Polyamines, IAA and ABA during germination in two recalcitrant seeds: Araucaria angustifolia (Gymnosperm) and Ocotea odorifera (Angiosperm)

Background and Aims Plant growth regulators play an important role in seed germination. However, much of the current knowledge about their function during seed germination was obtained using orthodox seeds as model systems, and there is a paucity of information about the role of plant growth regulators during germination of recalcitrant seeds. In the present work, two endangered woody species with recalcitrant seeds, Araucaria angustifolia (Gymnosperm) and Ocotea odorifera (Angiosperm), native to the Atlantic Rain Forest, Brazil, were used to study the mobilization of polyamines (PAs), indole-acetic acid (IAA) and abscisic acid (ABA) during seed germination. Methods Data were sampled from embryos of O. odorifera and embryos and megagametophytes of A. angustifolia throughout the germination process. Biochemical analyses were carried out in HPLC. Key Results During seed germination, an increase in the (Spd + Spm) : Put ratio was recorded in embryos in both species. An increase in IAA and PA levels was also observed during seed germination in both embryos, while ABA levels showed a decrease in O. odorifera and an increase in A. angustifolia embryos throughout the period studied. Conclusions The (Spd + Spm) : Put ratio could be used as a marker for germination completion. The increase in IAA levels, prior to germination, could be associated with variations in PA content. The ABA mobilization observed in the embryos could represent a greater resistance to this hormone in recalcitrant seeds, in comparison to orthodox seeds, opening a new perspective for studies on the effects of this regulator in recalcitrant seeds. The gymnosperm seed, though without a connective tissue between megagametophyte and embryo, seems to be able to maintain communication between the tissues, based on the likely transport of plant growth regulators.

Global Gene Expression Analysis during Germination in the Chytridiomycete Blastocladiella emersonii

Blastocladiella emersonii is an aquatic fungus of the Chytridiomycete class. During germination, the zoospore, a motile nongrowing cell, goes through a cascade of morphological changes that culminates with its differentiation into the germling cell, capable of coenocytic vegetative growth. Transcriptome analyses of B. emersonii cells were carried out during germination induced under various environmental conditions. Microarray data analyzing 3,563 distinct B. emersonii genes revealed that 26% of them are differentially expressed during germination in nutrient medium at at least one of the time points investigated. Over 500 genes are upregulated during the time course of germination under those conditions, most being related to cell growth, including genes involved in protein biosynthesis, DNA transcription, energetic metabolism, carbohydrate and oligopeptide transport, and cell cycle control. On the other hand, several transcripts stored in the zoospores are downregulated during germination in nutrient medium, such as genes involved in signal transduction, amino acid transport, and chromosome organization. In addition, germination induced in the presence of nutrients was compared with that triggered either by adenine or potassium ions in inorganic salt solution. Several genes involved in cell growth, induced during germination in nutrient medium, do not show increased expression when B. emersonii zoospores germinate in inorganic solution, suggesting that nutrients exert a positive effect on gene transcription. The transcriptome data also revealed that most genes involved in cell signaling show the same expression pattern irrespective of the initial germination stimulus.

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.

Germination and mycelial growth of Bipolaris euphorbiae Muchovej & Carvalho as influenced by herbicides and surfactants

