The carbohydrate-binding specificity and molecular modelling of Canavalia maritima and Dioclea grandiflora lectins

The carbohydrate-binding specificity of lectins from the seeds of Canavalia maritima and Dioclea grandiflora was studied by hapten-inhibition of haemagglutination using various sugars and sugar derivatives as inhibitors, including N-acetylneuraminic acid and N-acetylmuramic acid. Despite some discrepancies, both lectins exhibited a very similar carbohydrate-binding specificity as previously reported for other lectins from Diocleinae (tribe Phaseoleae, sub-tribe Diocleinae). Accordingly, both lectins exhibited almost identical hydropathic profiles and their three-dimensional models built up from the atomic coordinates of ConA looked very similar. However, docking experiments of glucose and mannose in their monosaccharide-binding sites, by comparison with the ConA-mannose complex used as a model, revealed conformational changes in side chains of the amino acid residues involved in the binding of monosaccharides. These results fully agree with crystallographic data showing that binding of specific ligands to ConA requires conformational chances of its monosaccharide-binding site.

Inflammatory cutaneous reaction induced by the lectin of Dioclea grandiflora (Mart. )

The lectin from Dioclea grandiflora (Mart.) that selectively binds glucose and mannose, when subcutaneously injected in mouse induces an inflammatory cutaneous reaction whose histological analysis reveals an hemorrhagic ulceration with exudative reaction accompanied by an influx of polymorphonuclear leukocytes and giant cells. The presence of lymphocytes and plasma cells in the lesion was insignificant. In order to characterize the in vivo action of inflammatory factors generated by this lesion, distinct lines of mice were used: high and low antibody responder mice; the genetically selected mice to the acute phase of inflammatory reaction; lines of mice deficient in C5, a protein of the complement system. It is shown that the lectin of D. grandiflora acts as an inflammatory agent probably promoting exocytosis and release of mediators.

In vivo lymphocyte activation and apoptosis by lectins of the Diocleinae subtribe

This paper reports the overall effects of three lectins, extracted from Canavalia brasiliensis, Dioclea violacea, and D. grandiflora, on BALB/c mice popliteal draining lymph nodes. These lectins have presented high stimulatory capacity on lymph node T cells. Additionally, they were able to induce apoptosis and inflammation (frequently associated with high endothelial venule necrosis). The data presented here suggest that the Diocleinae lectins studied can stimulate in vivo T cell activation and apoptosis, as well as present important side effects.

In vitro and in vivo antimalarial activity and cytotoxicity of extracts, fractions and a substance isolated from the Amazonian plant Tachia grandiflora (Gentianaceae)

Tachia sp. are used as antimalarials in the Amazon Region and in vivo antimalarial activity of a Tachia sp. has been previously reported. Tachia grandiflora Maguire and Weaver is an Amazonian antimalarial plant and herein its cytotoxicity and antimalarial activity were investigated. Spectral analysis of the tetraoxygenated xanthone decussatin and the iridoid aglyone amplexine isolated, respectively, from the chloroform fractions of root methanol and leaf ethanol extracts was performed. In vitro inhibition of the growth of Plasmodium falciparum Welch was evaluated using optical microscopy on blood smears. Crude extracts of leaves and roots were inactive in vitro. However, chloroform fractions of the root and leaf extracts [half-maximal inhibitory concentration (IC50) = 10.5 and 35.8 µg/mL, respectively] and amplexine (IC50= 7.1 µg/mL) were active in vitro. Extracts and fractions were not toxic to type MRC-5 human fibroblasts (IC50> 50 µg/mL). Water extracts of the roots of T. grandiflora administered by mouth were the most active extracts in the Peters 4-day suppression test in Plasmodium berghei-infected mice. At 500 mg/kg/day, these extracts exhibited 45-59% inhibition five to seven days after infection. T. grandiflora infusions, fractions and isolated substance have potential as antimalarials.

