sponge tissue, endothelial layer cells, and stomatal bands, and the terpenoids in palisade tissue, sponge tissue, and the edge of the resin duct. This work reveals the relation between anatomy and chemistry in J. rigida needles, contributes to the quality control of its ethno-medicine, and provides the evidence to develop the commercial cultivation.

Significance of leaf morphoanatomical markers for the authentication of adultered drugs sold in herbal markets of district Lahore, Pakistan.

A number of herbal plants sold in herbal markets of Lahore are under adulteration threat which can pose harmful health effects to end-user. This adulteration problem of medicinal plants can be resolved by the implication of some valuable taxonomic parameter such as leaf epidermal anatomical characteristics. Hence, this research was aimed to provide viable anatomical markers in order to resolve this adulteration issue persisted in some common marketed medicinal plants of district Lahore, that is, Cinnamomum verum Presl., Cinnamomum tamala (Buuch.-Ham.) T.Nees&Eberm., Gymnema sylvestre (Retz.) R.Br.ex Sm., Sphaeranthus indicus Linn., Artemisia maritima Linn., Achillea millifolim L., Adhatoda vasica Nees, Butea monosperma (Lam.) Taub, and Morus nigra L. Overall multiple anatomical variations (epidermal cell shape, their length and width, type of stomata, length and width of guard cells along with presence or absence of trichomes) had been reported in the study that could be worthwhile for the correct identification of medicinal plants. Irregular shapes of epidermal cells were observed in Cinnamomum verum and Achillea millifolium while pentagonal and polygonal cells were found in their adultaerants, that is, Canella winterana and Adhatoda vasica, respectively. Types of stomata were also strikingly varied among genuine plant and its adulterant, for example, anisocytic stomata were observed in Artemisia maritima while in its adulterant (Artemisia absinthium) anomocytic stomata were found. Similarly, paracytic stomata were observed in Butea monosperma, whereas its adulterant plant (Averrhoa carambola) characteristically possessed anisocytic stomata. Hence, anatomical characteristics were proved to be a valuable taxonomic tool in resolving the adulteration issue of medicinal plants.

Metabolomic profiling delineate taste qualities of tea leaf pubescence.

The amount of pubescence on leaf epidermis is an important morphological marker for the quality of green tea, and the tea with plenty of pubescence is generally recognized as having a better taste. However, there is no systematic study on chemical compositions of tea leaf pubescence. The contributions of pubescence to taste properties are far from clear. In this research, 114 components were identified from the tea leaf pubescence of yunkang 10, a broad-leaf tea cultivar with plenty leaf pubescence, for the first time with a non-targeted metabolomics approach using ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UHPLC-Q-TOF-MS). Compared to the tea leaf with the pubescence removed (non-pubescent leaf), the pubescence obtained from the same shoots had relatively higher contents of amino acids and lower contents of polyphenols. It was also found that the umami of pubescence was elevated, while the bitterness and astringency were significantly declined. Partial least-squares (PLS) analysis suggested that the polyphenols and amino acids accounted for the taste quality. To the best of our knowledge, this is the first time that the metabolites in tea leaf pubescence were profiled. The results offer the direct concrete evidence on the contributions of pubescence to the tea taste properties.

Ultrastructure of the Epidermal Cell Wall and Cuticle of Tomato Fruit (Solanum lycopersicum L.) during Development.

The epidermis plays a pivotal role in plant development and interaction with the environment. However, it is still poorly understood, especially its outer epidermal wall: a singular wall covered by a cuticle. Changes in the cuticle and cell wall structures are important to fully understand their functions. In this work, an ultrastructure and immunocytochemical approach was taken to identify changes in the cuticle and the main components of the epidermal cell wall during tomato fruit development. A thin and uniform procuticle was already present before fruit set. During cell division, the inner side of the procuticle showed a globular structure with vesicle-like particles in the cell wall close to the cuticle. Transition between cell division and elongation was accompanied by a dramatic increase in cuticle thickness, which represented more than half of the outer epidermal wall, and the lamellate arrangement of the non-cutinized cell wall. Changes in this non-cutinized outer wall during development showed specific features not shared with other cell walls. The coordinated nature of the changes observed in the cuticle and the epidermal cell wall indicate a deep interaction between these two supramolecular structures. Hence, the cuticle should be interpreted within the context of the outer epidermal wall.

