Leaf Cross Sections Research Articles

Study of the Relationship between Leaf Color Formation and Anthocyanin Metabolism among Different Purple Pakchoi Lines.

Purple pakchoi (Brassica rapa ssp. Chinensis) is particularly appreciated due to its high edible quality and ornamental value, but there are few studies on the underlying mechanisms of leaf color formation. To comprehensively assess the differences in purple formation in pakchoi, four lines of pakchoi with different purple leaves were used in this experiment to determine the pigment content and to investigate the distribution and components of anthocyanin using LCMS (Liquid Chromatography Mass Spectrometry) and leaf cross-sections. Moreover, the expression levels of anthocyanin synthesis-related genes in four lines were calculated by qRT-PCR. The results showed that three new purple lines rich in anthocyanin and of high-quality were bred, and the anthocyanin were mainly distributed in both the upper epidermis and lower epidermis of leaves. Thirteen anthocyanin components were separated and identified, all the anthocyanins were acylated and glycosylated cyanidins; the main anthocyanins in purple pakchoi were a diacylated form of cyanidin 3-trans-(feruloyl)diglucoside-5-(malonyl)glucoside. Both the ratio of non-aromatic acylated cyanidin to aromatic acylated cyanidin and the ratio of anthocyanin content to chlorophyll content were responsible for the color formation in different purple pakchoi lines. When the ratio was high, the leaf appeared reddish purple, and when the ratio was low, the leaf appeared deep purple, even blackish purple. The expression level of BrF3H was significantly correlated with the content of anthocyanin through the correlation coefficient, which was speculated to be the main anthocyanin synthesis-related gene resulting in color differences among the four purple pakchoi lines. These results will enhance our understanding for the cultivation of new purple pakchoi varieties.

Comparative analysis of the DNA isolated from thyme leaves using different methods

Background. The base for a molecular analysis is DNA of high quality. For DNA isolation, different kits or classical methods are used. For mass analysis, isolation with kits is a very expensive process. So, the objective of our investigation was to find a cheap method for high-quality DNA isolation from leaves of various thyme cultivars.Materials and methods. Leaves cut from thyme accessions (Thymus mastichina L. cv. ‘Svetliachok’, T. striatus Vahl. cv. ‘Jubileiniy’, T. vulgaris L. cv. ‘Fantasia’, and T. vulgaris cv. ‘Jalos’.) maintained ex situ in the collection of the Nikita Botanical Gardens were used as the material for the analysis. Light microscopy was used to study leaf anatomy and localize essential oil on leaf cross sections. Essential oil was extracted on Ginsberg devices, and phenolic content was measured with The Folin–Ciocâlteu reagent (FCR). Commercial kits (DiamondDNATM, PureLink® Plant Total DNA Purification Kit) and classical methods (CTAB, CTAB with 2% polyvinylpyrrolidone) were used for DNA isolation. DNA quality was evaluated spectrophotometrically, with electrophoresis (horizontal, automated system Agilent 4200 TapeStation) and PCR.Results. The analysis showed that the leaf blade mesophyll of four thyme cultivars had inclusions with essential oil. The content of essential oil and phenolic compounds was measured biochemically. Since the plants were characterized by the presence of secondary metabolites, DNA was isolated by different methods. Spectrophotometry demonstrated that the classical CTAB method and CTAB with 2% PVP provided the best results. Using an automated electrophoresis system, the presence of high-molecularweight DNA (more than 52000 bp) in significant amounts was detected in the samples isolated with DiamondDNATM kit and CTAB + 2% PVP.Conclusion. Among the tested kits and methods, CTAB + 2% PVP provided thyme DNA suitable for PCR and, presumably, for genome library preparation. The low cost of reagents for this technique makes it applicable for future mass analysis of plant material.

