Development and validation of a high-performance liquid chromatography method for quantification of egonol and homoegonol in Styrax species

Styrax camporum Pohl, known in Brazil as estoraque do campo or cuia de brejo, has been used in the treatment of gastrointestinal diseases. The therapeutic action of S. camporum has been attributed to the ethyl acetate fraction, although the chemical composition of this fraction has not yet been analyzed. In this study, a high-performance liquid chromatography photodiode array detection (HPLC-PAD) method for analysis of Brazilian Styrax species has been developed. The compounds egonol (1) and homoegonol (2) were found to be present in all the samples investigated by HPLC. These compounds were isolated by open column chromatography followed by preparative TLC, and were identified by 1H NMR. Compounds 1 and 2 were thus proposed as phytochemical markers for Styrax, owing to their biological properties and presence in other Styrax species. The developed method has been validated and successfully applied for quantification of 1 and 2 in S. camporum dried leaves and crude ethanolic extracts from S. ferrugineus and S. pohlii aerial parts. Copyright (c) 2011 John Wiley & Sons, Ltd.

Germination, Carbohydrate Composition and Vigor of Cryopreserved Caesalpinia echinata Seeds

The present study investigated the germination and vigor of Caesalpinia echinata (Brazilwood) seeds stored at negative temperatures. Recently harvested seeds were cryopreserved at -18 degrees or -196 degrees C and periodically evaluated for germination, seed vigor and carbohydrate composition. The temperatures did not influence the germination percentages or vigor. The germination percentage decreased from 88% in recently harvested seeds to 60% after 730 days of storage. The different temperature and storage times tested did not affect the vigor seed germination as indicated by the measures of plant growth and survival. The different temperatures used did not cause changes in the carbohydrate composition. The tegument cell walls were rich in lignin, arabinose and xylose. The cytoplasm of the cotyledons and embryos had high levels of glucose, fructose, and sucrose. The cryopreservation technique here presented was effective in the conservation of Brazilwood seeds for the medium term.

Biochemical Aspects of a Serine Protease from Caesalpinia echinata Lam. (Brazilwood) Seeds: A Potential Tool to Access the Mobilization of Seed Storage Proteins

Several proteins have been isolated from seeds of leguminous, but this is the first report that a protease was obtained from seeds of Caesalpinia echinata Lam., a tree belonging to the Fabaceae family. This enzyme was purified to homogeneity by hydrophobic interaction and anion exchange chromatographies and gel filtration. This 61-kDa serine protease (CeSP) hydrolyses H-D-prolyl-L-phenylalanyl-L-arginine-p-nitroanilide (K-m 55.7 mu M) in an optimum pH of 7.1, and this activity is effectively retained until 50 degrees C. CeSP remained stable in the presence of kosmotropic anions (PO43-, SO42-, and CH3COO-) or chaotropic cations (K+ and Na+). It is strongly inhibited by TLCK, a serine protease inhibitor, but not by E-64, EDTA or pepstatin A. The characteristics of the purified enzyme allowed us to classify it as a serine protease. The role of CeSP in the seeds cannot be assigned yet but is possible to infer that it is involved in the mobilization of seed storage proteins.

Germination, carbohydrate composition and vigor of cryopreserved Caesalpinia echinata seeds

The present study investigated the germination and vigor of Caesalpinia echinata (Brazilwood) seeds stored at negative temperatures. Recently harvested seeds were cryopreserved at -18º or -196ºC and periodically evaluated for germination, seed vigor and carbohydrate composition. The temperatures did not influence the germination percentages or vigor. The germination percentage decreased from 88% in recently harvested seeds to 60% after 730 days of storage. The different temperature and storage times tested did not affect the vigor seed germination as indicated by the measures of plant growth and survival. The different temperatures used did not cause changes in the carbohydrate composition. The tegument cell walls were rich in lignin, arabinose and xylose. The cytoplasm of the cotyledons and embryos had high levels of glucose, fructose, and sucrose. The cryopreservation technique here presented was effective in the conservation of Brazilwood seeds for the medium term.

