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Vanilla bean is the second most expensive spice in the world. While widely cultivated for centuries due to its importance for industry and gastronomy, its dispersers are unknown. In the present study, I investigated the system of seed dispersal of Neotropical vanillas based on experimental observations, the morpho-anatomy of fruits and seeds, and the effect of dormancy breakdown on seed germination. My results show the central cavity of the fruit contains copious amounts of seed with sclerified coats that are consumed by birds. The combination of gradually opening dehiscent fruits and a well-developed mesocarp rich in raphide idioblasts reinforces the idea that seed dispersal in Vanilla is more adapted to non-chewing animals. The digestive acids of birds sclerify the hard seed coats, breaking dormancy and promoting germination at the beginning of the wet season, which is fundamental for seedling survival and for the establishment of plants in the early stages of development during the rainy season. Zoochory in Vanilla is show for the first time. The chemical scarification of the seed coat is crucial for synchronizing the biological processes involved in seed germination. My evidence on endozoochory and the processes involved in seed germination of Neotropical vanillas provides new insights into understanding of the early evolution of seed dispersal in orchids.

Objective: This study was undertaken to carry out genetic barcoding, pharmacognostical and phytochemical studies on leaves of Pothos scandens L. belonging to the family Araceae. Methods: P. scandens, was collected from Cochin, Kerala, during the month of August 2019. Its leaves were examined for pharmacognostical characters and physicochemical, qualitative analysis and HPTLC studies was carried out following standard protocols recommended by the Ayurvedic Pharmacopoeia of India. Genetic barcoding of the fresh tender leaves samples was carried out through following standard methods. Results: Leaves are simple, alternate, broadly winged petiole. Transverse section through winged petiole portion showed U-shaped, epidermal cell along with cluster crystals. Transverse section through midrib boat-shaped structure, mesophyll undifferentiated, consist parenchyma cell, epidermis with thick cuticle along with waxy coating. Leaves powder shows green in color and microscopy reveals the presence of prismatic crystals, raphides, fibres, acicular crystals and monocot type 1 stomata. The plant is identified as Pothos scandens based on sequence homology and phylogenic analysis. Qualitative results show the presence of carbohydrates, protein, alkaloids and tannins. HPTLC profile shows 7 spots at both short UV and long UV and ferulic acid and kaempferol were identified through quantitative HPTLC. Conclusion: Pothos scandens is a climber with simple, alternate leaves. Acicular crystals and raphides, mucilaginous cavities, undifferentiated mesophyll are key characters of the identification of its leaf. DNA Barcoding results helps in correct identification of Pothos scandens. The results obtained from the phytochemical study will be helpful in the determination of the strength of purity and also for further standardization of the plant.

An attempt for study of two terrestrial orchids i.e. Eulophia nuda (Lindl.) and Geodorum densiflorum (Lam.) Schltr. were carried out. Main focus was on the corms of both the orchids due to its huge medicinal properties like anticancerous, antioxidant, antidiabetic, antimicrobial, phytotoxic, etc. This is one of the prime reasons behind their threatened status along with endemic habitat. The parameters which were analysed during the study include macro- and micro-morphology, preliminary phytochemistry, and antioxidant activity (DPPH activity). In macro-morphology of both the orchids, slight difference was reported and, thus sometimes Geodorum densiflorum is being used as an adulterant in place of Eulophia nuda. During micro-morphological analysis, the oxalate crystals, raphides, mucilage ducts, starch grains, and oil granules were observed from cortical region of corm and in the mesophyll tissue of leaves in both the orchids. Stomatal Index of leaf surface was calculated along with the type of stomata, i.e. Cyclic II type. Preliminary analysis of phytochemical constituents in both the orchids was performed with three different solvent systems. A comparative antioxidant activity between leaves extract of both the orchids was carried out.

The research aims to observe the influence of harvesting time on the change of calcium oxalate (CaOx) content and crystal density in Porang corms. The corms were harvested at different times, i.e., (1) two weeks before the plants shed (R0-1), (2) when the plants shed (R0), and (3) two weeks after the plants shed (R0+1). CaOx was obtained using the modified extracting method. Microscopic observations were obtained from the slices of the edge and center part of porang corms. Parameter observed including CaOx content, corm weight, shape, and density of CaOx crystal. CaOx content and crystal density in corms were analyzed using One way ANOVA. If the results are significant, it will be followed by Tukey Test α 0.05. In the meantime, the relation between CaOx content and corm weight was analyzed using Correlation Test Bivariate. The results showed that CaOx content was relatively higher in porang corms, i.e., 15.98 ± 0.60g/100g. On the other hand, the increasing of CaOx content might improve corm weight. The total density of druse, styloid, and prism crystal was pretty high in corms obtained when the plants shed compared to another harvest time, i.e., 1,494 ± 286; 31,280 ± 17,406 and 6,256 ± 1,533 crystals/cm2 . Raphide crystal density, by contrast, increased in corms obtained after the plants shed, i.e.,1,656 ± 368 crystals/cm2 . Total CaOx crystal density in the edge parts of corms harvested when the plants shed was proportionately higher than in the other harvest times, i.e., 12,292 ± 4,687.89 crystals/cm2 . In contrast, CaOx crystal densities in the center parts of corms were not much different at three harvesting times. The density of druse and prism crystals was somewhat higher in the center part of corms than in the edge parts. In opposition to, the density of raphide and styloid crystals was fairly higher in the edge part of corms than it was in the center parts. However, only raphide crystal density found in the edge and center part of corms was significantly affected by harvest time from all these results

Graphic abstract Unprocessed dried tuberous root of Pinellia ternata (Pinellia Tuber) has been used as a component of traditional Japanese Kampo medicine, while this crude drug is usually used after processing with Ginger in traditional Chinese medicine (TCM). It is known that the raphides contained in unprocessed Pinellia Tuber can induce severe acrid irritation of the oral and laryngopharynx mucosa when it is boiled insufficiently. However, the mechanism of the inducing irritation by the raphides nor that of the detoxification by heat or processing with Ginger have been left unclear, mainly because of the difficulties in the extraction and the purification of the raphides and in the assays of acridity. In this study, we developed an easy protocol that can be used to collect raphides from unprocessed Pinellia Tuber and an assay protocol that can be used to evaluate the acridity of the raphides in vitro. The raphides of Pinellia Tuber were discovered to have a lipophilic character and to be collected easily by the extraction using petroleum ether. It was also found that the denaturation of the raphides could be assayed by the dispersity of them in petroleum ether layer of the water/petroleum ether partition, and the acridity of the raphides was found to be in correlation with the assayed denaturation. The raphides were denatured by heat, methanol, or Ginger extract and the denaturing activity of Ginger on raphides was found to be attributable to its lipophilic and thermostable components, which may explain the meaning of the processing of Pinellia Tuber with Ginger in TCM, and may lead to the development of an easier and safer protocol to administer Pinellia Tuber. In addition, it was found that rinsing the mouth with salad oil can effectively relieve irritation of the oral mucosa caused by unprocessed Pinellia Tuber, probably due to the lipophilicity of the raphides. Electronic supplementary material The online version of this article (10.1007/s11418-020-01425-6) contains supplementary material, which is available to authorized users.

