A comparative study on biological activities of different solvent extracts from whole seed, seed coat and cotyledon of two Lathyrus species

Abstract The present study was conducted to assess the phenolic content, and antibacterial and antioxidant activities of Lathyrus L. species. The extraction of phenolic compounds from whole seeds, seed coat and cotyledon of Lathyrus hierosolymitanus Boiss. and Lathyrus annuus L. seeds was performed employing different solvents. Total phenolic content (TPC) was measured by Folin- Ciocalteau assay, while the antioxidant activity was determined by DPPH radical scavenging activity, and reducing power assay. It was found that TPC of extracts ranged from 0.12 mg to 6.53 mg GAE/gdw. For each solvent, seed coat extracts were generally observed to render higher TPC and antioxidant activities. There was a correlation between TPC and antioxidant activity. In addition, all extracts were also examined for their antimicrobial activity against Bacillus cereus, Escherichia coli and Pseudomonas aeruginosa. Methanol extracts showed the highest antibacterial activity which is consistent with TPC, but there was no correlation between TPC and antibacterial activity. Solvents were observed to have effects on gallic acid, caffeic acid, and epicatechin extractions. HPLC analysis results of extracts confirmed methanol and ethanol as preferred solvents for phenolic extraction from Lathyrus sp. Phenolic content in the extracts could be suggested to contribute to their antioxidant and antibacterial activity.

Optimization of callus cultures at Echinacea purpurea L. for the amount of caffeic acid derivatives

BACKGROUND: In order to produce an effective callus in Echinacea purpurea L.; determination of the explant type and growth regulators that best respond to callus induction and the optimization of the culture conditions to increase the amount of caffeic acid derivatives (CADs) in the obtained callus. CADs contents of callus cultures of E. purpurea were evaluated by establishing an effective callus induction system in vitro. RESULTS: Various medium containing different growth regulators were tested using leaf, petiole, cotyledon and root as the explants. The best callus development was achieved in MS medium with 1.0 mg l 1 2,4- D + 2.0 mg l 1 BAP in leaf, 1.0 mg l 1 NAA + 0.5 mg l 1 TDZ in petiole, 2.0 mg l 1 NAA + 1.0 mg l 1 TDZ in cotyledon and 0.5 mg l 1 NAA + 0.5 mg l 1 BAP in roots. Upon optimisation of callus growth, each type of explant was cultured for 4, 6, 8 and 10 weeks in medium for the analyses of caftaric acid, chlorogenic acid, caffeic acid and chicoric acid contents. The highest amounts of caftaric acid (4.11 mg/g) and chicoric acid (57.89 mg/g) were found from petiole explants and chlorogenic acid (8.83 mg/g) from root explants at the end of the 10-week culture time. CONCLUSIONS: As a result of the present study, the production of caffeic acid derivatives was performed by providing the optimization of E. purpurea L. callus cultures. Effective and repeatable protocols established in this study may offer help for further studies investigating the production of caffeic acid derivatives in vitro.

Morphoanatomy of Serjania communis Cambess. seedling (Sapindaceae) / Morfoanatomia da plântula de Serjania communis Cambess. (Sapindaceae)

Serjania communis Cambess. (Sapindaceae) is a plant with climbing habit and occurs relatively often in Paraná State, Brazil. The fruits were collected at the 'Parque dos Pioneiros' a remnant of subtropical forest in Maringá, Paraná State, Brazil. The seedlings obtained in the greenhouse were described according to traditional techniques in plant morphology. Seedlings were embedded in historesin and sectioned in rotation microtome. The fruit is the samaroid type, the seeds have about 3 mm in length and brown color. Seedlings are epigeal phanerocotylar. The seedlings have a hairy hypocotyl, foliaceous cotyledons, reduced epicotyl, and two opposite eophylls. The root is diarch, the hypocotyl shows root-stem transition structure, stem epicotyl, and dorsiventral and hypostomatous cotyledons and eophylls. 'Tirodendros' with 45 days of age do not develop cambial variant.