RELATIONSHIP AMONG EXTRACTIVES, LIGNIN AND HOLOCELLULOSE CONTENTS WITH PERFORMANCE INDEX OF SEVEN WOOD SPECIES USED FOR BOWS OF STRING INSTRUMENTS

In this paper we investigate the influence of extractives, lignin and holocellulose contents on performance index (PI) of seven woods used or tested for violin bows. Woods with higher values of this index (PI = root MOE/rho, where MOE is modulus of elasticity and rho is density) have a higher bending stiffness at a given mass, which can be related to bow wood quality. Extractive content was negatively correlated with PI in Caesalpinia echinata, Hanclroanthus sp. and Astronium lecointei. In C. echinata holocellulose was positively correlated with PI. These results need to be further explored with more samples and by testing additional wood properties. Although the chemical constituents could provide an indication of quality, it is not possible to establish appropriate woods for bows solely by examining their chemical constituents.

Soil-mediated effects on potential Euterpe edulis (Arecaceae) fruit and palm heart sustainable management in the Brazilian Atlantic Forest

Euterpe edulis is an endangered species due to palm heart overharvesting, the most important non-timber forest product of the Brazilian Atlantic Forest, and fruit exploitation has been introduced as a low impacting alternative. However, E. edulis is a keystone species for frugivores birds, and even the impact of fruit exploitation needs to be better investigated. Since this species occurs over contrasting habitats, the establishment of site-specific standards and limits for exploitation may also be essential to achieve truly sustainable management. In this context, we sought to investigate how soil chemical composition would potentially affect E. edulis (Arecaceae) palm heart and fruit exploitation considering current standards of management. We studied natural populations found in Restinga Forest and Atlantic Rainforest remnants established within Natural Reserves of Sao Paulo State, SE Brazil, where 10.24 ha permanent plots, composed of a grid of 256 subplots (20 m x 20 m), were located. In each of these subplots, we evaluated soil chemical composition and diameter at breast height of E. edulis individuals. Additionally, we evaluated fruit yield in 2008 and 2009 in 20 individuals per year. The Atlantic Rainforest population had a much higher proportion of larger diameter individuals than the population from the Restinga Forest, as a result of habitat-mediated effects, especially those related to soil. Sodium and potassium concentration in Restinga Forest soils, which have strong negative and positive effect on palm growth, respectively, played a key role in determining those differences. Overall, the number of fruits that could be exploited in the Atlantic Rainforest was four times higher than in Restinga Forest. If current rules for palm heart and fruit harvesting were followed without any restriction to different habitats, Restinga Forest populations are under severe threat, as this study shows that they are not suitable for sustainable management of both fruits and palm heart. Hence, a habitat-specific approach of sustainable management is needed for this species in order to respect the demographic and ecological dynamics of each population to be managed. These findings suggest that any effort to create general management standards of low impacting harvesting may be unsuccessful if the species of interest occur over a wide range of ecosystems. (C) 2012 Elsevier B.V. All rights reserved.

FRUGIVORY OF NEOSILBA SPECIES (DIPTERA: LONCHAEIDAE) AND THEPYTUS ECHELTA (LEPIDOPTERA: LYCAENIDAE) ON PSITTACANTHUS (SANTALALES: LORANTHACEAE) IN ECOTONAL CERRADO-SOUTH PANTANAL, BRAZIL

