Osmotic potentials on water uptake and germination of Guazuma ulmifolia Lam. (Sterculiaceae) seeds

Osmotic potentials on water uptake and germination of Guazuma Ulmifolia Lam. (Sterculiaceae) seeds. This work was carried out in the Germination Lab. of the Department of Botany, Institute of Biosciences, São Paulo State University (UNESP), Botucatu, São Paulo State, Brazil. The aims of this work were to determine the water uptake curve and to evaluate the germination of Guazuma ulmifolia seeds subjected to different water potentials. For the water uptake curve, seven replicates of 50 pre-scarified seeds were placed onto paper moistened with 15 mL PEG 6000 solution under the potentials 0 (control), -0.3 and -0.6 MPa at 25o C in the darkness. For the germination assay, four replicates of 50 seeds were subjected to the same above-described conditions; however, one lot of seeds was modified when there was variation in the refractometric index, whereas the remaining ones were kept in the same solutions until the end of the experiment. All three phases of water uptake were detected under 0 and -0.3 MPa; however, phase II was prolonged under -0.6 MPa and germination was not observed. For 0 and -0.3 MPa, the adopted statistical models consisted of asymptotic (phases I and II) and exponential (phase III) functions, y = a*[1 - b*exp (-c*t) + exp (-d + e*(t - t0)]. For -0.6MPa, only the asymptotic function y = a* [1 - b* exp (-c*t)] was used since there was no evidence of germination. The germination final percentage and speed index were lower under -0.3 MPa, mainly when solutions were not replaced; besides, germination was not detected under -0.6 MPa, with or without solution replacement.

Study of Venturia inaequalis sensitivity to fungicides through molecular and biological methodologies

This study was aimed to correlate the results of relative germination from in vitro tests by trifloxystrobin with those of qPCR on a wide range of V. inaequalis populations and monoconidial isolates. Samples were collected in Italian and Turkish distinct locations from orchards with different scab management. In this study, an allele-specific qPCR with primer sets designed was successfully developed to quantitatively determine the frequency of QoI-resistant allele G143A in populations and monoconidial isolates of V. inaequalis. qPCR followed a similar pattern to that obtained using in vitro conidial germination test in very sensitive and very resistant populations. However, the variability between two test results was observed in hetereogenous populations. Therefore, the results of correlations between in vitro and qPCR showed a positive but not very high correlation for Venturia inaequalis populations (R2=0.70). On the contrary, this correlation between two assessment methods was very high for monoconidial isolates (R2=0.92). Results obtained in quantitative PCR and from traditional spore germination assay differed for the same fungal population and in some cases, it is difficult to assess the resistance in the field by only qPCR. It was concluded that it is not always possible to correlate the frequency of detection of the mutation with biological assessment. In such situations, monitoring by molecular techniques must be supported by standard in vitro resistance assessments and observation of field performance in order to have correct conclusions.

Efecto de la temperatura, escarificación y concentraciones de calcio en la germinación de Gymnocalycium schickendantzii (F.A.C. Weber) Britton & Rose (Cactaceae)

Gymnocalycium schickendantzii (F.A.C. Weber) Britton & Rose (Cactaceae) es una especie endémica de Argentina. En la provincia de Mendoza las poblaciones de este cactus todavía no están siendo afectadas por las urbanizaciones y cultivos que ponen en peligro su supervivencia. Conocer aspectos sobre su germinación podría ayudar a explicar el porqué de la presencia de esta especie en su hábitat natural. El objetivo del presente trabajo fue determinar en un ensayo de germinación de G. schickendantzii el efecto de tratamientos de temperaturas de 20 y 30°C durante 25 días en laboratorio usando 3 concentraciones de calcio (1, 10 y 20 meq/l Ca), y agua como testigo con y sin escarificación, y bajo condiciones de luz blanca. Se observó que a 30°C los porcentajes de germinación, con o sin escarificación, fueron significativamente mayores que a 20°C. El efecto de la escarificación sólo ayudó a incrementar los valores de germinación cuando fueron tratadas a 20°C. Las concentraciones de calcio tienen un débil efecto en la germinación de las semillas en las mismas condiciones. El tiempo de inicio de germinación (IG) y para obtener el 50% de ella (T50) fueron mayores a 30°C tanto en las semillas escarificadas como sin escarificar.

