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.

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.

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.

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 ofFouquieria 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.

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.

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.

Histochemical and Phytochemical Analysis of Lamium album subsp. album L. Corolla: Essential Oil, Triterpenes, and Iridoids

The aim of this study was to conduct a histochemical analysis to localize lipids, terpenes, essential oil, and iridoids in the trichomes of the L. album subsp. album corolla. Morphometric examinations of individual trichome types were performed. Light and scanning electron microscopy techniques were used to show the micromorphology and localization of lipophilic compounds and iridoids in secretory trichomes with the use of histochemical tests. Additionally, the content of essential oil and its components were determined using gas chromatography-mass spectrometry (GC-MS). Qualitative analyses of triterpenes carried out using high-performance thin-layer chromatography (HPTLC) coupled with densitometric detection, and the iridoid content expressed as aucubin was examined with spectrophotometric techniques. We showed the presence of iridoids and different lipophilic compounds in papillae and glandular and non-glandular trichomes. On average, the flowers of L. album subsp. album yielded 0.04 mL/kg of essential oil, which was dominated by aldehydes, sesquiterpenes, and alkanes. The extract of the L. album subsp. album corolla contained 1.5 × 10−3 ± 4.3 × 10−4 mg/mL of iridoid aucubin and three triterpenes: oleanolic acid, β-amyrin, and β-amyrin acetate. Aucubin and β-amyrin acetate were detected for the first time. We suggest the use of L. album subsp. album flowers as supplements in human nutrition.

Anatomy of the sporophyte of Anemia phyllitidis var. phyllitidis (Anemiaceae) from a riparian forest (Tucumán, Argentina)

Background and Aims: In Argentina seven species of the genus Anemia are recognized; one of them, A. phyllitidis, has two varieties. The anatomical records for A. phyllitidis var. phyllitidis are scarce and refer to the ontogeny of stomata, epidermis structure and type of petiole stele. This variety, which occurs in riparian forests, is affected by the increase of seasonal ecological pressures. There is scarce information about its sporophyte anatomy; hence, the aim of this work was to characterize it.Methods: Five specimens were collected from the margin of the El Parque stream (Tucumán-Argentina). A part was herborized and the other part was fixed in FAA. Subsequently, conventional histological techniques were used and stoma density and size of stomata and trichomes were determined. Histochemical tests were performed to detect starch (lugol) and phenolic compounds (10% ferric chloride).Key results: Anemia phyllitidis var. phyllitidis shows diarch roots and dictyostelic rhizome covered with glandular trichomes. The petiole is covered by glandular trichomes, epidermis and subepidermis composed of lignified fibers, vascular bundle with phloem surrounding xylem, two-layered pericycle surrounded by endodermis and cortical tissue with starch and phenolic compounds. Sterile pinnae have pericytic, anomocytic, and desmocytic stomata (mean size 49.8 µm × 38.7 µm); glandular trichomes; dorsoventral hypostomatic lamina, vascular bundles with pericycle and endodermis. Fertile pinnae are anatomically similar to the rachis. Silica is present in some epidermal cells of the different organs.Conclusions: The sporophyte of A. phyllitidis var. phyllitidis showed anatomical, morphological and physiological traits that reveal its adaptation to the riparian habitat where it grows. At the same time, the description of this variety is deepened and unpublished data for the genus are provided, such as the presence of stegmata in the fertile pinnae.