Leaf anatomy of Araucaria angustifolia (Bertol.) Kuntze (Araucariaceae)

Immature and mature leaves of juvenile and adult plants of Araucaria angustifolia (Araucariaceae) were observed with the objective of updating the morphoanatomical data of the leaves of this species, which were restricted to basic descriptions in previous studies. The observations, made in optical allowed to establish anatomical differences among mature leaves of juvenile and adult plants in relation to the number of palisade parenchimal layers, the number of compartmented cells and the transfusion tissue development. Epidermis, the albuminous cells, the phloem, and the transfusion tissue descriptions are in disagreement with the data obtained data by different authors. The epidermal tissue and the hypodermis differ entirely when the plant is still juvenile, being inferred that these tissues would soon perform the protection function against mechanical damages and water loss, the vital characteristics during the first development months of young offspring.

Leaf anatomy variation within and between three "restinga" populations of Erythroxylum ovalifolium Peyr: (Erythroxylaceae) in Southeast Brazil

Erythroxylum ovalifolium is a woody shrub widespread in the "restinga", i.e. the open scrub vegetation of the Brazilian coastal sandy plains. We examined leaf anatomy variation of this species both within populations and between populations of three "restingas" in the state of Rio de Janeiro. Sites were ca.100 km far from each other and differed in regard to rainfall and vegetation structure: a dry, open site; a wet, dense site and an intermediate one. Microhabitats within sites were: (i) exposed to full irradiance, outside vegetation islands; (ii) partially exposed to full irradiance, at the border of vegetation islands; (iii) shaded, inside vegetation islands. Leaf anatomy parameters were measured for five leaves collected in each of five plants per microhabitat, in each population; they were thickness of the leaf blade, of the palisade and spongy parenchyma, and of the adaxial and abaxial epidermis. Leaves from the dry, open site had narrower abaxial epidermis and a smaller contribution of spongy parenchyma to total leaf blade thickeness than the other two sites, which we attributed to water stress. Adaxial epidermis and leaf are thicker in more exposed microhabitats (i and ii, above), irrespective of site. We proposed that between-site anatomical variation in traits related to water stress, and within-site anatomical variation in traits related to light-use are indicative of ecological plasticity and might help explain the high abundance of E. ovalifolium in the studied populations and along the State of Rio de Janeiro coast.

Brazilian soybean Glycine max (L.) Merr. cultivars adapted to low latitude regions: seed composition and content of bioactive proteins

Among the goals of the Brazilian soybean improvement programmes, the breeding strategies for cultivars adapted to low latitudes have been included to extend crop areas and to increase production. Seeds of nine Brazilian soybean cultivars adapted to low latitudes were investigated regarding to their composition, and amino acid and antinutritional/toxic protein contents. Protein (394.5 ± 13.1 to 445.3 ± 8.0 g kg-1 dry matter) and oil (200.6 ± 1.2 to 232.3 ± 4.7 g kg-1 dry matter) contents showed low correlation to each other (r = -0.06). The total carbohydrate (141.7 ± 6.1 to 211.1 ± 15.0 g kg-1 dry matter) and ash contents (48.2 ± 4.2 to 52.2 ± 0.5 g kg-1 dry matter) were similar to data available for other soybean cultivars. All soybean cultivars presented low levels of tryptophan and sulphur amino acids. The lectin (1,152 to 147,456 HU kg-1 flour), trypsin inhibitor (34.45 ± 2.28 to 77.62 ± 2.63 g trypsin inhibited kg-1 flour), toxin (6,210 ± 134 to 34,650 ± 110 LD50 kg-1 flour) and urease (0.74 ± 0.02 to 1.22 ± 0.10 g kg¹ flour) presented variations in their contents amongst the cultivars. Compared to other soybean cultivars, urease was higher, the acute toxicity lower and the lectin and trypsin inhibitor contents similar to data available. In general, soybean cultivars showed similar biochemical composition to those developed in different geographic regions. The relevance of these findings to the agronomic features and to choice of soybean cultivars to be used as food or feed is discussed.

Harvesting of Protium heptaphyllum (Aubl.) March. seeds (Burseraceae) by the leaf-cutting ant Atta sexdens L. promotes seed aggregation and seedling mortality

