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The changing window of conditions that promotes germination of two fire ephemerals, Actinotus leucocephalus (Apiaceae) and Tersonia cyathiflora (Gyrostemonaceae).

BACKGROUND AND AIMS: Following a period of burial, more Actinotus leucocephalus (Apiaceae) and Tersonia cyathiflora (Gyrostemonaceae) seeds germinate in smoke water. The main aim of this study was to determine whether these fire-ephemeral seeds exhibit annual dormancy cycling during burial. This study also aimed to determine the effect of dormancy alleviation on the range of light and temperature conditions at which seeds germinate, and the possible factors driving changes in seed dormancy during burial. METHODS: Seeds were collected in summer, buried in soil in mesh bags in autumn and exhumed every 6 months for 24 months. Germination of exhumed and laboratory-stored (15 degrees C) seeds was assessed at 20 degrees C in water or smoke water. Germination response to light or dark conditions, incubation temperature (10, 15, 20, 25 and 30 degrees C), nitrate and gibberellic acid were also examined following burial or laboratory storage for 24 months. In the laboratory seeds were also stored at various temperatures (5, 15, 37 and 20/50 degrees C) for 1, 2 and 3 months followed by germination testing in water or smoke water. KEY RESULTS: The two species exhibited dormancy cycling during soil burial, producing low levels of germination in response to smoke water when exhumed in spring and high levels of germination in autumn. In autumn, seeds germinated in both light and dark and at a broader range of temperatures than did laboratory-stored seeds, and some Actinotus leucocephalus seeds also germinated in water alone. Dormancy release of Actinotus leucocephalus was slow during dry storage at 15 degrees C and more rapid at higher temperatures (37 and 20/50 degrees C); weekly wet/dry cycles further accelerated the rate of dormancy release. Cold stratification (5 degrees C) induced secondary dormancy. By contrast, no Tersonia cyathiflora seeds germinated following any of the laboratory storage treatments. CONCLUSIONS: Temperature and moisture influence dormancy cycling in Actinotus leucocephalus seeds. These factors alone did not simulate dormancy cycling of Tersonia cyathiflora seeds under the conditions tested.

Analysis of Variance↗

Predicting germination response to temperature. I. Cardinal-temperature models and subpopulation-specific regression.

BACKGROUND AND AIMS: The purpose of this study was to compare the relative accuracy of different thermal-germination models in predicting germination-time under constant-temperature conditions. Of specific interest was the assessment of shape assumptions associated with the cardinal-temperature germination model and probit distribution often used to distribute thermal coefficients among seed subpopulations. METHODS: The seeds of four rangeland grass species were germinated over the constant-temperature range of 3-38 degrees C and monitored for subpopulation variability in germination-rate response. Subpopulation-specific germination rate was estimated as a function of temperature and residual model error for three variations of the cardinal-temperature model, non-linear regression and piece-wise linear regression. The data were used to test relative model fit under alternative assumptions regarding model shape. KEY RESULTS: In general, optimal model fit was obtained by limiting model-shape assumptions. All models were relatively accurate in the sub-optimal temperature range except in the 3 degrees C treatment where predicted germination times were in error by as much as 70 d for the cardinal-temperature models. CONCLUSIONS: Germination model selection should be driven by research objectives. Cardinal-temperature models yield coefficients that can be directly compared for purposes of screening germplasm. Other model formulations, however, may be more accurate in predicting germination-time, especially at low temperatures where small errors in predicted rate can result in relatively large errors in germination time.

Germination↗

Dormancy and the fire-centric focus: germination of three Leucopogon species (Ericaceae) from South-eastern Australia.

