Characterization of AFN1, a Gene Associated with Cereal Grain Germination

The AFN1 gene is transiently expressed in germinating oat grains. As AFN1 is not expressed in dormant oat grains during imbibition, we hypothesize that AFN1 may be involved in stimulating the germination process. Sequence analysis of an AFN1 cDNA clone indicates that the AFN1 polypeptide is similar to a previously identified abscisic acid (ABA) glucosyl transferase. This suggests that AFN1 may be acting to glucosylate ABA, thereby inactivating it. As the hormone ABA is known to inhibit germination, ABA glucosylation/inactivation could lead to germination in grains expressing AFN1. To test this hypothesis, we have constructed an expression plasmid that encodes an MBP::AFN1 (maltose binding protein) fusion protein. E. coli cells carrying the expression plasmid were found to produce the MBP::AFN1 fusion protein as a substantial fraction of total protein. We are currently in the process of purifying the MBP::AFN1 fusion protein by affinity chromatography, so that it can be assayed for ABA glucosyl transferase activity. We also wish to test the effect of AFN1 gene expression during grain imbibition on the germination behavior of the grains. To this end, we have constructed plasmids for the overexpression and RNAi-based suppression of AFN1 in transgenic plants. These plasmids have been introduced into oat cells by particle bombardment and we are in the process of regenerating transgenic plants for study.

Plant or fungal sequences? An alternative optimized PCR protocol to avoid ITS (nrDNA) misamplification

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

Controle alternativo de Colletotrichum acutatum agente causal da queda prematura dos frutos cítricos

A queda prematura dos frutos cítricos (QPFC), causada por Colletotrichum acutatum, dados os grandes prejuízos que têm causado aos produtores, constitui-se numa doença de grande importância econômica. O controle da doença é feito predominantemente mediante uso de fungicidas, que eleva o custo de produção e afeta negativamente o meio ambiente. Diante disso, este trabalho teve por objetivo buscar um método alternativo de controle da QPFC, mediante o uso de agentes de biocontrole ou de biofertilizantes. Diferentes concentrações de biofertilizantes (originários de duas fontes distintas e denominados de Bio1 e Bio 2); três isolados de Bacillus subtilis (ACB-69; 72 e 77) e três isolados de Trichoderma spp. (ACB-14; 37 e 39) foram testados, isoladamente ou em combinação, sob condições de laboratório, quanto à capacidade inibitória da germinação de conídios de C. acutatum. Estudaram-se, ainda, a produção de metabólitos termoestáveis por B. subtilis e o efeito sobre a germinação do patógeno. Quinze isolados de B. subtilis foram testados quanto à capacidade de prevenir a infecção por C. acutatum em flores destacadas de lima- ácida 'Tahiti' e, no campo, foram instalados dois experimentos, visando a testar ACBs e biofertilizantes no controle da doença. Verificou-se que o isolado ACB-72 (B. subtilis) e ACB-37 (T. pseudokoningii) foram os que mais inibiram a germinação do patógeno. Quanto à produção de metabólitos termoestáveis, ACB-69 e 77 foram os mais eficientes em produzir substâncias antifúngicas, e em quantidades suficientes para inibirem a germinação do patógeno. A mistura dos quatro isolados de Bacillus (ACBs: 69; 72; 77 e AP3) foi o que apresentou maior porcentagem de inibição (73%). Os biofertilizantes (Bio1 e Bio2), em concentrações acima de 10% e, quando em associação com isolados de Trichoderma spp., promoveram maiores inibições na germinação de C. acutatum. em testes com flores destacadas, verificou-se que, onde foram aplicados os ACBs 69; 76; 74 e 77, as porcentagens de pétalas sem sintomas de infecção por C. acutatum foram de 83; 92; 92 e 97%, respectivamente. Mediante avaliações a campo, verificou-se a potencialidade de B. subtilis e de biofertilizantes em controlar a doença.

Comparison of microbial numbers in soils by using various culture media and temperatures

The influence of different media and incubation temperatures on the quantification of microbial populations in sorghum, eucalyptus and forest soils was evaluated. Microbial growth was compared by using complex (tryptone soybean agar, TSA, casein-starch, CS, and Martin) and saline (Thorton, M3, Czapeck) media and incubation temperatures of 25 and 30° C. Higher numbers of total bacterial. and fungal colony-forming units (CFU) were observed in sorghum soils, and of spore-forming and Gram-negative bacteria in forest soils than other soils. Actinomycetes counts were highest in forest soil when using CS medium at 30° C and in sorghum soil at 25° C in M3 medium. Microorganism counts were dependent on the media and incubation temperatures. The counts at temperatures of 30° C were significantly higher than at 25° C. Microbial quantification was best when using TSA medium for total. and spore-forming bacteria, Thorton for Gram-negative bacteria, M3 for actinomycetes, and Martin for fungi. © 2005 Elsevier GmbH. All rights reserved.