The role played by leaf-cutting ants as seed dispersers of non-myrmecochorous plants remains poorly understood. Here we document the harvesting of Protium heptaphyllum (Aubl.) March. seeds (Burseraceae) by the leaf-cutting ant Atta sexdens L. and its consequences for (1) seed deposition pattern; (2) seed germination; and (3) seedling mortality. The study was carried out at Dois Irmãos, a 390 ha reserve of Atlantic forest, northeast Brazil. Ant-seed harvesting on the ground was detected in 18.5% of all fruiting trees and ants harvested 41.1% ± 19.7% of the seed crop (mean ± s). In average, ants piled seeds 3.4 ± 2.2 m away from the trunk of parent trees and seed density in these piles reached 128.8 ± 138.8 seeds 0.25 m² during the peak of seed discarding by ants. During a 13 month period, mean seedling mortality varied from 0.54% up to 10.6% in ant-made seed piles vs. 0.05-4.2% in control samples, what resulted in a total seedling mortality of 97.7% vs. 81%. Ants systematically cut seedling epicotyls, accounting for 55% of seedling mortality in seed piles, whereas only 14 seedlings (4.2%) were cut by ants in the control samples. Our results suggest that seed harvesting by A. sexdens (1) affects approximately 20% of fruiting P. heptaphyllum trees and their seed crops; (2) promotes short-distance seed dispersal and high levels of seed aggregation; and (3) reduces seedling survival beneath parents.

Leaf anatomy of two Anemia Sw. species (Schizaeaceae-Pteridophyte) from a rocky outcrop in Niterói, Rio de Janeiro, Brazil

The ferns Anemia tomentosa (Sav.) Sw. var. anthriscifolia (Schrad.) Mickel and Anemia villosa Humb. & Bonpl. ex Willd. are widely associated with vegetation islands on rocky outcrops in Rio de Janeiro. Both species are desiccation tolerant. The leaf anatomy of these species was examined aiming to identify morphological characteristics that would allow the establishment of these species in water-scarce environments. The plants were harvested on "Pedra de Itacoatiara" and prepared according to the usual procedures. The petiole has a uniseriate epidermis with lignified cell walls, conical stegmata, and uniseriate multicelular and glandular trichomes. In A. villosa, the stomata protrude in a respiratory line. Under the epidermis the cells have thick, lignified walls. The parenchyma has phenolic compounds and starch grains. The petiole vascular bundles are surrounded by endodermis with Casparian strips and the xylem is V-shaped (A. villosa) or arc-shaped (A. tomentosa var. anthriscifolia). The leaf blades have a uniseriate epidermis with sinuous anticlinal and convex periclinal walls, conical stegmata and chloroplasts on both surfaces. The leaf margins of A. villosa have lignified cells. The guard cells of the stomata on the abaxial surface are on the same level or are raised above ordinary epidermal cells. Multicelular uniseriate trichomes and glandular hairs were observed. The dorsiventral mesophyll has loosely packed chlorenchyma with arm-shaped and H-shaped cells. The vascular bundles are surrounded by endodermis with Casparian strips and with parenchymatic extensions towards the epidermis. Anatomical results were analyzed considering the interaction of these plants with abiotic factors.

Anatomical and developmental aspects of leaf galls induced by Schizomyia macrocapillata Maia (Diptera: Cecidomyiidae) on Bauhinia brevipes Vogel (Fabaceae)

Samples of healthy leaves and galls induced by Schizomyia macrocapillata Maia on Bauhinia brevipes Vogel were submitted to routine techniques to investigate gall anatomy and development. Pouch galls are induced on the abaxial surface of unfolded immature leaves, and become spheroid with long reddish hairs covering their external surface. Galls occur isolated or coalesce when in larger numbers. Gall development was divided into six phases: 1) initiation; 2) tissue re-arrangement; 3) tissue differentiation; 4) maturation; 5) growth phase; and 6) dehiscence. This last phase corresponds to gall senescence, which takes place just after the larva exits the chamber to pupate. An important developmental phase of tissue reorientation was recorded after the initiation phase. The presence of hyphae close to the covering layer characterizes this gall as an ambrosia gall and the feeding mode of the gall migde is discussed. Few hyphae were found during the first developmental phases and fungi may play an important role during gall morphogenesis. Neoformed trichomes may provide not only photoprotection but also protection against natural enemies and water loss. The neoformation of phloematic bundles suggests host manipulation and indicates the establishment of a deviating sink.

Leaf damage in a mangrove swamp at Sepetiba Bay, Rio de Janeiro, Brazil

Leaf damage to Rhizophora mangle L., Avicennia schaueriana Stapf. & Leechman, and Laguncularia racemosa L. was studied in a two hectare mangrove swamp site in Sepetiba Bay. Seventeen arthropod morphospecies were identified as being responsible for the damage, and their species diversity was highest on A. schaueriana, followed by R. mangle and L. racemosa. Damage in terms of relative area was greatest in L. racemosa. Almost 9% of mangrove canopy leaf area demonstrated some damage. Loss of leaf area to herbivory was 12.1%, 8.3% and 6.2% in L. racemosa, A. schaueriana and R. mangle respectively. L. racemosa and A. schaueriana showed high percentages of leaf-margin damage in terms of the total damaged leaf area (82.2% and 56.3% respectively), while the most important type of damage in R. mangle was necrosis, representing 58.1% of the total damaged leaf area.

