Use of microprojectile bombardment in transient expression assays to analyze protochlorophyllide reductase gene expression in greening maize seedling leaf cells

In young cells of leaf meristems the progenitors of chloroplasts are small organelles known as proplastids, which divide and differentiate into chloroplasts. However, in the absence of light, proplastids undergo a different sequence of development and become etioplasts. When light is supplied to etiolated plants during the "greening" process, etioplasts differentiate into chloroplasts containing chlorophyll. An important light dependent step in chlorophyll biosynthesis is the photoreduction of protochlorophyllide to chlorophyllide by the NADPH:protochlorophyllide reductase (PCR) enzyme. This enzyme is present at high activity only in etiolated tissue and during early stages of light-induced chlorophyll synthesis. The enzyme and its corresponding mRNAs decrease dramatically with prolonged exposure to light. We have investigated the light-dependent transcriptional regulation of a PCR gene in greening maize leaf cells using a transient expression assay based on microprojectile bombardment. The promoter region was isolated and cloned into a ?-glucuronidase (GUS) reporter gene expression plasmid. We have used this chimeric plasmid in tungsten particle bombardment of both etiolated and greening maize seedling leaves to determine whether the cloned promoter region contains regulatory sequences that control light-responsive PCR gene expression.

The establishment mode choise of German multinational enterprises in Brazil: a comparative approach

We examine the drivers behind the establishment mode choice of German multinational enterprises (MNEs) in the sectors of Automotive, Chemicals and Mechanical Engineering in Brazil for the years 1993-2013 using a novel sample of primary data obtained directly from German MNEs. Based on prevalent theories found in the literature, we test the most common hypotheses on our sample. Firms with high R&D activities and firms with prior market knowledge in Brazil in from of previous sales offices are more likely to enter Brazil by a Greenfield investment. We also show that it is the specific private ownership of the German so-called hidden champions that drive those specific SMEs to enter Brazil by Greenfield, a sneaking suspicion that has been made before. Finally, we show that the establishment mode choice between Brazil and the USA only deviates to a low extent, with German MNEs preferring to enter Brazil by Greenfield and the USA by M&A. Thereby, we provide valuable insights for future research in this field.

The effects of gender segregation at the establishment level: an empirical analysis using a panel of matched employer-employee data

This paper studies the relationship between segregation of women across establish- ments and the wages of males and females. To investigate this issue empirically we use a panel of matched employer-employee data from Brazil. Various longitudinal models are used to assess the wage impact of establishment gender segregation. Overall, the results indicate that the e ect of establishment female proportion on the wages of males and females is negative. We also compare these longitudinal results with cross-section estimates, which are the usual ones obtained in the related literature. This com- parison suggests that unmeasured, time-invariant worker- and establishment-speci c e ects are correlated with the establishment female composition.

Headspace solid-phase microextraction-gas chromatography-quadrupole mass spectrometric methodology for the establishment of the volatile composition of Passiflora fruit species

Dynamic headspace solid-phase microextraction (HS-SPME) followed by thermal desorption gas chromatography-quadrupole mass spectrometry analysis (GC-qMS), was used to investigate the aroma profile of different species of passion fruit samples. The performance of five commercially available SPME fibres: 65 μm polydimethylsiloxane/divinylbenzene, PDMS/DVB; 100 μm polydimethylsiloxane, PDMS; 85 μm polyacrylate, PA; 50/30 μm divinylbenzene/carboxen on polydimethylsiloxane, DVB/CAR/PDMS (StableFlex); and 75 μm carboxen/polydimethylsiloxane, CAR/PDMS; was evaluated and compared. Several extraction times and temperature conditions were also tested to achieve optimum recovery. The SPME fibre coated with 65 μm PDMS/DVB afforded the highest extraction efficiency, when the samples were extracted at 50 °C for 40 min with a constant stirring velocity of 750 rpm, after saturating the sample with NaCl (17%, w/v — 0.2 g). A comparison among different passion fruit species has been established in terms of qualitative and semi-quantitative differences in volatile composition. By using the optimal extraction conditions and GC-qMS it was possible to tentatively identify seventy one different compounds in Passiflora species: 51 volatiles in Passiflora edulis Sims (purple passion fruit), 24 in P. edulis Sims f. flavicarpa (yellow passion fruit) and 21 compounds in Passiflora mollissima (banana passion fruit). It was found that the ethyl esters comprise the largest class of the passion fruit volatiles, including 82.8% in P. edulis variety, 77.4% in P. edulis Sims f. flavicarpa variety and 39.9% in P. mollissima. The semi-quantitative results were then submitted to principal component analysis (PCA) in order to establish relationships between the compounds and the different passion fruit species under investigation.

