Associação micorízica em espécies arbóreas, atividade microbiana e fertilidade do solo em áreas degradadas de cerrado

The recuperation of areas used during the construction of the hydroelectric plant, especially in 'borrowed areas', is a difficult and long process since all vegetation and the fertile layer of soil were removed. Interventions in these degraded areas could accelerate the revegetation process. The objective of this research was to evaluate the association of arbuscular mycorrhizal fungi (AMF) in tree species, microbial activity (basal respiration) and fertility of 'cerrado' degraded areas. Soil from two areas, pasture soil and exposed subsoil, were utilized. Organic and mineral fertilization, and liming, were added to the pit for better seedlings' initial growth, where 50 mL of preserved cerrado soil was applied as inoculum of microorganisms. Seedlings of 11 tree species were planted: Anadenanthera falcata (Benth.) Speg ('angico-preto'), Acacia polyphylla D. C. ('monjoleiro'), Stryphnodendron adstringens (Mart.) Coville ('barbatimao'), Dimorphandra mollis Benth ('faveiro'), Hymenaea stigonocarpa Mart. ex Hayne ('jatoba-de-cerrado'), Dipteryx alata Vog. ('baru'), Machaerium acutifolium Vogel ('jacaranda-do-campo'), Schinus terebinthifolia Raddi ('aroeirapimenteira'), Magonia pubescens St. Hil. ('tingui'), Lafoensia pacari St. Hil. ('dedaleira') and Tabebuia aurea (Manso) Benth. & Hook ('ipe-amarelo'). Twelve months later, root samples were colleted at the depth of 0-0.10 m and used for evaluations. The subsoil, as compared to pasture soil, was poor in organic matter and presented less microbial activity. The highest mycorrhizal colonization was seen in the species Acacia polyphylla D. C. (monjoleiro), Magonia pubescens St. Hil. (tingui), Hymenaea stigonocarpa Mart. ex Hayne (jatoba-de-cerrado) and Schinus terebinthifolia Raddi (aroeira-pimenteira). These species could be indicated in revegetation projects in 'cerrado' degraded areas. Plants from both areas showed seedlings form high mycorrhizal colonization and low numbers of spores.

Sobrevivência e crescimento inicial de espécies arbóreas nativas do Cerrado em consórcio com mandioca

O objetivo deste trabalho foi avaliar a sobrevivência e o crescimento inicial de seis espécies arbóreas do Cerrado em consórcio com mandioca (Manihot esculenta), com ou sem adubação fosfatada. Utilizou-se o delineamento experimental de blocos ao acaso, em arranjo fatorial 6x2x2, com quatro repetições. As variáveis consideradas foram as espécies: Anadenanthera colubrina var. cebil (angico-vermelho), Anacardium occidentale (caju), Dipteryx alata (cumbaru), Hymenaea stigonocarpa (jatobá), Hancornia speciosa (mangaba) e Sclerolobium paniculatum var. rubiginosum (taxi-branco), em monocultivo ou em consórcio com mandioca, com ou sem adubação fosfatada. Uma distância fixa de 3x3 m foi usada para as espécies arbóreas e de 1,00x0,60 m para a mandioca. Até a idade de 20 meses, as espécies arbóreas foram avaliadas quatro vezes quanto à altura e ao diâmetro do coleto. A produção de biomassa da mandioca foi avaliada aos 20 meses. A taxa média de sobrevivência das espécies arbóreas foi crescente na seguinte ordem: cumbaru (79%), taxi-branco (86%), jatobá (95%), mangaba (98%), angico (99%) e caju (100%). O taxi-branco apresentou maiores taxas de crescimento relativo em diâmetro e altura, enquanto jatobá e cumbaru apresentaram os menores valores. A adubação fosfatada favoreceu apenas ao taxi-branco. A produtividade de mandioca não foi afetada pelo consórcio. Caju, angico-vermelho e taxi-branco são as espécies mais indicadas para o consórcio com mandioca no Cerrado.