Serjania communis Cambess. (Sapindaceae) é uma planta de hábito lianescente e ocorre com relativa frequência no Estado do Paraná, Brasil. A coleta dos frutos para a obtenção de sementes foi realizada no parque florestal dos Pioneiros (bosque II), um remanescente de floresta subtropical de Maringá, Estado do Paraná, Brasil. As plântulas, obtidas em casa de vegetação foram descritas conforme técnicas usuais em morfologia vegetal. Após a germinação, as plântulas foram descritas morfoanatomicamente e também foi verificada se ocorria a instalação de variante cambial nesta fase inicial de desenvolvimento. O fruto é do tipo samaroide. As sementes têm cerca de 3 mm de comprimento, de cor marrom e com germinação epigeia e as plântulas são fanerocotiledonares. As plântulas foram emblocadas em historresina e secionadas em micrótomo de rotação. As plântulas apresentam o hipocótilo piloso, cotilédones foliáceos, o epicótilo é reduzido, os dois primeiros eofilos são opostos e os demais eofilos apresentam filotaxia alterna helicoidal. A raiz é diarca. O hipocótilo é uma região de transição raiz-caule. O epicótilo tem estrutura caulinar eustélica. Os cotilédones e os eofilos são hipostomáticos e dorsiventrais. Tirodendros com 45 dias de idade não desenvolvem variante cambial.

Cambios microestructurales en los granos de Phaseolus vulgaris endurecidos / Microstructural changes in hardened beans (Phaseolus vulgaris)

El endurecimiento de los granos de Phaseolus vulgaris almacenados a alta temperatura y alta humedad relativa es una de las principales limitantes para su consumo. El objetivo de esta investigación fue evaluar por microscopia de barrido electrónico los cambios estructurales ocurridos en los cotiledones y en la testa de los granos endurecidos. Los granos recién cosechados se almacenaron durante doce meses bajo dos condiciones: 5°C-34% HR y 37°C-75% HR. Esta última con el fin de propiciar el endurecimiento. Los granos almacenados crudos y cocidos se liofilizaron y se fracturaron. Las secciones de testa y cotiledones se observaron en un microscopio electrónico JSM-6390. Al cabo de doce meses se constató que los granos almacenados a 37°C-75% HR aumentaron su dureza en un 503%, mientras que los granos almacenados a 5°C-34% HR no incrementaron su dureza significativamente. A nivel microestructural, en los cotiledones de los granos crudos se notaron claras diferencias en el aspecto de la pared celular, en el tamaño del espacio intercelular y en la textura de la matriz proteica. Mientras que en la testa de los granos crudos se evidenciaron diferencias en la compactación de la empalizada y de la capa sub-epidérmica. En los granos cocidos se observó una total separación entre las células de los cotiledones de los granos blandos y una muy limitada separación en los granos duros. Se concluye que las diferencias observadas en los granos duros y blandos, demostraron una participación importante de ambas estructuras, cotiledones y testa, en el endurecimiento de los granos.

The hardening of Phaseolus vulgaris beans stored at high temperature and high relative humidity is one of the main constraints for consumption. The objective of this research was to evaluate by scanning electron microscopy, structural changes in cotyledons and testa of the hardened beans. The freshly harvested grains were stored for twelve months under two conditions: 5 ° C-34% RH and 37 ° C-75% RH, in order to promote hardening. The stored raw and cooked grains were lyophilized and fractured. The sections of testa and cotyledons were observed in an electron microscope JSM-6390. After twelve months, grains stored at 37 ° C-75% RH increased their hardness by 503%, whereas there were no significant changes in grains stored at 5 ° C-34% RH. At the microstructural level, the cotyledons of the raw grains show clear differences in appearance of the cell wall, into the intercellular space size and texture matrix protein. There were also differences in compaction of palisade and sub-epidermal layer in the testa of raw grains. After cooking, cotyledon cells of the soft grains were well separated while these of hard grains were seldom separated. In conclusion, the found differences in hard and soft grains showed a significant participation of both structures, cotyledons and testa, in the grains´ hardening.

Potencial de preparados de cavalinha (Equisetum sp. ) na síntese de metabólitos de defesa em cotilédones de soja (Glycine max L. ) e o efeito sobre o crescimento de Rhizoctonia solani Kuhn, in vitro / Potential of horsetail (Equisetum sp. ) derivatives on the synthesis of defense metabolites using soybean ( Glycine max L. ) cotyledons and their effect on the in vitro growth of Rhizoctonia solani Kuhn

