Histochemical Tests Research Articles

Pathomorphological alterations in the kidneys in acute and chronic alcohol intoxication

The literature review presents data on pathomorphological changes in the kidneys in acute and chronic alcohol intoxication. A lack of consensus among researchers on the direct toxic damage to renal tissue by ethanol is demonstrated. Analysis of the literature showed that the issue of attributing microscopic changes detected in the kidneys to characteristic or specific signs remains controversial. Some authors identify several characteristic microscopic patterns in the kidney tissue in alcohol intoxication: thickening of glomeruli basal membranes, narrowing of glomerular capsule tubular lumen. Fatty dystrophy of the proximal tubules is most often indicative of concomitant conditions related to alcohol intoxication, such as ketoacidosis. In alcohol intoxication, some changes in the kidneys occur variably, such as dyscirculatory, dystrophic, pigmentary, necrotic, and autolytic changes. Data on the causes of their occurrence are often contradictory from different authors. The authors of this review believe that this issue can be resolved using up-to-date histochemical, immunohistochemical, and morphometric test methods.

Morphology, histochemical test, potential, and conservation effort of Alpinia warburgii K. Schum., a native species to Sulawesi

Alpinia warburgii is a species of the Zingiberaceae family native to Sulawesi. This study aims to characterize the plant morphology; analyze the anatomical structure, histochemical localization, and secretory cell structure of the rhizome; also discuss the potential and conservation effort of the species. Anatomical study and histochemical tests were observed microscopically using the fresh preparation method and wet mounted slide. Histochemical tests carried out include starch, protein, lipid, tannin, alkaloid, and flavonoid. This study is considered the first report of histochemical tests on A. warburgii. The main morphological characteristics of A. warburgii are perennial herbaceous; inflorescence terminal and raceme; flowers white, labellum obovate with red lines in the middle, and flanked by light green edges; fruit globose, green when young and yellow when ripe. The rhizome anatomical structure comprises the epidermis, endodermis, hypodermis, parenchyma, cortex, and vascular bundles, classified as closed collateral type. Histochemical tests showed positive localization for starch, protein, alkaloids, flavonoids, and tannins. The density of flavonoids secretory cells is higher than tannins and alkaloids, which indicates its potential as medicine to be associated with antioxidant, antimicrobial, antiinflammation, and anticancer. In addition to in-situ, ex-situ conservation effort through propagation is also important to maintain the species.

Callus of Curculigo latifolia Dryand. ex W.T. Aiton: initiation, regeneration, secretory structure and histochemistry

In Indonesia, Curculigo latifolia, popularly known as lemba, is used as traditional medicine for antihypertension, antidiabetic, and as a food and drink additive. Despite the high benefits, this plant is difficult to propagate due to its low seed viability. Tissue culture offers an alternative for plant propagation, thus providing a continuous source of bioactive substances. The study aimed to establish efficient methods of explant sterilisation, callus induction and its regeneration from leaf tissues for plant multiplication. The study also investigated histochemically secondary metabolites produced by the callus. Our results showed that sterilising agents in the lowest concentrations and the shortest duration was the most suitable for getting aseptic cultures. BAP (3 mg L−1) and IBA (5 mg L−1) were the highest callus initiation, shoots, and roots regeneration. The histological sections of fresh callus showed secretory structure in the form of secretory cavities surrounded by a layer of epithelial cells and idioblasts around the secretory cavities. The histochemical tests identified phenolic compounds and essential oils in organogenic callus. Idioblasts were detected containing essential oils too. The results will contribute to C. latifolia propagation and the development of callus as a producing source of secondary metabolites.

Phytochemical composition and biological screening of two Lophophytum species

Lophophytum species are holoparasites that grow on tree roots. The objectives of the work were to explore the chemical composition of the tubers of two Lophophytum species and to analyze the antioxidant, anti-inflammatory and antilithiatic activity of their extracts using in vitro methods. The chemical composition was determined by histochemical, phytochemical and TLC tests. In addition, the profile of phenolic compounds was determined by HPLC-MS. The presence of secondary metabolites of recognized activity was demonstrated. The results of the HPLC-MS/MS allowed the tentative identification of catechin, luteolin and glycosides of eriodictyol, naringenin and luteolin in the extract of Lophophytum leandri and eriodictyol, naringenin, luteolin and their glycosylated derivatives in Lophophytum mirabile. The extracts showed promising antioxidant (DPPH, ABTS and β-carotene-linoleic acid), anti-inflammatory (inhibition of 5-LOX) and anti-urolytic (by bioautographic TLC) activity. It is noteworthy that these are the first results of the phytochemical composition and biological activity of L. mirabile. However, in vivo studies are required to corroborate these activities.