In the study, Puschkinia scilloides Adams, P. bilgineri Yıldırım, and P. peshmenii Rix et B. Mathew were compared anatomically. Although the stem vascular bundles were arranged in two rows in P. peshmenii, they can observed in three rows in P. scilloides and P. bilgineri. In addition, some differences were found, such as the metaxylem number in the root, number of vascular bundles in the scape, presence of raphide crystals, and number of protruding epidermis cells at the tip of the leaf. These differences have been found to be taxonomically useful for distinguishing these species. Moreover, similarities and differences between Puschkinia Adams and the taxonomically closely related Scilla L. section Chionodoxa Boiss. and Scilla bifolia L. taxa were also discussed.

The calcium oxalate content from two Amazonian amilaceous roots, Xanthosoma sagittifolium and Pachyrhizus tuberosus, before and after cooking were studied in the present work. Calcium oxalate raphides were observed by light microscopy in longitudinal sections from fresh tubers. The functional properties and physicochemical characteristics of isolated starches from both species were also investigated. Starches granules were observed with scanning electronic microscopy and presented a typical oval-circular shape, smooth surfaces and size ranging from 3.6 μm to 14 μm. Lower gel clarity, slightly high water-binding capacity, A-type starch structure and low values of crystallinity degree were determined in the isolated starches. High intensity bands related to vibrations of glucosidic bonds of polysaccharides were observed by infrared spectroscopy in both starches. Thermal degradation behavior data of the isolated starches reveled high initial decomposition temperatures. Thus, the studied cooked tubers presented low levels of calcium oxalates and are safe to be consumed as a boiled vegetable. Their isolated starches possess good functional characteristics to be applied as an ingredient to products in food industry.

International audience; In the context of a symbiotic plant-fungus interaction study concerning Cattleya purpurata, we focused on some aspects of seed morphology and biology, and the early stages of seedling development. Seed morphology was characterized using light and scanning electron microscopy. In vitro seed germination capability was evaluated, comparing symbiotic and asymbiotic methods. The morphology of the seeds was overall comparable to that of other congeneric species, showing classical adaptations related to the aerodynamic properties and to the wettability of seeds, but calcium oxalate druses were identified inside the suspensor cells. Asymbiotic seed germination was successful in all tested media (17.1–46.5%) but was higher on 1/2 Murashige & Skoog. During symbiotic interaction with the fungal strain MUT4178 (Tulasnella calospora), germination rate was significantly lower than that obtained with the best three asymbiotic media, suggesting a low fungal compatibility. Seedling morphology was in line with other taxa from the same genus, showing typical characteristics of epiphytic species. Our observations, in particular, highlighted the presence of stomata with C-shaped guard cells in the leaves, rarely found in Cattleyas (where usually they are reniform), and confirm the presence of tilosomes in the roots. Idioblasts containing raphides were observed in both roots and leaves.

Anatomical attributes are important for taxonomic studies of plants. This study investigated foliar and petiole anatomy of some members of the genera Colocasia and Xanthosoma. Similar and diagnostic characters critical for the taxonomy of the two genera were identified. The similar characters include, polygonal epidermal cell shape, straight adaxial anticlinal wall pattern, brachyparacytic stomata type, elliptic shaped stomata and unmodified raphide type. The presence of papillae on the adaxial surfaces of the members of genus Colocasia but not in the Xanthosoma taxa; lamellar collenchyma type in Xanthosoma mafaffa (Red), and unicellular non-glandular trichomes in Xanthosoma mafaffa (White) were recorded as diagnostic characters. Key words: Brachyparacytic stomata, collenchyma, druses, foliar anatomy, petiole anatomy, papillae, raphides, trichome.

Summary Seasonal changes in the content of Ca and Mg in leaf blades collected from mature kiwifruit [Actinidia deliciosa (A. Chev.) C. F. Liang et A. R. Ferguson var. deliciosa] vines in a high-producing orchard were measured by a sequential fractionation procedure (14 M acetic acid, 0.25 M HC1 and residual). Total Ca and Mg concentrations decreased within the first 4 weeks of growth, but increased linearly thereafter to be 1322 and 197 μmol g−1 dry weight, respectively, by leaf fall. Total quantities of Ca and Mg accumulated at this time were approximately 2500 and 380 μmol per blade. Calcium oxalate, dissolved in the HC1 extract, was the predominant fraction at every harvest, accounting for up to 79 % of the total Ca content shortly after leaf emergence, and decreasing to 42 % at leaf fall. The quantity of physiologically active Ca (acetic acid-soluble) ranged from a minimum of 15 % (8 weeks after emergence) to a maximum of 41 % at the end of the season. Forms of Ca remaining in the solid residue after extraction made only a minor contribution (3 to 21 %) to the total content at any time during the season. In contrast, however, 89 to 96% of the total Mg was extracted by acetic acid at every harvest, with the remainder dissolved by HC1. Scanning electron microscopy and X-ray microanalysis of the lamina of mature leaves indicated the presence of two crystalline, Ca-rich products which differed in their morphology, location and distribution. Large styloids (120 to 270 μm in length) were concentrated in idioblasts adjacent to vascular tissue, while bundles of four-sided raphides, containing wedge-shaped apices, were located predominantly between the major transport vessels of the vascular tissue. X-ray diffraction analysis of crystalline isolates confirmed that both morphological forms are calcium oxalate monohydrate (whewellite).

Passiflora cincinnata is a species of wild passion fruit whose fruit has potential for use by the food industry. In this context, an anatomical study of the pericarp may help in understanding the changes which occur during the postharvest period. The objective of this work was to anatomically characterize the P. cincinnata pericarp, focusing on the structural changes which occur during refrigerated storage. The selected fruits were kept in net-like packaging at 5°C and evaluated every 15 days for a period of 90 days. Pericarp samples were hand cut on these dates for structural description and to detect starch, phenolic compounds, alkaloids, polysaccharides, proteins, lipids and lignin through histochemical tests. Other samples were analyzed by scanning electron microscope. The fruit is composed of three anatomically distinct regions: exocarp, mesocarp and endocarp. The mesocarp stands out for the presence of sclerified cells and the lax parenchyma, which increased its number of layers during the evaluated period. Idioblasts with prismatic crystals, drusen, silica crystals and raphides were identified, but the amounts reduced over time. This reduction and enlargement of the lax parenchyma were the main changes observed in the postharvest period of P. cincinnata pericarps.

While this book has introduced the most abundant and potentially informative microparticles that will be found in archaeological contexts, there are a few that have not yet been discussed. This chapter includes brief descriptions and pictures of some of the other microparticles that the reader may encounter.