Efeito de Brachiaria decumbens na herbivoria e no desenvolvimento de duas leguminosas nativas de cerrado

Este estudo avaliou o impacto da presença da gramínea exótica (Brachiaria decumbens) na herbivoria e no desenvolvimento de plântulas de espécies nativas (Hymenaea stigonocarpa e Dipteryx alata) de Cerrado. Para avaliar o impacto da presença da gramínea, 10 blocos em pastagem (antiga área de Cerrado) na Embrapa Gado de Corte (Campo Grande-MS) foram demarcados em janeiro de 2009. As gramíneas foram mantidas intactas em metade dos blocos e, na outra metade, foram removidas. Em cada tratamento, foram avaliados a herbivoria, a altura e o diâmetro na altura do solo (DAS) de 25 plântulas de cada espécie ao longo do ano. O conteúdo de carbono, nitrogênio e água das folhas também foi quantificado. Plântulas de Hymenaea stigonocarpa nos tratamentos com gramínea tiveram maior taxa de crescimento em diâmetro que as plântulas no tratamento sem gramíneas. As plântulas de Dipteryx alata que estavam no tratamento sem gramíneas tiveram maiores taxas de crescimento em altura que as do tratamento com gramíneas. Houve diferença na taxa de herbivoria de D.alata entre os tratamentos. As plântulas das duas espécies que estavam no tratamento com gramínea mostraram maiores concentrações de carbono e menores de água. O conteúdo de nitrogênio foliar foi maior no tratamento com gramíneas que no sem gramíneas em D.alata; já o nitrogênio em H. stigonocarpa não variou entre os tratamentos. Esses resultados mostram que cada espécie nativa possui distintas respostas fisiológicas e suscetibilidade à herbivoria em ambientes sob competição de recursos com a gramínea exótica, B. decumbens.

Germination, initial growth, and biomass allocation in three native Cerrado species

The parameters of germination, initial growth, and biomass allocation of three native plant species of Cerrado (Copaifera langsdorffii, Dipteryx alata and Kielmeyera coriacea) were established. The species had germination percentages above 88% and average germination times longer than 139 hours. The average time for the opening of the first leaf pair was more than 538 hours for all three species. The average root length of C. langsdorffii and D. alata seedlings after 80 days of growth was around 40cm, four times larger than the average shoot length (<10cm), although the root and shoot biomasses were similar for both species. The average root length (>20cm) of K. coriacea seedlings was four times larger than the average shoot length (<5cm), and the root biomass was 243% greater than the shoot biomass. Increase in seedling biomass was sustained primarily by the cotyledons in C. langsdorffii and D. alata, which acted as reserve organs and showed progressive weight reductions. Increase in seedling biomass in K. coriacea was sustained primarily by photosynthesis, since the cotyledons showed no significant weight reduction, acting primarily as photosynthetic organs. The length of the root systems was at least four times larger than the length of the shoots parts in all three species. Higher investment in root length rather than in root biomass suggest that the initial growth of these species is primarily to ensure access to water resources, apparently putting off the function of the radicular system as storage organ.

Antioxidant And Anti-diabetic Potential Of Passiflora Alata Curtis Aqueous Leaves Extract In Type 1 Diabetes Mellitus (nod-mice).

Leaves of Passiflora alata Curtis were characterized for their antioxidant capacity. Antioxidant analyses of DPPH, FRAP, ABTS, ORAC and phenolic compounds were made in three different extracts: aqueous, methanol/acetone and ethanol. Aqueous extract was found to be the best solvent for recovery of phenolic compounds and antioxidant activity, when compared with methanol/acetone and ethanol. To study the anti-inflammatory properties of this extract in experimental type 1 diabetes, NOD mice were divided into two groups: the P. alata group, treated with aqueous extract of P. alata Curtis, and a non-treated control group, followed by diabetes expression analysis. The consumption of aqueous extract and water ad libitum lasted 28 weeks. The treated-group presented a decrease in diabetes incidence, a low quantity of infiltrative cells in pancreatic islets and increased glutathione in the kidney and liver (p<0.05), when compared with the diabetic and non-diabetic control-groups. In conclusion, our results suggest that the consumption of aqueous extract of P. alata may be considered a good source of natural antioxidants and compounds found in its composition can act as anti-inflammatory agents, helping in the control of diabetes.