Seed Dispersal and Reproduction Patterns Among Everglades Plants

In this study three aspects of sexual reproduction in Everglades plants were examined to more clearly understand seed dispersal and the allocation of resources to sexual reproduction— spatial dispersal process, temporal dispersal of seeds (seedbank), and germination patterns in the dominant species, sawgrass (Cladium jamaicense). Community assembly rules for fruit dispersal were deduced by analysis of functional traits associated with this process. Seedbank ecology was investigated by monitoring emergence of germinants from sawgrass soil samples held under varying water depths to determine the fate of dispersed seeds. Fine-scale study of sawgrass fruits yielded information on contributions to variation in sexually produced propagules in this species, which primarily reproduces vegetatively. It was hypothesized that Everglades plants possess a set of functional traits that enhance diaspore dispersal. To test this, 14 traits were evaluated among 51 species by factor analysis. The factorial plot of this analysis generated groups of related traits, with four suites of traits forming dispersal syndromes. Hydrochory traits were categorized by buoyancy and appendages enhancing buoyancy. Anemochory traits were categorized by diaspore size and appendages enhancing air movement. Epizoochory traits were categorized by diaspore size, buoyancy, and appendages allowing for attachment. Endozoochory traits were categorized by diaspore size, buoyancy, and appendages aiding diaspore presentation. These patterns/trends of functional trait organization also represent dispersal community assembly rules. Seeds dispersed by hydrochory were hypothesized to be caught most often in the edge of the north side of sawgrass patches. Patterns of germination and dispersal mode of all hydrochorous macrophytes with propagules in the seedbank were elucidated by germination analysis from 90 soil samples collected from 10 sawgrass patches. Mean site seed density was 486 seeds/m2 from 13 species. Most seeds collected at the north side of patches and significantly in the outer one meter of the patch edge (p = 0.013). Sawgrass seed germination was hypothesized to vary by site, among individual plants, and within different locations of a plant’s infructescence. An analysis of sawgrass fruits with nested ANOVAs found that collection site and interaction of site x individual plant significantly affect germination ability, seed viability, and fruit size (p < 0.050). Fruit location within a plant’s infructescence did not significantly affect germination. As for allocation of resources to sexual reproduction, only 17.9% of sawgrass seeds germinated and only 4.8% of ungerminated seeds with fleshy endosperm were presumed viable, but dormant. Collectively, only 22% of all sawgrass seeds produced were viable.

Spatially and genetically distinct control of seed germination by phytochromes A and B.

Phytochromes phyB and phyA mediate a remarkable developmental switch whereby, early upon seed imbibition, canopy light prevents phyB-dependent germination, whereas later on, it stimulates phyA-dependent germination. Using a seed coat bedding assay where the growth of dissected embryos is monitored under the influence of dissected endosperm, allowing combinatorial use of mutant embryos and endosperm, we show that canopy light specifically inactivates phyB activity in the endosperm to override phyA-dependent signaling in the embryo. This interference involves abscisic acid (ABA) release from the endosperm and distinct spatial activities of phytochrome signaling components. Under the canopy, endospermic ABA opposes phyA signaling through the transcription factor (TF) ABI5, which shares with the TF PIF1 several target genes that negatively regulate germination in the embryo. ABI5 enhances the expression of phytochrome signaling genes PIF1, SOMNUS, GAI, and RGA, but also of ABA and gibberellic acid (GA) metabolic genes. Over time, weaker ABA-dependent responses eventually enable phyA-dependent germination, a distinct type of germination driven solely by embryonic growth.