Rindyastuti R, Nurfadilah S, Rahadiantoro A, Hapsari L, Abywijaya IK. 2018. Leaf anatomical characters of four epiphytic orchids of Sempu Island, East Java, Indonesia: The importance in identification and ecological adaptation. Biodiversitas 19: 1906-1918. Leaf anatomy features are important characters to support species identification and classification, and they are related to ecological adaptation of species. The aims of the present study were: (i) to investigate leaf anatomical characters of four epiphytic orchids of Sempu Island (Ascochilus emarginatus, Dendrobium subulatum, Thrixspermum subulatum, and Thrixspermum acuminatissimum) in relation to the significance in species identification and ecological adaptation in coastal habitats of Sempu Island, (ii) to compare the adaptive ability of the four species in coastal habitats based on adaptive anatomical characters. The procedure of leaf anatomical studies as follows: orchid leaves were fixed in ethanol 70% and sliced into thin pieces with a microtome, and stained with 1% Safranin. The leaf anatomical organization of orchids (stomata, epidermis, mesophyll, vascular bundles, and other characters such as hypodermis, fibre bundles, raphide bundles, and spiral thickenings) was observed under light microscope. The results showed the comparable data of leaf anatomical characters among the orchids. There was distinct variation in the anatomical characters of the orchids including stomata anomocytic, tetracytic, and cyclocytic; the presence or absence of hypodermis, spiral thickenings, fibre bundles, raphide bundles, and bundle sheaths; homogenous and heterogenous mesophyll; and variation in vascular bundle arrangement. Detailed leaf anatomical characters can be used to distinguish a species from others, which are important to support species identification. The similarity of anatomical characters among these orchids were they possessed relatively thick cuticle and other specific anatomical characters as a structural adaptation to coastal habitat with high irradiation to reduce leaf transpiration. D. subulatum can be considered as the most adaptive orchid species to coastal habitats based on adaptive anatomical characters as it possessed the largest number of adaptive anatomical characters. The implication of this study is the importance of leaf anatomical features to support species identification and to increase understanding of orchid biology and ecology which are important in orchid conservation.

In this study, anatomical properties of Biarum marmarisense (P. C. Boyce) P. C. Boyce and Biarum pyrami (Schott) Engl. var. pyrami were examined. The specimens were collected from Antalya province, South West of Anatolia. Degree of spathe tube margin connection, distribution of pistillodes and staminate-pistillate flower zone interstice length characters are important in Biarum Schott. taxonomy at specific level. The cross-sections of root, stem and leaf parts of the plant were examined and demonstrated by photographs. Although thicknesses of the cell wall of the endodermis and xylem strands are distinguishing features, raphide crystals are same feature in both species. B. marmarisense has observed paracytic stoma types while B. pyrami var. pyrami has anomocytic stoma types. Most of the anatomical properties were similar to the other members of the Araceae family.

Piper caldense C. DC., Piperaceae, commonly known as “pimenta-d’água”, “pimenta-darda” or “paguarandy” in Brazil, is a shrub that grows mainly in humid and shaded habitats. The present study investigates the anatomy of the leaves and stems of P. caldense by light and scanning electron microscopy in order to provide supporting data for correct identification of the species. The leaves are hypostomatic, have a 2-layered hypodermis, and posses pearl glands. The midrib shows a ‘U’-shaped stele comprised of about ten collateral vascular bundles. The main anatomical marker of the stem is the presence of a continuous sclerenchymatous sheath in the pith. Two forms of calcium oxalate crystals, namely crystal sand and raphides, are observed in this species. Keywords: Anatomy, Pimenta-d’água, Piperaceae, Light and scanning electron microscopy

Crepidium is a large genus of mainly pantropical orchids. The lips of its flowers are upwardly directed and do not serve as landing platforms for pollinators. This role is assumed by the dorsal sepal and/or gynostemium. Information about the pollination and floral morphology of this genus is scarce. To date, no papers have been published on these topics. Field observations have revealed that the flowers are visited by small flies, midges, fruit flies, other small dipterans, ants, spiders, and mites. Preliminary observations revealed at least two forms of small liquid droplets secreted on the lip surface of Crepidium species: simple secretions from epidermal cells, and cell sap released upon the rupturing of raphide-producing cells. Further research revealed that this was the first time liquid secretion was recorded in this genus. Floral secretions were subjected to sequential organic solvent extraction and gas chromatography–mass spectrometry (GC–MS). Floral parts were investigated by means of scanning (SEM) and transmission electron microscopy (TEM), and histochemical tests. The presence of liquid droplets on the lip of Crepidium, the presence of a food reward, and the sequence of raphide development are reported here for the first time.

Micromorphological studies revealed the developmental changes in micropropagated plants of Morinda citrifolia L. from in vitro conditions to field environment. The lower relative humidity, higher light intensity and septic stressful conditions in the field environment could make gradual changes in the micropropagated plants so as to adapt the external environment. Arrested stomatal development, single guard cells and indistinct subsidiaries were observed in vitro with highest stomatal density (52.0±0.11) obtained in the ambient in vitro environment. The development of tissues, epidermal ornamentation, efficient stomatal functionality and vein-islets numbers (12.0±0.0) during in vivo transfer may help in acclimation of micropropagated plants under field conditions. Raphides were observed in the in vitro propagated as well as in vivo transferred plantlets. The gradual acclimatisation and ex vitro rooting technique increased the survival rate of plantlets in the field. The micromorphological changes resultin...

This study provides detailed information of the anatomical attributes of the epidermis and the three regions of the petiole of four members of the genus Dieffenbachia Schott. Fresh samples of the leaves of Dieffenbachia picta Schott, Dieffenbachia oerstedii Schott, Dieffenbachia senguine (Jacq) Schott and Dieffenbachia senguine cultivar ‘Candida’ Schott were used. Epidermal peels and transverse sections were made following standard procedures. Generic characters revealed uniform epidermal cell shape on the abaxial surface, wavy to undulating and straight to wavy anticlinal wall patterns on the adaxial and abaxial surfaces respectively, brachyparacytic stomata types, round abaxial petiole outline, the presence of raphides and druses in the petioles of all the taxa. Delimiting features include irregular epidermal cell shape on the adaxial surfaces of D . senguine and D . senguine cv. ‘Candida’, additional anomocytic stomata types on the abaxial surfaces of D . picta , D . oerstedii and adaxial surface of D . senguine , cuticular striations on the abaxial surfaces of D . oerstedii and D . senguine cv. ‘Candida’, druses and raphide bundle on the epidermal surface of D . senguine only, flat adaxial petiole outline and slightly concave adaxial petiole outline in the proximal and median regions of D . senguine cv. ‘Candida’ and the presence of lamellar collenchyma cells in the petiole of D . picta . Data for both quantitative and qualitative characters were subjected to Principal Components Analysis and Single Linkage Cluster Analysis. Interestingly, anomocytic stomata complex, cuticular striations, raphide bundles and druses and the adaxial petiole outline separated D. senguine and D. senguine cv. ‘Candida’.