This paper presents a survey of the insects that feed on fruits of Psittacanthus Martius (Santalales: Loranthaceae), a hemiparasitic mistletoe genus that infects trees in Brazil and other neotropical countries. The aim of the study was to identify candidate insects for biological control of Psittacanthus mistletoes. Unripe and mature fruits were collected in several localities of Cerrado, bordering South Pantanal, Southwestern Brazil, from 29 Apr 1998 to 30 Jul 2000. A total of 24,710 fruits (54 samples) of Psittacanthus acinarius infecting 15 species from 10 plant families were evaluated. Psittacanthus acinarius (Mart.) was the most abundant and frequent species of mistletoe parasitizing trees in the ecotonal Cerrado-Pantanal. From 24,710 fruits of Psittacanthus acinarius were obtained 1,812 insect larvae including 1,806 Neosilba McAlpine (Diptera: Lonchaeidae) species and 6 Thepytus echelta (Hewitson) (Lepidoptera: Lycaenidae). From these emerged 1,550 Neosilba spp. adults and 6 T. echelta. Neosilba pantanense Strikis was described from this research. Larvae of T. echelta occurred in fruits of P. acinarius parasitizing Cecropia pachystachya Trecul (Urticaceae) and Anadenanthera colubrina (Vellozo) Brenan (Fabaceae). Larvae of Neosilba caused no adverse effects on the germination of infected fruits of Psittacanthus, because they do not eat the embryo or viscin tissues. This differs from the larvae of T. echelta that interrupted the germination of seeds by feeding on those tissues. Thepytus echelta may be a promising insect for the biological control of P. acinarius in the ecotonal Cerrado-Pantanal, although its abundance and frequency were low throughout the sampling period.

Soil-nutrient availability modifies the response of young pioneer and late successional trees to elevated carbon dioxide in a Brazilian tropical environment

The aim of this work was to determine the impact of three levels of [CO2] and two levels of soil-nutrient availability on the growth and physiological responses of two tropical tree species differing in their ecological group: Croton urucurana Baillon, a pioneer (P), and also Cariniana legalis (Martius) Kuntze, a late succession (LS). We aimed to test the hypothesis that P species have stronger response to elevated [CO2] than LS species as a result of differences in photosynthetic capacity and growth kinetics between both functional groups. Seedlings of both species were grown in open-top-chambers under high (HN) or low (LN) soil-nutrient supply and exposed to ambient (380 mu mol mol(-1)) or elevated (570 and 760 mu mol mol(-1)) [CO2]. Measurements of gas exchange, chlorophyll a fluorescence, seedling biomass and allocation were made after 70 days of treatment. Results suggest that elevated [CO2] significantly enhances the photosynthetic rates (A) and biomass production in the seedlings of both species, but that soil-nutrient supply has the potential to modify the response of young tropical trees to elevated [CO2]. In relation to plants grown in ambient [CO2], the P species grown under 760 mu mol mol(-1) [CO2] showed increases of 28% and 91% in A when grown in LN and HN, respectively. In P species grown under 570 mu mol mol(-1) [CO2], A increased by 16% under HN, but there was no effect in LN. In LS species, the enhancement of A by effect of 760 mu mol mol(-1) [CO2] was 30% and 70% in LN and HN, respectively. The exposure to 570 mu mol mol(-1) [CO2] stimulated A by 31% in HN, but was no effect in LN. Reductions in stomatal conductance (g(s)) and transpiration (E), as a result of elevated [CO2] were observed. Increasing the nutrient supply from low to high increased both the maximum rate of carboxylation (V-cmax) and maximum potential rate of electron transport (J(max)). As the level of [CO2] increased, both the V-cmax and the J(max) were found to decrease, whereas the J(max)/V-cmax ratio increased. In the LS species, the maximum efficiency of PSII (F-v/F-m) was higher in the 760 mu mol mol(-1) [CO2] treatment relative to other [CO2] treatments. The results suggest that when grown under HN and the highest [CO2], the performance of the P species C. urucurana, in terms of photosynthesis and biomass enhancement, is better than the LS species C. legalis. However, a larger biomass is allocated to roots when C. legalis seedlings were exposed to elevated [CO2]. This response would be an important strategy for plant survival and productivity of the LS species under drought stresses conditions on tropical environments in a global-change scenario. (C) 2011 Elsevier B.V. All rights reserved.