BACKGROUND AND AIMS: Germination studies of species from fire-prone habitats are often focused on the role that fire plays in breaking dormancy. However, for some plant groups in these habitats, such as the genus Leucopogon (Ericaceae), dormancy of fresh seeds is not broken by fire cues. In the field, these same species display a flush of seedling emergence post-fire. Dormancy and germination mechanisms therefore appear complex and mostly unknown. This study aimed to identify these mechanisms by establishing dormancy class and testing the effects of a set of typical germination cues, including those directly related to fire and entirely independent of fire. METHODS: To classify dormancy, we assessed seed permeability and embryo morphology, and conducted germination experiments at seasonal temperatures in incubators. To test the effects of fire cues on germination, factorial combinations of smoke, heat and dark treatments were applied. Ageing treatments, using burial and seasonal incubation, were also tested. Germination phenology was established. KEY RESULTS: Seeds were dormant at release and had underdeveloped embryos. Primary dormancy of the study species was classified as morphophysiological. Seasonal temperature changes overcame primary dormancy and controlled timing of germination. Fire cues did not break primary dormancy, but there was a trend for smoke to enhance germination once this dormancy was overcome. CONCLUSIONS: Despite the fact that fire is a predominant disturbance and that many species display a flush of emergence post-fire, seasonal temperatures broke the primary physiological dormancy of the study species. It is important to distinguish between fire being responsible for breaking dormancy and solely having a role in enhancing levels of post-fire germination for seeds in which dormancy has been overcome by other factors. Biogeographical evidence suggests that morphological and physiological factors, and therefore seasonal temperatures, are likely to be important in controlling the dormancy and patterns of post-fire germination of many species in fire-prone regions.

Australia↗

Effects of fire and fire intensity on the germination and establishment of Acacia karroo, Acacia nilotica, Acacia luederitzii and Dichrostachys cinerea in the field.

BACKGROUND: While fire has been used in some instances to control the increase of woody plants, it has also been reported that fire may cause an increase in certain fire-tolerant Acacia tree species. This study investigated germination of Acacia karroo, A. luederitzii and Dichrostachys cinerea, thought to be increasing in density, as well as the historically successful encroaching woody species, A. nilotica, in savanna grassland, Hluhluwe-iMfolozi Park, South Africa. A. karroo is thought to be replacing A. nilotica as the dominant microphyllous species in the park. We tested the hypothesis that observed increases in certain woody plants in a savanna were related to seed germination and seedling establishment. Germination is compared among species for burnt and unburnt seeds on burnt and unburnt plots at three different locations for both hot and cool fires. RESULTS: Acacia karroo showed higher germination (A. karroo 5.1%, A. nilotica 1.5% and A. luederitzii 5.0%) levels and better establishment (A. karroo 4.9%, A. nilotica 0.4% and A. luederitzii 0.4%). Seeds of the shrub Dichrostachys cinerea did not germinate in the field after fire and it is thought that some other germination cue is needed. On average, burning of A. karroo, A. nilotica and A. luederitzii seeds did not affect germination. There was a significant difference in the germination of burnt seeds on burnt sites (4.5%) and burnt seeds on unburnt plots (2.5%). Similarly, unburnt seeds on unburnt sites germinated better (4.9%) than unburnt seeds on burnt sites (2.8%). CONCLUSION: We conclude that a combination of factors may be responsible for the success of A. karroo and that fires may not be hot enough or may occur at the wrong time of year to control A. karroo establishment in HiP. Although germination and establishment of A. karroo was higher than for A. nilotica a competitive advantage after fire could not be shown.

Acacia↗

In vitro pollen germination in Hypericum perforatum L. and Hypericum rumeliacum Boiss.

In vitro pollen germination and pollen tube growth investigations are valuable tools used in identification of the effects of environmental factors and genotypic differences on pollen viability, pollen germination and tube elongation. In this study pollen viability, in vitro pollen germination capacity, abnormality ratios and tube length in germinated pollens of Hypericum perforatum L. and H. rumeliacum Boiss. were investigated. Both of these species has spheroid-shaped and tricolporate pollen grains. The diameters of Hypericum perforatum and H. rumeliacum pollens were found as 24 +/- 3 microm and 19 +/- 2 microm, respectively. Pollen viability of H. perforatum and H. rumeliacum was found as 83% and 72%, respectively. The germination percentages were found as 12.85% for H. perforatum and 64.42% for H. rumeliacurm. Tube lengths in germinated pollens of both taxa were measured approximately as 95.25 +/- 38 microm in H. perforatum and 165.92 +/- 53 microm in H. rumeliacium 4 h after inoculation. In germinated pollen grains of H. perlbratum and H. rumeliacumn abnormality percentages were determined as 13.23% and 43.97%, respectively. In germinated pollens of these two species, highly significant (P < 0.00001) differences in in vitro germination percents and abnormality percents were observed. Abnormalities such as swollen tube tip, branched tube, spiralled tube and excessive tube formation were observed in pollen tubes. The results of this study showed that there were obvious differences in pollen germinability between these two species growing under the same environmental conditions.