In Gachantivá, Colombia, Rubus alutaceus has been used for commercial purposes for more than one decade but so far with unknown taxonomic identity. For its identification, the anatomical and morphological characteristics of its flowers, fruits, leaves, stems, and seeds, as well as the physicochemical characteristics of its fruits such as color, equatorial and polar diameter, fresh weight, pH and total soluble solids (TSS), were studied. Qualitative and quantitative morphological parameters were evaluated, and longitudinal and transverse cuts were made for histological studies. The plants were found to have trifoliate and pentafoliate leaves; whitish–yellow subglobose polydrupes, fully mature fruits with 8.8°Brix TSS; elliptical, subglobose seeds with ruminated endocarps; anatomical calcium oxalate crystals in the form of druses and raphides in the leaves and branches; and abundant hairs, simple and glandular (colleters) addition to vascular tissue with a more or less continuous appearance. High value in TSS, positively influences fruit agribusiness and the presence of simple hair and colleters can reduce the attack of pests, these variables identify the new species and give it agronomic potential to strengthen its cultivation and marketing. Resumo No município de Gachantivá, Colômbia, Rubus alutaceus é usada com fins comerciais, ha mais de uma década, sem que até o momento se conheça a sua identidade taxonômica. Com o propósito de identifica-la, estudaram–se as características anatômicas e morfológicas das suas flores, frutos, folhas, caules, sementes; características físico-químicas dos frutos, sua cor, segundo a escala CIELAB, o diâmetro equatorial e polar, peso fresco, pH e sólidos solúveis totais (SST). Avaliaram-se as variáveis morfológicas qualitativas e quantitativas, e realizaram-se cortes longitudinais e transversais com o objetivo de fazer estudos histológicos. Encontraram-se plantas que apresentam folhas trifoliadas e pentafoliadas; polidrupas subglobosas de cor amarela–esbranquiçada, SST de 8,8 °Brix em frutos completamente maduros; sementes elípticas, subglobosas com endocarpo ruminado; anatomicamente apresentam-se cristais de oxalato cálcico em forma de drusas e ráfides nas folhas e ramos e abundantes pelos simples e coléteres, além do tecido vascular de aspecto mais ou menos contínuo. O alto teor em TSS influencia positivamente o agronegócio dos frutos e a presença de estruturas secretoras incluindo tricomas e coléteres simples podem reduzir o ataque de pragas; essas variáveis caracterizam a nova espécie e conferem potencial agronômico para fortalecer seu cultivo e sua comercialização.

Among other taxa of Araceae, the genus Anthurium is the largest of the family and one of the most important taxa. Some species of this genus have been used as a condiment, and its plants as ornaments and medicines for over a century. Anthurium maricense is an endemic species from “restingas” of Rio de Janeiro, Brazil, and it is used as ornamental and medicinal plant. This paper aimed an anatomical and toxicological study of its vegetative organs. A. maricense has subterraneous rhizome, adventitious roots and complete leaves. The anatomical features of vegetative organs are similar to other species of Araceae and genus Anthurium, especially the velamen in roots and calcium oxalate crystals in all organs. Under scanning electron microscopy (SEM), longitudinal grooves were observed in the raphides. For toxicological analyses, whole (with crystals) and centrifuged (free from crystals) fractions of the juices of each organ were administered via mouthwash and plantar inoculation to groups of five mice. The formation of edema was observed at 1, 3, 6, 24 and 96 hours and 7, 14, 21 and 28 days. The oral inoculations presented negative results for all groups, without edema. The plantar inoculations showed differing results: with centrifuged juices, mild edema formed but regressed; with whole juices, some animals presented severe edema. Our results indicated distribution and morphology of raphides and diverse chemical substances were related to the edematogenic process. A. maricense may potentially induce more chronic edema if compared to other species of Araceae. Keywords : Anatomy. Crystals. Restinga. Toxicity.

Introducción y Objetivos: Heterophyllaea pustulata (Rubiaceae) crece en el noroeste argentino. Tiene importancia ganadera debido a que produce fotosensibilización en el ganado que la consume. Es por ello que se plantea estudiar la morfoanatomía foliar y caulinar de la especie con el objeto de determinar caracteres estructurales que faciliten la identificación de la planta. M&M: Se tomaron y estudiaron hojas y tallos de plantas adultas jóvenes y plántulas obtenidas a través de cultivos in vitro, mediante técnicas histológicas convencionales. Se presentan imágenes obtenidas con microscopio óptico y electrónico de barrido. Resultados: Los resultados indican que la especie estudiada comparte caracteres morfológicos y anatómicos con otras especies de Rubiaceae, aunque presenta otras que no fueron descritas anteriormente, tales como la epidermis con cutícula estriada y la anatomia del tallo. Conclusiones: Los caracteres diagnósticos que facilitan la identificación de la especie son: margen microdenticulado de la lámina, epidermis con cutícula estriada en hojas y tallos jóvenes, tricomas simples eglandulares, mesófilo con rafidios y drusas, además de las cavidades lisígenas en todos los órganos de la planta. Background and aims: Heterophyllaea pustulata (Rubiaceae) grows in northwestern Argentina. It has livestock importance because it produces photosensitization in the cattle that consume it. That is why it is proposed to study the foliar and caulinar morpho-anatomy of the species in order to determine structural characters that facilitate the identification of the plant.M&M: Leaves and stems of young adult plants and seedlings obtained through in vitrocultures were taken and studied by conventional histological techniques. Images obtained with optical and scanning electron microscopy are presented.Results: The results indicate that the studied species have morphological and anatomical characters similar to other species of Rubiaceae, although it presents others that were not previously described, such as the structure of the epidermis with striated cuticle and the anatomy of the stem.Conclusions: It is found that the diagnostic characters that facilitate the identification of the plant are: microdenticulated margin of the lamina, epidermis with striated cuticle in leaves and young stems, simple eglandular trichomes, mesophyll with raphides and druses, in addition to the lysigenous cavities in all the organs of the plant. Instituto de Investigación Animal del Chaco Semiárido Fil: Gallardo, Silvana C. Universidad Nacional de Salta. Facultad de Ciencias Naturales; Argentina Fil: Alemán, María M. Universidad Nacional de Salta. Facultad de Ciencias Naturales; Argentina Fil: Avilés, Zulma J. Universidad Nacional de Salta. Facultad de Ciencias Naturales; Argentina Fil: Colque Caro, Luis Adrián. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Investigación Animal Chaco Semiarido. Área de Investigación en Salud Animal; Argentina. Universidad Católica de Salta. Facultad de Ciencias Agrarias y Veterinarias; Argentina Fil: Micheloud, Juan Francisco. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Investigación Animal Chaco Semiarido. Grupo de Patología, Epidemiología e Investigación Diagnóstica-Área de Sanidad Animal; Argentina. Universidad Católica de Salta. Facultad de Ciencias Agrarias y Veterinarias; Argentina Fil: Martínez, Olga Gladys. Universidad Nacional de Salta. Facultad de Ciencias Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Bio y Geociencias del NOA; Argentina

Crystals in selected medical plants growing naturally in Place palayam reserve forest, Thiruvallur District were studied with Polarised light microscope. Four types of crystals were observed: druses, prismatics, raphide and rosettes crystal. Druses more in cortical parenchyma and ground parenchyma cells in the stem & petiole region. In contrast prismatic, raphide and rosettes present in cortical, ground, axial and phloem parenchyma in the roots and stems. The Preliminary results show the presence of four types of calcium oxalate crystals in the stems, Petiole & roots of 10 Species: Achyranthes aspera, Aerva lanata, Atalantia monophylla, Bridelia Crenulata presence of Druses crystal, Plumeria rubra, Adhatoda zeylanica presence of prismatic crystal, Asparagus racemosus, Monochoria vaginalis presence of raphide crystal and Carica papaya and Carissa spinarum presence of rosette type of crystal. These observations indicate that there is relationship between the distribution of calcium oxalate crystals and the medicinal parts of the plant and supports the view that the presence of calcium oxalate crystals may relate to the adulterants.