NEW MICROSATELLITE LOCI FOR WATER YAM (DIOSCOREA ALATA, DIOSCOREACEAE) AND CROSS-AMPLIFICATION FOR OTHER DIOSCOREA SPECIES

Premise of the study: Dioscorea alata L. is one of the most widely distributed species of the genus in the humid and semihumid tropics and is associated with traditional agriculture. Only a few microsatellite markers have been developed so far for this and other Dioscorea species. Methods and Results: We isolated 14 codominant polymorphic microsatellite markers using a microsatellite-enriched genomic library technique. Ten microsatellite loci were selected, and 80 D. alata accessions from different regions in Brazil were evaluated with nine polymorphic loci. The polymorphism information content (PIC) varied from 0.39 to 0.78 and the power discrimination (PD) ranged from 0.15 to 0.91. Six of the markers showed transferability for the species D. bulbifera, D. cayenensis-D. rotundata, and D. trifida. Conclusions: The SSR markers obtained are an important tool for further studies aiming to characterize the genetic diversity in D. alata and other Dioscorea spp. accessions.

In vitro organogenesis of Passiflora alata

Passiflora alata in vitro organogenesis was studied based on explant type, culture medium composition, and incubation conditions. The results indicated that the morphogenic process occurred more efficiently when hypocotyl segment-derived explants were cultured in media supplemented with cytokinin and AgNO(3) incubated under a 16-h photoperiod. The shoot bud elongation and plant development were obtained by transferring the material to MSM culture medium supplemented with GA(3) and incubated in flasks with vented lids. Histological analyses of the process revealed that the difficulties in obtaining plants could be related to the development of protuberances and leaf primordia structures, which did not contain shoot apical meristem. Roots developed easily by transferring elongated shoots to 1/2 MSM culture medium. Plant acclimatization occurred successfully, and somaclonal variation was not visually detected. The efficiency of this organogenesis protocol will be evaluated for genetic transformation of this species to obtain transgenic plants expressing genes that can influence the resistance to Cowpea aphid borne mosaic virus.

Stability Testing of Spray- and Spouted Bed-Dried Extracts of Passiflora alata

The aim of this research was to perform a stability testing of spray- and spouted bed-dried extracts of Passiflora alata Dryander (Passion flower) under stress storage conditions. Spouted bed- and spray-dried extracts were characterized by determination of the average particle diameter (dP), apparent moisture content (XP), total flavonoid content (TF), and vitexin content. Smaller and more irregular particles were generated by the spouted bed system due to a higher attrition rate (surface erosion) inside the dryer. The SB dryer resulted in an end product with higher concentration of flavonoids (approximate to 10%) and lower moisture content (1.6%, dry basis) than the spray dryer, even with both dryers working at similar inlet drying air temperature and ratio between the extract feed flow rate to drying air flow rate (Ws/Wg). Samples of the spouted bed- and spray-dried extracts were stored at two different temperatures (34 and 45 degrees C) and two different relative humidities (52 and 63% RH for 34 degrees C; 52 and 60% RH for 45 degrees C) in order to perform the stability testing. The dried extracts were stored for 28 days and were analyzed every 4 days. The flavonoid vitexin served as the marker compound, which was assayed during the storage period. Results revealed shelf lives ranging from 9 to 184 days, depending on the drying process and storage conditions.

Structure and distribution of N-glycans on the S-7-allele stylar self-incompatibility ribonuclease of Nicotiana alata

S-RNases are the stylar products of the self-incompatibility (S)-locus in solanaceous plants (including Nicotiana alata), and as such, are involved in the prevention of self-pollination. All cDNA sequences of S-RNase products of functional S-alleles contain potential N-glycosylation sites, with one site being conserved in all cases, suggesting that N-glycosylation is important in self-incompatibility. In this study, we report on the structure and localization of the N-glycans on the S-7-allele RNase of N, alata, A total of nine N-glycans, belonging to the high-mannose- and xylosylated hybrid-classes, were identified and characterized by a combination of electrospray-ionization mass-spectrometry (ESI-MS), H-1-NMR spectroscopy, and methylation analyses. The glycosylation pattern of individual glycosylation sites was determined by ESI-MS of the glycans released from isolated chymotryptic glycopeptides, All three N-glycosylation sites showed microheterogeneity and each had a unique complement of N-glycans, The N-glycosylation pattern of the S-7-RNase is significantly different to those of the S-1- and S-2-RNases.

