Anatomy and Histochemistry of Leaf and Stem of Brazilian Endemic Species Mollinedia clavigera Tul

Abstract This study aimed to investigate the anatomy and histochemistry of Mollinedia clavigera leaves and stems through photonic microscopy and scanning electron microscopy. Noteworthy features of leaves were: presence of paracytic stomata on both surfaces; simple as well as bifurcate non-glandular trichomes; prismatic calcium oxalate crystals; flat-convex midrib with a central and two dorsal bundles; concave-convex petiole with a single vascular bundle in open archh. Stems were cylindrical and showed prismatic and styloid crystals in the pith. Histochemical analysis detected lipophilic and phenolic compounds, starch grains and lignified elements such as brachysclereids and fibers. These features may assist in future identifications and quality control of M. clavigera, avoid misidentification between other genus members, once species and genus studies are scarce.

Leaf area estimation for Psychotria carthagenensis and Psychotria hoffmannseggiana as a function of linear leaf dimensions

Determining leaf area is important for studies involving plant growth and development. The aim of the present study was to obtain models for estimating leaf area of Psychotria carthagenensis and Psychotria hoffmannseggiana using linear measurements of leaf blades (length and width). Two hundred leaf blades of each species were collected in Parque Estadual Mata do Pau-Ferro in the municipality of Areia, Paraíba, Northeast Brazil. The equations evaluated for producing potential models included the following: linear, quadratic, potential and exponential. The criteria used to determine the best model(s) were as follows: high coefficient of determination (R²), low root-mean-square error (RMSE), low Akaike information criterion (AIC), high Willmott concordance index (d) and a BIAS ratio close to zero. All evaluated models satisfactorily estimated leaf area for the two species, but the equation ŷ = 0.6373 * LW0.9804 was the most appropriate for P. carthagenensis, while ŷ = 0.6235 * LW0.9712 was the most appropriate for P. hoffmannseggiana.

Origin of the subepidermal tissue in Piper L. leaves / Origem do tecido subepidérmico em folhas de Piper L.

Studies on the anatomy of Piper leaves demonstrate the presence of a subepidermal tissue distinct from the adjacent epidermis, which cells show thin walls and hyaline contents. Some authors consider such cells a hypodermal tissue, while others refer to them as components of a multiple epidermis. In this study, the nature of this subepidermal tissue was investigated through the analysis of leaf ontogeny in three Piper species. The analysis showed that the referred tissue originates from the ground meristem and, thus, should be considered a hypodermis. The studied species suggests that the role of the hypodermis would be to protect the photosynthetic apparatus from excess light, regulating the intensity of light reaching the chlorophyll parenchyma.

Os estudos de anatomia foliar de espécies de Piper revelam a presença de um tecido subepidérmico distinto da epiderme e cujas células apresentam paredes finas e conteúdo hialino. Estas células são referenciadas por alguns autores como um tecido hipodérmico e por outros, como sendo constituintes de uma epiderme múltipla. Nesse estudo verificou-se a natureza deste tecido subepidérmico a partir da análise da ontogênese foliar de três espécies de Piper. A análise revelou que o referido tecido tem origem do meristema fundamental e, portanto, deve ser denominado de hipoderme. Para as espécies avaliadas, sugere-se que a hipoderme teria função de, proteger o aparato fotossintético do excesso de luminosidade, regulando a intensidade luminosa que atinge o parênquima clorofiliano.

Micromorfología de la epidermis foliar de cultivares de arroz Venezolano (Poaceae) asociado con el daño mecánico de sogata Tagosodes orizicolus (Homoptera: Delphacidae) / Micromorphology of leaf epidermis of some Venezuelan rice cultivars (Poaceae) associated with the mechanical damage of Sogata T. orizicolus (Homoptera: Delphacidae)

Rice cultivars are affected directly and indirectly by the insect sogata. The mechanical damage or direct loss, is produced after feeding and ovoposition on the young leaves tissues, while the indirect damage is produced after the transmission of the Rice hoja blanca virus. We studied the morpho-anatomic structures associated with the resistance of the mechanical damage produced by the insect, in six rice cultivars, including controls for resistance and susceptibility (Makalioka and Bluebonnet 50), during August 2011, in Fundacion Danac, Venezuela. Samples were taken from leaf 3, where cuticle thickness, presence of macrohair, microhair and silica bodies in the second third of the leaf was evaluated. A significant difference to thickness of the cuticle, the presence of microhair in the leaves, and presence of silica bodies was observed among cultivars, determining a significant correlation among the number of macrohair and microhair in the adaxial leaf blade with the presence of silica bodies, and thickness of the cuticle and number of posed insects. Thickness of the cuticle and presence of silica bodies in the intercostals space of microhair and macrohair showed to be the variables most related to mechanical damage and resistance mechanism.