Morphological, anatomical and biochemical studies on the foliar galls of Alstonia scholaris (Apocynaceae)

Morphological, anatomical and biochemical alterations in foliar galls of Alstonia scholaris R. Br. induced by the insect Pauropsylla tuberculata (Psyllidae) are described and quantified. Galls occur isolated or agglomerated on the abaxial surface of the leaf. The insect along with the egg deposits some physiologic fluid which act as a stimulant for the induction of the gall. This stimulus brings about hypertrophy followed by hyperplasia of cells next to the location of the deposited eggs. The psyllid presents three nymphal instars, from eclosion of the egg to the adult. Hyperplasia in the palisade cells is very distinctly noticed. Hypertrophy followed by hyperplasia takes place and brings about elevation of hypodermal and palisade parenchyma which undergoes repeated anticlinal divisions. Neoformation of phloematic bundles were distinctly noticed close to the site of infection. With an increase in the growth of the gall, chlorophyll content in the gall tissue decreases. A steady increase of sugar content is noticed. The immature galled tissue showed almost two fold increases in the protein content. The mature galled tissue showed a very high increase in the proline content compared to the immature galled tissue indicating a stressed condition of the galled tissue.

Allelopathic interference of Sapindus saponaria root and mature leaf aqueous extracts on diaspore germination and seedling growth of Lactuca sativa and Allium cepa

Sapindus saponaria (soapberry) is a species that presents a great diversity of chemical compounds, such as saponins; however, few studies have examined the allelopathic effect of this species. Therefore, this study provides an evaluation of the allelopathic potential of aqueous extracts of the roots and mature leaves of S. saponaria on the germination of diaspores and seedlings growth of lettuce (Lactuca sativa) and onion (Allium cepa). The aqueous extract was prepared in the proportion of 100 g of dry plant material in 1,000 mL of distilled water (a concentration of 10% w v-1), and diluted with distilled water to 7.5, 5.0 and 2.5% concentrations. The mature leaf extracts caused delay and decrease in the germination process of the lettuce and onion diaspores, with inhibitory effect concentration-dependent, while the root extracts showed no allelopathic effects on the germination process. Both extracts caused abnormalities and inhibited the growth of shoot and root seedlings.

Leaf and calycine colleters in Odontadenia lutea (Apocynaceae - Apocynoideae - Odontadenieae): their structure and histochemistry

The structure and histochemistry of colleters found on the vegetative and floral apices of Odontadenia lutea are described. Colleters occur on vegetative apices starting at the fourth node, with 68 to 80 colleters being found at each node. Each leaf primordium has only one colleter of axillary origin, 3-5 intra-petiolar, and 12-16 inter-petiolar (intra-stipular). There are four types of colleters: standard, bipartite standard, sessile, and bipartite sessile. Colleters on the reproductive apices alternate with the sepals and are sessile, reduced sessile, tripartite laminar sessile, or asymmetrical. All of the colleters have a central nucleus of parenchymatous cells covered by a palisade uniseriate secretory epidermis and a thin cuticle. Secretory idioblasts were observed in the parenchymatous axis. Vascularization was observed only in standard axillary and laminar colleters. Crystals were observed in the parenchyma of the axillary colleter. Histochemical tests demonstrated that there was no rupturing or distension of the cuticle during the secretion process. Mucilage was identified using the PAS reaction as well as by Mayer's reagent and Ruthenium red staining. The calycine colleters had two distinct secretory phases, the first synthesizing mucilage and the second producing phenolic compounds.

Relationships between crown architecture and available irradiance in two cerrado species with different leaf phenologies

Structural differences between cerrado species with different leaf phenologies are linked to crown architecture, leaf production, and biomass allocation to shoots and leaves. The present study characterized crown structures and the patterns of biomass allocation to leaves and shoots in two woody cerrado species with contrasting leaf phenologies and quantified the irradiance reaching their leaves to determine the best period during the day for photosynthetic activity. The shoots and leaves of five individuals of both Annona coriacea (deciduous) and Hymenaea stigonocarpa (evergreen) were collected along a 50 m transect in a cerrado fragment within the urban perimeter of Catalão - GO, to determine their patterns of biomass allocation in their crowns. The evergreen H. stigonocarpa had significantly higher mean values of shoot inclination (SI), petiole length (PL), leaf area (LA), leaf display index (LDI), and individual leaf area per shoot (ILA), while the deciduous species A. coriacea had significantly higher leaf numbers (LN). The more complex crown of H. stigonocarpa had shoots in more erect positions (orthotropic), with intense self-shading within shoots; A. coriacea, on the other hand, had slanting (plagiotropic) shoots in the crown, allowing similar irradiance levels to all leaf surfaces. The production of plagiotropic shoots by the deciduous species (A. coriacea) is a strategy that enables its use of incident sunlight early in the morning and preventing excessive water loss or excessive irradiance. Hymenaea stigonocarpa (an evergreen), by contrast, had orthotropic shoots and uses intense self-shading as a strategy to avoid excessive irradiance, especially at midday. Differences in crown architectures between evergreen and deciduous species of cerrado sensu stricto can therefore be viewed as adaptations to the environmental light regime.