Fecundidade e longevidade de Aphis gossypii Glover, 1877 (Hemiptera, Aphididae) em diferentes temperaturas e cultivares comerciais de crisântemo (Dendranthema grandiflora Tzvelev)

Fecundity and longevity of Aphis gossypii Glover, 1877 (Hemiptera, Aphididae) at different temperatures and commercial chrysanthemum cultivars (Dendranthema grandiflora Tzvelev). The aphid A. gossypii is one of the main pests in a number of crops both under field and protected conditions. The objective of the present study was to evaluate the fecundity and longevity of A. gossypii under different temperatures and commercial chrysanthemum cultivars (Yellow Snowdon, White Reagan and Dark Splendid Reagan) with different trichomes densities (11.3; 16.6 and 21.6 trichome/mm² of the leaf, respectively) The trials were carried out in climatic chambers, at four temperatures (15, 20, 25 and 30 ±1 °C), 70 ± 10% RH and photophase 10h. The reproductive period significantly decreased with increase of temperature in the three cultivars. In Yellow Snowdon cultivar average duration of the reproductive period was 14.3 days at 25 °C. The maximum fecundity was obtained at the temperature of 25 ºC with 3,1; 2,8 and 3,6 nymphs/female/day in the Yellow Snowdon, White Reagan and Dark S. Reagan cultivars, respectively. The total fecundity was reduced by extreme temperatures (15 and 30 °C), and was obtained at 25 °C with 35,9 nymphs/female. Females maintained in Yellow Snowdon cultivar significantly showed superiority (30,7 nymphs/female) in total fecundity in relation to White Reagan (22,1 nymphs/female) and Dark S. Reagan (22,9 nymphs/female). The Yellow Snowdon cultivar (with a lower trichome density) had a significant influence in daily and total capacity of nymphs production, showing a higher fecundity of A. gossypii females. The aphid's longevity was affected by cultivars and temperature, and this longevity decreased whit increase of temperature. The results showed that there was an interaction between the temperature and host plant on reproductive parameters of A. gossypii.

químicos das folhas de Qualea grandiflora: atribuição dos dados de RMN de dois flavonóides glicosilados acilados diastereoisoméricos

Phytochemical investigation from leaves of the Qualea grandiflora (Vochysiaceae) resulted in the isolation and identification of kaempferol-3-O-α-L-(4"-E-p-coumaroyl)-rhamnoside, kaempferol-3-O-α-L-(4"-Z-p-coumaroyl)-rhamnoside, squalene, phytol, lupeol, α-amyrin, β-amyrin, sitosterol, sitostenone, sitosterol-3-O-β-D-glucopyranoside, ursolic and oleanolic acids. The structures of the compounds were identified by 1D- and 2D-NMR experiments, mass and UV spectrometry and comparison with literature data.

Alcaloides β-indolopiridoquinazolínicos de Esenbeckia grandiflora mart. (Rutaceae)

The chemical composition of two specimens of Esenbeckia grandiflora, collected in the south and northeast regions of Brazil, was investigated. In this study, three β-indolopyridoquinazoline alkaloids from the leaves (rutaecarpine, 1-hydroxyrutaecarpine) and roots (euxylophoricine D) were isolated for the first time in this genus. In addition, the triterpenes α-amyrin, β-amyrin, α-amyrenonol, β-amyrenonol, 3α-hydroxy-ursan-12-one, and 3α-hydroxy-12,13-epoxy-oleanane, the coumarins auraptene, umbelliferone, pimpinelin, and xanthotoxin, the furoquinoline alkaloids delbine and kokusaginine, and the phytosteroids sitosterol, stigmasterol, campesterol and 3β-O-β-D-glucopyranosylsitosterol were also isolated from the leaves, twigs, roots and stems of this species. Structures of these compounds were established by spectral analysis.

Determination of flavonoids in stamen, gynoecium, and petals of Magnolia grandiflora L. and their associated antioxidant and hepatoprotection activities

Chromatographic analysis of flavonoids in ethyl acetate fractions of the stamen, gynoecium, and petal of Magnolia grandiflora L. by HPLC-PDA-MS/MS-ESI in the negative ionization mode was performed in this study. The results revealed the presence of eight flavonoids: apigenin 8-C-glucoside, luteolin 8-C-glucoside, quercetin 3-O-rutinoside, quercetin 3-O-galactoside, quercetin, 3-O-glucoside, kaempferol 3-O-rutinoside, isorhamnetin 3-O-glucoside, and isorhamnetin. Their quantification revealed that luteolin 8-C-glucoside is the major flavonoid and that the total phenolic content is concentrated primarily in the stamen. The antioxidant and hepatoprotective effects of ethanolic extract of the flower organs were evaluated against hepatotoxicity induced by CCl4, compared with the effects of silymarin.