Foram desenvolvidos dois experimentos com objetivo de avaliar o potencial de preparados de cavalinha (Equisetum sp.) na síntese de metabólitos de defesa em cotilédones de soja (Glycinemax L.) e o efeito sobre o crescimento de Rhizoctonia solani, in vitro. O delineamento experimental utilizado para os experimentos foi inteiramente casualizado em esquema fatorial 3x5 (formas de extração x concentrações), com quatro repetições. As formas de extração foram extrato alcoólico, infusão e maceração, nas concentrações de zero; 1; 10, 20 e 40%. No primeiro experimento foi avaliada a indução de compostos de defesa vegetal em cotilédones de soja em resposta aos derivados a base de cavalinha, sendo quantificada a atividade da enzima fenilalanina amônia-liase (FAL), via espectofotometria, a fitoalexina gliceolina, e o teor de fenóis totais. No segundo experimento, in vitro, a unidade experimental foi uma placa de Petri, sendo os preparados de cavalinha incorporados ao meio BDA (Batata-dextrose e Agar) e avaliado o crescimento micelial de R. Solani. Os preparados de extrato alcoólico, infusão e maceração de cavalinha apresentaram capacidade de indução das fitoalexinas gliceolinas em cotilédones de soja, bem como, ativaram o metabolismo de compostos fenólicos. Entre os preparados, o extrato alcoólico e a maceração, se sobressaem sobre a infusão. Os preparados de extrato alcoólico, infusão e maceração de cavalinha em todas as suas concentrações inibem o crescimento do fungo R. solani, in vitro. .

Two experiments were carried out in the Federal Technological University of Paraná - Dois Vizinhos Campus - with the aim to evaluate the potential of horsetail (Equisetum sp.) derivatives for the synthesis of defense metabolites in soybean (Glycine max L.) cotyledons and their effect on the in vitro growth of Rhizoctonia solani. The experimental design was completely randomized in a 3 x 5 factorial design (extraction form x concentration), with four replications. The extraction forms were alcoholic extract, infusion and maceration and the concentrations tested were zero, 1, 10, 20 and 40%. In the first experiment, we evaluated the induction of plant defense in soybean cotyledons as a response to horsetail derivatives through spectrophotometry according to phytoalexin glyceollin, phenylalanine ammonia lyase enzyme activity (PAL) and total phenols. In the second experiment, in vitro, the experimental unit was a Petri dish, and the horsetail derivatives were incorporated into medium culture (potato dextrose agar), and we evaluated the mycelial growth of R. solani. The alcoholic extract, infusion and maceration of horsetail derivatives presented phytoalexin glyceolin induction in soybean cotyledons, in addition to activating the metabolism of phenolic compounds. Among the derivatives, the alcoholic extract and the maceration form of extraction were superior in relation to the infusion. The alcoholic extract, infusion and maceration of horsetail derivatives inhibited the in vitro growth of R. solani in all concentrations.

Morphology and anatomy of the seedling and the tirodendro of Calophyllum brasiliense Cambess. (Clusiaceae) / Morfoanatomia da plântula e do tirodendro de Calophyllum brasiliense Cambess. (Clusiaceae)

Calophyllum brasiliense Cambess. is a tree species that is presented as an alternative to replace endangered species of hardwood. The morphology and anatomy of the seedling of this species is the object of the present study. Seedlings at different stages of development were obtained in greenhouse and analyzed fresh and fixed in FAA (Formalin- Acetic-Alcohol) 50. The anatomical analysis was done by the freehand and microtome sections, according to standard techniques in plant anatomy. The seedling and/or tirodendro is cryptocotylar and hypogeal, has cataphylls, and presents eophylls and metaphylls simple with venation pinnate craspedodromous simple. The root is polyarch, the hypocotyl is very short, the cotyledons have an oily and starchy reserve, the epicotyl has stem structure, and eophylls and metaphylls are dorsiventral. The seedling may be classified in the Horsfieldia type/subtype.

Calophyllum brasiliense Cambess. é uma planta arbórea que se apresenta como possível alternativa para substituir espécies em extinção com madeira nobre. A morfologia e anatomia da plântula desta espécie é objeto do presente trabalho. As plântulas, em diferentes fases de desenvolvimento, foram obtidas em casa de vegetação e analisadas a fresco e fixadas em FAA 50. A análise anatômica foi feita mediante seções a mão livre e microtômicas, de acordo com técnicas usuais em anatomia vegetal. A plântula e/ou tirodendro é criptocotiledonar e hipogeia, possui catafilos, e apresenta eófilos e metafilos simples de venação pinada craspedódroma simples. A raiz é poliarca, o hipocótilo é muito reduzido, os cotilédones possuem reserva amilácea e oleaginosa, o epicótilo tem natureza caulinar e os eofilos e metafilos são dorsiventrais. A plântula enquadra-se no tipo e subtipo Horsfieldia.