Environment↗

[Effects of light and temperature on seed germination of Picea asperata and Betula albo-sinensis].

The effects of different light and temperature conditions on seed germination of two keystone species (Picea asperata and Betula albo-sinensis) in subalpine coniferous forests were studied by simulating environmental conditions in climatic chambers. The results indicated that the optimal temperature of seed germination for both species was 25 degrees C, with the germination rate of 88.8 +/- 8.4% and 30.8 +/- 10.4%, respectively. Through multiple comparisons with Tukey's HSD, B. albo-sinensis was more sensitive to temperature than P. asperata. Seeds of P. asperata could germinate when the temperature reached 10 degrees C, but those of B. albo-sinensis could not, which implied that the latter as a pioneer species of subalpine coniferous forest areas needed higher lowest temperature for seed germination. Day and night temperature difference (15 degreesC/10 degrees C, 20 degrees C/15 degrees C and 20 degrees C/10 degrees C) had significant effect on the germination rates of tested seeds, which was the highest under 20 degrees C/15 degrees C. A wide range of day and night temperature difference seemed to be unfavorable to the seed germination of B. albo-sinensis. Light intensity also had a marked effect on the germination rates of test seeds. Under 20% transmittance, the germination rate of P. asperata and B. albo-sinensis was the highest, being 83.0 +/- 6.6% and 29.2 +/- 5.5%, respectively, and the initiation time and regularity of germination were the best as well. Furthermore, B. albo-sinensis was less sensitive to light than P. asperata, which was determined by the life behavior of tree itself.

Betula↗

Seed germination in response to chemicals: effect of nitrogen and pH in the media.

Seed germination generally presents a peak in the next growing season after a fire. Among other factors associated with fire are the increase of soil nitrogen and changes in the pH of the soil. In this study, we addressed the question, whether or not the germination response of eight species is linked with the increase in pH and nitrogenous compounds in the germination media? We assessed the separate and combined effects of nitrogenous compounds and pH on the percentage and rate of germination of seeds of Medicago arabica (L.) Hudson, Epilobium hirsutum L., Foeniculum vulgare Miller, Daucus carota L., Thapsia villosa L., Cynosurus cristatus L., Dactylis glomerata L. and Rumex crispus L. All these species are well represented in the Mediterranean ecosystems of the central-west Spain. Water and CaCl2 were used as controls. Nitrogenous compounds increased percent germination (level) and rate in three of the species studied. High pH negatively affected the germination rate of seeds from most species, but had no effect on the per cent germination of any of the species. The higher concentration of the nutritious solutions affected negatively the germination level and rate. The different germination responses of seeds of the studied species could not be exclusively attributed to pH values in the media, whereas the amount and form of Nitrogen in the media has a greater effect on it. These differences in germination are species dependent.

Apiaceae↗

Mouse CD38 is down-regulated on germinal center B cells and mature plasma cells.

Germinal center formation is the result of antigenic stimulation of B cells in a T cell-rich area. B cells cycle through the germinal centers, and a small percentage survive to become plasma cells or memory B cells. The transformation from a mature B cell into a germinal center B cell and finally into a terminally differentiated B cell is not well understood. Human CD38 is highly expressed on both germinal center B cells and plasma cells, and is useful in delineating these B cell subsets and in understanding the signaling events involved in the development of these B cells. To determine whether CD38 expression on activated germinal center B cells and postgerminal center B cells influences germinal center differentiation, we studied the expression of CD38 in the mouse. CD38 is expressed on follicular B cells in the Peyer's patches but is down-regulated on germinal center B cells located within the Peyer's patches. CD38dim/-B220+ germinal center B cells are also found in the spleens of immunized but not control mice, suggesting that Ag-stimulated germinal center formation is involved in the production of CD38dim/-B220+ B cells. Furthermore, mature plasma cells isolated from in vitro LPS cultures do not express CD38, but do contain high levels of cytoplasmic Ig. These results are in contrast to studies in humans in which CD38 is not found on follicular B cells but is highly expressed on germinal center B cells and plasma cells.