WOS: 000393617600006, Amorphophallus titanum (Becc.) Becc. ex Arcang., the titan arum, is popularly known for having the world's largest unbranched inflorescence. The enormous flower-like inflorescence is also called 'corpse flower' or 'carrion flower' due to the characteristic putrid odor of the bloom. The present study illustrates detailed anatomy and micromorphology of the inflorescence by light and scanning electron microscopy. Collenchyma was found to be the major tissue providing mechanical support in the spathe. The appendix consisted of a dense parenchymatous cortex enclosing a hollow center traversed by loose networks of aerenchyma. Anthocyanin pigments found in the epidermises and cortical tissues imparted a purple color to various parts of the inflorescence. Two types of raphide crystals, Psychotria type and Lemna type, and cluster crystals were observed. The volatile compounds from different parts of the inflorescence were collected by a modified headspace-solid phase microextraction (HS-SPME) technique and the samples were analyzed by gas chromatography-mass spectrometry (GC-MS). The major volatile compounds found by this method included isovaleric acid (21.6%), butyric acid (17.0%), benzyl alcohol (16.2%), and g-butyrolactone (12.1%) in the appendix; g-butyrolactone (27.0%), tetradecane (13.4%), and 4-hydroxy-4-methyl-2-pentanone (10.5%) in the male flowers; tetradecane (19.9%), 4-hydroxy-4-methyl-2-pentanone (13.0%), and 3-hydroxy-2-butanone (10.6%) in the female flowers; and butyl acetate (44.5%) and 3-hydroxy-2-butanone (12.2%) in the spathe. Use of different types of SPME fibers and multiple analytical methods is necessary in order to obtain a complete picture of the volatile composition of titan arum blooms., USDA-ARS, a Deployed War-Fighter Protection Research Program grant - the U.S. Department of Defense through the Armed Forces Pest Management Board [56-6402-1-612], We appreciate Dr Louis Ricciardiello, in Gilford, New Hampshire, for his generous donation of the titan arum seeds. The plants were raised and cared for by Aruna Weerasooriya, Edward Lowe, Derek Oglesby, and Lal Jayaratna at the Maynard W. Quimby Medicinal Plant Garden, University of Mississippi. This research was supported by USDA-ARS Grant No. 56-6402-1-612, a Deployed War-Fighter Protection Research Program grant funded by the U.S. Department of Defense through the Armed Forces Pest Management Board.

Four Caladium bicolor variants collected from different parts of Nigeria were subjected to anatomical comparison to enhance the taxonomic status of the species. Fresh samples (leaf and petiole) of these variants were fixed in formalin, acetic acid and alcohol (FAA), dehydrated in alcohol series, peeled or sectioned. Peeled specimens were stained with safranin, while sectioned ones were stained with Alcian blue and counterstained with safranin. Good preparations were mounted on slides, viewed and photographed with Optika B-1000 FL LED microscope. Epidermal cells from the variants are mainly pentagonal-hexagonal but rarely heptagonal while the anticlinal cell walls are mainly straight and partly arced/curved. Variants A, B and D are amphistomatic while variant C is hypostomatic. Isotricytic, anisocytic, tetracytic, anomocytic and contiguous stomata were observed among the taxa. The stomata index (SI) varied from 4.35 to 11.76 on the adaxial surface, and from 6.25 to 47.62 on the abaxial surface. Calcium oxalate crystals (druses, 8.18 to 19.09 µm and raphides, 21.82 to 68.18 µm) occur in all variants. Raphides are predominantly found in the petiole while druses and raphides are found in the midrib and petiole. The shapes of the adaxial surface of the midrib are relatively different from each other, and include curved (convex) and flat surfaces. The leaf lamina comprised one layer of palisade and spongy mesophylls each. The number of vascular bundles varied from 11 to 23 in the midrib. These characters can be used to distinguish these taxa especially when combined with the existing data on the species. The similarities among these variants point towards the same evolutionary origin; however may suggest that intraspecific or interspecific hybridizations may have produced the variants. Key words: Amphistomatic, Caladium, druses, hypostomatic, midrib, petiole, raphides.

S ocratea exorrhiza (Mart.) H. Wendl. is a palm tree that occurs in significant abundance in lowland forests of the Amazon region due to its root system adapted to flooded environments. However, with the sheet the organ that more changes in response to environmental change is proposed in this study to characterize the leaf structure of S. exorrhiza , aimed at generating useful information to better understand the anatomical characteristics of this body, in the occurrence of natural conditions of species. For this, samples were submitted to the usual techniques in plant anatomy. The regions of the leaf have different structures. The sheath is presented dorsiventral with papillose epidermis, air spaces, canals with septa and crystal raphides, the petiole and rachis exhibit a peripheral region and other spinal cord, with vascular bundles surrounded by sheaths amiliferous. The pinna is amphistomatic, with waxy streaks on the walls of guard cells, trichomes in the form of T, filamentous and branched. In the mesophyll, three types of vascular bundles: primary, secondary and tertiary, the border along the pinna has tapered form, orthorhombic and revolute. From the results it is concluded that, in general, the leaves of S. exorrhiza have characteristics common to Arecaceae. However, striking feature have been observed that may be unique to the species. Keywords : Palm; pinna; trichomes; secretory ducts; floodplain forest.

In this study, calcium oxalate (CaOx) crystals in the tissues and organs of 18 species that belong to tribe Cardueae (Asteraceae), Arctium minus, Silybum marianum, Cirsium vulgare, Cirsium creticum, Cirsium arvense, Carduus nutans, Carduus candicans, Carduus pycnocephalus, Tyrimnus leucographus, Jurinea consanguinea, Centaurea diffusa, Centaurea solstitialis, Centaurea iberica, Centaurea salonitana, Centaurea cyanus, Carthamus lanatus, Carthamus dentatus and Xeranthemum annuum , were investigated, and their morphologies and distribution were determined using a light microscope. Two morphological types of crystals, styloids and prismatics, were the most found crystals in the species examined. Raphides were only observed in the stem cortex and pith parenchyma cells of Silybum marianum, while druses were only viewed in the stem pith cells of Carduus pycnocephalus. Styloids were observed (if present) in the leaf, corolla, anther, filament, and style tissues. Prismatic crystals were seen almost in all the ovaries of the species investigated, except for Xeranthemum annuum. Prismatics, styloids, raphides, and druses were present in the stems of the species. It is considered that crystal formation within the cell is under genetic control. Thus the type, and presence or absence of crystals may be represented as a taxonomic character. In conclusion, it is suggested that the shape and location of CaOx crystals in the Cardueae constitute a diagnostic character and they may have taxonomic significance in this tribe.