Leaf Structure and Histochemical Analysis of Camphor Tree (Cinnamomum Camphora), a Plant Used in Traditional Remedies

ABSTRACT An integrated microscopical investigation of the secretive activity in the above ground parts such as the leaves, petioles, and young stems of the camphor tree (Cinnamomum camphora) was performed. Different kinds of cells were localized accumulating different metabolites as this was demonstrated by histochemical tests. The wide distribution of oil and mucilage cells and other idioblasts was set out. Virtually all palisade cells appeared to be involved in secondary metabolite production which may explain the complexity of the plant extracts and their wide range of therapeutic anti-inflammatory and anti-oxidative effects they have in traditional medicine.

Foliar microstructure and histochemical analysis of the lavender tree (Heteropyxis natalensis Harv.)

Heteropyxis natalensis Harv. is a native South African tree used in traditional medicine among Venda and Zulu communities. In this study, micromorphological investigations using light and scanning electron microscopy identified long, tapered non-glandular trichomes on the abaxial and adaxial surfaces of the leaves. The total number of trichomes appeared to be greater on emergent leaves and decreased as leaves matured. In addition, schizolysigenous secretory cavities and druse crystals were found within the leaves. Swollen subdermal secretory cavities were also distributed in the midrib of the leaves. Transmission electron microscopy confirmed the presence of nuclei, plastids, mitochondria, vesicles, rough endoplasmic reticulum cisternae and Golgi bodies in the secretory epithelia of these cavities. Various histochemical tests revealed the presence of alkaloids, phenolics, lipids, proteins, essential oils, resin acids and trace amounts of unesterified pectins and polysaccharides in the leaves. This study provides new findings and contributes to the existing research regarding H. natalensis with respect to the micromorphology, ultrastructure and histochemical composition of this species.

Crepidium sect. Crepidium (Orchidaceae, Malaxidinae)-Chemical and Morphological Study of Flower Structures in the Context of Pollination Processes.

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.

Morpho-anatomical features of the leaves and stems of Baccharis notosergila (Asteraceae) and their relationship with the environment and chemical control

Background and aims: Baccharis notosergila is an aggressive weed inhabiting the Salado river basin, Buenos Aires province, Argentina. The aims of this work were: to analyze the morpho-anatomy and histochemistry of aerial vegetative organs in order to understand the adaptation strategies that ensure its survival, as well as to expand knowledge on traits determining resistance to the control methods applied. M&M: The material collected was prepared and examined with conventional techniques of microscopy. Histochemical tests to identify starch, resins, polyphenols, and lipophilic substances were performed. Results: The major features found were small and deciduous leaves; uniseriate epidermis with massive and striate cuticle; stomata at level or slightly above the other epidermal cells and glandular trichomes secreting oily substances; stomata on both surfaces and isobilateral mesophyll. Tannins, starch and lipophilic substances were identified in leaves and stems; polyphenols, resins and lipids in ducts, and calcium oxalate crystals in leaves, stems and capitate trichomes. Conclusions: The aerial vegetative organs features of B. notosergila explain its tolerance to the unfavorable conditions of the Salado river basin area, as well as its high competitive ability over others species of the natural prairie. The reduced and deciduous leaves, the epidermal traits, and chemical substances found constitute a physical and chemical barrier reducing dehydration as well as the penetration of the herbicides applied for its control. Botanical knowledge of B. notosergila is the basis for the design and development of new and appropriate management methods for this species.

Stinging Trichomes in Apocynaceae and Their Evolution in Angiosperms.