Growth and enzymatic responses of phytopathogenic fungi to glucose in culture media and soil

The effect of inoculation of Aspergillus flavus, Fusarium verticillioides, and Penicillium sp. in Dystrophic Red Latosol (DRL) and Eutroferric Red Latosol (ERL) soils with or without glucose on the total carbohydrate content and the dehydrogenase and amylase activities was studied. The fungal growth and spore production in culture medium with and without glucose were also evaluated. A completely randomized design with factorial arrangement was used. The addition of glucose in the culture medium increased the growth rate of A. flavus and Penicillium sp. but not of F. verticillioides. The number of spores increased 1.2 for F. verticillioides and 8.2 times for A. flavus in the medium with glucose, but was reduced 3.5 times for Penicillium sp. The total carbohydrates contents reduced significantly according to first and second degree equations. The consumption of total carbohydrates by A. flavus and Penicillium sp. was higher than the control or soil inoculated with F. verticillioides. The addition of glucose to soils benefited the use of carbohydrates, probably due to the stimulation of fungal growth. Dehydrogenase activity increased between 1.5 to 1.8 times (p <0.05) in soils with glucose and inoculated with the fungi (except F. verticillioides), in relation to soil without glucose. Amylase activity increased 1.3 to 1.5 times due to the addition of glucose in the soil. Increased amylase activity was observed in the DRL soil with glucose and inoculated with A. flavus and Penicillium sp. when compared to control.

Patogenicidade de isolados de Beauveria bassiana para ovos, larvas e ninfas ingurgitadas de Rhipicephalus sanguineus

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

Effect of temperature on percentage of germination of canistel seeds (Pouteria campechiana)

O canistel é uma fruteira originada no México e América Central, sendo introduzida no Brasil em 1986. As plantas apresentam porte médio, porém podem atingir até 15 metros de altura; as folhas medem cerca de 10 a 25 cm; as flores são completas e pequenas, e o fruto apresenta coloração amarela quando maduro, com polpa esbranquiçada e sabor doce. A propagação pode ser realizada por semente ou por enxertia. em vista da quase total ausência de informações sobre a cultura e a possibilidade de cultivo comercial, realizou-se o presente trabalho, no qual se avaliou o efeito da temperatura na porcentagem de germinação de sementes. Foi verificado que as melhores taxas foram obtidas à temperatura de 30ºC e as menores a 15ºC, 20ºC e 40ºC, sendo esta última a pior delas.

Germination of Albizia hassleri seeds at different temperatures, under laboratory conditions

The objective of this study was to analyze the germination of seeds of Albizia hassleri under different temperatures. A completely random design arranged as a split plot for temperatures regimes, with 11 seed lots and four replications of 15 seeds was used. The plot was represented by the various lots and the sub plots for different temperatures. The means were compared by Scott-Knott test at 5% probability. The temperatures used were: a) constant: 20, 25 and 30 degrees C, and b) alternating: 20-30 and 25-35 degrees C. For all 11 seed lots the mean germination was 90%, speed germination index (IVG) was 5.059, fresh matter of seedlings (MMF) was 0.0628 g and dry matter (MMS) 0.0499 g. The variation coefficient (CV) between plots ranged from 8.48% for germination to 51.71% for dry matter of seedlings and sub plot of 6.77% to 60.45% for germination and MMS. These high values of CV, tested for MMS and MMF, indicate low repeatability of results within each treatment. In general, the IVG obtained at temperatures of 20 and 25 degrees C was lower than those in temperatures of 30, 20-30 and 25-35 degrees C. The best temperature for IVG was the alternating 25-35 degrees C and constant 30 degrees C. The germination test can be conducted at 30, 20-30 and 25-35 degrees C for 19 days.

Effects of substrate and mechanical scarification on the germination of Syagrus oleracea (Arecaceae) seeds

The slow and uneven germination of Syagrus oleracea seeds represents a problem for its propagation, especially for commercial production. This work aimed to study the effects of substrate and mechanical scarification on the germination of S. oleracea seeds. Germination (% G), emergence rate (ER), leaf area (cm(2)), root length (cm), leaf dry mass (g) and root dry mass (g) were also determined. Non-scarified seeds sown in vermiculite germinated faster and showed higher germination (65%), as well as greater root length (16.55 cm). leaf dry mass (5.07 g) and root dry mass (12.41 g), when compared to the scarified seeds.

Temperature and Scarification Effects on the Germination of Syagrus picrophylla Seeds, a Brazilian Native Palm

Syagrus picrophylla belongs to the Arecaceae family. Its propagation is made by seeds; however there is a lack of information in the literature about its seed germination. The objective of this work was to evaluate the effects of temperature and scarification on the germination of S. picrophylla seeds. The experiment was carried out at the State University of São Paulo, Jaboticabal Campus, Brazil. The experimental design was entirely randomized in a factorial arrangement 5 x 2 (five temperature conditions combined with or without mechanical scarification. The number of germinated seeds that showed primary root protrusion was noted daily, and when the germination was stabilized, the germination (%G) and germination rate (GR) were calculated. The analysis of variance (ANOVA) was carried out for means comparing by the Tukey test at 5% confidence level. It was concluded that the greatest %G were obtained at 20-30 degrees C (62%) and 25-35 degrees C (59%) alternated temperatures; %G and GR were similar for both scarified and non-scarified seeds, and there was a faster germination at 20-30 degrees C alternated temperature.