Germinação de sementes e otimização de técnicas de micropropagação de umbuzeiro (spondias tuberosa, arr.) anacardiaceae

The brazilian-plum (Spondias tuberosa, His) is a tropical fruit tree that has been consolidated in the market for agribusiness processing, due to its characteristic flavor of fruit. Accordingly, studies to optimize the propagation of plants are necessary for production of seedlings with agronomic and quality assurance measures. This study aimed at determining the efficient techniques for uniform seed germination, as brazilian-plum seed present mechanical dormancy, and establish optimal culture media for multiplication of shoots from the in vitro micropropagation. Firstly, in a greenhouse at the Universidade Federal do Rio Grande do Norte, was evaluated the influence of different methods of breaking dormancy in the emergence of seedlings of brazilian-plum and speed of germination (IVG) of seeds. After 60 days of cultivation, it was found that splay in the distal portion of the seed was the best treatment, with rates of 85.33% in germinability and 3.415 of IVG, compared with the treatment of seed-soaking in water for 12h + humus and the control group. Subsequently, new sources of seedling explants were obtained in studies of tissue culture. Laboratory of Plant Biotechnology that the university, was used stem apex, nodal segments and internodes in search of decontamination with various concentrations of calcium hypochlorite [Ca(OCl)2] and micropropagation, inoculating them in half WPM (1980) with various concentrations of 6-benzylaminopurine (BAP). We used 10 sample units with three replications for different concentrations of [Ca(OCl)2], BAP and explants type. After thirty days, which was observed for the control of contamination, during the establishment in vitro, concentrations of [Ca(OCl)2] between 0.5% and 2.0% were effective in combating exogenous contamination of the apex. In nodal segments and internodes, concentrations of [Ca(OCl)2] between 1.0% and 2.0% and 1.5% and 2.0% were respectively, sufficient to reduce the percentage of losses in these infestations explants. For micropropagation, the culture medium supplemented with 0.1 mg.L-1 BAP promotes better development of multiple shoots per explants from nodal segment. However, success does not get to shoot training in internodal segment

A simple approach to integrate the ecotoxicological and chemical data for the establishment of environmental risk levels

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Electrical conductivity of the seed soaking solution and soybean seedling emergence

Vigor of soybean [Glycine max (L.) Merrill] seeds can be evaluated by measuring the electrical conductivity (EC) of the seed soaking solution, which has shown a satisfactory relationship with field seedling emergence, but has not had aproper definition of range yet. This work studies the relationship between EC and soybean seedling emergence both in the field and laboratory conditions, using twenty two seed lots. Seed water content, standard germination and vigor (EC, accelerated aging and cold tests) were evaluated under laboratory conditions using -0.03; -0.20; -0.40 and -0.60 MPa matric potentials, and field seedling emergence was also observed. There was direct relationship between EC and field seedling emergence (FE). Under laboratory conditions, a decreasing relationship was found between EC and FE as water content in the substrate decreased, Relationships between these two parameters were also found when -0.03; -0.20 and -0.40 MPa matric potentials were used. EC tests can be used successfully to evaluate soybean seed vigor and identify lots with higher or lower field emergence potential.

Electrical conductivity and soybean seedling emergence

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Automated system of seedling image analysis (SVIS) and electrical conductivity to assess sun hemp seed vigor

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeito do vigor de sementes de soja sobre o seu desempenho em campo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Chemical and physical attributes of two substratum for eucalypt seedling production

This work evaluated the viability of one commercial substrate with pinus bark, vermiculite and 30% of coconut fiber for Eucalyptus grandis seedlings production under different water management: 6, 8, 10, 12 e 14 mm day(-1). In relation to the substrates, it was evaluated, before and after the seedlings production, the total content of carbon, nitrogen and organic substances, pH and EC of the extract, the content of macronutrients (P, K, Ca, S, Mg) and macronutrients (B, Fe, Mn, Cu, Zn), as well as total porosity, density and water retention. It was evaluated the macro and mcronutrients accumulation in the roots and in the aerial part of the seedlings, 105 days after sowing. It was concluded that the substrates presented some distinct properties of those cited as ideal for production in containers with irrigation and fertilization. At the end of the production process, both substrates registered nutrient content little influenced by water management what would guarantee the initial development of the plants in the field.