ADP-ribosyl Cyclase↗

Functional anatomy of germinal centers.

This year we celebrate the first centennial of the discovery of germinal centers by Flemming in 1884. The present paper reviews and adds new data to the functional anatomy of a germinal center. Emphasizing its reactive nature, we first describe a germinal center reaction and then deal with its infrastructural aspects and constituent cell populations, both lymphoid and nonlymphoid. Elements involved in the de novo formation of a germinal center, like antigen, T cells, and the mysterious germinal-center-precursor cell, are discussed. Next, attention is paid to the requirements for lymphoid cells to migrate into germinal centers, and novel features of germinal-center-seeking cells are presented. Subsequently, we discuss kinetic aspects of the high proliferative activity in a germinal center; and finally a description of the functional capacities of germinal-center-derived cells, such as B memory cells and IgM-antibody-forming cell precursors, completes this picture of present-day knowledge of the germinal center, a structure which has yet to reveal its last secrets.

Animals↗

Effects of exogenous nutrients on conidial germination and virulence against the silverleaf whitefly for two hyphomycetes.

Exogenous protein and sugar sources were tested for their impact on conidial germination of two silverleaf whitefly (Bemisia argentifolii) pathogens: Beauveria bassiana and Paecilomyces fumosoroseus. In liquid culture, sugars stimulated only 5-27% germination of B. bassiana and < or =11% germination of P. fumosoroseus, whereas, yeast extract or peptone stimulated 95-100% germination. In the absence of additional nutrients, agar alone stimulated approximately 50% germination. Storing spores for different periods of time did not alter their general response to exogenous nutrients. When spores were germinated before being applied to third instar B. argentifolii, mortality was as much as 2.45 times greater and occurred more rapidly than that for fresh spores. For ungerminated conidia, the mean time to death from infection was 5.45 (SE = 0.16) and 4.74 (SE = 0.08) days for application rates of 37 and 144 conidia x mm(-2), respectively. When conidia were germinated before application, infection times dropped to 4.58 (SE = 0.16) and 4.45 (SE = 0.10) days, respectively. A likely explanation for the greater pathogenicity and virulence of germinated over ungerminated B. bassiana conidia is that only a fraction of the spores applied to whitefly nymphs actually germinate on the cuticle. For some specialized applications, such as greenhouse production systems, it may be beneficial to germinate spores immediately prior to application.

Animals↗

Differential gene expression during germination and after the induction of adventitious bud formation in Norway spruce embryos.

A pulse treatment of embryos of Norway spruce with cytokinin suppresses germinative development and induces the coordinate formation of adventitious buds from subepidermal cell layers. To analyse the patterns of gene expression associated with germination and the alterations induced by the bud induction treatment, we have isolated cDNA clones corresponding to genes that are differentially expressed in cytokinin-treated and untreated in vitro germinating embryos. One category of 14 clones hybridized to transcripts that were abundant specifically during germination. The expression of 8 of these genes was reduced by the bud induction treatment. Four clones, including one identified as a histone H2A gene, recognized transcripts that showed an increased abundance in bud-induced versus in vitro germinating embryos. A second category of 13 clones hybridized to transcripts that increased in abundance during post-germinative development of the seedling. Among these a subset of 8 clones, including an alpha-tubulin clone, corresponds to genes suppressed by the bud induction treatment, whereas 5 clones, including a gene with sequence similarity to polyubiquitin, were unaffected by the treatment. One clone hybridized to a message abundant in the seed, during early germination as well as in the vegetative bud, and showed 60% partial sequence identity to a barley (1----3)-beta-glucanase gene. Genes expressed exclusively in bud-induced or in vitro germinating embryos were not found. The results show that a major difference in gene expression between treated and untreated embryos is related to the shift from extensive cell proliferation to elongation and differentiation that occurs at the transition from germination to post-germinative development, and which is suppressed in the bud-induced embryos.

Amino Acid Sequence↗

Nutritional requirements of keratinophilic fungi and dermatophytes for conidial germination.