Antioxidant flavonol glycosides from Nectandra grandiflora (Lauraceae)

The bioassay-guided fractionation of the ethanol extract from Nectandra grandiflora leaves led to the isolation of two flavonol glycosides which inhibited the bleaching of beta -carotene on the TLC assay. Both compounds had their molecular structures elucidated by means of extensive use of uni- and bidimensional NMR techniques and were identified as 3-O-beta -rhamnosylkaempferol and 3-O-beta -rhamnosylquercetine.

Allelopathic potential and systematic evaluation of secondary compounds in extracts from roots of Canavalia ensiformis by capillary electrophoresis

Organic extracts were obtained from roots of Canavalia ensiformis and evaluated for allelopathic potential on the germination of the weed seeds: Mimosa pudica, Cassia tora and Cassia occidentalis showing a strong allelopathic potential. After that, a systematic study of these crude extracts was made using specific protocols developed in capillary electrophoresis (CE) in order to determine some classes of secondary metabolites. Capillary electrophoresis protocols were highly specific, which makes it possible to identify 5 classes of compounds using the same crude extract samples and analyze them fartly. Some of the compounds identified show activity in the inhibition of seeds germination.

REPRODUCTIVE SYSTEM AND POLLEN FLOW IN PROGENIES OF Qualea grandiflora Mart., A TYPICAL SPECIES OF THE BRAZILIAN CERRADO

This study analyzed the reproductive system and the pollen dispersion pattern of Qualea grandiflora progenies. This is a typical species from the Brazilian Cerrado about which there are not too many studies from the genetics point of view. The study was conducted in an area of 2.2 hectares located in the Conservation Unit managed by the Forest Institute of the state of São Paulo, Brazil (Assis State Forest). Total genomic DNA of 300 seeds from 25 plants (12 seeds from each plant) was extracted and amplified using specific primers to obtain microsatellite markers. Results showed that selfing is frequent among adults and progenies, and the species reproduces by outcrossing between related and unrelated individuals (0.913). The single-locus outcrossing rate was 0.632, which indicates that mating between unrelated individuals is more frequent than between related plants. The selfing rate was low (0.087), that is, the species is allogamous and self-fertilization is reduced. About 35% of the plants in the progenies were full-sibs, and about 57%, half-sibs. Besides, about 8% of the progenies were selfing siblings. The genetic differentiation coefficient within progenies was 0.139, whereas the fixation rate was about 27%. The estimate of the effective size revealed that the genetic representativeness of descent was lower than expected in random mating progenies: The analyzed samples corresponded to only 13.2 individuals of an ideal panmictic population. In environmental recovery programs, seeds, preferably from different fruits, should be collected from 95 trees to preserve the genetic diversity of the species.

ANÁLISE DO EFEITO DA SAZONALIDADE SOBRE O RENDIMENTO DO ÓLEO ESSENCIAL DAS FOLHAS DE Nectandra grandiflora Nees1

Este estudo teve por objetivos avaliar o efeito da sazonalidade sobre o teor de óleo essencial (OE) presente nas folhas de Nectandra grandiflora Nees e determinar se a secagem do material vegetal à temperatura ambiente influencia o rendimento do OE. Para tanto, foi coletado material vegetal de 12 indivíduos ao longo de um ano, e o OE foi extraído separadamente de folhas frescas e folhas secas, pelo método de hidrodestilação. Paralelamente, 20 g de folhas frescas foram levados à estufa a 40 °C até peso constante, para determinação do teor de umidade, procedimento esse realizado para cada coleta, individualmente. O rendimento de OE das folhas (% m/m) foi calculado para o material vegetal fresco com base em massa de matéria seca, determinado considerando o teor de umidade, bem como para o material seco à temperatura ambiente. Os dados de rendimento foram analisados através do teste de Tukey (P<0,05). Os resultados mostraram que o teor de OE das folhas da espécie apresentou variabilidade sazonal, com a maior produção detectada na primavera (0,75 ± 0,06%). Esse valor foi estatisticamente diferente dos rendimentos encontrados nas outras estações do ano, sendo o menor rendimento obtido no inverno (0,39 ± 0,02%). Desse modo, indica-se que a coleta de folhas de N. grandiflora seja realizada durante a primavera, visando obter maior teor de OE. Adicionalmente, verificou-se que não há perda de teor OE durante o processo de secagem à temperatura ambiente e, dessa forma, esse procedimento pós-colheita poderá vir a ser adotado sem comprometer o rendimento do extrativo.