Mechanism and control of Genipa americana seed germination

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influence of light, plant density and disinfection in spore germination of fern blechnum brasiliense desv. blechnaceae

The present assay had as objective evaluating spore germination of Blechnum brasiliense in relation to light, plant density and disinfection. The assay was carried out at Jaboticabal, Sào Paulo State, Brazil, from February, 22 to June, 30, 1996. The experimental design was randomized blocks on a factorial scheme (3x2x2), consisting of 12 treatments, three environments (shade-house, dark-house and germination camera), 2 densities (0.005 grs and 0.010 grs of spores/treatment) and presence or absence of disinfection. The leaf coverage area (130 days) and the number of days necessary to germinate were evaluated. The germination camera data were not analysed because they were insignificant; consequently, the remining data were analysed on a 2×2×2 scheme. The shade-house provided larger green covering area and a faster germination. The density of 0.0 lOg of spore/treatments presented the largest green covering area. The supply of partial light was necessary for good germination. The interaction between the environment and the density had significant effect on the green covering area.

Environmental assay on the effect of poultry manure application on soil organisms in agroecosystems

This paper reports the effects produced on the organisms of the soil (plants, invertebrates and microorganisms), after the application of two types of poultry manure (sawdust and straw bed) on an agricultural land. The test was made using a terrestrial microcosm, Multi-Species Soil System (MS3) developed in INIA. There was no difference in the germination for any of the three species of plants considered in the study. The biomass was increased in the wheat (Triticum aestivum) coming from ground treated with both kinds of poultry manure. Oilseed rape (Brasica rapa) was not affected and regarding vetch (Vicia sativa) only straw poultry manure showed significant difference. For length only Vicia sativa was affected showing a reduction when straw was exposed to poultry manure. When the effect on invertebrates was studied, we observed a reduction in the number of worms during the test, especially from the ground control (13.7%), higher than in the ground with sawdust poultry manure (6.7%), whereas in the ground with straw poultry manure, there was no reduction. The biomass was affected and at the end of the test it was observed that while the reduction of worms in the ground control was about 48%, the number of those that were in the ground with sawdust poultry manure or straw poultry manure decreased by 41% and 22% respectively. Finally, the effects on microorganisms showed that the enzymatic activities: dehydrogenase (DH) and phosphatase and basal respiration rate increased at the beginning of the test, and the differences were statistically significant compared with the values of the control group. During the test, all these parameters decreased (except DH activities) but they were always higher than in the ground control. This is why it is possible to deduce that the contribution of poultry manure caused an improvement in the conditions of fertilization and also for the soil.

Action spectra for phytochrome A- and B-specific photoinduction of seed germination in Arabidopsis thaliana.

We have examined the seed germination in Arabidopsis thaliana of wild type (wt), and phytochrome A (PhyA)- and B (PhyB)-mutants in terms of incubation time and environmental light effects. Seed germination of the wt and PhyA-null mutant (phyA) was photoreversibly regulated by red and far-red lights of 10-1,000 micromol m-2 when incubated in darkness for 1-14 hr, but no germination occurred in PhyB-null mutant (phyB). When wt seeds and the phyB mutant seeds were incubated in darkness for 48 hr, they synthesized PhyA during dark incubation and germinated upon exposure to red light of 1-100 nmol m-2 and far-red light of 0.5-10 micromol m-2, whereas the phyA mutant showed no such response. The results indicate that the seed germination is regulated by PhyA and PhyB but not by other phytochromes, and the effects of PhyA and PhyB are separable in this assay. We determined action spectra separately for PhyA- and PhyB-specific induction of seed germination at Okazaki large spectrograph. Action spectra for the PhyA response show that monochromatic 300-780 nm lights of very low fluence induced the germination, and this induction was not photoreversible in the range examined. Action spectra for the PhyB response show that germination was photoreversibly regulated by alternate irradiations with light of 0.01-1 mmol m-2 at wavelengths of 540-690 nm and 695-780 nm. The present work clearly demonstrated that PhyA photoirreversibly triggers the germination upon irradiations with ultraviolet, visible and far-red light of very low fluence, while PhyB controls the photoreversible effects of low fluence.

Evaluation Of The Mutagenicity And Antimutagenicity Of Ziziphus Joazeiro Mart. Bark In The Micronucleus Assay.