Stinging trichomes are rare in plants, occurring only in angiosperms, where they are reported for a few genera belonging to six families. Although there is no report of stinging trichomes in Apocynaceae, previous fieldwork collections of Fischeria and Matelea caused us a mild allergic reaction on the skin when we contacted the dense indumentum of the plants. This fact associated with the well-known presence of glandular trichomes with acute apex in both genera raised suspicions that stinging trichomes could be present in the family. Hence, this study aimed to investigate the likely occurrence of stinging trichomes in Fischeria and Matelea. We analyzed vegetative shoots and leaves of Fischeria stellata and Matelea denticulata through the usual procedures of light and scanning electron microscopy. We also performed several histochemical tests to investigate the chemical composition of trichome secretion. We detected that glandular trichomes occur throughout the surface of the leaf and stem. They are multicellular, uniseriate with an apical secretory cell, which has a dilated base and a needle-shaped apex. The secretion is compressed into the acuminate portion of the apical cell by a large vacuole, and crystals are deposited in the cell wall in a subapical position, providing a preferential site of rupture. The secretion, composed of amino acids and/or proteins, is released under mechanical action, causing skin irritation. Based on our detailed morphological and anatomical analyses, and in the functional aspects observed, we concluded that the glandular trichomes in Fischeria and Matelea can indeed be classified as stinging. Thus, Apocynaceae is the seventh family for which this type of trichome has been reported. We also compiled information on stinging trichomes in all families of angiosperms. Their phylogenetic distribution indicates that they have evolved at least 12 times during angiosperm evolution and may represent an evolutionary convergence of plant defense against herbivory.

Biochemical and morphological characterization of freshwater microalga Tetradesmus obliquus (Chlorophyta: Chlorophyceae)

Tetradesmus is a microalgal genus with biotechnological potential due to its rapid production of biomass, which is plenty in proteins, carbohydrates, lipids, and bioactives. However, its morphology and physiology need to be determined to guide better research to optimize the species cultivation and biocompounds processing. Thus, this study describes the biochemistry and morphology of the strain Tetradesmus obliquus BR003, isolated from a sample of freshwater reservoirs in a Brazilian municipality. In the T. obliquus BR003 dry biomass, we identified 61.6% unsaturated fatty acids, and 3.4% saturated fatty acids. Regarding other compounds, 28.50 ± 1.47g soluble proteins/100g, 0.14 ± 0.009g carotenoids/100g, 0.76 ± 0.013g chlorophyll a/100g, and 0.42 ± 0.015g chlorophyll b/100g with a chlorophyll a/b ratio of 1.8 were detected. The main chemical elements found were S, Mg, and P. The cells of BR003 were elliptically curved at the ends and without appendages. Histochemical tests showed carbohydrates distributed in the cytoplasm and pyrenoids, some lipid droplets, and proteins. The cytoplasm is rich in vacuoles, rough endoplasmic reticulum, mitochondria, and chloroplasts. The nucleus has a predominance of decondensed chromatin, and the cell wall has three layers. Chloroplasts have many starch granules and may be associated with a spherical central pyrenoid. To the best of our knowledge, this was the first biochemical description combined with ultrastructural morphological characterization of the strain T. obliquus BR003, grown under standard conditions, to demonstrate specific characteristics of the species.

Bone Marrow Mesenchymal Stem Cell-Derived Exosome-Educated Macrophages Promote Functional Healing After Spinal Cord Injury.

The spinal cord injury is a site of severe central nervous system (CNS) trauma and disease without an effective treatment strategy. Neurovascular injuries occur spontaneously following spinal cord injury (SCI), leading to irreversible loss of motor and sensory function. Bone marrow mesenchymal stem cell (BMSC)–derived exosome-educated macrophages (EEM) have great characteristics as therapeutic candidates for SCI treatment. It remains unknown whether EEM could promote functional healing after SCI. The effect of EEM on neurovascular regeneration after SCI needs to be further explored. We generated M2-like macrophages using exosomes isolated from BMSCs, which were known as EEM, and directly used these EEM for SCI treatment. We aimed to investigate the effects of EEM using a spinal cord contusive injury mouse model in vivo combined with an in vitro cell functional assay and compared the results to those of a normal spinal cord without any biological intervention, or PBS treatment or macrophage alone (MQ). Neurological function measurements and histochemical tests were performed to evaluate the effect of EEM on angiogenesis and axon regrowth. In the current study, we found that treatment with EEM effectively promoted the angiogenic activity of HUVECs and axonal growth in cortical neurons. Furthermore, exogenous administration of EEM directly into the injured spinal cord could promote neurological functional healing by modulating angiogenesis and axon growth. EEM treatment could provide a novel strategy to promote healing after SCI and various other neurovascular injury disorders.