Bipolaris euphorbiae Muchovej & Carvalho é um forte candidato para o controle de Euphorbia heterophylla L. (amendoim bravo). Este fungo pode ser aplicado em combinação com herbicidas para controlar um maior espectro de espécies daninhas. Para tanto, experimentos laboratoriais foram realizados para verificar a possibilidade da utilização de mistura de tanque de esporos de B. euphorbiae e herbicidas ou surfatantes recomendados para a cultura da soja. Crescimento micelial e germinação de conídios foram avaliados em meio BDA acrescido dos herbicidas, nas concentrações recomendadas dos produtos comerciais, oxasulfuron (80 g/ha), glifosato (4 L/ha), bentazon (1.5 L/ha), fomesafen (1 L/ha), chlorimuron-ethyl (80 g/ha), lactofen (1 L/ha) e imazetaphyr (1 L/ha) e dos surfatantes Energic (2 ml/L), Aterbane (2,5 ml/L), Silwet L-77Ag (1 ml/L), Herbitensil (2 ml/L) e Natur L'óleo (10 ml/L). Diluições dos herbicidas de 50% e 25% foram avaliadas com um consumo de calda equivalente a 300 L/ha. Os surfatantes foram somente utilizados nas concentrações recomendadas. O crescimento micelial não foi afetado por bentazon e fomesafen e apenas levemente por oxasulfuron. Porém, glifosato, chlorimuron-ethyl, lactofen, Energic, Herbitensil, Silwet, e Aterbane o reduziram drasticamente. A redução observada com imazetaphyr foi intermediária e Natur L' óleo promoveu o crescimento micelial. Na presença dos surfatantes, observou-se que todos permitiram uma porcentagem de germinação equivalente àquela alcançada na presença de água. Energic e Herbitensil causaram um retardamento expressivo. Com Herbitensil, o processo germinativo iniciou somente aos 120 minutos. Com herbicidas, foi observado que somente na presença de glifosato e imazetaphyr a germinação dos conídios não seguiu a tendência observada com água, como ocorreu com os outros produtos testados.

Effect of Fusarium graminearum and infection index on germination and vigor of maize seeds

Patógenos em sementes de milho (Zea mays) causam sérios problemas, como a perda de sua capacidade germinativa. O objetivo do trabalho foi determinar qual o melhor tempo para infecção das sementes de milho com Fusarium graminearum, para posterior avaliação dos danos causados pelo fungo na germinação e vigor das mesmas. As sementes foram colocadas sobre meio de BDA contendo o patógeno e incubadas por 4, 8, 16 e 32 h. Após os respectivos períodos de incubação, estas foram submetidas ao teste de sanidade (papel de filtro), com duas variações, sem e com assepsia superficial, usando hipoclorito de sódio a 1% de cloro ativo, por 3 min. Determinado o melhor tempo para infecção, outras sementes foram infetadas com o patógeno, para realização dos testes de germinação e vigor (envelhecimento acelerado e teste de frio) com uma mistura de sementes sadias (colocadas sobre o meio BDA) e sementes inoculadas, resultando em 0, 20, 40, 60, 80 e 100% de sementes infetadas com o fungo em estudo. Os resultados obtidos mostraram que o período de incubação de 32 h foi suficiente para se obter sementes infetadas. Com relação à germinação, não houve diferenças significativas entre os diferentes níveis de infecção, provavelmente devido ao alto vigor das sementes de milho testadas. Quanto aos testes de vigor, os níveis de infecção diferiram significativamente da testemunha, apesar de não terem diferido entre si.

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.

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.

Influence of the temperature in germination of seeds of jabuticaba tree

A jabuticabeira (Myrciaria spp.) é uma fruteira originada do centro-Sul do Brasil e apresenta tipos diferentes. É uma árvore de tamanho médio, com tendência a formar copa com grande número de ramos desde pouco acima do solo. Uma característica considerada como limitante para a cultura comercial dessa frutífera é o grande período juvenil, aconselhando-se então produzir bons porta-enxertos de pés-francos e enxertar as variedades desejadas, ou ainda por outros processos vegetativos. Objetivando-se avaliar a influência da temperatura na germinação de sementes de três clones de jabuticabeira, realizou-se um experimento em condições de laboratório. Observou-se que há influência da temperatura na porcentagem de germinação, sendo que, nas mais baixas temperaturas (15ºC e 20ºC), foi verificada uma maior porcentagem de germinação para os três clones em estudo. Quando do uso da temperatura de 35ºC, dois clones tiveram apenas 8% de germinação, enquanto, para o outro, foi verificado 35%. Estes dados mostram a possibilidade de ocorrência de variabilidade entre clones de jabuticabeira.

Mechanism and control of Genipa americana seed germination

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