Essential oils from Vitex trifolia as an effective repellent for Aedes aegypti

Abstract. Arpiwi NL, Muksin IK, Kriswiyanti E. 2020. Essential oils from Vitex trifolia as an effective repellent for Aedes aegypti. Biodiversitas 21: 4536-4544. Vitex trifolia is a shrub or small tree grown wild from family Verbenaceae commonly found in the sandy shore habitat of the tropicals and sub-tropical countries. This research aims to measure the essential oils yield of V. trifolia leaves, to investigate the secretory cells of the leaf, and to measure repellent activity of the essential oils against Aedes aegypti. Leaves and soil samples were taken from Panjer, Sanur, and Sidakarya Villages of Denpasar, Bali Indonesia. Oil was extracted from fresh leaf samples using steam distillation. The essential oils were analyzed for its constituents using Gas Chromatography-Mass Spectrometry. The leaf cross-sections were prepared using paraffin embedding method and stained with safranin. The essential oils of V. trifolia at different concentrations (0, 2, 3, 4, 5, 6% w/w) were formulated into lotions. The yield of essential oils was significantly different from the three locations. The main constituent of the oil was cis-ocimene. The essential oil yield was positively correlated with the sand content. The secretory structures of the leaf were idioblast and glandular trichome. Lotions formulated with the essential oils were soft, not greasy, and not sticky upon application to the skin and non-irritant. Lotions with 5 and 6% essential oils gave 100% protection for 3 hours from Ae. aegypti.

Water Adsorption to Leaves of Tall Cryptomeria japonica Tree Analyzed by Infrared Spectroscopy under Relative Humidity Control.

Leaf water storage is a complex interaction between live tissue properties (anatomy and physiology) and physicochemical properties of biomolecules and water. How leaves adsorb water molecules based on interactions between biomolecules and water, including hydrogen bonding, challenges our understanding of hydraulic acclimation in tall trees where leaves are exposed to more water stress. Here, we used infrared (IR) microspectroscopy with changing relative humidity (RH) on leaves of tall Cryptomeria japonica trees. OH band areas correlating with water content were larger for treetop (52 m) than for lower-crown (19 m) leaves, regardless of relative humidity (RH). This high water adsorption in treetop leaves was not explained by polysaccharides such as Ca-bridged pectin, but could be attributed to the greater cross-sectional area of the transfusion tissue. In both treetop and lower-crown leaves, the band areas of long (free water: around 3550 cm−1) and short (bound water: around 3200 cm−1) hydrogen bonding OH components showed similar increases with increasing RH, while the band area of free water was larger at the treetop leaves regardless of RH. Free water molecules with longer H bonds were considered to be adsorbed loosely to hydrophobic CH surfaces of polysaccharides in the leaf-cross sections.

Pharmacognostic Profile of Ageratum conyzoides L Plant and Simplicia

Ageratum conyzoides Linn. is known as billy goat weed, including the family Asteraceae. This plant has been used empirically as a medicine for diarrhea, scratches, antinflammatory, snakebite, insecticide and nemasiticide. This study aims to identify plants based on pharmacognitive profiles including macroscopy, microscopy and non-parametric characters from plant parts of Ageratum conyzoides to determine the standardization of herbs. The results showed that the leaves were ovoid, pale green, soft hairy and purple, bluish or white flowers flowers. Microscopic observations of cross section of stem and leaf showed the presence of parenchyma cells, collenchyma cells and multicellular non-glandular trichomes, upper and lower epidermal cells, stomata in the upper and lower epidermis of the leaf. The highest value of water content was in the radix. The highest value of ash content was in the stem. The highest value of acid soluble ash content was in the radix. The highest value of water-soluble extract was in the folium. The highest value of ethanol-soluble extract was in the flower. The results of this study can be used as authentication of plant parts and a basis for the analysis of the standard pharmacopoeia A. conyzoides.

Wettability, water absorption and water storage in rosette leaves of the dragon tree (Dracaena draco L.)

Main conclusionLeaf surfaces of Dracaena draco are wettable and can absorb water. The thick, basal leaf part could act as a water reservoir that changes in volume with plant hydration.Rosettes of leaves of Dracaena draco play an important role in directing fog water through leaf axils into the stem tissues, where it can be stored for further use. However, how water is intercepted and collected by the leaves remains unclear, especially since leaf blade surfaces are considered hydrophobic. Based on the observations of D. draco individuals growing in Spain and in glasshouse conditions in Poland, we hypothesised that their long leaves (~ 70 cm) are able to absorb water along the whole leaf blade, but leaf age affects this process. We used water droplet contact angle measurements, anatomical analyses of leaf cross sections along the age gradient and dye tracer experiments to test this hypothesis. The data showed that the leaf surfaces of D. draco are wettable. In general, the mature leaves of the rosette are more wettable than the young ones. Water can be absorbed both through the adaxial and abaxial surfaces. The hydrenchyma is not uniformly distributed along the leaf, it is especially abundant towards the leaf base where it forms a massive water reservoir, which changes in volume depending on plant water status. The results of these studies shed light on the role of rosettes in water absorption by D. draco, and broaden our understanding of the functioning of this vulnerable species.

