Effect of small, acid-soluble proteins on spore resistance and germination under a combination of pressure and heat treatment

To find the range of pressure required for effective high-pressure inactivation of bacterial spores and to investigate the role of alpha/beta-type small, acid-soluble proteins (SASP) in spores under pressure treatment, mild heat was combined with pressure (room temperature to 65 degrees C and 100 to 500 MPa) and applied to wild-type and SASP-alpha(-/)beta(-) Bacillus subtilis spores. On the one hand, more than 4 log units of wild-type spores were reduced after pressurization at 100 to 500 MPa and 65 degrees C, On the other hand, the number of surviving mutant spores decreased by 2 log units at 100 MPa and by more than 5 log units at 500 MPa. At 500 MPa and 65 degrees C, both wild-type and mutant spore survivor counts were reduced by 5 log units. Interestingly, pressures of 100, 200, and 300 MPa at 65 degrees C inactivated wild-type SASP-alpha(+)/beta(+) spores more than mutant SASP-alpha(-)/beta(-) spores, and this was attributed to less pressure-induced germination in SASP-alpha(-)/beta(-) spores than in wild-type SASP-alpha(+)/beta(+) spores. However, there was no difference in the pressure resistance between SASP-alpha(+)/beta(+) and SASP-alpha(-)/beta(-) spores at 100 MPa and ambient temperature (approximately 22 degrees C) for 30 min. A combination of high pressure and high temperature is very effective for inducing spore germination, and then inactivation of the germinated spore occurs because of the heat treatment. This study showed that alpha/beta-type SASP play a role in spore inactivation by increasing spore germination under 100 to 300 MPa at high temperature.

Polyamines, amino acids, IAA and ABA contents during Ocotea catharinensis seed germination

The aim of this work was to Study biochemical variations of IAA (indole-3-acetic acid), ABA (abscisic acid). PAs (polyamines) and amino acids at endogenous levels, during seed germination in Ocotea catharinensis. Seeds were germinated in a vermiculite substratum (100%), samples being collected after 15, 30 and 60 days. Total amino acid levels decreased during the first 15 days. Followed by all increment at the end of germination. Among amino acids, higher concentration was observed in asparagine, this being the predominant amino acid during the whole germination period. Total PAs (free + conjugated) content increased during the first 15 days, followed by a decrease and stabilization between 30 and 60 days of germination. Among the PAs, free putrescine levels rose during the first 15 days, followed by a drop and Stabilization up to 60 days of germination, while spermidine and spermine (spm) contents diminished during the period. Only spin was detected in a conjugated form, with increasing concentrations starting from 30 days on. IAA levels increased during tire first 15 days. followed by a decrease and stabilization until the end of germination (60 days), while ABA contents dwindled during the first 15 days, with similar Values until the end of germination.

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.

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.

A scanning electron microscopy study of the seed and post-seminal development in Angelonia salicariifolia Bonpl. (Scrophulariaceae)

The external morphology of seeds and post-germination developmental stages of Angelonia salicariifolia Bonpl. (Scrophulariaceae) were investigated using scanning electron microscopy. Some structural features of the seed exotesta and seedling in Angelonia are presented for the first time and are of potential taxonomic value for this neotropical genus. The seeds are very small (0.9-1.7 mm long and 0.5-0.9 mm wide), ovate, with a reticulate-crested exotesta, reticules arranged uniformly in longitudinal rows, with a high density of microcilia-like projections on the cell wall of the reticule base and on the edge of the crests. The hilum is located beside the micropyle at the narrow end of the seed. Germination is epigeal. During germination the radicle develops, followed by elongation of the hypocotyl and primary root. At this stage dense root hairs develop on the lower part of the hypocotyl. The apical bud-located between the cotyledons-begins to develop after the cotyledons have unfolded. The cotyledons are equal in size, sessile and ovate. The seedlings have two types of trichomes, one characteristic of the cotyledons and first pair of leaves (glandular, sessile, four-celled head with quadrangular shape) and the other characteristic of the hypocotyl and epicotyl (stalked, erect, elongate and three-celled with dome-shaped unicellular head). (C) 2001 Annals of Botany Company.

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.

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.

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.

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.

Germination and early growth of soybean, dry bean and turnip grown under Digitaria horizontalis straw

The study aimed to evaluate the possible inhibitory effects of different concentrations of crabgrass (Digitaria horizontalis Willd.) dry mass incorporated to the soil over the germination and early growth of soybean (Glycine max (L.) Merril.), dry bean (Phaseolus vulgaris L.), and turnip (Raphanus raphanistrum L.). The experimental design adopted was completely random, with four replications where, each one was consisted of a 2.5 L capacity pot. Dry mass of crabgrass at equivalent amounts of 0, 2.5, 5.0 and 10 t ha(-1) were incorporated into the soil. Crops seedling emergence was checked daily, and germination, speed germination index, mean germination time, relative frequency and synchronization index of germination were computed at the final of 10 days. The height and dry mass of plants were evaluated at 35 days after sowing. The incorporation to the soil of D. horizontalis dry mass caused significant reduction of the height and dry weight of soybean, dry bean and turnip, but were not observed consistent influence over the germination of these species.

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

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Seletividade de herbicidas aplicados em pós-emergência sobre capim-colonião e efeito na qualidade das sementes: effects on seed germination

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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.