Atividade potencialmente alelopática de extratos brutos e hidroalcoólicos de feijão-de-porco (Canavalia ensiformis)

Extratos hidroalcoólicos de parte aérea, raízes e sementes e extratos brutos de sementes de Canavalia ensiformis foram preparados, visando identificar e caracterizar os efeitos potencialmente alelopáticos sobre a germinação de sementes e o alongamento da radícula das plantas daninhas Mimosa pudica, Urena lobata, Senna obtusifolia e Senna occidentalis. Os trabalhos foram desenvolvidos em condições controladas de 25 ºC de temperatura e fotoperíodo de 12 horas, para o bioensaio de germinação, e 24 horas, para o de alongamento da radícula. Os efeitos foram aquilatados tendo por contraste (testemunha) a água destilada. Os resultados variaram em função da espécie receptora, da concentração e da parte da planta utilizada no preparo dos extratos. A inibição da germinação das sementes e do alongamento da radícula foi diretamente proporcional à concentração do extrato, com as mais intensas inibições observadas na concentração de 4%. Independentemente da espécie receptora, as sementes, seguidas das raízes, foram as principais fontes de substâncias químicas com atividades potencialmente alelopáticas no feijão-de-porco. A análise dos diferentes extratos brutos revelou que as substâncias químicas com atividades potencialmente alelopáticas presentes nas sementes do feijão-de-porco têm polaridade compreendida entre o acetato de etila e o metanol. Para o extrato bruto metanólico, concentrações a partir de 0,4% inibiram completamente a germinação das espécies receptoras, enquanto para M. pudica e S. occidentalis concentrações de 0,6 e 0,8% proporcionaram inibições da ordem de 100% para a germinação das sementes dessas espécies. A sensibilidade das espécies aos efeitos potencialmente alelopáticos variou na seguinte ordem decrescente: M. pudica > S. occidentalis > S. obtusifolia > U. lobata.

Development of bean plants in soil contaminated with trifloxysulfuron-sodium after Stizolobium aterrimum and Canavalia ensiformis cultivation

Phytoremediation, the use of plants to decontaminate soils and water resources from organic pollutants such as herbicides, is economically and environmentally a promising technique applied in many areas, including agriculture. The objective of this work was to evaluate the development of bean plants cultivated in the field, in soil with different levels of trifloxysulfuron-sodium contamination, following cultivation of two green manure species, as well as to evaluate the possibility of recontamination of the area by such herbicide with the straw permanence on the soil. The experiment was carried out in Coimbra, MG, Brazil, on a sandy clayey Red - Yellow Argisol from March to November 2003. Four levels of soil contamination with trifloxysulfuron-sodium (0.00; 3.75; 7.50; and 15.00 g ha-1) were used as well as the following five types of cultivation prior to bean sowing in the area after herbicide application: black velvet beans (Stizolobium aterrimum) followed by removal of straw; S. aterrimum, followed by permanence of straw; jack bean (Canavalia ensiformis), followed by removal of straw; C. ensiformis followed by permanence of straw; and without prior cultivation, weed-free (weeded control). The leguminous plants were kept in the area for 65 days, cut close to the soil, and with its aerial part left or not on the surface of the experimental plot, depending on the treatment. Fifteen days after the species were cut, bean was sown in the area. At 45 days after emergence (DAE) of the bean plants, plant height and dry mass of the aerial part were evaluated. Grain productivity was determined during harvest. Height, dry matter of the aerial part and grain productivity of the bean plants, cultivated in an area previously contaminated with trifloxysulfuron-sodium at any of the levels tested, were higher with prior cultivation of S. aterrimum or C. ensiformis. At the lowest level of herbicide contamination, prior cultivation of C. ensiformis was found to be more efficient than that of S. aterrimum in mitigating the harmful effects of trifloxysulfuron-sodium on bean grain production. The permanence of the straw of the green manure species during the bean cycle did not harm the development of the plants or caused culture productivity losses, indicating that straw permanence in the area does not promote recontamination of the area.