Harvest time is assumed to potentially influence shape and size variation of calcium oxalate (CaOx) crystals; therefore it needs to be observed microscopically. This research used porang corms from the second growing period which were planted to produce the vegetative phase of third growing period. These corms were obtained based on the harvest time had determined. The harvest time was determined, i.e. 1) at two weeks before the plants shed (R0 - 1); 2) when the plants shed (R0) and 3) at two weeks after the plants shed (R0 + 1). Slides for microscopic observation were obtained from slices on the edge and center of porang corms. Organ slices were cleared using modified clearing method. Parameters observed were the shape and the size of CaOx crystal. The variations of styloid, prism, druse and raphide crystals found in porang corms at the three harvest time were 1, 2, 3 and 37 variations respectively. The variation of CaOx crystals tended to be same in porang corms at three harvest times. The variation of these crystals tended to be static. It is also known that raphide crystal has the greatest variation amount. On the contrary, styloid crystal has the fewest variation amounts. This abundance of a number of raphide crystal variations is possibly due to its role as a defense mechanism in porang corms.

This study showed anatomical differences related to Piper lepturumvar. lepturum and P. lepturum var. angustifolium species, sometimes considered varieties and in other cases synonyms. For histological analysis, fully expanded leaves were collected and for analysis by scanning electron microscope (SEM), fragments from the midrib were fixed on both leaf surfaces. The two species revealed differences in plant anatomy and it was observed that the stem of P. lepturum var. lepturum showed persistent wings and papillary epidermal cells, and these characters are absent in P. lepturum var. angustifolium. There was also the presence of raphides only in the lamina leaf and petiole of P. lepturum var. angustifolium, differentiating the two species. Recent studies have shown the importance of plant anatomy with species where there are issues related to taxonomic delimitation.

Calcium oxalate crystals (COC) are one of the most prevalent and widely distributed biomineralizations in plants. The aim of this work is to analyze and compare the data previously reported about the presence and production of COC in leaves of plant species from forests, wetlands and agroecosystems of the southeast of the Pampean Plain. Diaphanization, clearing of tissues with 50% sodium hypochlorite and cross sectioning of the leaves were realized. The material was mounted with gelatin-glycerin and COC were identified and described with optical, polarization and scanning electron microscopes. Crystal size and density were calculated. Calcification mainly occurred in leaf mesophyll. In terrestrial species, crystals were closely associated to vascular bundles, instead in aquatic species they were associated to aerenchyma. Druses, prisms and raphides were observed in the leaves of all species analyzed. Average crystals size was smaller in terrestrial species than aquatic ones (12 and 80μm respectively), but average crystal density was higher (246 and 23 crystals/mm2, respectively). These different patterns in COC production and distribution maybe related to taxonomical characteristics, the types of cells where crystals precipitate, their function and the differential transpiration rates, among other factors. Fil: Altamirano, Stella Maris. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Geología de Costas y del Cuaternario. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto de Geología de Costas y del Cuaternario; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Borrelli, Natalia Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Geología de Costas y del Cuaternario. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto de Geología de Costas y del Cuaternario; Argentina Fil: Benvenuto, María Laura. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Geología de Costas y del Cuaternario. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto de Geología de Costas y del Cuaternario; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Fernández Honaine, Mariana. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Geología de Costas y del Cuaternario. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto de Geología de Costas y del Cuaternario; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina Fil: Osterrieth, Margarita Luisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Geología de Costas y del Cuaternario. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto de Geología de Costas y del Cuaternario; Argentina

We investigated the involvement of guttation and calcium oxalate crystals in the maintenance of calcium homeostasis in leaf blades and petioles of eddo in hydroponic solution containing calcium at different concentrations. Under scanning electron microscopy, two types of crystals were observed in leaf blades and petioles: raphides (bundles of needle-shaped crystals) and druses (multifaceted conglomerate crystals). The number and size of crystals of leaf blades and petioles in 1 mMcalcium treatment (control) were smaller than that in 15 mM calcium treatments and larger than that in 0 mM calcium treatment. Calcium contents of leaf blades, petioles and whole plants increased with the increase of calcium concentration in the treatment solution. In addition, calcium-mapping images demonstrated a positive correlation between the amount of calcium in crystal idioblasts and concentration of calcium in treatment solutions. On the other hand, the weight percentage of calcium per mesophyll cell (spongy cell and palisade cell) of leaf blades and per normal parenchyma cell of petioles was stable irrespective of calcium treatment conditions. These results suggest that calcium accumulates in crystals under calcium-excessive conditions and is released from crystals under calcium-deficient conditions to stabilize calcium levels in leaf tissues other than the idioblasts. A positive correlation was observed between the calcium concentration of guttation fluid (μg mL−1), the total amount of calcium in guttation fluid (μg leaf−1night−1) and the calcium concentration of treatment solutions. These results suggest that guttation eliminates excess calcium and would be involved in maintaining calcium ion homeostasis in eddo.

Aim of this is to examine the Pharmacognastical and Phytochemical properties of Asparagus racemosus. The qualitative microscopy, Phytochemical screening, physicochemical evolution, fluorescence analysis and HPTLC studies were carried out according to the standard procedure recommended in WHO guidelines. The root had compactly arranged, uniseriate, polygonal to radially elongated, thick walled cell represent the outermost piliferous layer. Immediately laying below the epidermis is extensively developed, several layers of thick cortex made up of parenchymatous cells. The cortical cells contain raphide bundles. The innermost one or two layer of cortex immediately outside the endodermis comprises thick-walled cells, with the numerous oval or circular pits on their walls. Endodermis is composed of a single layer of compactly arranged, barrel-shaped parenchymatous cells. Inner to endodermis is a single layer of thin-walled, parenchymatous cells constituting the pericycle in the form of a ring, which surrounds a central stele. Phytochemical investigation of root shows total ash (6.5 %w/w), acid insoluble ash (1.3% w/w), and water-soluble ash (2.2% w/w). Loss on drying is (5.5% w/w). Cold extractive value in pet, ether (0.5%% w/w) in chloroform (3.4% w/w), methanol (8.0 % w/w) and in water (10.2% w/w). Hot extractive value in pet. ether (0.9% w/w), in chloroform (4.6 % w/w), methanol (10.40 % w/w) and in water (11.2% w/w). In methanolic extract and aqueous extract, tannins and saponins present in a major amount. Other constituents include carbohydrate, glycosides and phenolic compounds were present. Various pharmacognistical parameters evaluated in this study help in identification and standardization of the root of Asparagus racemosus. Keywords: Asparagus racemosus Willd, Satavar; Phytochemical investigation, Phytochemistry

In Siddha system of medicine dried roots of Aloe barbadensis Mill. Synonyms Aloe vera Tourn ex Linn. Aloe indica Royle belonging to family Liliaceae is known as Kumari Ver. Aloe vera is the oldest medicinal plant ever known and the most applied medicinal plant worldwide. The plant produces at least six antiseptic agents such as lupeol, salicylic acid, urea nitrogen, cinnamonic acid, phenols and sulphur. All of these substances are recognized as antiseptics because they kill or control mold, bacteria, fungus and viruses, hence has the ability to eliminate many internal and external skin infections. The morphological and anatomical characteristics of the root were investigated by hand sectioning and then camera lucida diagrams to ascertain the relevance of these characters in establishment of interspecific similarities and differences in the taxa. The results showed that there is many layered cork cells followed by cortex and a vascular bundle with radial arrangement and exarch xylem. Although, the size and number of vascular bundles are varying the presence of one or two raphide bundles and starch grains is prominent. Phytochemical screening reveal the presence of tannin, saponin and flavonoids. These features both anatomy and phytochemistry clearly help in identification of this plant.