Substrate and Temperature Effects on the Germination of Dypsis leptocheilos (Hodel) Beentje & J. Dransf. (Arecaceae) Seeds

Dypsis leptocheilos is highly valued as an ornamental palm. Its propagation is done by seeds; however, there is little information about this process. The objective of this work was to study the substrate and temperature effects on the germination of D. leptocheilos seeds. The experiments were carried out at FCAV/UNESP, Campus of Jaboticabal, São Paulo State, Brazil. In order to study the substrate effects, four treatments were arranged (coconut fiber, sand, vermiculite and Plantmax (R)) at 30 degrees C. For the temperature effects, six treatments were performed (temperature at laboratory conditions (21.5 degrees C and 72% RU), 25 degrees C, 30 degrees C, 35 degrees C, 20-30 degrees C and 25-35 degrees C), having the coconut fiber as the substrate. The experimental design was completely randomized, with four replicates of 25 disseminules per plot (seeds with attached endocarp). Water replacement was managed to maintain 100% of the retention capacity of the substrates. The germination tests were observed every two days, and conducted over 148 days for the substrate effects, and over 152 days for the temperature effects, when no more seed germination was noted. In order to determine the germination percentage (% G) and the Germination Speed Index (GSI), the seed was considered germinated when the germinative button appeared. At the end of the experiments, leaf area (cm(2)), root and shoot length (cm), root and shoot dry mass (g) were also obtained. Statistical analysis was performed and means were compared by the Tukey test. Germination rate and speed in coconut fiber was higher at 25 and 30 degrees C. However, when other substrates were tested at 30 degrees C, the highest germination percentage was observed in vermiculite, which also sustained better results for the seed germination and for the characteristics related to shoots and roots.

Entomopathogenic fungal activity against pupae and adult Haematobia irritans (Diptera: Muscidae)

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

Influence of the substrate in germination of lychee seeds

A lichia é considerada em todo o mundo como a rainha das frutas, por sua aparência e sabor delicado. Atualmente, a cultura vem despertando interesse, principalmente no Estado de São Paulo, onde, em virtude dos problemas enfrentados pelos agricultores, destacando-se os de laranja e cana-de-açúcar, a procura por alternativas agrícolas vem se intensificando ainda mais. A propagação através de sementes não é recomendada comercialmente, pois as plantas de pé-franco são geneticamente muito desuniformes e apresentam período juvenil muito longo, demorando até 10 anos para entrar em produção; no entanto, a semente pode ser utilizada para a obtenção de porta-enxertos. Observou-se que a germinação de sementes de lichia ocorre rapidamente, reforçando a importância do uso de um substrato adequado. Pela análise dos resultados obtidos, verifica-se que o substrato que aparentemente permitiu as melhores combinações de água e disponibilidade de oxigênio para as sementes - casca de arroz carbonizada e areia lavada, foi o que proporcionou valores de germinação mais rápidos e mais altos.

Seed anatomy and germination of Phoenix roebelenii O'Brien (Arecaceae)

O objetivo desse trabalho foi estudar a morfologia, anatomia e o processo germinativo de sementes de Phoenix roebelenii. Para o levantamento dos dados biométricos foram utilizadas 100 sementes de frutos recém-colhidos, deixados secar ao ar por um dia. Para a germinação, quatro repetições de 50 sementes tratadas com Vitavax-Thiran foram semeadas em bandejas de plástico, contendo Sphagnum sp. como substrato e mantidas sob condições ambientais de laboratório. Detalhes da morfologia da semente foram documentados com o uso de microscopia eletrônica de varredura e esquematizados com auxílio de câmara clara, acoplada ao estereomicroscópio. Foram confeccionadas lâminas permanentes com cortes do embrião, para o estudo de sua anatomia. As dimensões médias das sementes foram: 10,32mm comprimento, 5,21mm largura e 3,91mm espessura. O peso de 1000 sementes foi de 151,1g e 1kg continha 6600 unidades. O início da germinação variou entre 27 e 58 dias. As sementes são albuminosas, com endosperma duro e o embrião é pouco diferenciado, lateral e periférico. A germinação inicia-se pela abertura de um opérculo, através do qual é emitido o pecíolo cotiledonar com o eixo embrionário na extremidade. O pecíolo funciona, internamente, como um haustório, digerindo gradativamente o endosperma. Na sua parte posterior, desenvolve-se a plúmula, que emerge através de uma fenda. Nota-se o aparecimento de raízes secundárias na porção anterior da raiz primária.