A comparative anatomical investigation of three taxa of Brassica L. from Bangladesh

In the present study, micromorphology and comparative anatomy of young root, stem and leaf of three economically important taxa of Brassica L. viz., B. rapa L. subsp. campestris (L.) Clapham, B. juncea (L.) Czern. and B. napus L. are investigated in detail for the first time in Bangladesh by using light microscopy (LM). In anatomical studies, cross sections of young root, stem and leaf have been examined and biometric measurement of cell and tissues are presented. The micromorphological studies are related to the epidermal surface. In addition, the stomatal index and stomatal index ratio of these taxa have been calculated and presence of simple, non-glandular, unicellular trichome on stems and leaves also been observed except on the stem of B. rapa L. subsp. campestris (L.) Clapham. The anatomical study reveals that the investigated taxa have primary growth in roots and stems as well as amphistomatic and bifacial leaves with anisocytic stomata have been noticed. Finally, presence of tetrarch or single strand exarch xylem in vascular bundle of root, the size, shape and presence or absence of trichome over stem, shape of midrib of leaf have been considered to provide reliable features for identification of the Brassica species.
 Bangladesh J. Plant Taxon. 27(1): 15-26, 2020 (June)

Dietary palaeoecology of an Early Cretaceous armoured dinosaur (Ornithischia; Nodosauridae) based on floral analysis of stomach contents.

The exceptionally well-preserved holotype of the armoured dinosaur Borealopelta markmitchelli (Ornithischia; Nodosauridae) from the Early Cretaceous (Clearwater Formation) of northern Alberta preserves a distinct mass within the abdominal cavity. Fourteen independent criteria (including: co-allochthony, anatomical position, gastroliths) support the interpretation of this mass as ingested stomach contents—a cololite. Palynomorphs in the cololite are a subset of the more diverse external sample. Analysis of the cololite documents well-preserved plant material dominated by leaf tissue (88%), including intact sporangia, leaf cross-sections and cuticle, but also including stems, wood and charcoal. The leaf fraction is dominated (85%) by leptosporangiate ferns (subclass Polypodiidae), with low cycad–cycadophyte (3%) and trace conifer foliage. These data represent the most well-supported and detailed direct evidence of diet in an herbivorous dinosaur. Details of the dietary palaeoecology of this nodosaur are revealed, including: selective feeding on ferns; preferential ingestion of leptosporangiate ferns to the exclusion of Osmundaceae and eusporangiate ferns such as Marattiaceae; and incidental consumption of cycad–cycadophyte and conifer leaves. The presence of significant (6%) charcoal may represent the dietary use of recently burned conifer forest undergoing fern succession, early evidence of a fire succession ecology, as is associated with many modern large herbivores.

Anatomical structure of Nitraria spp. leaves from different habitats in Southern Xinjiang, China

The leaf anatomy of the Nitraria roborowskii Kom., Nitraria sibirica Pall., and Nitraria tangutorum Bobr. populations from southern Xinjiang, China, were studied. Cross-sections of the leaves were made, and the structural characteristics were determined. Seven major traits of the leaf cross-sections were analyzed by cluster analysis. The leaf morphology and anatomical structure of Nitraria leaves from different populations significantly differed, but the coefficient of variation between different populations was not significant. Leaf palisade and spongy tissues were positively correlated with the mean temperature, average sunshine, and annual precipitation of the habitat. Populations were divided into three main categories: Class I (XBC-2 and XBC-3), which included plants that grew at higher altitudes and were exposed to longer than average sunshine time; Class II (XBC-1, BC-1, and DBC-1), which included plants that grew at lower latitudes and had longer than average sunshine time; and Class III (BC-2, BC-3, DBC-1, and DBC-2), which included plants that grew at intermediate latitudes. Habitat and climate were the main factors associated with phenotype differences among the populations of the three Nitraria species.