Calcium oxalate crystals in higher plants occur in five major forms namely raphides, styloids, prisms, druses and crystal sand. The form, shape and occurrence of calcium oxalate crystals in plants are species- and tissue-specific, hence the presence or absence of a particular type of crystal can be used as a taxonomic character. So far, four different types of needle-like raphide crystals have been reported in plants. The present work describes two new and unusual forms of raphide crystals from the tubers of Dioscorea polystachya--six-sided needles with pointed ends (Type V) and four-sided needles with beveled ends (Type VI). Both of these new types of needles are distinct from the other four types by each having a surrounding membrane that envelopes a bundle of 10-20 closely packed thin crystalline sheets. The previously known four types of needles have solid or homogenous crystalline material, surrounded by a membrane or lamellate sheath called a crystal chamber. Only the Type VI crystals have beveled ends and the needles of the other five types have pointed ends.

The skin is the physical barrier between the fruit and the environment in which it develops. Environmental conditions during fruit development have a large influence on fruit quality, both at the time of harvest and during subsequent storage. It is hypothesized that some features of the skin and sub-epidermal tissues could provide information about the past growing conditions to which the fruit was exposed and therefore be of predictive value for storage quality. In this study, five commercial kiwifruit cultivars (‘Hayward’, ‘Hort16A’, ‘G3’, ‘G9’ and ‘G14’) were studied, and ‘Hayward’ fruit were manipulated during growth with different light or cultural practices. After harvest at horticultural maturity, X-ray micro computed tomography (µCT) was used to investigate features of the skin and the immediate parenchyma tissue. Despite orchard management practices (crop load and girdling) being observed to effect macro fruit quality parameters (mass, firmness, SSC, and DM), differences in microstructure (e.g. porosity) caused by these practices were not observed. However, porosity and pore size were found to be highly variable between cultivars. The thickness of dense sub-epidermal tissue could be readily measured and the 3-D distribution of raphide bundles was visible as high density particles distributed within the parenchyma. Overall, µCT was found to be a powerful technique to explore fruit epidermal and sub-epidermal structures in three dimensions at a micro level. However, the length of time required for data capture and analysis and the large number of samples required to overcome natural variation within horticultural products need to be considered. Future work may define the impact of differences in porosity or sub-epidermal anatomy on kiwifruit physiology (e.g. firmness change or sensitivity to low oxygen storage atmospheres). With this information, µCT could be used as a screening tool during plant breeding, or to determine the response to agronomic treatments, without conducting lengthy storage trials. publisher: Elsevier articletitle: Microstructural characterisation of commercial kiwifruit cultivars using X-ray micro computed tomography journaltitle: Postharvest Biology and Technology articlelink: http://dx.doi.org/10.1016/j.postharvbio.2014.01.012 content_type: article copyright: Copyright © 2014 Elsevier B.V. All rights reserved. ispartof: Postharvest Biology and Technology vol:92 pages:79-86 status: published

The aim of this research was to know the effects of nitrogen fertilizer to the shape and density of calcium oxalate (CaOx) crystals in porang tuber. Bulbil derived from porang plant in the first growth period seeded until the leaves fully open. Seedlings were transferred to a polybag after most of them has height 50 cm. After adapting for three weeks, the plants were treated with nitrogen fertilizer (urea) with a dose 0, 1, and 5 g/polybag. The plants were grown until the end of the growing period. The slides were prepared by slicing tuber as thin as possible using a sliding microtome. Sliced tuber of porang were cleared by using the modified clearing method. The slides were observed using light microscope to calculate the density of CaOx crystals. The densities of calcium oxalate crystals were analyzed using ANOVA (α = 0,05). The results showed that porang tuber has four kind of crystals i.e. raphide, druse, stiloid, and prism. Nitrogen fertilization had no significant effect on the densities of calcium oxalate crystals.

The stems of Dendrobium officinale Kimura et Migo, named Tie-pi-shi-hu, is one of the most endangered and precious species in China. Because of its various pharmacodynamic effects, D. officinale is widely recognized as a high-quality health food in China and other countries in south and south-east Asia. With the rising interest of D. officinale, its products have a high price due to a limited supply. This high price has led to the proliferation of adulterants in the market. To ensure the safe use of D. officinale, a fast and convenient method combining normal and fluorescence microscopy was applied in the present study to distinguish D. officinale from three commonly used adulterants including Zi-pi-shi-hu (D. devonianum), Shui-cao-shi-hu (D. aphyllum), Guang-jie-shi-hu (D. gratiosissimum). The result demonstrated that D. officinale could be identified by the characteristic “two hat-shaped” vascular bundle sheath observed under the fluorescence microscopy and the distribution of raphides under normal light microscopy. The other three adulterants could be discriminated by the vascular bundle differences and the distribution of raphides under normal light microscopy. This work indicated that combination of normal light and fluorescence microscopy is a fast and efficient technique to scientifically distinguish D. officinale from the commonly confused species.

We assessed the petiole anatomy of 8 species of Sri Lankan Cinnamomum (C. capparu-coronde, C. citriodorum, C. dubium, C. litseaefolium, C. ovalifolium, C. rivulorum and C. sinharajaense and C. verum) belonging to the family Lauraceae using light microscopy in the petiole anatomy for identification and classification of species. This is the first report on comprehensive petiole anatomy in Cinnamomum species. The distinctive, important and key taxonomic characteristics of the petiole include the shape and outline of the petiole, the presence of winged extensions, surface grooves on upper surface, presence or absence of trichomes, structure of vascular bundle and stone cell characters (shape, structure, pits and fissure size). These identified petiole characters provide the additional information for the identification of Cinnamomum species. Almost all the characters studied had little difference within a species but vary among the species. However, shape of epidermal cells was relatively constant for all species. All polymorphic characters were used for Principal Component Analysis (PCA) and species were divided into three main clusters. The PCA revealed that acicular crystals, cluster of crystals, trichomes and trichome abundance contribute to the first component that account for 31.76% total variance. Shape of the petiole, upper surface wings and raphides are the major loading characters to the second component. Therefore, anatomical structures of the petiole are useful in the identification and may help for crop improvement, conservation, management and future usefulness of germplasm resources of Cinnamomum species. Keywords. Cinnamomum, Lauraceae, PCA, petiole anatomy, taxonomy