Seedling structure of Euphorbia L. and Chamaesyce Gray species / Estrutura da plântula de espécies de Euphorbia L. e Chamaesyce Gray

Chamaesyce prostrata (Aiton) Small, Euphorbia heterophylla L. and Euphorbia graminea Jacq. are weedy species that occur with relative frequency in the region of Maringá, Paraná State, Brazil. Data about the morphology and anatomy of seedlings of these species were obtained, with emphasis on significant structural differences of seedlings between species of Euphorbia and Chamaesyce. Seedlings are phanerocotylar and epigeal. Root is axial and triarch. Cotyledons are leaf-like and dorsiventral. Eophylls and prophylls are simple and dorsiventral. Morphological variations occur in the cotyledon shape of Euphorbia seedlings. It is remarkable the Kranz structure of leaves that occurs only in Chamaesyce.

Chamaesyce prostrata (Aiton) Small, Euphorbia heterophylla L. e E. graminea Jacq. (Euphorbiaceae) são espécies invasoras frequentes na região de Maringá, Paraná. Foram obtidos dados sobre a morfologia e anatomia das plântulas dessas espécies, com ênfase se há caracteres estruturais significativos das plântulas entre as espécies de Euphorbia e Chamaesyce. As plântulas são fanerocotiledonares e epígeas. A raiz é axial e triarca. Os cotilédones são foliáceos e dorsiventrais. Os eofilos e profilos são folhas simples e dorsiventrais. As variações mofológicas ocorrem no formato dos cotilédones das plântulas de Euphorbia. Sob o aspecto anatômico é especialmente notável a estrutura Kranz das folhas que ocorrem somente em Chamaesyce.

Establishment of an in vitro plantlet regeneration protocol for unique varieties of brinjal (Solanum melongena L.) var. Mattu Gulla and Perampalli Gulla.

Brinjal (Solanum melongena L.) var. Mattu Gulla (MG) and var. Perampalli Gulla (PG) are unique varieties with distinct flavour cultivated in Udupi, Karnataka State, and are exposed to several biotic and abiotic stresses. An efficient and reproducible in vitro regeneration method is required to expedite the manipulation of these brinjal varieties to cope up with stress by tissue culture and gene transfer methods. The present study, reports a rapid and efficient in vitro regeneration protocol for these two varieties. The in vitro growth response was studied on Murashige and Skoog (MS) medium supplemented with 2, 4-D, BAP and IAA, and the plantlets were regenerated efficiently from callus cultures of leaf, cotyledon and hypocotyl explants. Among the three explants, the hypocotyl explants were found to have better callus induction and multiple shoot regeneration. High frequency of shoot initiation was achieved from hypocotyl derived calluses in MS media with 2.0 mg/L BAP and 0.5 mg/L IAA in MG and PG. Efficient and rapid shoot proliferation, and elongation were noted in MS medium with 1.0 mg/L BAP and 0.3 mg/L GA3. The in vitro regenerated shoots produced healthy roots when they were cultured on MS medium supplemented with 0.5 mg/L IBA. A significant difference was observed in percentage of callus induction, number of shoots per callus, shoot elongation and number of hardened plantlets of MG and PG. MG showed maximum response in all stages of culture than PG. Hardening of plantlets in tissue culture was achieved in three weeks. The hardened plantlets were grown in pots for further acclimatization in green house and finally transplanted to experimental garden where they developed into flowering plants and produced mature fruits with viable seeds.

Inhibition of TNF-alpha-Mediated NF-kappaB Transcriptional Activity by Dammarane-Type Ginsenosides from Steamed Flower Buds of Panax ginseng in HepG2 and SK-Hep1 Cells

Panax ginseng is a medicinal herb that is used worldwide. Its medicinal effects are primarily attributable to ginsenosides located in the root, leaf, seed, and flower. The flower buds of Panax ginseng (FBPG) are rich in various bioactive ginsenosides, which exert immunomodulatory and anti-inflammatory activities. The aim of the present study was to assess the effect of 18 ginsenosides isolated from steamed FBPG on the transcriptional activity of NF-kappaB and the expression of tumor necrosis factor-alpha (TNF-alpha)-stimulated target genes in liver-derived cell lines. Noticeably, the ginsenosides Rk3 and Rs4 exerted the strongest activity, inhibiting NF-kappaB in a dose-dependent manner. SF and Rg6 also showed moderately inhibitory effects. Furthermore, these four compounds inhibited the TNF-alpha-induced expression of IL8, CXCL1, iNOS, and ICAM1 genes. Consequently, ginsenosides purified from steamed FBPG have therapeutic potential in TNF-alpha-mediated diseases such as chronic hepatic inflammation.