Efeito da umidade do solo sobre a capacidade de Canavalia ensiformis e Stizolobium aterrimum em remediar solos contaminados com herbicidas

Objetivou-se, neste trabalho, avaliar os efeitos da umidade do solo sobre a capacidade de Canavalia ensiformis e Stizolobium aterrimum em remediar solos contaminados com os herbicidas tebuthiuron e trifloxysulfuron-sodium. O trabalho foi realizado em duas etapas, sendo na primeira avaliado o crescimento de C. ensiformis e S. aterrimum em solo com diferentes níveis de umidade, contaminados ou não com herbicidas. Os tratamentos foram compostos pela combinação entre os herbicidas tebuthiuron, trifloxysulfuron-sodium e ausência de herbicida, associados a quatro teores de água do solo (0,287, 0,358, 0,431 e 0,575 kg kg-1), dispostos em esquema fatorial 3 x 4, sob delineamento inteiramente casualizado, com três repetições, para cada espécie remediadora. Após o preparo do substrato e enchimento dos vasos, aplicou-se à superfície do solo o herbicida trifloxysulfuron-sodium ou tebuthiuron. Um dia após essa aplicação, procedeu-se à semeadura das espécies remediadoras. Nessa mesma época, utilizaram-se, como testemunha, vasos sem planta remediadora, porém com os mesmos níveis de umidade e com aplicação do herbicida mantido nas mesmas condições daqueles com plantas remediadoras, as quais foram colhidas 60 dias após semeadura. Nessa ocasião, foram avaliadas a altura e a massa seca da parte aérea (MSPA) dessas espécies. Todo o material colhido foi triturado e incorporado ao solo dos seus respectivos vasos. Na segunda etapa, avaliou-se a capacidade remediadora de C. ensiformis e S. aterrimum. Os tratamentos foram compostos pela combinação entre quatro níveis de umidade e cinco tipos de cultivo prévio: cultivo de C. ensiformis e S. aterrimum na presença e ausência do herbicida e um tratamento sem cultivo prévio e com aplicação do herbicida, dispostos em esquema fatorial 4 x 5, sob delineamento inteiramente casualizado, com três repetições, para cada herbicida avaliado. Amostras de 0,5 kg de solo foram retiradas dos vasos (6 L) utilizados na primeira etapa e colocadas em vasos de 0,5 L. Em seguida, cultivaram-se as espécies indicadoras de resíduo dos herbicidas no solo: sorgo (Sorghum bicolor) para o trifloxysulfuron-sodium e soja (Glycine max) para o tebuthiuron. Estas plantas foram colhidas 20 dias após a semeadura, época em que se avaliaram a altura, a MSPA e o grau de intoxicação delas pelos herbicidas. Canavalia ensiformis não sobreviveu até a época de avaliação (60 DAS) em solo contaminado pelo tebuthiuron, independentemente do nível de umidade mantido no solo. S. aterrimum sobreviveu quando cultivada em solo com teor de água entre 0,287 e 0,358 kg kg-1 e, quando comparada a C. ensiformis, foi mais eficiente na descontaminação do solo com residual do tebuthiuron. Nos solos contaminados com trifloxysulfuron-sodium ou com tebuthiuron, com o cultivo prévio das espécies remediadoras, o crescimento do sorgo e da soja foi melhor se comparado ao daquelas plantas cultivadas no solo onde não foi feito o cultivo das espécies remediadoras. De maneira geral, a variação da umidade não interferiu no processo de remediação, sendo os efeitos observados no desenvolvimento das espécies remediadoras. O melhor desenvolvimento de C. ensiformis e S. aterrimum foi observado em solo com teor de água mantido em torno de 0,431 kg kg-1; contudo, nesse teor de água, o tebuthiuron é mais facilmente disponibilizado para a solução do solo.