Plectranthus neochilus Schltr.: Anatomic and cytogenetic analyses and chemical characterization of its essential oil

Although used for medicinal purposes, there are many inconsistencies in the literature regarding the identification of Plectranthus neochilus Schltr. due to the morphological similarities with other species belonging to the so-called “P. caninus complex”. Studies that develop tools for the correct identification of medicinal plants are of vital importance as the use of the wrong species can lead to serious health problems. Thus, the objective of the study was to determine a profile to identify individuals of P. neochilus based on anatomic, chemical and cytogenetic analyses. The cytogenetic analysis showed that all the individuals used in the present study belonged to the same species (2n = 30), P. neochilus, in agreement with literature. The presence of diacytic stomata, absence of digitiform trichome, distribution of peltate glandular trichomes on both sides of the leaves are also in agreement with previous reports for P. neochilus. However, the leaf mesophyll and the layout and morphology of glandular trichomes were different from those previously described in the literature. Histochemical tests showed the absence of phenolic compounds but presence of total and neutral terpenes and lipids. The essential oil obtained by hydrodistillation contained β-caryophyllene as the major compound (20.11%) which also coincides with other studies of P. neochilus in Brazil. These results show the cytogenetic, anatomical and the chemical profile of the essential oil of P. neochilus in Brazil and can be used as a tool for its identification.

Histochemical Test of root, petiole and leaf of Kelembak (Rheum officinale Baill.)

Kelembak (Rheum officinale Baill.) is perennial shrub with pharmacological activities, and well-known as one of traditional herbal medicine used widely in the world. The phytochemical study, as well as histochemical test, is pivotal to determine the potency of herbs used for disease treatment. Based on its secondary metabolite content. Histochemical test of R.officinale vegetative organs had not been reported. This study aims to determine the content of compounds in the vegetative organs of kelembak and provide information on the anatomical position of each compound in root, petiole and leaf. The preparation of histochemical test used freehand section method. Each sample was made in three replications for this test. A total of four secondary metabolites were examined in this study, i.e. alkaloid, tannin, lipid and flavonoid. The positive content of each compound was indicated by the coloration change. The specimens were then observed and photographed using light microscope. The results showed that the root, petiole and leaf showed positive content of alkaloids, tannins, lipids and flavonoids. However, the anatomical position of each secondary metabolites are different among root, petiole and leaf. The results of this study provide the first information of histochemistry vegetative organs of Rheum officinale.

Anatomical and histochemical characterization of leaves of Spondias purpurea L.

The Anacardiaceae family has species rich in secondary metabolites. They are widely used in popular medicine. Among them, Spondias purpurea L. stands out for containing several secondary metabolites with important biological activities. To aid in the precise identification of the species, this work aims to perform an anatomical and histochemical characterization of leaves of S. purpurea. Microscope slides containing cross-sections of the petiolule and leaflets, in addition to paradermal sections of the leaflets, were prepared and analyzed in an optical and polarized microscope. Histochemical tests were performed on fresh leaflets. Through microscopic analysis, it was possible to identify the anatomical structures that allow the diagnosis of the studied species, such as petiolule with concave-convex shape, non-glandular and glandular trichomes, druses in phloem; leafletsamphistomatic, with non-glandular trichomes on the adaxial face and glandular trichomes on the abaxial face, midrib with concave-convex shape, two collateral vascular bundles, one layer of palisade parenchyma, druses in phloem and mesophyll. Through histochemistry, the presence of phenolic compounds, alkaloids, tannins, triterpenes, steroids, lipophilic compounds, essential oils, lignin, starch and calcium oxalate crystals were evidenced in the leaflets. The results are important for the quality control of plant material.

Comparative Histochemical analysis of above-ground parts of Filipendula vulgaris and Filipendula ulmaria growing in Central Kazakhstan

The histochemical analysis of medicinal plants and raw materials makes it possible to establish the presence of certain groups of active and related substances or products of their metabolism, as well as to study their localization in tissues. The article presents the results of histochemical analysis of leaves, flowers, petioles and stems of Filipendula vulgaris and Filipendula ulmaria. These types of plant are used in traditional medicine as an anti-inflammatory, antipyretic and bactericidal agent. Our research objective - to determine the peculiarities of the localization features of individual groups of secondary metabolites in the aerial part of meadowsweet and meadowsweet growing in the territory of Central Kazakhstan at the microscopic level. Materials and Methods. Transverse sections of petioles, stem and whole flower; transverse and superficial sections of leaves of Filipendula vulgaris and Filipendula ulmaria were fixed in Strauss-Fleming solution, after which histochemical reactions were carried out to locate and identify essential oils, phenolic acids, flavonoids, sesquiterpene lactones, polysaccharides and alkaloids. The study of microscopic signs, as well as histochemical tests were carried out according to the methods of the State Pharmacopoeia of the Republic of Kazakhstan using a light microscope Biomed-4. Results and Discussion. The localization of biologically active substances in the studied types of plant raw materials was determined by histochemical methods using light microscopy. The presence of phenolic compounds, flavonoids, traces of essential oil, alkaloids and sesquiterpene lactones was established. The absence of polysaccharides in the investigated micropreparations was established. Conclusions. For the first time, the localization of secondary metabolites in the tissues of Filipendula vulgaris and Filipendula ulmaria was studied by histochemical tests. The results of histochemical studies can be used to confirm the authenticity, identification and standardization of the aerial parts of Filipendula vulgaris and Filipendula ulmaria.