Comparative morphological and leaf anatomical analysis of the species Crocus danubensis and Crocus variegatus (Iridaceae)

Based on previous molecular analysis, it has been established that Crocus reticulatus is not a separate species with wide heterogeneity and distribution, but a species aggregate. The aim of this research is to present, for the first time, a comparative overview of the morphological and leaf anatomical characters of two species from the Crocus reticulatus aggregate, Crocus danubensis and Crocus variegatus, and to indicate the taxonomic significance of the leaf anatomical characters within this taxa. We included 17 quantitative and 13 qualitative characters in the morphological analysis and 20 quantitative parameters in the anatomical analysis, measured on cross sections of the leaves. The degree of variability between the species is displayed using statistical PCA and CDA analyses. The key morphological differences between the species are the color of the perigone and the perigone throat, while the values of all the characters measured are higher for C. variegatus. The key anatomical differences between the species are the size of the largest vascular bundles of leaves (area of the xylem, phloem and sclerenchyma) and the size and shape of the leaf cross sections. Cross sections of the leaves of C. variegatus are smaller, with mostly two ribs and a smaller parenchyma area. The leaf cross sections of C. danubensis are bigger, with four ribs and a larger parenchyma area. The results confirm our hypothesis that the leaf anatomical features are very significant for differentiating these taxa.

MORPHO-ANATOMICAL AND PALYNOLOGICAL STUDIES ON ENDEMIC CRENOSCIADIUM SIIFOLIUM BOISS. & HELDR. FROM TURKEY

The genus Crenosciadium (Apiaceae) is represented by C. siifolium. which is a monotypic endemic species, in the Flora of Turkey. In the current study, morphological, anatomical and palynological features of C. siifolium was described in detail. Expanded descriptions and images of C. siifolium were given. Cross-sections of stem, leaf and fruit are examined and a detailed anatomical description is presented and supported by photographs. Mericarps are almost is pentagonal in transverse section. Each mericarp has 3-5 vittae on the valleculae and 6-8 vittae on the commissural. The pollen morphology of Crenosciadium was studied by SEM. The palynological results confirmed the stenopalynous characteristic of the family Apiaceae, and revealed that the pollen grains of the genera is perprolate in shape. Also fruit surface ornamentation of genera was rugulate-striate at equator and irregular striate at poles.

<p><strong>New morphological and anatomical data on the genus <em>Fuernrohria</em> (Apiaceae)</strong></p>

In this study, morphological and anatomical data (including fruit micromorphological characters) of the genus Fuernrohria were studied and compared with the related genera Grammosciadium, Caropodium and Vinogradovia. The morphological description of Fuernrohria setifolia, which is the only known species in the genus, is expanded. Photos of inflorescences, infructescences and leaves of the species taken from the wild are provided. Leaf segment and mericarps are illustrated, and also the distribution area of the genus is mapped. Anatomical features of root, stem, leaf sheath, leaf segment and fruit are presented with photographs of cross sections for each of them. Vegetative anatomical characteristics of the species and micromorphological description of fruit are given and exhibited for the first time in the present study. The results show that two important diagnostic characters for Fuernrohria are determined for the first time as “shape and size of commissural vittae in mericarps” and “number of vascular bundles in leaf cross section”.

Comparative Anatomical Studies of Some Species of Family Malvaceae from Saudi Arabia and its Systematic significance

In this study, the micromorphological characters of 17 species belonging to seven genera of family Malvaceae present in Saudi Arabia were examined. These characters used in analyzing the phylogenetic relationship among the studied species viz., shape of stem, petiole and leaf in cross section, trichomes, crystals, ducts, star- shape idioblasts, secondary vascular tissue system in stem and petiole vasculature. The anatomical features are used for the preparation of an identification key. Depending on the presence of trichomes, the studied species separate in two main groups; first group contain the species of Althaea, Hibiscus and Malva (their classification depend on the stem vascular system), second group contain the four remaining genera where their classification depend on many attributes such as, cross section of stem, petiole and leaf, arrangement of trichomes, ducts.

