Renata Maria Strozi Alves Meira

Plant Origin of Green Propolis: Bee Behavior, Plant Anatomy and Chemistry

Propolis, a honeybee product, has gained popularity as a food and alternative medicine. Its constituents have been shown to exert pharmacological effects, such as anti-microbial, anti-inflammatory and anticancer. Shoot apices of Baccharis dracunculifolia (alecrim plant, Asteraceae) have been pointed out as sources of resin for green propolis. The present work aimed (i) to observe the collecting behavior of bees, (ii) to test the efficacy of histological analysis in studies of propolis botanical origin and (iii) to compare the chemistries of alecrim apices, resin masses and green propolis. Bee behavior was observed, and resin and propolis were microscopically analyzed by inclusion in methacrylate. Ethanol extracts of shoot apices, resin and propolis were analyzed by gas chromatography/mass spectroscopy. Bees cut small fragments from alecrim apices, manipulate and place the resulting mass in the corbiculae. Fragments were detected in propolis and identified as alecrim vestiges by detection of alecrim structures. Prenylated and non-prenylated phenylpropanoids, terpenoids and compounds from other classes were identified. Compounds so far unreported for propolis were identified, including anthracene derivatives. Some compounds were found in propolis and resin mass, but not in shoot apices. Differences were detected between male and female apices and, among apices, resin and propolis. Alecrim apices are resin sources for green propolis. Chemical composition of alecrim apices seems to vary independently of season and phenology. Probably, green propolis composition is more complex and unpredictable than previously assumed.

Leaf anatomy as an additional taxonomy tool for 16 species of Malpighiaceae found in the Cerrado area (Brazil).

This work describes the leaf anatomy of 16 species belonging to three genera of the Malpighiaceae family found in the Cerrado (Minas Gerais State, Brazil). The scope of this study was to support the generic delimitation by contributing to the identification of the species and constructing a dichotomous identification key that includes anatomical characters. The taxonomic characters that were considered to be the most important and used in the identification key for the studied Malpighiaceae species were as follows: the presence and location of glands; presence of phloem in the medullary region of the midrib; mesophyll type; presence and type of trichomes; and presence, quantity, and disposition of accessory bundles in the petiole. It was also possible to indicate promising characters for future taxonomic and phylogenetic studies in the Malpighiaceae family, especially for the Banisteriopsis, Byrsonima, and Heteropterys genera.

Articulated laticifers in the vegetative organs of Mandevilla atroviolacea (Apocynaceae, Apocynoideae)

The occurrence of nonarticulated laticifers has been recorded in the literature for Apocynaceae, but little infor- mation is available on their structure and the chemical compounds in their latex. The objective of this study was to de- scribe, anatomically and histochemically, the laticifers and latex found in the roots, stems, and leaves of Mandevilla atroviolacea (Stadelm) Woodson, a genus in the Apocynaceae. Laticifer anatomy was examined by light microscopy, and the chemical nature of the exuded latex was studied using cytochemistry. Articulated laticifers were present in M. atroviolacea. The lactiferous system in stems and leaves originated from both procambium and ground meristem, and it was determined to be functional, as indicated by changes in the cytoplasm immediately subsequent to its formation. However, in tuberous roots, laticifers originated from the vascular cambium, phellogen, and parenchymatous cortical cells. The primary wall of laticifers consists of pectins and cellulose and is thicker than that of adjacent cells. The laticifer secre- tion contained an emulsion of lipophilic substances rich in neutral lipids, resin oils, and possibly rubber. The presence of articulated laticifers is reported for the first time for Apocynoideae, contributing to the knowledge and understanding of these structures in the family Apocynaceae.

Leaf anatomy and morphometry in three eucalypt clones treated with 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.

Anatomical and histochemical characterization of extrafloral nectaries of Prockia crucis (Salicaceae)

Besides being vital tools in taxonomic evaluation, the anatomy of plant secretory structures and the chemical composition of their secretions may contribute to a more thorough understanding of the roles and functions of these secretory structures. Here we used standard techniques for plant anatomy and histochemistry to examine secretory structures on leaves at different stages of development of Prockia crucis, to evaluate the origin and development of the structures, and to identify the disaccharides and monosaccharides in the exudates. Fructose, glucose, and sucrose constituted up to 49.6% of the entire secretion. The glands were confirmed to be extrafloral nectaries (EFNs); this is the first report of their presence in the genus Prockia. These EFNs are globular, sessile glands, with a central concavity occurring on the basal and marginal regions of the leaf. The epidermis surrounding the concavity is secretory, forming a single-layered palisade that strongly reacts with periodic acid-Schiffs reagent (PAS) and xylidine Ponceau, indicators of total polysaccharides and total proteins, respectively, in the exudate. On the basis of the similarity of these glands to the salicoid teeth in Populus and Salix, we suggest that these three taxa are phylogenetically close.

Efeito do glyphosate sobre a morfoanatomia das folhas e do caule de Commelina diffusa e C. benghalensis

The purpose of this work was to evaluate the effect of glyphosate in the anatomical and morphologic structures of leaves and stems of two species of Commelina, and to investigate the involvement of the starch reservation in larger tolerance to glyphosate of C. diffusa in relation to C. benghalensis. From 10 pots for each species, 9 were applied with 1440 g ha-1 of glyphosate and one was used as check. Fragments of stems and leaves were collected and fixed at time zero (before the application of glyphosate), 15, 30, and 50 days after the application (DAA). The histological slides were obtained according to the usual methodology, with emphasis to the application of the lugol reagent to check starch. Grades from 1 to 5 were given according to the growing intensity of the reaction. In leaves, the epidermal cells and the vascular tissues are just a little bit affected. How seer the mesophyll is completely disorganized, resulting in cell death. Injures are more obvious stems and leaves of C. benghalensis. Morphologically, clorotic regions and deceased spots all over the leaf surface lead to leaf fall after 15° DAA in C. benghalensis. In C. diffusa, the leaf abscission is delayed, despite the injuries are almost the same. Both species presented higher quantities of starch in the node region than the internode. C. benghalensis has few and small starch grains, compared to many and big grains in C. diffusa. In response to glyphosate application , there was a variation in the number of starch grains in the stem to the time after the application. C. diffusa will always have larger possibilities to recover after glyphosate application because of its larger reservation of starch.