Germination of spores of Chrysosporium crassitunicatum, Nannizzia fulva (+), Nannizzia fulva (-) and Trichophyton equinum was studied in the presence of various carbon and nitrogen sources. Effect of different temperatures on spore germination was also determined. Maximum spore germination within 24 hours was recorded when glucose was used as a carbon source for all the test fungi. Except sodium nitrate all the inorganic nitrogen sources enhanced the spore germination at 0.05% concentration. Most of the organic nitrogen sources used were found to be stimulatory for the spore germination of test fungi. Optimum temperature i.e. 28 degrees C supported maximum spore germination of all the test fungi within 24 hours. C. Crassitunicatum, N. fulva (+), N. fulva (-) could germinate upto 35 degrees C but beyond that no spore germination was noticed in these fungi. T. equinum could germinate at a higher temperature of 40 degrees C but the percentage of germination was very low.

Arthrodermataceae↗

Germination of cysts in acetabularia mediterranea.

Techniques for the growth of uniformly reacting populations of cysts of Acetabularia mediterranea and for quantitative measurement of cyst germination have been developed. Cysts of A. mediterranea can be induced to germinated by exposure to the atmosphere. Germination rates are very low in young cysts. They increased during exposure to total darkness. This "maturation of cysts" is found to be completed after a period of 12-15 weeks. Germination rates of cysts that have passed the maturation period exceed 90 percent in continuous white light and 80 percent in darkness. Cysts germinate in less than two days in darkness and less than four days in light. The influence of temperature at a range of 15 degrees C to 25 degrees C on germination kinetics is studied in light and darkness. Germination is accelerated with increasing temperature up to 21 degrees C. At higher temperature germination is delayed in light but the time of germination remains constant in darknesss. Rates of germination are not altered by the influence of temperature in light while in darkness there is a dramatic decrease at temperatures higher than 21 degrees C. From these findings it is concluded that cyst germinationA. mediteranea does not need any light but is influenced by light dependent systems. The influence of light is strongest at elevated temperatures.

Acetabularia↗

A gene encoding alanine racemase is involved in spore germination in Bacillus thuringiensis.

Alanine racemase is a major component of the exosporium of Bacillus cereus spores. A gene homologous to that of alanine racemase (alrA) was cloned from Bacillus thuringiensis subsp. kurstaki, and RT-PCR showed that alrA was transcribed only in the sporulating cells. Disruption of alrA did not affect the growth and sporulation of B. thuringiensis, but promoted L-alanine-induced spore germination. When the spore germination rate was measured by monitoring DPA release, complementation of the alrA disruptant reduced the rate of L-alanine-induced spore germination below that of even wild-type spores. As previously reported for spores of other Bacillus species, D-alanine was an effective and competitive inhibitor of L-alanine-induced germination of B. thuringiensis spores. D-cycloserine alone stimulated inosine-induced germination of B. thuringiensis spores in addition to increasing L-alanine-induced germination by inhibiting alanine racemase. D-alanine also increased the rate of inosine-induced germination of wild-type spores. However, D-alanine inhibited inosine-induced germination of the alrA disruptant spores. It is possible that AlrA converted D-alanine to L-alanine, and this in turn, stimulated spore germination in B. thuringiensis. These results suggest that alrA plays a crucial role in moderating the germination rate of B. thuringiensis spores.

Alanine Racemase↗

Effect of heat treatment on nutritional quality of germinated legume seeds.

The effect of heat treatment of germinated soybean, lupin, and black bean on chemical composition and protein utilization in rats was evaluated. Heat treatment caused complete inactivation of trypsin inhibitors whereas it did not affect phytic acid levels. Proximate components, minerals, and amino acids did not change, but low molecular weight sugars were affected by heat treatment differently for each germinated legume. The sugar digestibility ratio (total digestible sugars/total nondigestible sugars) in germinated black beans doubled after heat treatment. True protein digestibility (TD) increased with heat treatment only in germinated soybean. Net protein utilization was markedly improved (20%) with heat treatment in germinated soybean and lupin. Utilizable protein of heat-treated germinated legumes was 289, 236, and 132 g/kg of legume dry weight for soybean, lupin, and black bean, respectively. Supplementation with methionine did not alter TD but improved all other indices of protein utilization in the germinated legumes, particularly in black bean. All three germinated legumes become equivalent in protein quality when heating and supplementation with methionine are combined with germination. The use of germinated heat-treated soybean, lupin, and black bean on their own and/or as food ingredients is nutritionally advantageous due to the low content of nondigestible oligosaccharides and the high protein utilization.