MORPHOLOGICAL, ANATOMICAL AND GEOGRAPHICAL DISTRIBUTION STUDIES OF SPECIES Horwoodia dicksoniae (Turrill) IN IRAQ

This research aims to study the morphological anatomical characteristics and geographical distribution of the species Horwoodia dicksoniae (Turrill) belong to Brassicaceae family in Iraq. The results were revealed the morphological characteristics of the root, stem, leaves, fruits and seeds, abaxial surface of the leaf characterized by covered with simple hairs. The fruits were circular in shape with wide wing and have one seed. The seeds were conical shape and pustular surface configuration. The anatomical characteristics of the root, stem, and leaf were studied and the cross-section of the root was characterized by thick periderm layer and cortex layer consisting primarily of parenchyma cells, stem cross section was characterized by thick layer of cuticle, as well as epidermis layer that followed the cuticle layer was double epidermis, but the layer of phloem is composed of rectangular cells and arranged in a continuous ring. Cross section of leaf was bifacial , leaf contains palisade cells in the form of a rectangle at the top surface of the leaf and irregular spongy cells at the low surface and distinguish the middle vein of the leaf as a circular shape. The epidermis was characterized by an Anisocytic type. The study of geographical distribution has been shown that the species is widespread in the desert region of Iraq.

MORPHOLOGICAL CHARACTERISTICS AND POLLINIA OBSERVATION OF 10 INDONESIA NATIVE DENDROBIUM ORCHIDS

Characterization of native orchid (species) in Indonesia is important for preservation of germplasm and selection process to obtain dominant character of orchids as potential parental. Morphological characterization is a method to obtain scientific information in phenotypic manner. Characterization of plants was carried out on 10 native orchids of Dendrobium genus to obtain specific character. Characterization process based on UPOV guidebook and morphological observation of pollinia was also carried out on 10 native orchids to obtain information about the structure and physiology of pollinia. The results of morphological characterization on 10 native orchid showed several qualitative characters such as flowering position, cross section of leaf, pseudobulb firmness, and leaf arrangement in D. Canaliculatum was different from 9 other native orchid. Morphological pollen of 10 native orchid shows the diversity structure of pollinia and the physiological conditions of mature pollen which are ready to pollinate are pollinia with a golden yellow colored. Knowledge and information about parental relationships is the main point for developing ornamental plant breeding programs. Selection of native orchids as potential elder is important where selected elders will be used as a source of genetic diversity. The results of this study are expected to provide scientific information and phenotypic selection for the development of hybrid orchids.

A mechanistic assessment of the potential ecological risk to seagrass meadows posed by marine echosounders

Motivated by experimental evidence, a mechanistic model is used to investigate the impact of marine echosounder transmissions on seagrass leaves. The interaction of the ultrasonic field with the seagrass is solved within the leaf cross-section. The leaf tissue is assumed to be elastic, homogeneous and isotropic, with embedded cavities representing lacunae. A standard Galerkin finite element method is employed to numerically solve the resulting equations. Natural vibration frequencies are computed for leaves of Halodule wrightii and Syringodium filiforme. Strong perturbations in the leaf structure are expected when natural and echosounder frequencies match. The results reveal that a significant number of natural frequencies are within or near the typical frequency bands of echosounder operations. In addition, stresses in the leaf tissues of seagrass at 20 m depth were computed in a typical scenario for echosounder operations. Leaf-tissue damage in S. filiforme could result under these conditions. Within the framework of the theoretical assumptions, the results highlight the potential and, until now, unnoticed risks to seagrass meadows posed by marine echosounders.

<p><strong><em>Festuca devesae </em></strong><strong>(Poaceae), a new species from northern Spain</strong></p>

Festuca devesae (Poaceae), a new species from the low mountains of northern Iberian Peninsula (Burgos and Álava provinces, Spain), is described. According to its morphology, it belongs to Festuca subsect. Festuca, the most diverse group within the genus. The new species differs from other known consectional taxa by having a continuous layer of abaxial subepidermal sclerenchyma in the leaf cross-sections, and by the leaf sheaths closed to the mouth. Both traits are found together only in F. segimonensis and F. hystrix, from which F. devesae clearly differs in the leaf apex shape, the size of the auricles, and the leaf anatomy in cross-section, among other traits. A detailed morphometric description, photographs, and data on leaf anatomy, phenology, ecology, and distribution are given. A comparison with morphologically similar species and an identification key are also provided.