Structure of a putative ancestral protein encoded by a single sequence repeat from a multidomain proteinase inhibitor gene from Nicotiana alata

Background: The ornamental tobacco Nicotiana alata produces a series of proteinase inhibitors (Pls) that are derived from a 43 kDa precursor protein, NaProPl. NaProPl contains six highly homologous repeats that fold to generate six separate structural domains, each corresponding to one of the native Pls. An unusual feature of NaProPl is that the structural domains lie across adjacent repeats and that the sixth Pl domain is generated from fragments of the first and sixth repeats. Although the homology of the repeats suggests that they may have arisen from gene duplication, the observed folding does not appear to support this. This study of the solution structure of a single NaProPl repeat (aPl1) forms a basis for unravelling the mechanism by which this protein may have evolved, Results: The three-dimensional structure of aPl1 closely resembles the triple-stranded antiparallel beta sheet observed in each of the native Pls. The five-residue sequence Glu-Glu-Lys-Lys-Asn, which forms the linker between the six structural domains in NaProPl, exists as a disordered loop in aPl1. The presence of this loop in aPl1 results in a loss of the characteristically flat and disc-like topography of the native inhibitors. Conclusions: A single repeat from NaProPl is capable of folding into a compact globular domain that displays native-like Pl activity. Consequently, it is possible that a similar single-domain inhibitor represents the ancestral protein from which NaProPl evolved.

The solution structure of C1-T1, a two-domain proteinase inhibitor derived from a circular precursor protein from Nicotiana alata

A two-domain portion of the proteinase inhibitor precursor from Nicotiana alata (NaProPI) has been expressed and its structure determined by NMR spectroscopy. NaProPI contains six almost identical 53 amino acid repeats that fold into six highly similar domains; however, the sequence repeats do nut coincide with the structural domains. Five of the structural domains comprise the C-terminal portion of one repeat and the N-terminal portion of the next. The sixth domain contains the C-terminal portion of the sixth repeat and the N-terminal portion of the first repeat. Disulphide bonds link these C and N-terminal fragments to generate the clasped-bracelet fold of NaProPI. The three-dimensional structure of NaProPI is not known, but it is conceivable that adjacent domains in NaProPI interact to generate the circular bracelet with the N and C termini in close enough proximity to facilitate formation of the disulphide bonds that form the clasp The expressed protein, examined in the current study, comprises residues 25-135 of NaProPI and encompasses the first two contiguous structural domains, namely the chymotrypsin inhibitor C1 and the trypsin inhibitor T1, joined by a five-residue linker, and is referred to as C1-T1. The tertiary structure of each domain in C1-T1 is identical to that found in the isolated inhibitors. However, no nuclear Overhauser effect contacts are observed between the two domains and the five-residue linker adopts an extended conformation. The absence of interactions between the domains indicates that adjacent domains do not specifically interact to drive the circularisation of NaProPI. These results are in agreement with recent data which describe similar PI precursors from other members of the Solanaceae having two, three, or four repeats. The lack of strong interdomain association is likely to be important for the function of individual inhibitors by ensuring that there is no masking of reactive sites upon release from the precursor. (C) 2001 Academic Press.

The Three-dimensional Solution Structure of NaD1, a New Floral Defensin from Nicotiana alata and its Application to a Homology Model of the Crop Defense Protein alfAFP

NMR spectroscopy and simulated annealing calculations have been used to determine the three-dimensional structure of NaD1, a novel antifungal and insecticidal protein isolated from the flowers of Nicotiana alata. NaD1 is a basic, cysteine-rich protein of 47 residues and is the first example of a plant defensin from flowers to be characterized structurally. Its three-dimensional structure consists of an a-helix and a triple-stranded anti-parallel beta-sheet that are stabilized by four intramolecular disulfide bonds. NaD1 features all the characteristics of the cysteine-stabilized up motif that has been described for a variety of proteins of differing functions ranging from antibacterial insect defensins and ion channel-perturbing scorpion toxins to an elicitor of the sweet taste response. The protein is biologically active against insect pests, which makes it a potential candidate for use in crop protection. NaD1 shares 31% sequence identity with alfAFP, an antifungal protein from alfalfa that confers resistance to a fungal pathogen in transgenic potatoes. The structure of NaD1 was used to obtain a homology model of alfAFP, since NaD1 has the highest level of sequence identity with alfAFP of any structurally characterized antifungal defensin. The structures of NaD1 and alfAFP were used in conjunction with structure - activity data for the radish defensin Rs-AFP2 to provide an insight into structure-function relationships. In particular, a putative effector site was identified in the structure of NaD1 and in the corresponding homology model of alfAFP. (C) 2002 Elsevier Science Ltd. All rights reserved.