Amino Acids↗

Temperature and moisture requirements for conidial germination of an isolate of Beauveria bassiana, pathogenic to Rhodnius prolixus.

The effects of temperature, relative humidity and water activity on germination of conidia of an isolate of Beauveria bassiana (Bals.) Vuill. pathogenic to the triatomine vector of Chagas' disease, Rhodnius prolixus Stål., were investigated in vitro. Germination occurred at temperatures between 15 degrees C and 35 degrees C under saturated atmosphere and the optima ranged from 25 degrees C to 30 degrees C. At the extreme temperatures tested (15 degrees C and 35 degrees C) the germination process was delayed, but germination rates reached more than 95%. Germination of B. bassiana conidia was strongly affected by moisture conditions. The availability of water, in both atmospheric and liquid conditions, caused changes in germination times as well as in germination rates. For example, at 25 degrees C + 0.5 degrees C, germination took place within 20 h at 95.5% RH, whereas it needed 72 h of incubation at 90% RH. Germination times increased as the water activity declined from 0.96 a(w) to 0.92 a(w). Below 0.92 a(w), no germination was observed after a 72 h incubation time.

Journal Article↗

Ecological determinants for germination and growth of some Aspergillus and Penicillium spp. from maize grain.

This study compared the effect of temperature (5-45 degrees C), water availability (water activity, aw; 0.995-0.75) and their interactions on the temporal rates of germination and mycelial growth of three mycotoxigenic strains of Aspergillus ochraceus and one isolate each of A. flavus, A. niger, Penicillium aurantiogriseum and P. hordei in vitro on a maize extract medium. Germination was very rapid at > 0.90 aw with an almost linear increase with time for all species. However, at < 0.90 aw, the germination rates of A. flavus and P. hordei were slower. The aw minima for germination were usually lower than for growth and varied with temperature. The effect of aw x temperature interactions on the lag phases (h), prior to germination, and on the germination rates (h(-1)), were predicted for the first time for these fungi using the Gompertz model modified by Zwietering. This showed that A. flavus, A. niger and the two Penicillium spp. had very short lag times between 0.995-0.95 aw over a wide temperature range. At marginal temperatures, these were significantly higher, especially at < 10 degrees C for Aspergillus spp. and > 30 degrees C for Penicillium spp. There were also statistically significant differences between lag phases and germination rates for three different isolates of A. ochraceus. The Aspergillus spp. also germinated faster than the Penicillium spp. The temperature x aw profiles for mycelial growth varied considerably between species, both in terms of rates (mm d(-1)) and tolerances. Predictions of the effects of important environmental factors such as temperature, aw and their interactions on lag times to germination, germination rates and mycelial growth are important in the development of hurdle technology approaches to predicting fungal spoilage in agricultural and food products.

Aspergillus↗

Variability in spore germination response by strains of proteolytic Clostridium botulinum types A, B and F.

AIMS: The objective of the study was to evaluate the variability of germination response of 10 strains of proteolytic Clostridium botulinum. METHODS AND RESULTS: An automated turbidometric method was used to follow the fall in optical density. Spores of proteolytic Cl. botulinum germinated in response to l-alanine alone, with rate and extent of germination increased by addition of l-lactate or bicarbonate ions. Other hydrophobic amino acids also triggered germination of spores of proteolytic Cl. botulinum but not AGFK and inosine, germinants for Bacillus subtilis or B. cereus. CONCLUSIONS: Unlike spores of nonproteolytic Cl. botulinum, all proteolytic Cl. botulinum germinate in hydrophobic l-amino acids without l-lactate. However, a great variability of response to germinant is evidenced between the species. SIGNIFICANCE AND IMPACT OF THE STUDY: The selection of a model strain to study germination of Cl. botulinum spores should consider the variability in sensitivity to germinants shown in this work. In particular, the sequenced strain ATCC 3502 may not be the most appropriate model for germination studies.

Alanine↗