Background and Aims Recently, molecular approaches have been used to investigate the phylogeny of Oncidiinae. This has resulted in the transfer of taxa previously considered to be species of Oncidium Sw. into Gomesa R. Br. and the re-circumscription of both genera. In this study, the structure of the floral elaiophore (oil gland) is described and compared for Gomesa echinata (Barb. Rodr.) M.W. Chase & N.H. Williams, G. ranifera (Lindl.) M.W. Chase & N.H. Williams, Oncidium amazonicum (Schltr.) M.W. Chase & N.H. Williams and O. oxyceras (Koniger & J.G. Weinm.) M.W. Chase & N.H. Williams in order to determine whether phylogenetic revision is supported by differences in its anatomy. Methods The floral elaiophore structure was examined and compared at three developmental stages (closed bud, first day of anthesis and final stage of anthesis) for all four species using light microscopy, fluorescence microscopy, scanning electron microscopy, transmission electron microscopy and histochemistry. Key results In all species investigated, the floral elaiophore occurs on the labellar callus and is of the epithelial type, comprising cuboidal to palisade-like, secretory epidermal cells and a layer of sub-epidermal cells, both tissues enclosing ground parenchyma supplied with collateral vascular bundles and containing idioblasts, often with raphides or phenolic contents. A bi-layered cuticle comprising an outer, lamellate and an inner, reticulate layer is present, and sub-cuticular accumulation of secreted material results in distension of the cuticle. Secretion-filled cavities are present at anthesis in the elaiophore cell walls and, in most species, the outer, tangential walls of the elaiophore have small, peg-like projections that protrude into the cytoplasm. In all taxa examined, the elaiophore organelle complement, especially the smooth endoplasmic reticulum (SER), is typical of lipid-secreting cells. Conclusions In terms of location, morphology, anatomy and ultrastructure, the floral elaiophores of both Gomesa and Oncidium species examined are very similar, and distinction between these genera is not possible based on elaiophore features alone. Furthermore, many of these elaiophore characters are shared with representatives of other clades of Oncidiinae, including the Ornithocephalus clade. Consequently, elaiophores are considered homoplasious and of limited value in investigating the phylogeny of this subtribe.

Leea species was originally placed in the family Vitace ae but was later placed in the Leeaceae due to the anatomical differences between the two families and between species. A study on the anatomical differe nce in leaves of four species in the Vitaceae ( Tetrastigma cruciatum , Pterisanthes miquelii , Cissus hastata and Nothocissus spicifera ) and two species from Leeaceae ( Leea rubra and Leea indica ) was undertaken. Show that species from the two families differ in the sh ape of the transverse sections of the petiole and m idrib. Druses and raphides are present in the petioles and midribs of Tetrastigma cruciatum , Pterisanthes miquelii , Cissus hastata and Nothocissus spicifera , while in Leea rubra and Leea indica only druses are present. Starch grains are present in the pith of the Vitaceae spec ies studied but are absent in both L. rubra and L. indica .

This research had aimed to determine variations in form of calcium oxalate (CaOx) crystals in porang. Slides for microscopic observation were prepared from sliced leaf, petiole, and tuber of porang. Sliced organs were cleared by using modified clearing method. The parameters that were observed included form and variety of morphology type of CaOx crystals. Microscopic observation showed there were four basic forms of crystals, i.e., druse, raphide, prism and styloid with some variation of diverse types, each of which had 49, 10, 8, and 5 variations. CaOx crystals, based on the size, were grouped into large (20 - 250 µm) and small (1 - 15 µm) crystals.

The structure of septal nectaries in Asphodelus aestivus flowers was investigated by using light microscopy (LM), scanning electron microscopy (SEM) and transmission electron microscopy (EM). It was found that the outlets of the three parts of the nectary were situated on the ovary surface at 2/3 of its height and had the shape of elongated openings. The nectariferous tissue was in the septa of the lower part of the ovary. The secretory tissue cells formed 1-3 layers surrounding the nectary slits. They contained thin cell walls with the cuticle layer from the slit side, large cell nuclei, numerous mitochondria and plastids characterised by various shapes. In plastids, small starch grains occurred sporadically. At the beginning of anthesis, the cells were poorly vacuolized. ER cisternae and secretory vesicles were located near the outer cell wall. Fibrous substance was present in the nectary slits. In the subglandular tissue, numerous starch grains occurred at the beginning of anthesis. In this zone, cells containing raphides and xylem elements were observed. Based on the ultrastructure of the nectary it can be stated that granulocrine nectar secretion occurs in A. aestivus.

Premise of the study: This is the first large-scale study comparing leaf crystal macropatterns of the species-rich sister genera Piper and Peperomia. It focuses on identifying types of calcium oxalate crystals and their macropatterns in leaves of both genera. The Piper results are placed in a phylogenetic context to show evolutionary patterns. This information will expand knowledge about crystals and provide specific examples to help study their form and function. One example is the first-time observation of Piper crystal sand tumbling in chlorenchyma vacuoles. Methods: Herbarium and fresh leaves were cleared of cytoplasmic content and examined with polarizing microscopy to identify types of crystals and their macropatterns. Selected hydrated herbarium and fresh leaf punches were processed for scanning electron microscopy and x-ray elemental analysis. Vibratome sections of living Piper and Peperomia leaves were observed for anatomical features and crystal movement. Key results: Both genera have different leaf anatomies. Piper displays four crystal types in chlorenchyma-crystal sand, raphides, styloids, and druses, whereas Peperomia displays three types-druses, raphides, and prisms. Because of different leaf anatomies and crystal types between the genera, macropatterns are completely different. Crystal macropattern evolution in both is characterized by increasing complexity, and both may use their crystals for light gathering and reflection for efficient photosynthesis under low-intensity light environments. Conclusions: Both genera have different leaf anatomies, types of crystals and crystal macropatterns. Based on Piper crystals associated with photosynthetic tissues and low-intensity light, further study of their function and association with surrounding chloroplasts is warranted, especially active crystal movement.

The shining appearance of the flowers of Oncidium cheirophorum and Ornithocephalus kruegeri results from the presence of lipids on the flower surface. The lipids are produced by elaiophores - secretory structures situated symmetrically at the base of the labellum or upon the callus. In O. cheirophorum, the elaiophores are epithelial type. They consist of one layer of cuboidal secretory cells and subsecretory parenchyma. The thick cuticle covering the outer, tangential wall of epithelial cells becomes distended and wrinkled as secreted oil accumulates beneath its surface. Oil secretion begins at the bud stage and lasts till the end of anthesis, that is 22 days, on average. Pollination does not influence oil production. In O. kruegeri, trichomatous elaiophores are situated on the central part of the callus. Unicellular trichomes project from the epidermis cells. Their outer walls are covered by a thin cuticle. In the dense cytoplasm of the trichomes, small plastids with few starch grains occur, whereas subsecretory parenchyma cells contain amyloplasts with large starch grains and raphides. The oil is already produced at the bud stage, about one week before flower opening, and lasts till the end of anthesis.

Problem statement: Leea species was originally placed in the family Vitace ae but was later placed in the Leeaceae due to the anatomical differ ences between the two families. Approach: A comparative study on the stem and leaf anatomy of Cayratia mollissima and Pterisanthes caudigera (Vitaceae) and Leea indica (Leeaceae) was conducted to investigate anatomical differences which could provide additional differentiating characters for supporting this treatment. Results: The Transeverse Sections (TS) revealed species differen ces with respect to the shape of the stems, the lea f margin, midrib and petiole. Druses were present in the leaves and stems of both species, while raphide s were present in the lamina. Conclusion: Starch grains and trichomes were present only in Cayratia mollissima and Pterisanthes caudigera .