TRATAMENTOS PRÉ-GERMINATIVOS EM SEMENTES FLORESTAIS DA AMAZÔNIA: VI - MUIRAJUBA Apuleia leiocarpa (VOG.) MACBRIDE VAR. molaris SPR. ex BENTH. (LEGUMINOSAE)

Com o objetivo de superar a dormência de sementes de muirajuba (Apuleia leiocarpa(Vog.) Macbride var. molarisSpr. ex Benth), foi conduzido um ensaio experimental com 12 tratamentos pré-germinativos: testemunha (sem tratamento); imersão em ácido sullürico concentrado (96%) por diferentes períodos (5, 10, 15, 20 e 30 minutos), imersão em água quente a 80"C (2 e 10 minutos); desponte do lado oposto ao hilo; escarificação manual por 2 minutos; e escarificação manual seguida por imersão em água (6 e 12 horas). A germinação foi realizada em germinador Jacobsen sob temperatura constante (30"C), e acompanhada por 30 dias. Os efeitos dos tratamentos foram avaliados através da porcentagem de germinação e do índice de Velocidade de Germinação (I.V.G.). Todos os tratamentos aplicados proporcionaram uma maior germinação em relação a testemunha, cuja germinação foi de somente 13%. Porcentagens de germinação elevadas foram obtidas com as imersões em ácido sulfúrico, desponte do lado oposto ao hilo e escarificação manual por 2 minutos (com e sem imersão em água por 12 horas), os quais não diferiram entre sí. Dentre os tratamentos testados, o maior índice de vigor das sementes (I.V.G.) foi obtido com a escarificação manual por dois minutos, seguida por imersão em água por 12 horas. Os tratamentos onde as sementes foram submetidas a água quente não foram eficientes para estimular a germinação das sementes de muirajuba.

Influência do sombreamento no crescimento de mudas de cumaru (Dipteryx odorata (Aubl.) Willd. - Fabaceae), cultivadas em viveiro

Este trabalho apresenta os resultados de um experimento realizado com mudas de Dipteryx odorata (Aubl.) Willd. no viveiro da Estação Experimental de Silvicultura Tropical, do Inpa, em Manaus, Amazonas. O objetivo foi avaliar o desempenho da espécie cultivada em viveiro sob diferentes níveis de sombreamento. O experimento foi instalado em blocos ao acaso, com cinco tratamentos: dois tratamentos com sombra temporária, passando a pleno sol e três tratamentos com sombreamento de 30%, 50% e 70%, obtidos com o uso de telas de poliolefinas, de cor preta. Aos 3 e 5 meses foram avaliados após a semeadura, diâmetro do colo, altura, comprimento das raízes, peso de matéria seca de raízes, caule e folhas e as relações altura/ diâmetro do colo e parte aérea/sistema radicular. Tanto aos 3 quanto aos 5 meses, o sombreamento não afetou o crescimento em altura e o comprimento das raízes. O diâmetro do colo foi maior sob 0% de sombreamento, sendo superior estatisticamente ao tratamento com 70% em ambos períodos. Com relação ao peso de matéria seca, o tratamento com 70% de sombreamento foi inferior aos demais na primeira medição. Na segunda medição verificou-se uma tendência de aumento do peso de matéria seca com a diminuição dos níveis de sombreamento. Os menores valores das relações altura/diâmetro do colo e parte aérea/sistema radicular foram obtidos a pleno sol. Dipteryx odorata, uma espécie típica das matas altas de terra-firme, cresce melhor a plena luz, quando cultivada em viveiro.

Morfo-anatomia de sementes de Dipteryx odorata (Aubl.) Will. (Fabaceae) como contribuição ao estudo Farmacognóstico de plantas da região Amazônica

Dipteryx odorata (Aubl.) Willd. (Fabaceae) é popularmente conhecida como cumaru. Suas sementes são utilizadas pela população, por seus efeitos terapêuticos, dados pela cumarina, hoje comercializada para distúrbios vasculares e linfáticos e pela produção de óleo. Realizou-se um estudo morfo-anatômico de sementes de seis indivíduos desta espécie através da caracterização morfológica, determinação do peso da matéria fresca e das dimensões de 100 sementes, além de cortes histológicos transversais e longitudinais, considerando-se o tegumento e o embrião. A semente é oblonga, levemente comprimida na região do hilo. O tegumento seminal apresenta cutícula delgada e lisa, macroesclereídeos, osteoesclereídeos, mesofilo interno e membrana basal. O embrião constitui-se de dois cotilédones e o eixo embrionário retilíneo, formado por plúmula, epicótilo e radícula.