The aim of this study was to evaluate the mutagenicity (clastogenicity/aneugenicity) of a glycolic extract of Ziziphus joazeiro bark (GEZJ) by the micronucleus assay in mice bone marrow. Antimutagenic activity was also assessed using treatments associated with GEZJ and doxorubicin (DXR). Mice were evaluated 24-48 h after exposure to positive (N-nitroso-N-ethylurea, NEU - 50 mg.kg(-1) and DXR - 5 mg.kg(-1)) and negative (150 mM NaCl) controls, as well as treatment with GEZJ (0.5-2 g.kg(-1)), GEZJ (2 g.kg(-1)) + NEU and GEZJ (2 g.kg(-1)) + DXR. There were no significant differences in the frequencies of micronucleated polychromatic erythrocytes in mice treated with GEJZ and GEJZ + DXR compared to the negative controls, indicating that GEZJ was not mutagenic. Analysis of the polychromatic:normochromatic erythrocyte ratio revealed significant differences in the responses to doses of 0.5 g.kg(-1) and 1-2 g.kg(-1) and the positive control (NEU). These results indicated no systemic toxicity and moderate toxicity at lower and higher doses of GEZJ. The lack of mutagenicity and systemic toxicity in the antimutagenic assays, especially for treatment with GEZJ + DXR, suggested that phytochemical compounds in Z. joazeiro bark attenuated DXR-induced mutagenicity and the moderate systemic toxicity of a high dose of Z. joazeiro bark (2 g.kg(-1)). Further studies on the genotoxicity of Z. joazeiro extracts are necessary to establish the possible health risk in humans and to determine the potential as a chemopreventive agent for therapeutic use.

Carotenoids Are Effective Inhibitors Of In Vitro Hemolysis Of Human Erythrocytes, As Determined By A Practical And Optimized Cellular Antioxidant Assay.

β-Carotene, zeaxanthin, lutein, β-cryptoxanthin, and lycopene are liposoluble pigments widely distributed in vegetables and fruits and, after ingestion, these compounds are usually detected in human blood plasma. In this study, we evaluated their potential to inhibit hemolysis of human erythrocytes, as mediated by the toxicity of peroxyl radicals (ROO•). Thus, 2,2'-azobis (2-methylpropionamidine) dihydrochloride (AAPH) was used as ROO• generator and the hemolysis assay was carried out in experimental conditions optimized by response surface methodology, and successfully adapted to microplate assay. The optimized conditions were verified at 30 × 10(6) cells/mL, 17 mM of AAPH for 3 h, at which 48 ± 5% of hemolysis was achieved in freshly isolated erythrocytes. Among the tested carotenoids, lycopene (IC(50) = 0.24 ± 0.05 μM) was the most efficient to prevent the hemolysis, followed by β-carotene (0.32 ± 0.02 μM), lutein (0.38 ± 0.02 μM), and zeaxanthin (0.43 ± 0.02 μM). These carotenoids were at least 5 times more effective than quercetin, trolox, and ascorbic acid (positive controls). β-Cryptoxanthin did not present any erythroprotective effect, but rather induced a hemolytic effect at the highest tested concentration (3 μM). These results suggest that selected carotenoids may have potential to act as important erythroprotective agents by preventing ROO•-induced toxicity in human erythrocytes.

Post Hoc Analysis Of The Patricia Randomized Trial Of The Efficacy Of Human Papillomavirus Type 16 (hpv-16)/hpv-18 As04-adjuvanted Vaccine Against Incident And Persistent Infection With Nonvaccine Oncogenic Hpv Types Using An Alternative Multiplex Type-specific Pcr Assay For Hpv Dna.