El arroz es afectado directa e indirectamente por el insecto sogata. El daño directo o mecánico es debido a la postura de huevos y alimentación del tejido foliar por parte del insecto y el daño indirecto que ocurre es por la trasmisión del virus de la hoja blanca del arroz. El ensayo fue conducido en la Fundación DANAC en agosto 2011, con la finalidad de evaluar las estructuras morfoanatómicas asociadas con la resistencia al daño mecánico en seis cultivares de arroz que incluyen a los testigos de resistencia y susceptibilidad (Bluebonnet 50 y Makalioka). En el tercio medio de la tercera hoja, se evaluaron las variables grosor de la cutícula, presencia de macropelos, micropelos y cuerpos silíceos. Se observaron diferencias significativas entre los cultivares para el grosor de cutícula, número de micropelos y presencia de cuerpos silíceos, se determinó una correlación significativa entre el número de macro-pelos y micropelos en la cara adaxial con la presencia de cuerpos silíceos y el grosor de la cutícula y el número de insectos posados a las 96h. El grosor de la cutícula y la presencia de cuerpos silíceos en los espacios intercostales de los micropelos y macropelos mostraron ser los caracteres más relacionados con la resistencia al daño mecánico y los mecanismos de resistencia al insecto.

An analysis of vascular system in the compound tendrilled afila leaf in Pisum sativum.

Recent work on the venation patterning and morphogenesis of leaf/leaflet has posed the question how different are these in tendrils, which are another type of vegetative lateral organ. Here, the venation patterns of leaflets, stipules and tendrils were compared in the model species, P. sativum. Unlike reticulated venation in leaflets and stipules, venation in tendrils comprised of one or more primary veins. A few secondaries were attached to a primary vein, mostly distally. Bilaterally symmetrical secondary veins were rare. The primary veins in tendrils were daughter strands from dichotomously divided mother veins in rachis, connected finally to vascular strands in stem. A tendril received primary vein from one or more mother strands. Some mother strands contributed primary veins to proximal, distal and terminal domain tendrils of af leaf. The tendrils shared the multi-primary vein character with stipules. Vein redundancy provided a mechanism for survival of tendril/leaf against injury to some of the veins/mother veins. The presence of aborted primary veins that did not reach apex, rows of cambium cells attached to primary vein(s) at apex, the pattern of attachment of primary veins to mother veins and cessation of vein growth in apical direction in aborted tendrils of af lld genotype indicated that the growth of primary veins and tendril was acropetal. Loss-of-function of AF extended the repression of TL and MFP genes on leaflet development from distal and apical domains to proximal domain of leaves in af mutants.

Establishment of an in vitro plantlet regeneration protocol for unique varieties of brinjal (Solanum melongena L.) var. Mattu Gulla and Perampalli Gulla.

Brinjal (Solanum melongena L.) var. Mattu Gulla (MG) and var. Perampalli Gulla (PG) are unique varieties with distinct flavour cultivated in Udupi, Karnataka State, and are exposed to several biotic and abiotic stresses. An efficient and reproducible in vitro regeneration method is required to expedite the manipulation of these brinjal varieties to cope up with stress by tissue culture and gene transfer methods. The present study, reports a rapid and efficient in vitro regeneration protocol for these two varieties. The in vitro growth response was studied on Murashige and Skoog (MS) medium supplemented with 2, 4-D, BAP and IAA, and the plantlets were regenerated efficiently from callus cultures of leaf, cotyledon and hypocotyl explants. Among the three explants, the hypocotyl explants were found to have better callus induction and multiple shoot regeneration. High frequency of shoot initiation was achieved from hypocotyl derived calluses in MS media with 2.0 mg/L BAP and 0.5 mg/L IAA in MG and PG. Efficient and rapid shoot proliferation, and elongation were noted in MS medium with 1.0 mg/L BAP and 0.3 mg/L GA3. The in vitro regenerated shoots produced healthy roots when they were cultured on MS medium supplemented with 0.5 mg/L IBA. A significant difference was observed in percentage of callus induction, number of shoots per callus, shoot elongation and number of hardened plantlets of MG and PG. MG showed maximum response in all stages of culture than PG. Hardening of plantlets in tissue culture was achieved in three weeks. The hardened plantlets were grown in pots for further acclimatization in green house and finally transplanted to experimental garden where they developed into flowering plants and produced mature fruits with viable seeds.