Inhibition of TNF-alpha-Mediated NF-kappaB Transcriptional Activity by Dammarane-Type Ginsenosides from Steamed Flower Buds of Panax ginseng in HepG2 and SK-Hep1 Cells

Panax ginseng is a medicinal herb that is used worldwide. Its medicinal effects are primarily attributable to ginsenosides located in the root, leaf, seed, and flower. The flower buds of Panax ginseng (FBPG) are rich in various bioactive ginsenosides, which exert immunomodulatory and anti-inflammatory activities. The aim of the present study was to assess the effect of 18 ginsenosides isolated from steamed FBPG on the transcriptional activity of NF-kappaB and the expression of tumor necrosis factor-alpha (TNF-alpha)-stimulated target genes in liver-derived cell lines. Noticeably, the ginsenosides Rk3 and Rs4 exerted the strongest activity, inhibiting NF-kappaB in a dose-dependent manner. SF and Rg6 also showed moderately inhibitory effects. Furthermore, these four compounds inhibited the TNF-alpha-induced expression of IL8, CXCL1, iNOS, and ICAM1 genes. Consequently, ginsenosides purified from steamed FBPG have therapeutic potential in TNF-alpha-mediated diseases such as chronic hepatic inflammation.

Assessment of somatic embryogenesis potency in Indian soybean [Glycine max (L.) Merr.] cultivars.

Majority of the Indian soybean cultivars are recalcitrant to tissue culture regeneration. The present communication reports the development of somatic embryogenesis in a liquid culture medium from immature cotyledons of G. max. Following induction with 2,4-dichlorophenoxyacetic acid (2,4-D) or naphthalene acetic acid (NAA), the number of somatic embryos and percentage of explants that responded were higher with 45.24 µM 2,4-D. The proliferation of somatic embryos for three successive cycles was achieved in 22.62 µM 2,4-D. Histodifferentiation of somatic embryos under NAA (10.74 µM) indicated that better embryo development and maturation was achieved without any growth regulator. The amino acids such as L-glutamine favoured the somatic embryo induction and histodifferentiation at 20 and 30 mM respectively, where as L-asparagine at 10 mM concentration enhanced the somatic embryo proliferation. In addition, somatic embryos that were desiccated (air-drying method) for 5 days showed better germination (40.88%). The Indian soybean cultivars also showed strict genotypic influence and cv. Pusa 16 was emerged as a best responding cultivar for somatic embryo induction with 74.42% of response.

Efeito de extratos de cogumelos na indução de fitoalexinas e no controle de oídio da soja em casa de vegetação / Mushrooms extracts effect on the induction of phytoalexins and on the control of soybean powdery mildew in greenhouse

O oídio (Erysiphe diffusa) tornou-se uma das principais doenças que afeta a cultura da soja (Glycine max), podendo causar redução significativa no rendimento da cultura. Esse trabalho teve por objetivo avaliar o efeito dos extratos aquosos de Agaricus blazei, Lentinula edodes e Pycnoporus sanguineus no controle do oídio em casa-devegetação e na indução de fitoalexinas em cotilédones de soja. O delineamento experimental utilizado foi inteiramente casualizado com cinco repetições. Como controle utilizou-se fungicida (enxofre ­ 3g/L) e testemunha - sem aplicação. Os tratamentos utilizados foram: extratos aquosos de Agaricus blazei, Lentinula edodes e Pycnoporus sanguineus na concentração de 20 %v/v sendo estes aplicados aos 61 e 68 dias após a semeadura. Avaliou-se a incidência e severidade aos 59, 65, 69, 73 e 77 dias após a semeadura em todos os folíolos de cada planta. Com os dados da severidade, calculouse a área abaixo da curva de progresso da doença (AACPD). Para a incidência, detectou-se diferença significativa entre o fungicida e os demais tratamentos. Para severidade e AACPD, observou-se que os extratos diferiram da testemunha, sendo que L. edodes mostrou ser mais eficiente que os demais tratamentos. Para o bioensaio de fitoalexinas foi aplicado sobre cada cotilédone de soja uma alíquota de 75 µL de cada extrato aquoso nas concentrações de 1%, 2%, 4% e 10%. Após 20 horas, os mesmos foram transferidos para erlenmeyers contendo 15 mL de água destilada esterilizada e deixados em agitação por 1 h para extração da fitoalexina formada. A absorbância foi determinada a 285 nm em espectrofotômetro. Observou-se um maior acúmulo de fitoalexina nas maiores concentrações dos extratos aquosos.