Preparation of mitotic chromosomes of leaf-cutting ants from the genera Atta and Acromyrmex

Some modifications were made to the methodology of Imai et al. (Jpn. J. Genet. 63: 159-185, 1988) for cytogenetic analysis of the leaf-cutting ants Atta sexdens piriventris and Acromyrmex heyeri (Hymenoptera, Formicidae), shortening preparation time and improving chromosomal preparations. The brain ganglia of prepupae were dissected in a 0.0025% hypotonic solution of colchicine, placed on a glass slide on a cold plate (4 ± 1oC) for 20 min. The material was fixed directly on the cold slide (with cold fixative I), macerated with a histological needle and fixed again with fixative I, followed by fixatives II and III, all of them cold. The slide was flame-dried right after the use of fixative III, and it was allowed to air-dry at room temperature for 2 h. The resulting metaphases presented less contracted chromosomes, with separated and well defined sister chromatids at a high frequency, when the material was processed in the manner described and stained with 3% Giemsa in phosphate buffer (pH 6.8) for 15 min.

Identification of reciprocal hybrids in citrus by the broadness of the leaf petiole wing

Broadness of leaf petiole wing (WB) was investigated as a morphological marker for screening hybrids of the very narrow-winged species Citrus limonia and C. sunki with broad-winged species C. aurantium and C. sinensis. Controlled polinizations produced over 500 reciprocal hybrids with potential in the ongoing rootstock breeding program identified by the Pgi-1 and Prxa-1 isozyme loci. Measurement ratios WB/leaf length, WB/leaf broadness and WB/petiole length identified 86 to 91% of the reciprocal hybrids produced. However, visual classification of WB was an equally efficient but much easier and faster method. It can be very useful in breeding programs when large number of plants have to be screened or when isozyme, RFLP or RAPD laboratories are not available.

Changes in N2 fixation in Stylosanthes scabra derived from tissue culture

Plants were regenerated from leaf-derived callus culture of Stylosanthes scabra, a polyploid legume tolerant to drought and adapted to acid soils. A total of 168 regenerants were planted out in Leonard jars in a complete randomized design. Nitrogen fixation and vegetative growth were indirectly evaluated by shoot dry weight, root dry weight, shoot N content and acetylene reduction activity. The results showed higher variation in the regenerants than in controls not submitted to tissue culture. Significant differences were found for all nitrogen fixation related-traits

Dose-dependent effects of 17-ß-estradiol on pituitary thyrotropin content and secretion in vitro

We studied the basal and thyrotropin-releasing hormone (TRH) (50 nM) induced thyrotropin (TSH) release in isolated hemipituitaries of ovariectomized rats treated with near-physiological or high doses of 17-ß-estradiol benzoate (EB; sc, daily for 10 days) or with vehicle (untreated control rats, OVX). One group was sham-operated (normal control). The anterior pituitary glands were incubated in Krebs-Ringer bicarbonate medium, pH 7.4, at 37oC in an atmosphere of 95% O2/5% CO2. Medium and pituitary TSH was measured by specific RIA (NIDDK-RP-3). Ovariectomy induced a decrease (P<0.05) in basal TSH release (normal control = 44.1 ± 7.2; OVX = 14.7 ± 3.0 ng/ml) and tended to reduce TRH-stimulated TSH release (normal control = 33.0 ± 8.1; OVX = 16.6 ± 2.4 ng/ml). The lowest dose of EB (0.7 µg/100 g body weight) did not reverse this alteration, but markedly increased the pituitary TSH content (0.6 ± 0.06 µg/hemipituitary; P<0.05) above that of OVX (0.4 ± 0.03 µg/hemipituitary) and normal rats (0.46 ± 0.03 µg/hemipituitary). The intermediate EB dose (1.4 µg/100 g body weight) induced a nonsignificant tendency to a higher TSH response to TRH compared to OVX and a lower response compared to normal rats. Conversely, in the rats treated with the highest dose (14 µg/100 g body weight), serum 17-ß-estradiol was 17 times higher than normal, and the basal and TRH-stimulated TSH release, as well as the pituitary TSH content, was significantly (P<0.05) reduced compared to normal rats and tended to be even lower than the values observed for the vehicle-treated OVX group, suggesting an inhibitory effect of hyperestrogenism. In conclusion, while reinforcing the concept of a positive physiological regulatory role of estradiol on the TSH response to TRH and on the pituitary stores of the hormone, the present results suggest an inhibitory effect of high levels of estrogen on these responses