Leaf dimorphism of Microgramma squamulosa (Polypodiaceae): a qualitative and quantitative analysis focusing on adaptations to epiphytism

The epiphytic fern Microgramma squamulosa occurs in the Neotropics and shows dimorphic sterile and fertile leaves. The present study aimed to describe and compare qualitatively and quantitatively macroscopic and microscopic structural characteristics of the dimorphic leaves of M. squamulosa, to point more precisely those characteristics which may contribute to epiphytic adaptations. In June 2009, six isolated host trees covered by M. squamulosa were selected close to the edge of a semi-deciduous seasonal forest fragment in the municipality of Novo Hamburgo, State of Rio Grande do Sul, Brazil. Macroscopic and microscopic analyzes were performed from 192 samples for each leaf type, and permanent and semi-permanent slides were prepared. Sections were observed under light microscopy using image capture software to produce illustrations and scales, as well as to perform quantitative analyses. Fertile and sterile leaves had no qualitative structural differences, being hypostomatous and presenting uniseriate epidermis, homogeneous chlorenchyma, amphicribal vascular bundle, and hypodermis. The presence of hypodermal tissue and the occurrence of stomata at the abaxial face are typical characteristics of xeromorphic leaves. Sterile leaves showed significantly larger areas (14.80cm²), higher sclerophylly index (0.13g/cm²) and higher stomatal density (27.75stomata/mm²) than fertile leaves. The higher sclerophylly index and the higher stomatal density observed in sterile leaves are features that make these leaves more xeromorphic, enhancing their efficiency to deal with limited water availability in the epiphytic environment, compared to fertile leaves.

El helecho epífito Microgramma squamulosa se encuentra en el Neotrópico y tiene hojas estériles y fértiles dimorfas. El objetivo de este estudio fue describir y comparar cuantitativa y cualitativamente la organización estructural de las hojas de la M. squamulosa, investigando las características morfológicas y anatómicas, y señalando los factores que contribuyen a la adaptación al ambiente epífito. Los análisis macroscópicos y microscópicos se realizaron a partir de 192 muestras de hojas de cada tipo. Láminas permanentes y semi-permanentes fueron preparadas y las secciones fueron observadas en el microscopio de luz. Hojas fértiles y estériles no mostraron diferencias estructurales cualitativas, son hipostomáticas, presentan epidermis uniseriada, haz vascular anficrival e hipodermis. La presencia de hipodermis y aparatos estomáticos en la nsuperficie abaxial de la epidermis son típicas de hojas xeromórficas. Hojas estériles presentaran áreas significativamente mayores, tasa más alta de esclerofilia y mayor densidad de estomas que hojas fértiles. La mayor tasa de esclerofilia y mayor densidad de estomas son características que hacen las hojas estériles más xeromórficas, aumentando su eficiencia para hacer frente a la disponibilidad de agua limitada en el ambiente epífito, en comparación con las hojas fértiles.

Relación entre morfología foliar de antófitos y factores abióticos en las principales pluvisilvas de la Región Oriental cubana

Relationship between antophyte foliar morphology and abiotic factors in the main rainforests of Eastern Cuba. The foliar morphology of representative antophytes in four rainforest types of Eastern Cuba was studied in relation to the main abiotic factors. Although there are several leaf types in these forests, the microphyll type is the most important among endemic species in the ophiolites complex and the Montane rainforest. At the Lowland rainforest (metamorphic complex) the mesophyll leaf was the most important. Most foliar epidermis had structures normally found in mesomorphic plants, but xeromorphic and higromorphic morphologies were also present. Rev. Biol. Trop. 57 (1-2): 235-256. Epub 2009 June 30.

Se estudió la morfología foliar de los antófitos que son representativos de cuatro tipos de pluvisilvas de la Región Oriental de Cuba y se relacionó con los principales factores abióticos. Se encontró que existen varios tipos de hojas, pero predominó la micrófila en las especies endémicas y la mesófila, en las no endémicas. En la epidermis foliar se hallaron estructuras que son características de plantas mesomórficas y algunas xeromórficas, pero también se observaron otras que permiten una adaptación a las condiciones de alta humedad atmosférica, las que se consideraron higromórficas.