The decellularized ovary as a potential scaffold for maturation of preantral ovarian follicles of prepubertal mice

ABSTRACT In the current study, a decellularized ovary scaffold was used for the encapsulation and development of preantral follicles. Maturation and the expression of the related genes were evaluated and compared against in vivo conditions. Mature rat ovaries were decellularized and decellularization was confirmed using hematoxylin/eosin and Hoechst stains. Collagen and glycosaminoglycan (GAG) preservation and the extracellular matrix (ECM) architecture were evaluated by Masson’s trichrome staining, Alcian blue staining, and scanning electron microscopy, respectively. Raman confocal microscopy and DNA quantification were performed to show DNA and cell debris depletion and ECM retention. Preantral follicles of mice were cultured for 12 days in the decellularized (3D) and two-dimensional (2D) conditions. Finally, the survival rate, antrum formation rate and diameters of follicles, levels of estradiol (E2) and progesterone (P4), and expression of the ZP2, Gdf9, Bmp6, and Bmp15 genes were evaluated in the 2D and 3D culture systems. On day 13 of culture, oocyte maturation was induced by hCG. Although histochemical tests showed partial retention of ECM, Raman confocal microscopy showed some extent of ECM washing out along with the depletion of DNA and cell debris. However, SEM showed the preservation of the ovarian ECM’s ultrastructure. The survival rate of follicles was similar, but the antrum formation rate, concentrations of E2 and P4, and oocyte maturation rate were significantly higher in 3D compared with 2D. The level of ZP2, Gdf9, Bmp6, and Bmp15 gene expression in follicles matured in 2D, 3D, and in vivo was statistically similar. In all matured follicles, their gene expression levels significantly declined relative to preantral follicles. Ovarian follicles that matured in the decellularized ovaries demonstrated the same follicular growth and maturation gene expression levels as those that grew in the natural ovary. This is consistent with using the decellularized ovary as a scaffold suitable for ovarian reconstruction. Abbreviations GAG: glycosaminoglycan; ECM: extracellular matrix; 2D: two-dimensional; E2: estradiol; P4: progesterone; BMP15: bone morphogenetic protein 15; GDF9: growth differentiation factor 9; ZP2: zona pellucida 2; Gdf9: growth/differentiation factor-9; Bmp6: bone morphogenetic protein 6; Bmp15: bone morphogenetic protein 15.

Analisis Kadar Metabolit Sekunder, Histokimia, dan Aktivitas Antioksidan Akar Acalypha indica L.

Acalypha indica L. is a species of wild plant that has secondary metabolites are used as medicine, but the usage is not optimized yet. The objective of of this study was to determine secondary metabolites content, location and distribution of secondary metabolites, and also the capacity for antioxidant. Secondary metabolites content was determined included phenol, flavonoids, tannin of chloroform and methanol extracts from Acalypha indica L. root, using spectrophotometre. Histochemical test was done by making fresh sliced preparation which was reacted with reagent. Antioxidant activity was determined by using DPPH method. The result of this study showed that secondary metabolites content of chloroform extract, were phenol 9,89 ± 0,77 %GAE, flavonoids 5,87 ± 1,40 %QE, and tannin 3,33 ± 1,21 %GAE, while methanol extract were phenol 45,11 ± 4,86 %GAE, flavonoids 19,87 ± 0,61 %QE, and tannin 6,76 ± 0,31 %GAE. Histochemical test showed that phenol, flavonoids, tannin, and alkaloids were found in Acalypha indica L. root. Antioxidant activity showed the IC50 value was 161,81 ± 7,88 μg/mL for chloroform extract and 92,81 ± 4,33 μg/mL for methanol extract.