Expression of a Chloroplast-Targeted Cyanobacterial Flavodoxin in Tomato Plants Increases Harvest Index by Altering Plant Size and Productivity.

Tomato is the most important horticultural crop worldwide. Domestication has led to the selection of highly fruited genotypes, and the harvest index (HI), defined as the ratio of fruit yield over total plant biomass, is usually employed as a biomarker of agronomic value. Improvement of HI might then result from increased fruit production and/or lower vegetative growth. Reduction in vegetative biomass has been accomplished in various plant species by expression of flavodoxin, an electron shuttle flavoprotein that interacts with redox-based pathways of chloroplasts including photosynthesis. However, the effect of this genetic intervention on the development of reproductive organs has not been investigated. We show herein that expression of a plastid-targeted cyanobacterial flavodoxin in tomato resulted in significant reduction of plant size affecting stems, leaves, and fruit. Decreased size correlated with smaller cells and was accompanied by higher pigment contents and photosynthetic activities per leaf cross-section. Flavodoxin accumulated in green fruit but declined with ripening. Significant increases in HI were observed in flavodoxin-expressing lines due to the production of higher fruit number per plant in smaller plants. Therefore, overall yields can be enhanced by increasing plant density in the field. Metabolic profiling of ripe red fruit showed that levels of sugars, organic acids, and amino acids were similar or higher in transgenic plants, indicating that there was no trade-off between increased HI and fruit metabolite contents in flavodoxin-expressing plants. Taken together, our results show that flavodoxin has the potential to improve major agronomic traits when introduced in tomato.

Endemik Stachys pumila Banks & Sol. ve Stachys citrina Boiss. & Heldr. (Lamiaceae) Türleri Üzerine Karşılaştırmalı Anatomik ve Mikromorfolojik Bir Çalışma

Bu çalışmada, Türkiye’de yayılış gösteren Infrarosularis Bhattacharjee seksiyonuna ait endemik Stachys pumila Banks ve Sol., S. citrina subsp. citrina ve S. citrina subsp. chamaesideritis taksonları, anatomik ve mikromorfolojik olarak çalışılmış ve karşılaştırılmıştır. Her üç taksona ait Türkiye’nin farklı yerlerinden toplanan örnekler çalışmanın materyalini teşkil etmiştir. Toplanan örneklerin bir kısmı herbaryum materyali haline getirilirken, bir kısmı da %70’lik alkol içinde muhafaza edilmiştir. Anatomik çalışmalar için alkol içindeki örneklerden manuel olarak gövde ve yaprak enine kesitleri ile yaprak yüzeysel kesitleri alınmış ve daimi preparat haline getirilmiştir. Anatomik deskripsiyonlar ve tüy morfolojileri daimi preparatların incelenmesi ve çekilen fotoğrafların değerlendirilmesi ile yapılmıştır. Çalışmadan elde edilen sonuçlar, anatomik karakterler ve tüy morfolojisi karakterlerinin yakın akraba türler olan S. citrina ile S. pumila’nın taksonomik sınıflandırılmasında değere sahip olduğunu, fakat S. citrina’nın alt türlerinin ayrımında değerlerinin sınırlı olduğunu göstermiştir.

<p><strong>A new autumn flowering <em>Crocus</em> (Iridaceae) from SW Turkey: <em>C. terzioghluii</em></strong></p>

Crocus terzioghluii (Iridaceae) is described as a new species endemic to southwestern Anatolia, Turkey. It is related to Crocus babadagensis and differs from this species in having corm tunics with well-developed rings, and prominent stripes on outer segments, as well as in leaf number and flowering time. The corms and tunics of both species are illustrated. A phylogenetic tree obtained by Bayesian phylogenetic inference of the combined sequences of the nuclear rDNA ITS and ETS regions confirms the affiliation of C. terzioghluii to C. ser. Biflori and its close relationship to C. babadagensis. A microphotograph of the metaphase chromosomes (2n = 2x = 8) along with the idiogram and a leaf cross section of the new species are given.