Anthocyanin synthesis in native and wound periderms of potato.

Skin color of red potatoes is due to accumulation of anthocyanins in the tuber periderm, a protective tissue that replaces the epidermis at an early stage of tuber development. The periderm consists of external layers of suberized phellem cells making up the skin, and internal layers of parenchyma-like phelloderm cells. Red pigmentation is an important marketing factor for red-skinned potatoes. However, injuries to the tuber surface, which are common in the potato industry, result in the development of a wound periderm that is devoid of the characteristic red coloration. To study the reason for these differences in anthocyanin accumulation, the expression level of anthocyanin biosynthesis genes and regulators was monitored in native and wound periderm using microarray analysis and quantitative polymerase chain reaction. We found significantly higher expression of the anthocyanin pathway in the phelloderm cells compared with the skin and tuber-flesh samples. However, in wound periderm, the anthocyanin pathway was strongly downregulated relative to the native periderm. This was true for two developmental stages of the native periderm--'immature', when the skin is prone to skinning injuries, and 'mature', following skin set--suggesting that anthocyanin synthesis continues postharvest. Wound-induced expression of steroidal glycoalkaloid glycosyltransferases, suberin-related 3-ketoacyl-CoA synthase and actin indicated that downregulation of the anthocyanin-specific pathway does not reflect global repression of the wound-periderm transcriptome. Loss of pigmentation may result from reduced expression of the Myb-bHLH-WD40 anthocyanin regulatory complex--a possible candidate might be the bHLH transcription factor JAF13.

[Pharmacognostical study of Atropa belladonna].

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.

The epidermal cell structure of the secondary pollen presenter in Vangueria infausta (Rubiaceae: Vanguerieae) suggests a functional association with protruding onci in pollen grains.

Secondary pollen presentation is a well-known phenomenon in the Rubiaceae with particularly conspicuous pollen presenters occurring in the tribe Vanguerieae. These knob-like structures are formed by a modification of the upper portion of the style and stigma, together known as the stylar head complex. In the flower bud and shortly before anthesis, the anthers surrounding the stylar head complex dehisce and release pollen grains which adhere to the pollen presenter. The epidermal cells of the pollen presenter facing the anthers are radially elongated with a characteristic wall thickening encircling the anticlinal walls of each cell towards the distal end. These cells were studied in the pollen presenter of Vangueria infausta using electron and light microscopy in conjunction with histochemical tests and immunohistochemical methods. Other prominent thickenings of the cell wall were also observed on the distal and proximal walls. All these thickenings were found to be rich in pectin and possibly xyloglucan. The terms "thickenings of Igersheim" and "bands of Igersheim" are proposed to refer, respectively, to these wall structures in general and those encircling the anticlinal walls of each cell near the distal end. The epidermal cells have an intricate ultrastructure with an abundance of organelles, including smooth and rough endoplasmic reticulum, Golgi apparatus, mitochondria and secretory vesicles. This indicates that these cells are likely to have an active physiological role. The pollen grains possess prominent protruding onci and observations were made on their structure and development. Walls of the protruding onci are also rich in pectin. Pectins are hydrophilic and known to be involved in the dehydration and rehydration of pollen grains. We hypothesise that the thickenings of Igersheim, as well as the protruding onci of the pollen grains, are functionally associated and part of the adaptive syndrome of secondary pollen presentation, at least in the Vanguerieae.

Leaf anatomy and histochemistry of three species of Ficus sect. Americanae supported by light and electron microscopy.

In this work the leaf anatomy of three species of Ficus section Americanae (Miq.) Miq. from Brazil, whose leaves and latex are used in folk medicine is reported. The work was carried out using light and scanning electron microscopy in order to characterize these species and to evaluate their taxonomic significance, and also contribute to the quality control of their ethnodrugs. The three species (Ficus cyclophylla, Ficus elliotiana, and Ficus caatingae) showed hypostomatic leaves, anomocytic stomata, straight epidermal cell outlines, and a dorsiventral mesophyll. Some micro-morphological characters such as density and distribution of epicuticular waxes, glandular trichomes, the length and width of stomata, as well as the palisade of mesophyll and petiole outlines proved to be the most useful and distinctive characters for the separation of species. These may contribute as additional support for the taxonomy of the section and for the quality control of their ethnodrugs.