Morphoanatomical and histochemical study of seeds of four species of Swartzia (Fabaceae - Papilionoideae) at Tupé Sustainable Development Reserve AM/Brazil

Background: In Brazil, Swartzia Schreb. occurs in almost all phytogeographic domains, but the greatest richness of species is concentrated in the Amazon.
 Question: What are the morphological and anatomical variations of the seeds of species of the Swartzia ?
 Studied species: Swartzia laevicarpa , Swartzia macrocarpa , Swartzia recurva , and Swartzia sericea. 
 Study site and dates: The species were collected in the Sustainable Development Reserve of Tupé - AM Brazil, in 2017 and 2018.
 Methods: Cross-sections of the seed coat were cut, to observe the characteristics of the seed coat in the hilar region and opposite it. Histochemical tests were performed on the
 cross-sections of fresh samples, to verify ergastic substances present in the aril, seed coat and cotyledons.
 Results: The seeds have a smooth and membranous seed coat, partially covered by a spongy aril in Swartzia recurva , filamentous in S . laevicarpa , S . sericea , and S . macrocarpa . Linear hylum, imperceptible micropyle, cryptoradicular embryo, fleshy cotyledons, containing starch, lipids, alkaloids, protein and the rudimentary plumule.
 Conclusions: Morphological characteristics of the seed, such as shape, color, consistency of the aril and size of the hilum, are important diagnostic characters in the recognition of this species in the field, through its respective seeds; in the same way, the information on soil seed bank is expanded. The presence of fleshy cotyledons indicates and adaptation to the type of environment in which these species occurs, while the knowledge of ergastic substances present in the aril, seed coat and cotyledons, contributes to the understanding of their germinative processes.

Ovule development, megasporogenesis, and megagametogenesis of Fouquieria fasciculata (Willd. ex Roem. & Schult.) Nash (Fouquieriaceae)

Background: Fouquieriaceae is a small family that consists of a single genus, Fouquieria , with eleven species occurring mostly in arid and semiarid regions of Mexico and the southwestern United States. The development of the ovule and the female gametophyte has been poorly characterized in species of the family; thus, it is proposed to carry out a detailed study on the development of the ovule, megasporogenesis and megagametogenesis in Fouquieria fasciculata .
 Questions: How are the developmental patterns of the ovule and the female gametophyte in Fouquieria fasciculata ?
 Study species: Fouquieria fasciculata (Willd. ex Roem. & Schult.) Nash (Fouquieriaceae).
 Methods: Embryology, morphology and histochemical tests of floral buds and flowers at pre-anthesis of Fouquieria fasciculata were examined using light microscopy and scanning electron microscopy.
 Results: The ovules of Fouquieria fasciculata are anatropous, bitegmic and tenuinucellate, and the micropyle is formed by the inner integument. The innermost layer cells of the inner integument form an integumentary tapetum or endothelium. The cells of integumentary tapetum are characterized by very conspicuous wall thickenings on the inner tangential side, and are radially elongated surrounding the nucellus. The pattern of development of the female gametophyte is monosporic Polygonum -type, and the formation of a lateral haustorium was observed.
 Conclusions: This investigation describes in detail the development of the ovule and the female gametophyte of Fouquieria fasciculata . Our results show interesting features, such as the differentiation of an integumentary tapetum, a monosporic Polygonum -type pattern of development and the formation of lateral haustorial arm in the female gametophyte.

Contribution to the identification of the species Myrcia hatschbachii D. Legrand (Myrtaceae): anatomical and histochemical analyses

The genus Myrcia is used in folk medicine to treat diabetes. The plants used in folk medicine require morphological and anatomical references to attest to its authenticity. This is the first report of the microscopic study of Myrcia hatschbachii. In this sense, the aim of the present study was to describe the anatomical characteristics, in order to contribute to the species' identification. For anatomical and surface analyses free-hand sections were prepared and observed using optical microscopy, simultaneously some materials were processed and observed under scanning electron microscopy. In addition, histochemical tests were performed. The anatomical features described here correspond with previously reported features found in other species of the genus Myrcia. Furthermore, the following anatomical markers were observed in Myrcia hatschbachii: druses in the spongy parenchyma, concave-convex midrib, heart-shaped petiole, oval-shaped stem, C-shaped vascular bundle in the midrib and open arc shape with invaginated ends in the petiole; phenolic compounds in the phloem and lignified cells in the pith. The results of anatomical and histochemical analyses provide micromorphological and microchemical features that can help in the taxonomy and proper identification of the species.