The powdery mildew (Erysiphe diffusa) became one of the main diseases that effects soybean (Glycine max) crops. The aim was to evaluate the effect of Agaricus blazei, Lentinula edodes and Pycnoporus sanguineus aquous extracts on the control of powdery mildew in greenhouse and on the induction of phytoalexins in soybean cotyledons. The control treatments consisted of fungicide (Sulphur ­ 3 g/L) and control without application. The treatments were applied at 61 and 68 days after sowing (DAS). The incidence and severity was evaluated at 59, 65, 69, 73 and 77 DAS in all the folioles of each plant. With the severity values, the area under the disease progress curve (AUDPC) was estimated. To the incidence, a significative difference was detected between the fungicide and the other treatments. For severity and the AUDPC, the extracts and the fungicide differed from the control and L. edodes was the most efficient treatment. For the phytoalexins production was applied 75 mL to each soybean cotyledon of each aqueous extract at 1%, 2%, 4% and 10% concentrations. After 20 hours, they were transferred to erlenmayer flask containing 15 mL of sterile distilled water and remained stirring for 1 h for extraction of phytoalexin. The absorbance was measured at 285 nm in a spectrophotometer. The highest accumulation of the phytoalexins was observed on highests concentrations of the aqueous extracts.

Histochemical analysis of seed reserve mobilization in Passiflora edulis Sims fo. flavicarpa O. Deg. (yellow passion fruit) during germination / Análise histoquímica da mobilização de reservas em sementes de Passiflora edulis Sims fo. flavicarpa O. Deg (maracujá amarelo) durante a germinação

In the present work, we analyzed the histochemical aspects of Passiflora edulis seeds reserve mobilization during the first ten days of germination. Our results showed that mainly lipids present in the endosperm are used as a reserve source, and their levels reduce at the same time the radicle protrudes, between the fourth and sixth day of sowing. Furthermore, protein bodies are present in the cotyledons, which are degraded as germination occurs and are almost depleted by the time of radicle protrusion. Starch grains also appear in the late germination period, and it is not clear if there is any reserve wall polysaccharide consumption in the endosperm.

No presente trabalho analisamos os aspectos histoquímicos da mobilização de reservas das sementes de Passiflora edulis, durante os primeiros dez dias de germinação. Nossos resultados mostraram que principalmente lipídios presentes no endosperma são utilizados como reserva, com o seu nível começando a diminuir ao mesmo tempo em que ocorre a protrusão da radícula, entre o quarto e sexto dia do início da embebição. Corpos proteicos também estão presentes nos cotilédones, e são degradados à medida que ocorre a germinação e são consumidos quase totalmente quando da protrusão da radícula. Grãos de amido também aparecem no período tardio de germinação, e não está claro se há ou não consumo de polissacarídeos de reserva de parede no endosperma.

Morfologia da flor, fruto e plântula de Victoria amazonica (Poepp.) J.C. Sowerby (Nymphaeaceae) / Morphology of flower, fruit and seedling of Victoria amazonica (Poepp.) J.C. Sowerby (Nymphaeaceae)

Victoria amazonica (Poepp.) J.C. Sowerby é uma hidrófita que ocorre nas várzeas de águas brancas e igapós da Bacia Amazônica e na Bacia do rio Paraguai. A morfologia da flor, fruto e plântula/"tirodendro" é objeto do presente trabalho. O material botânico foi coletado em Parintins e Manaus, estado do Amazonas, Brasil. A análise morfológica foi feita em material fresco e fixado em FAA 50. O desenvolvimento das plântulas foi realizado no escuro em frascos com água com teor reduzido de oxigênio. As flores possuem pedicelo longo e são hemicíclicas, diclamídeas, monoclinas e com antese vespertina. Os frutos são carnosos, indeiscentes, com pseudossincarpia. As sementes apresentam arilo que atua na dispersão pela água. As plântulas se desenvolvem em condições de hipoxia e apresentam um cotilédone exposto acicular. O "tirodendro" apresenta eofilos com heterofilia. As flores apresentam caracteres morfológicos básicos da família, a definição do tipo de fruto exige estudo ontogenético e a heterofilia é um caráter típico de plântulas/"tirodendros"de Nymphaeaceae.

Victoria amazonica (Poepp.) J.C. Sowerby is a hydrophyte that occurs in the white water leas and igapos of the Amazonian and Paraguay Basin. The flower, fruit and seedling/"tirodendro" morphology is the object of the present work. The botanical material was collected at Parintins and Manaus, Amazonian state, Brazil. The morphological analysis was made in both fresh and fixed material. The seedling development was accomplished in flasks with water containing little oxygen and maintained in the darkness. Flowers present long pedicel and they are hemicyclic, dichlamydeous, bisexual with vespertine anthesis. Fruits are fleshy, indehiscent with pseudo-syncarpy. Seeds present aril that acts in the water dispersion. Seedlings grow in hypoxy conditions and they present an acicular and exposed cotyledon. The "tirodendro" stage presents eophylls with heterophylly. Flowers present basic morphologic characters of the family, the definition of the fruit type demands ontogenetic study and the heterophylly is a typical character of Nymphaeaceae seedlings/ "tirodendros".