[Comparative research on histological and microscopical leaves characters of five species in Chloranthus].

OBJECTIVE: To clarify the origin and provide pharmacognostical evidences for the leaves of 5 species in Chloranthus. METHOD: Histological observation and microscopic identification through different slice-making techniques were applied to the research. RESULT: There were subtle differences between the histological characteristics. In microscopical identification, the different structures of vascular bundles in veins were observed, appendages and non-glandular hairs were distinct. CONCLUSION: The method can be used to distinguish the features of 5 species in Chloranthus. This article offers information for the further research and exploitation of Chloranthus.

Comparative anatomy, morphology, and molecular phylogenetics of the African genus Satanocrater (Acanthaceae).

PREMISE OF THE STUDY: Anatomical and morphological features of Satanocrater were studied to test hypotheses of xeric adaptations in the genus, which is endemic to arid tropical Africa. These features, together with molecular data, were used to test the phylogenetic placement of Satanocrater within the large plant family Acanthaceae. METHODS: We undertook a comparative study of four species of Satanocrater. Carbon isotope ratios were generated to test a hypothesis of C(4) photosynthesis. Molecular data from chloroplast (trnG-trnS, trnG-trnR, psbA-trnH) and nuclear (Eif3E) loci were used to test the placement of Satanocrater within Acanthaceae. KEY RESULTS: Anatomical features reflecting xeric adaptations of species of Satanocrater included a thick-walled epidermis, thick cuticle, abundant trichomes and glandular scales, stomata overarched by subsidiary cells, tightly packed mesophyll cells, and well-developed palisade parenchyma on both leaf surfaces. Although two species had enlarged bundle sheath cells, a feature often implicated in C(4) photosynthesis, isotope ratios indicated all species of Satanocrater use the C(3) pathway. Molecular data resolved Satanocrater within tribe Ruellieae with strong support. Within Ruellieae, our data suggest that pollen morphology of Satanocrater may represent an intermediate stage in a transition series. CONCLUSIONS: Anatomical and morphological features of Satanocrater reflect adaptation to xeric environments and add new information about the biology of xerophytes. Morphological and molecular data place Satanocrater in the tribe Ruellieae with confidence. This study adds to our capacity to test hypotheses of broad evolutionary and ecological interest in a diverse and important family of flowering plants.

[Ecological adaptability of leaf epidermis of erosion-resistant plants in hilly-gully area of Loess Plateau, Northwest China].

By the temporary slide method of leaf epidermis, an observation was made on the morphological characteristics of the leaf epidermis of six erosion-resistant plant species in different soil erosion environments (gully, inter-gully, and inter-gully artificial Robinia pseudoacacia forest land) in hilly-gully area of Loess Plateau. Compared with those in the gully, the stomata aperture, stomata density, stomata index, stomata apparatus length/width plasticity, stomata apparatus area plasticity, epidermal hair density, and epidermal cell density of the leaf upper and lower epidermis of the plants in the inter-gully were 93.8% and 90.4%, 66.8% and 76.6%, 17.9% and 9.8%, 36.4% and 47.1%, 42.3% and 43.9%, 199.4% and 98.2%, and 46.5% and 50.1% higher, respectively; while in the inter-gully artificial R. pseudoacacia forest land, the same morphological indices of the leaf upper and lower epidermis of the plants were 66.7% and 106.7%, 20.5% and 45.8%, 11.9% and 11.9%, 37.9% and 41.3%, 19.8% and 21.2%, 113.1% and 52.2%, and 10.8% and 28.1% higher than those in the gully, respectively. The epidermal hair length and epidermal cell area of the leaf upper and lower epidermis of the plants in the inter-gully were 58.8% and 29.7%, and 40.3% and 37.0% lower than those in the gully, and the same morphological indices of the leaf upper and lower epidermis of the plants in the intergully artificial R. pseudoacacia forest land were respectively 25.0% and 23.6%, and 22.2% and 19.2% lower than those in the gully, respectively. The results suggested that the erosion-resistant plants in the study area were able to adapt to various soil erosion environments by increasing their leaf stomata aperture, stomata density, stomata index, stomata apparatus length/width plasticity, stomata apparatus area plasticity, epidermal hair density, and epidermal cell density, and by reducing their epidermal hair length and epidermal cell area.