Influence of leaflet age in anatomy and possible adaptive values of the midrib gall of Copaifera langsdorffii (Fabaceae: Caesalpinioideae)

Gall inducing insects most frequently oviposit in young tissues because these tissues have higher metabolism and potential for differentiation. However, these insects may also successfully establish in mature tissues as was observed in the super-host Copaifera langsdorffii. Among C. langsdorffii gall morphotypes, one of the most common is a midrib gall induced by an undescribed species of Cecidomyiidae. Following this ‘host plant and gall-inducing insect’ model, we addressed two questions: 1) Do the age of the tissues alter the gall extended phenotype? 2) Do gall morphological and anatomical features influence the adaptive value of the galling insect? For anatomical and histometrical studies, transverse sections of young and mature, galled and ungalled samples were prepared. Galls in young leaflets presented higher potential for cell division and greater nutritive reserves, whereas galls in mature leaflets perhaps provide more protection against natural predators and desiccation. Host organ age at the time of oviposition may influence plant cell fates and consequently the interpretation of the adaptive value of insect galls. Rev. Biol. Trop. 57 (1-2): 293-302. Epub 2009 June 30.

Leaf anatomy and morphometry in three eucalypt clones treated with glyphosate / Anatomia e morfometria foliar em clones de eucalipto tratados com glyphosate

This work aimed to evaluate the effects of simulated drift of glyphosate on the morphoanatomy of three eucalypt clones and to correlate the intoxication symptoms on a microscopic scale with those observed in this visual analysis. The effects of glyphosate drift were proportional to the five doses tested, with Eucalyptus urophylla being more tolerant to the herbicide than E. grandis and urograndis hybrid. The symptoms of intoxication which were similar for the different clones at 7 and 15 days after application were characterized by leaf wilting, chlorosis and curling and, at the highest rates, by necrosis, leaf senescence and death. Anatomically glyphosate doses higher than 86.4 g.ha-1 caused cellular plasmolysis, hypertrophy and hyperplasia, formation of the cicatrization tissue and dead cells on the adaxial epidermis. The spongy parenchyma had a decrease, and the palisade parenchyma and leaf blade thickness had an increase. The increased thickness in leaf blade and palisade parenchyma may be related to the plant response to glyphosate action, as a form of recovering the photosynthetically active area reduced by necroses and leaf senescence caused by the herbicide.

Este trabalho teve como objetivo avaliar os efeitos da deriva simulada de glyphosate na morfoanatomia de três clones de eucalipto e correlacionar os sintomas de intoxicação em escala microscópica com aqueles observados à vista desarmada. Os efeitos da deriva do glyphosate foram proporcionais às doses testadas, sendo Eucalyptus urophylla mais tolerante ao herbicida que E. grandis e o híbrido urograndis. Os sintomas de intoxicação foram semelhantes para os diferentes clones testados, tanto aos 7 quanto aos 15 dias após a aplicação, sendo caracterizados, morfologicamente, por murcha, clorose e enrolamento foliar e, no caso das maiores doses, por necrose, senescência foliar e morte das plantas de eucalipto. Anatomicamente, doses de glyphosate superiores a 86,4 g.ha-1 provocaram plasmólise, hipertrofia e hiperplasia celular, formação de tecido de cicatrização e morte das células da face adaxial da epiderme. Observou-se diminuição na espessura do parênquima lacunoso e aumento na espessura do parênquima paliçádico e da lâmina foliar. O aumento na espessura da folha e do parênquima paliçádico podem estar relacionados à resposta das plantas ao glyphosate, como forma de compensar a área fotossinteticamente reduzida pelas necroses e senescência causadas pelo herbicida.

Anatomía micrográfica del folíolo de la palma neotropical Bactris gasipaes (Arecaceae)

Leaf micrografic anatomy of the Neotropical palm Bactris gasipaes (Arecaceae). The economic importance of the palm Bactris gasipaes is growing in the Neotropoical region. We collected leaflets from plants under a chemical fertilization regime and a population of 5000 plants per hectare, in Costa Rica. The variety, Diamantes 10, has an ascendency fom the upper Amazon basin. We used Harries hematoxiline, eocine and standard light microscopy techniques. The presence of raphids and buliform cells was confirmed for the abaxial surface of the leaflets and for the hypodermic tissue on both sides. The absence of the Krantz anatomy was confirmed in consistence with former observations about the C3 photosynthesis in other species of Palmaceae. The average stomatal density on the abaxial surface was 96.87±16.31 stomata.mm-2 and 14.20±4.05 in the adaxial surface. Rev. Biol. Trop. 56 (2): 951-959. Epub 2008 June 30.