Establishing the regeneration system of sunflower / 生物工程学报

In order to establish a high efficient regeneration system of sunflower, we optimized the process of callus induction, differentiation and rooting by screening the optimum genotype, explant materials, hormone and cytokine concentration and additives. The results indicated that hybrid sunflowers were easier to regenerate than selfing ones; The best explant was four days cotyledon. The optimum induction medium was Murashige and Skoog (MS) + 2.0 mg/L 6-benzyladenine (6-BA) + 0.5 mg/L naphthaleneacetic acid (NAA) + 1.0 mg/L kinetin (KT). The maximum rate of callus induction was 100%. The optimum differentiation medium was MS +0.2 mg/L 6-BA + 0.5 mg/L NAA + 0.3 mg/L KT + 0.3 mg/L silver nitrate (AgNO3) + 0.2 g/L active carbon (AC), and the buds differentiation rate was up to 71%. The best rooting culture medium was 1/2 MS + 0.6 mg/L indolebutyric acid (IBA). The highest rooting rate was 77%. The analysis of variance showed that genotype, explants growth time, different kinds and concentration of hormone, AC concentration had a significant effect on sunflower regeneration.

Cytokinin preconditioning enhances multiple shoot regeneration in Pongamia pinnata (L) Pierre - a potential, non-edible tree seed oil source for biodiesel

An efficient, highly reproducible protocol for multiple shoot induction and plant regeneration of Pongamia pinnata has been successfully developed using cotyledonary node explants. This study also demonstrates that preconditioning of explant stimulates production of multiple shoots from cotyledonary nodes of P. pinnata. The highest direct shoot regeneration (90 percent) with an average of 18.4 +/- 3.1 shoots/explant were obtained when cotyledonary node explants were excised from seedlings germinated on Murashige and Skoog (MS) media supplemented with benzyladenine (BA) 1 mg l-1, and subsequently cultured on MS media with 1 mgl-1 thidiazuron (TDZ). Scanning electron microscope observations of cotyledonary node (CN) explants excised from pre-conditioned and normal seedlings, revealed larger buds with rapid development in BA-preconditioned CN explants. The addition of adenine sulphate significantly increased the average number of shoots per explant. The highest direct shoot regeneration (93 percent) with an average of 32.2 +/- 0.93 shoots/explant was obtained from BA-preconditioned CN when cultured on MS media supplemented with 1 mg l-1 TDZ and 200 mg l-1 adenine sulphate (ADS). Repeated shoot proliferation was observed from BA preconditioned CN explants up to 3 cycles with an average of 15 shoots/explant/cycle when cultured on MS media supplemented with 1 mg l-1 TDZ and 150 mg l-1 L-glutamine, thus producing 45 shoots/CN explant. Shoots were elongated on hormone free MS media and rooted on 1/2 MS media supplemented with 1 mg l-1 of IBA. Rooted shoots were successfully acclimatized and established in soil with 80 percent success. The highly regenerative system developed in this investigation for this important tree could be a useful tool for genetic transformation.

Simple method to prepare DNA templates from a slice of peanut cotyledonary tissue for polymerase chain reaction

An efficient DNA extraction method was developed for peanut seed, where only 3-5 mg cotyledonary tissue was enough for more than 50 PCR reactions with a reaction volume of 15 ul. Both low copy number and high copy number DNA sequences were successfully amplified. Processing one seed sample only took about half an hour. Sampling had no significant effects on germination and development. The DNA extraction method makes it possible to identify transformants and conduct molecular marker studies prior to sowing, and thus may greatly hasten research progress.

Indução de fitoalexinas em soja e sorgo por preparações de Saccharomyces boulardii / Induction of phytoalexins in soybean and sorghum by Saccharomyces boulardii