Pectin dynamic and distribution of exchangeable Ca2+ in Haemanthus albiflos hollow style during pollen-pistil interactions.

In this report, the localization and spatial distribution of two categories of pectin, high and low methylesterified, on the background of dynamic in loosely bound calcium (Ca(2+)) in Haemanthus hollow style were studied before and after pollination. In the style transmitting tract of unpollinated pistil, mainly high-methylesterified pectins were present, both in the transmitting tract epidermis and in the style canal. After pollination, an increase in the level of two investigated categories of pectin was observed, but the amount of high-methylesterified one in each period of time analyzed was permanently higher. Locally, in the regions of the style canal penetrated by pollen tubes, process of pectin de-esterification was initiated. However, pollination caused an increase of loosely bound Ca(2+) level in the style transmitting tract, this process appears to be not linked with pectin de-esterification and possible Ca(2+) release after the lysis of Ca(2+) cross-linked de-esterified pectin. Instead, it seems to be based on Ca(2+) exocytosis from the transmitting tract epidermis cells providing a source of Ca(2+) for pollen tubes growing in Haemanthus hollow style.

Comparative functional genomic analysis of Solanum glandular trichome types.

Glandular trichomes play important roles in protecting plants from biotic attack by producing defensive compounds. We investigated the metabolic profiles and transcriptomes to characterize the differences between different glandular trichome types in several domesticated and wild Solanum species: Solanum lycopersicum (glandular trichome types 1, 6, and 7), Solanum habrochaites (types 1, 4, and 6), Solanum pennellii (types 4 and 6), Solanum arcanum (type 6), and Solanum pimpinellifolium (type 6). Substantial chemical differences in and between Solanum species and glandular trichome types are likely determined by the regulation of metabolism at several levels. Comparison of S. habrochaites type 1 and 4 glandular trichomes revealed few differences in chemical content or transcript abundance, leading to the conclusion that these two glandular trichome types are the same and differ perhaps only in stalk length. The observation that all of the other species examined here contain either type 1 or 4 trichomes (not both) supports the conclusion that these two trichome types are the same. Most differences in metabolites between type 1 and 4 glands on the one hand and type 6 glands on the other hand are quantitative but not qualitative. Several glandular trichome types express genes associated with photosynthesis and carbon fixation, indicating that some carbon destined for specialized metabolism is likely fixed within the trichome secretory cells. Finally, Solanum type 7 glandular trichomes do not appear to be involved in the biosynthesis and storage of specialized metabolites and thus likely serve another unknown function, perhaps as the site of the synthesis of protease inhibitors.

Adaptações fisiológicas e anatômicas de melissa officinalis L. (Lamiaceae) cultivadas sob malhas termorrefletoras em diferentes intensidades luminosas / Physiological and morphological adaptations of melissa officinalis L. (Lamiaceae) cultivated under thermo-reflector shading nets at different luminous intensities

Objetivou-se, com a realização da pesquisa, avaliar modificações fisiológicas e anatômicas em plantas de melissa, cultivadas sob malhas termorrefletoras (Aluminet®), em diferentes níveis de sombreamento, visando conhecer a plasticidade fenotípica em resposta de adaptação a diferentes quantidades de luz. Os tratamentos foram caracterizados por plantas submetidas a pleno sol e a 20 e 60 por cento de intensidade luminosa, e arranjados conforme o delineamento inteiramente casualizado (DIC). As quantificações de clorofila foram feitas em quatro repetições, as medições das epidermes e parênquimas foram repetidas 15 vezes e utilizou-se 10 repetições para as avaliações das características de cloroplastos e grãos de amido destes. Plantas submetidas a 20 por cento de intensidade luminosa apresentaram maior quantidade de clorofila a e, portanto, maior razão clorofila a/b. Comparativamente, as folhas de melissa a pleno sol e a 60 por cento de luz apresentaram células da epiderme adaxial mais espessas, mas as células da epiderme abaxial mostraram características encontradas em folhas de sombra, ou seja, mais finas. Quanto maior a intensidade luminosa, maior o número de cloroplastos, porém, a pleno sol mostraram-se mais finos e com menor área. Os grãos de amido de plantas cultivadas sob ambientes sombreados tiveram maior área e ocuparam maior parte nos cloroplastos de plantas a 60 por cento de intensidade luminosa. Assim, plantas de melissa, quando submetidas ao sombreamento, tiveram plasticidade fenotípica.