Caracteres anatômicos de duas espécies de trapoeraba e a eficiência do glyphosate

The genus Commelina includes weed species of difficult control in several crops, chiefly when the herbicide glyphosate is used repeatedly. This work was conducted to evaluate the differences between the anatomic features of Commelina benghalensis and Commelina diffusa, grown under sun and shade conditions, which could influence herbicide absorption and translocation. The stomatal apparatus of the two species is similar and the leaf is amphistomatic. C. diffusa has a greater number of stomata in relation to C. benghalensis. The number of stomatas in the leaf epiderm is greater in C. diffusa (38/mm2) than in C. benghalensis (33.66/mm2), in the abaxial (54.86/mm2) rather than in the adaxial epiderm (16.80/mm2) and under sun (37.89/mm2) rather than shade conditions (33.77/mm2). The abaxial epiderm has a greater number of stomata under sun conditions. Similar secretory hair is present in both species, but in greater number in C. diffusa. Only C. benghalensis presented tector hair: (i) long with slender extremity and (ii) short with curved extremity. Long hair is concentrated in the abaxial epiderm and short hair in the adaxial epiderm. Although the presence of hair in the leaf epiderm is frequently associated to greater herbicide absorption, it is believed that the determinant factor of greater susceptibility of C. benghalensis to glyphosate in comparison to C. diffusa could be related to the starch stock of the stem. While C. benghalensis presents fewer and smaller starch grains in the pith parenchyma, C. diffusa presents larger and numerous starch grains, which may make herbicide simplastic translocation slower, reducing the herbicide accumulated in the growth points, and allowing it to regrowth even after total leaves loss.

Ontogenetic, anatomical and histochemical study of the extrafloral nectaries of Sapium biglandulosum (Euphorbiaceae)

The present paper aims to confirm the nature of secretory structures found on the petiole and leaf margins of Sapium biglandulosum Muell. Arg. The anatomy, ontogenesis and histochemistry were studied by light microscopy, whereas mono- and disaccharides in the exudates were detected by high performance liquid chromatography. The exudate from the petiole had a total sugar concentration of 32.5% (w/v), of which 38.1% was fructose, 43.7% glucose and 18.2% sucrose. The petiolar gland started its development from a group of meristematic cells that underwent asynchronous divisions. At the end of the ontogenesis, a well structured vascularised gland made up of a palisade secretory epidermis, secretory parenchyma and a secretory pore was observed. Leaf-margin glands showed a similar anatomy. Histochemical tests revealed the presence of proteins, pectins, carbohydrates, tannins and anthocyanins. On the basis of our results, there is compelling evidence that the studied glands are in fact extrafloral nectaries.

Inclusão de material herborizado em metacrilato para estudos de anatomia vegetal

The usual herbalized material processing techniques are limited due to the difficulties to produce permanent slide collections, and to register permanent results. In studies of taxonomy-applied anatomy, it is often necessary to use herbarium specimens. The objective of this study was to test methacrylate (Historesin, Leica) in the preparation and inclusion of herbalized materials to obtain a permanent slide collection. Herbalized leaves from the genus Senecio Tourn. ex L. (Asteraceae) were used, after being submitted to re-dehydration in ethanol 70%. The samples were dehydrated in an ethanol series and submitted to a mixture of AE 95% plus pure resin (1v:1v). The embbeding process in pure resin was submitted to a vaccum during 72 hours. The blocks were cut in an automatic progress microtome, using disposable steel razors and the cuts stained with Toluidine Blue pH 4.0. The slides were mounted in Permount. The permanent slide collection obtained showed high quality, compared with those obtained by the usual techniques, because the cuts were seriate; cut thickness could be determined allowing the use of few samples for the analysis of the material.

Danos visuais e anatômicos causados pelo glyphosate em folhas de Eucalyptus grandis

Symptomatology is one of the most important criteria used to determine the damages caused by biotic and abiotic factors in plants. However, microscopic aspects are needed to elucidate intoxication mechanisms and early injury diagnosis. This work aimed to evaluate the effects of four commercial glyphosate formulations under simulated spray drift (Scout®, Roundup NA®, Roundup transorb® and Zapp QI®) on the morphoanatomy of six Eucalyptus grandis clones (UFV01, UFV02, UFV03, UFV04, UFV05 and UFV06). Following glyphosate application (129.6 g ha-1), symptoms were monitored daily and, at 14 days, leaf samples from the regions without visible symptoms were collected for microscopic analysis. All clones presented chlorosis and necrosis at the fourth day of glyphosate exposure, regardless of the formulation used. The UFV04 clone did not show any anatomical damage. The other clones suffered plasmolysis, cellular collapse, hypertrophy and formation of cicatrisation tissue. However, changes in leaf blade thickness were not observed. Plants exposed to Roundup transorb® presented the greatest visible intoxication percentage. Anatomically, plants exposed to Roundup NA® showed a higher number of damages. Analysis considering both anatomic and visible damages showed that UFV06 clone was the most susceptible to the formulations tested. The results confirmed the diagnostic and prognostic value of visual and anatomical analysis, respectively, and showed that these parameters are essential to evaluate clone susceptibility and the the phytotoxic potential of herbicides.