The efficacy of the human papillomavirus type 16 (HPV-16)/HPV-18 AS04-adjuvanted vaccine against cervical infections with HPV in the Papilloma Trial against Cancer in Young Adults (PATRICIA) was evaluated using a combination of the broad-spectrum L1-based SPF10 PCR-DNA enzyme immunoassay (DEIA)/line probe assay (LiPA25) system with type-specific PCRs for HPV-16 and -18. Broad-spectrum PCR assays may underestimate the presence of HPV genotypes present at relatively low concentrations in multiple infections, due to competition between genotypes. Therefore, samples were retrospectively reanalyzed using a testing algorithm incorporating the SPF10 PCR-DEIA/LiPA25 plus a novel E6-based multiplex type-specific PCR and reverse hybridization assay (MPTS12 RHA), which permits detection of a panel of nine oncogenic HPV genotypes (types 16, 18, 31, 33, 35, 45, 52, 58, and 59). For the vaccine against HPV types 16 and 18, there was no major impact on estimates of vaccine efficacy (VE) for incident or 6-month or 12-month persistent infections when the MPTS12 RHA was included in the testing algorithm versus estimates with the protocol-specified algorithm. However, the alternative testing algorithm showed greater sensitivity than the protocol-specified algorithm for detection of some nonvaccine oncogenic HPV types. More cases were gained in the control group than in the vaccine group, leading to higher point estimates of VE for 6-month and 12-month persistent infections for the nonvaccine oncogenic types included in the MPTS12 RHA assay (types 31, 33, 35, 45, 52, 58, and 59). This post hoc analysis indicates that the per-protocol testing algorithm used in PATRICIA underestimated the VE against some nonvaccine oncogenic HPV types and that the choice of the HPV DNA testing methodology is important for the evaluation of VE in clinical trials. (This study has been registered at ClinicalTrials.gov under registration no. NCT00122681.).

Physiological and biochemical characterization of the assai palm (Euterpe oleracea Mart.) during seed germination and seedling growth under aerobic and anaerobic conditions

Physiological and biochemical aspects of assai palm during seed germination and early seedling growth were investigated. Seeds collected from plants growing in flooded and upland forests were used to determine the influence of normoxic (aerobic) and anoxic (anaerobic) conditions in germination and the initial and average time of development in the roots and shoots. After 75 days, seedlings germinated under normoxia were transferred to trays and submitted to flooding. Seed reserves (lipids, proteins, soluble sugars and starch) were monitored for quiescent and germinated seeds maintained under normoxic and anoxic conditions, as well as after 5, 10 and 20 days of seedling growth. Alcohol dehydrogenase (ADH) activity was quantified in roots and leaves of seedlings without or with flooding (partial and total). Seeds were not able to germinate under anoxia. Different strategies of storage mobilization of lipids, proteins, soluble sugars and starch were observed in seeds of each environment. ADH activity was induced by anoxia, with the highest level observed in the leaves. This study showed that, under normoxic conditions, the best developmental performance of assai palm seeds, from flooded or upland forest areas, during germination was associated with primary metabolites mobilization and seedling flooding tolerance with increased ADH activity. We conclude that the assai palm is well adapted to the anoxic conditions provoked by flooding.

Influence of light and temperature on seed germination of Cereus pernambucensis Lemaire (Cactaceae)

The objective of the present study was to evaluate the effects of light and temperature on germination of Cereus pernambucensis seeds, a species of columnar cactus native to Brazil and naturally incident in the restinga. Cereus pernambucensis seeds were incubated under different temperatures, from 5 to 45 °C, with 5 °C intervals, and under alternating temperatures of 15-20 °C, 15-30 °C, 20-25 °C, 20-30 °C, 20-35 °C, 25-30 °C, 25-35 °C, and 30-35 °C, both under continuous white light and dark. The seeds were also incubated in a gradient of phytochrome photoequilibrium at 25 °C. The highest percentage germination in this species was between 25 and 30 °C. The minimum temperature was between 15 and 20 °C and the maximum between 35 and 40 °C. Alternating temperatures did not affect the percentage of seed germination, but it did alter the rate and synchronization indexes. Seeds incubated in the dark did not germinate under any of the conditions tested, indicating that this species when cultivated present light sensitive seeds controlled by phytochrome. The seeds can tolerate a lot of shade conditions, germinating under very low fluence response of phytochrome.

Determinação eletroquímica da capacidade antioxidante de sucos de frutas industrializados usando o CRAC assay

Fruits juices are natural sources of several compounds that present antioxidant action. Together with the fruits, they contribute with almost 40% of the antioxidant capacity in a healthy diet avoiding and preventing diseases deriving from oxidative stress. The present study determined the antioxidant capacity of seven samples of industrialized fruits juices applying CRAC (Ceric Reducing/Antioxidant Capacity) assay, a new electrochemistry assay that evaluates, by means of chronoamperometric measurements, the ability of a sample in reducing species Ce4+ in acid media. At the end of the assay was obtained the following classification: cashew > guava > grape > mango > apple > orange > passion fruit.