O gênero Saccharomyces tem sido usado como indutor de resistência ou para controle biológico em muitos patossistemas. Neste trabalho objetivou-se a indução de fitoalexinas em mesocótilos de sorgo e cotilédones de soja pela levedura Saccharomyces boulardii na forma do produto comercial Floratil (Merck) (com 2 x 106 células/mg produto comercial - pc) e massa de células obtidas de meio líquido YEPG (primeiramente com 14 dias de cultivo e, posteriormente, com 7, 14, 21, 28 e 35 dias) ambos em concentrações de 0,005; 0,05; 0,5; 5; 15 e 25 mg/mL, além de filtrado desse meio nas concentrações de 0,01; 0,1; 1; 5; 10 e 20%. Como tratamentos controle utilizou-se água e S. cerevisiae (25 mg/mL de pc) para soja e água e acibenzolar-S-metil (ASM) (125 mg i.a./L) para sorgo. Em soja os três produtos apresentaram efeito dose-dependente, com ajustes de equações de 1° grau e R2 de 0,64; 0,94 e 0,98 não tendo efeito do tempo de cultivo da levedura na indução de fitoalexinas. Em sorgo apenas o filtrado e Floratil tiveram efeito dose-dependente com equação de 1° grau e R2 de 0,63 e 0,94 respectivamente e obteve-se nos diferentes dias de cultivo R2 de 0,62 com a massa de células somente. Portanto, pode-se evidenciar o potencial indutor de fitoalexinas dos produtos a base de S. boulardii para ensaios com indução de resistência em patossistemas envolvendo sorgo e soja.

Saccharomyces yeast compounds have been used as a resistance elicitor or for biological control in many pathosystems. Thus, the aim of this research was to verify the induction of phytoalexins in sorghum mesocotyls and soybean cotyledons by using Saccharomyces boulardii in the form of the commercial product Floratil (Merck) (with 2 x 106 cells/mg) and yeast-cell mass obtained from liquid culture in YEPG medium (with 7, 14, 21, 28 and 35 days old), both at concentrations of 0.005, 0.05, 0.5, 5, 15 and 25 mg/mL, as well as the filtrate of this medium in concentrations of 0.01, 0.1, 1, 5, 10 and 20%. The control treatments consisted of distilled water and S. cerevisiae (25 mg of commercial product per mL) for the soybean tests and distilled water and acibenzolar-S-methyl (125 mg of active ingredient per L) for the sorghum tests. In soybeans the three tested S. boulardii products presented a dose-dependent effect with R2 of 0.64, 0.94 and 0.98 for the culture filtrate, cell suspension and commercial product of S. boulardii, respectively, with no effect of culture time of yeasts on phytoalexin induction. In sorghum, only the culture filtrate and Floratil presented a dose-dependent effect, with R2 of 0.63 and 0.94, respectively, and the cell suspension of S. boulardii showed dependence of culture time with R2 of 0.62. Thus, S. boulardii and its derivates induce phytoalexins and have potential to be used as an elicitor for assays with induction resistance in pathosystems involving sorghum and soybean plants.

Effects of chromium on seed germination, root elongation and coleoptiles growth in six pulses

The ecotoxicological effects of Cr[2+] on germination and early seedling growth of six pulses were investigated. Seeds of these plants were exposed to seven different concentrations of Cr [0-3.2 mM]. The results indicated that root elongation and coleoptile growth of six pulse plants were more sensitive than seed germination for measurement of the toxic of Cr[2+] pollutions. Different species show different levels of tolerance to Cr[2+] pollution. Lablab purpureus and Glycine max are the most sensitive to Cr[2+], their germination percentage, root and coleoptile length were significantly lower than other tested species, by contrast, Lathyrus odoratus and Dumasia villasa are the most resist species, their germination and seedling growth almost were not influenced by Cr[2+] pollution significantly comparing the control. There were significantly negatively correlations between seedling growth and increasing concentration of Cr[2+] for Glvcine max, Vigna radiate and Lablab purpureus. The significantly negative correlations between germination and increasing concentration of Cr[2+] were shown for Glycine max, Vigna radiate and V. angularis

UV-B induced changes in antioxidant enzymes and their isoforms in cucumber (Cucumis sativus L.) cotyledons.

To assess the role of antioxidant defense system on exposure to ultra-violet-B (UV-B) radiation, the activities of antioxidant enzymes superoxide dismutase (SOD), ascorbic acid peroxidase (APX), glutathione reductase (GR) and guaiacol peroxidase (GPX), as well as the level of antioxidants ascorbic acid (AA) and alpha-tocopherol were monitored in cucumber (Cucumis sativus L. var long green) cotyledons. UV-B enhanced the activity of antioxidant enzymes as well as AA content, but decreased the level of alpha-tocopherol. Significant increase was observed in the activities of SOD and GPX. Analysis of isoforms of antioxidant enzymes by native-PAGE and activity staining revealed three isoforms of GPX in unexposed dark-grown cotyledons (control), and their intensity was enhanced by UV-B exposure. In addition, four new isoforms of GPX were observed in cotyledons after UV-B exposure. Although no new isoforms were observed for the other antioxidant enzymes, the activities of their existing isoforms were enhanced by UV-B.