Seed priming improves endosperm weakening, germination, and subsequent seedling development of Solanum lycocarpum St. Hil.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Fertilizer distribution mechanisms and side dress nitrogen fertilization in upland rice under no-tillage system

Algumas culturas têm pouca adaptação ao sistema plantio direto (SPD), em vista da alta compactação da camada superficial do solo. Nesse caso, o mecanismo utilizado na semeadora para a abertura dos sulcos para deposição do fertilizante pode ter grande importância no sentido de facilitar a penetração das raízes. Avaliouse a influência do mecanismo de distribuição de fertilizante e da adubação nitrogenada na cultura do arroz de terras altas (Oryza sativa) no sistema plantio direto. O experimento foi conduzido nos anos agrícolas 2001/2002 e 2002/2003, em Botucatu-SP, Brasil. O delineamento experimental foi em blocos casualizados, com parcelas subdivididas e quatro repetições. As parcelas foram constituídas por dois mecanismos de distribuição de fertilizantes (haste sulcadora e disco duplo). Nas subparcelas, quatro níveis de N foram aplicados em cobertura (0, 40, 80 e 120 kg ha-1). Avaliou-se a profundidade de abertura do sulco e de deposição das sementes, a população de plantas, a altura de plantas, o número de colmos e de panículas m-2, o número total de espiguetas por panícula, a fertilidade das espiguetas, a massa de 1.000 grãos, a matéria seca da parte aérea, a produtividade e o teor de N na folha bandeira. O sucesso no estabelecimento da cultura do arroz de terras altas no SPD nas regiões de inverno seco do Brasil está diretamente ligado ao mecanismo de distribuição das semeadoras-adubadoras. A haste sulcadora promoveu maior profundidade de deposição de sementes, conseqüentemente reduzindo o estande, o número de panículas por área e a produtividade. A aplicação de N em cobertura no arroz de terras altas em SPD proporciona maiores produtividades quando a semeadura é realizada com mecanismo de disco duplo.

Seed germination and seedling development of white oat affected by silicon and phosphorus fertilization

Os fertilizantes silicatados tem sido cada vez mais usados na agricultura devido a inúmeros benefícios, tais como correção da acidez de solos tropicais e efeitos positivos no desenvolvimento de gramíneas. A disponibilidade de nutrientes e a nutrição de plantas desempenham papel importante na produção de sementes e podem influenciar a qualidade fisiológica de sementes de aveia-branca (Avena sativa L.). Avaliou-se a germinação de sementes e o desenvolvimento de plântulas de aveia-branca em função da adubação com silício e fósforo. O delineamento experimental foi inteiramente casualizado, em esquema fatorial 2 x 4, com seis repetições. Os tratamentos consistiram de 20 e 200 mg dm-3 de P2O5, aplicados na forma de superfosfato triplo, combinados com 0, 150, 300 e 450 mg dm-3 de Si na forma de silicato de potássio. O experimento foi realizado em casa de vegetação, conduzindo-se sete plantas por vaso, com capacidade para 15 L de terra. As panículas foram colhidas e debulhadas manualmente e, as sementes, armazenadas em sacos de papel em condições normais de ambiente. As sementes foram avaliadas quanto ao teor de água, massa de sementes, germinação, condutividade elétrica, comprimento e massa da matéria seca de plântulas. Sementes de aveia-branca com qualidade superior são produzidas com 20 mg dm-3 de P2O5, independente da dose de Si. Sementes com maior germinação e vigor são obtidas com 300 e 450 mg dm-3 de K2SiO3, respectivamente. Os comprimentos da raiz e total das plântulas foram inferiores nas doses de Si até 300 kg ha-1, porém a dose de fósforo somente afetou o desenvolvimento das plântulas de maneira distinta quando aplicada junto com a maior dose de silício.