Se estudiaron hojas de plantas jóvenes cultivadas para producción de palmito de la variedad Diamantes 10 de ascendencia del Alto Amazonas. Se estudió la anatomía micrográfica foliar y la frecuencia de estomas en la superficie adaxial y abaxial de los foliolos de Bactris gasipaes provenientes de una plantación experimental en un régimen de fertilización química y una densidad de población de 5000 plantas por hectárea, en Guápiles, Costa Rica. Se confirmó la presencia de rafidios y células buliformes en la superficie abaxial de las láminas foliares y del tejido hipodermico en ambos lados. Confirmamos la ausencia de la anatomía Krantz en esta especie, reafirmando la fotosíntesis C3 como mecanismo metabólico en la familia Palmaceae. La densidad promedio de estomas en la superficie abaxial fue de 96,87±16,31 estomas.mm-2 y en la superficie adaxial fue de 14,20±4,05 estomas.mm-2.

Isolation and intracellular localization of insulin-like proteins from leaves of Bauhinia variegata

Evidence based on immunological cross-reactivity and anti-diabetic properties has suggested the presence of insulin-like peptides in plants. The objective of the present study was to investigate the presence of insulin-like proteins in the leaves of Bauhinia variegata ("pata-de-vaca", "mororó"), a plant widely utilized in popular medicine as an anti-diabetic agent. We show that an insulin-like protein was present in the leaves of this plant. A chloroplast protein with a molecular mass similar to that of bovine insulin was extracted from 2-mm thick 15 percent SDS-PAGE gels and fractionated with a 2 x 24 cm Sephadex G-50 column. The activity of this insulin-like protein (0.48 mg/mL) on serum glucose levels of four-week-old Swiss albino (CF1) diabetic mice was similar to that of commercial swine insulin used as control. Further characterization of this molecule by reverse-phase hydrophobic HPLC chromatographic analysis as well as its antidiabetic activity on alloxan-induced mice showed that it has insulin-like properties. Immunolocalization of the insulin-like protein in the leaves of B. variegata was performed by transmission electron microscopy using a polyclonal anti-insulin human antibody. Localization in the leaf blades revealed that the insulin-like protein is present mainly in chloroplasts where it is also found associated with crystals which may be calcium oxalate. The presence of an insulin-like protein in chloroplasts may indicate its involvement in carbohydrate metabolism. This finding has strengthened our previous results and suggests that insulin-signaling pathways have been conserved through evolution.

Morphology and anatomy of a leaf mine in Vismia guianensis (Aubl) Choisy (Clusiaceae) in a fragment of Brazilian Atlantic forest

Minas ou hiponomas são canais causados pelo consumo de tecidos internos da planta por larvas minadoras que habitam na folha e se alimentam desses tecidos. Minas em folhas são freqüentemente visíveis na folha em forma de serpentina, manchas ou outras formas características de tecido descolorido. Modificações morfológicas e anatômicas causadas por minadores em folhas de Vismia guianensis (Aubl.) Choisy são apresentadas neste trabalho, com o objetivo de elucidar a sua estrutura e contribuir com os estudos sobre interação inseto-planta. Folhas sadias e minadas de trinta indivíduos foram coletadas e levadas ao laboratório. Para cada ramo coletado foram mensurados o número total de folhas jovens e maduras, o número de folhas minadas e o número de minas por folha. Secções transversais de folhas sadias e minadas foram obtidas para observar aspectos morfo-anatômicos. Das 1.438 folhas maduras e 379 folhas jovens analisadas, 214 (14,9%) e 70 (18,5%) estavam minadas e muitas dessas folhas (67%) apresentavam uma única mina. Secções transversais das folhas minadas revelaram que a larva consome apenas tecido parenquimático do mesofilo e a epiderme permanece intacta. Não foi observado um padrão das minas foliares, mas a larva não consome as drusas observadas no mesofilo e os tecidos vasculares das nervuras central e laterais.