The aim of this study was to evaluate physiological and anatomical modifications in lemon balm plants, cultivated under thermo-reflector nets (Aluminet®) at different levels of shading, in order to understand the phenotypic plasticity in adaptation response to different light quantities. The treatments were characterized by plants subjected to full sun and 20 and 60 percent of luminous intensity, and arranged in completely randomized design (CRD). The quantifications of chlorophylls were done in four replicates, the measurements of epidermis and parenchymas were repeated 15 times and 10 replicates were used to evaluate characteristics of chloroplasts and their starch grains. Plants subjected to 20 percent of luminous intensity showed higher quantity of chlorophyll a and, therefore, higher chlorophyll a/b ratio. Lemon balm leaves under full sun and 60 percent of light showed thicker adaxial epidermis cells, but the abaxial epidermis cells showed characteristics found in shaded leaves, i.e., they were slender. The higher the light intensity, the larger the number of chloroplasts; however, under full sun, they were slender and had smaller area. The starch grains of leaves grown under shaded environments showed larger area and, at 60 percent of luminous intensity, occupied the largest part of chloroplasts. Thus, lemon balm plants, subjected to shading conditions, showed phenotypic plasticity.

Inner epidermis of onion bulb scale: As natural support for immobilization of glucose oxidase and its application in dissolved oxygen based biosensor.

Inner epidermal membrane of the onion bulb scales was studied as a natural polymer support for immobilization of the glucose oxidase (GOD) enzyme for biosensor application. Onion epidermal membrane was used for immobilization of glucose oxidase and was associated with dissolved oxygen (DO) probe for biosensor reading. Glucose was detected on the basis of depletion of oxygen, when immobilized GOD oxidizes glucose into gluconolactone. A wide detection range between 22.5 and 450 mg/dl was estimated from calibration plot. A single membrane was reused for 127 reactions with retention of approximately 90% of its initial enzyme activity. Membrane was stable for 45 days ( approximately 90% activity) when stored in buffer at 4 degrees C. Surface structure studies of the immobilized membranes were carried under SEM. To our knowledge, this is the first report on employing inner epidermal membrane of onion bulb scales as the solid support for immobilization of enzyme.

[Pharmacognostic identification of dried twigs and leaves of Taraphochlamys affinis (Giff) Bremekhu].

The pharmacognosy of Taraphochlamys affinis (Giff) Bremekhu was studied by microscopic observation to provide a scientific basis for the identification, development and utilization of its resources. As a result, obvious characteristics for its identification were revealed, which could be used to identify twigs and leaves of Taraphochlamys affinis (Giff) Bremekhu.

[Quantity study of cultivated and wild Rhizoma Polygonati Odorati].

METHODS: In this article, cultivated and wild Rhizoma Polygonati Odorati have been compared in macroscopical identificaation, microscopical identification, contents of odoratan of Rhizoma Polygonati Odorati. RESULTS: Cultivated and wild Rhizoma Polygonati Odorati are different in macroscopical identification, microscopical identification. Contents of odoratan of Rhizoma Polygonati Odorati: wild Rhizoma Polygonati Odorati 9.67%, cultivated Rhizoma Polygonati Odorait 8.05%, wild > cultivated.

[Character and microscopical identification of Xianglingcao (Crotalaria ferruginea Grah.), the herbal of Bai Nationality].

From its sources, shapes, microscopic characteristics of Xianglingcao (Crotalaria ferruginea Grah.), the herbal of Bai Nationality was studied. These studies provide reference formation for further development and identification of this crude drug.

[Morphological and microscopical identification of Murraya kwangsiensis].

OBJECTIVE: To identify medical plant Murraya kwangsiensis by its tissue characters. METHOD: Morphological characters and microstructure of this plant were described and illustrated. RESULT: Oil cell, fiber cell and calcium oxalate prism were found in its tissue. CONCLUSION: Tissue characters can be taken into account of developing basis for quality standard of this medical plant.