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PubMed1998

B cell responses to a peptide epitope. VII. Antigen-dependent modulation of the germinal center reaction.

Germinal center responses to two analogous peptides, PS1CT3 and G32CT3, that differ in sequence only at one position within the B cell epitopic region were examined. In comparison with peptide PS1CT3, peptide G32CT3 elicited a poor germinal center response. By demonstrating equal facility of immune complexes with IgM and IgG Ab isotypes to seed germinal centers, we excluded differences in isotype profiles of early primary anti-PS1CT3 and anti-G32CT3 Ig as the probable cause. Quantitative differences in germinal center responses to the two peptides were also not due to either qualitative/quantitative differences in T cell priming or variation in the frequency of the early Ag-activated B cells induced. Rather, they resulted from qualitative differences in the nature of B cells primed. Analysis of early primary anti-PS1CT3 and anti-G32CT3 IgMs revealed that the latter population was of a distinctly lower affinity, implying the existence of an Ag affinity threshold that restricts germinal center recruitment of G32CT3-specific B cells. The impediment in anti-G32CT3 germinal center initiation could be overcome by making available an excess of Ag-activated Th cells at the time of immunization. This resulted in the appearance of a higher affinity population of G32CT3-specific B cells that, presumably, are now capable of seeding germinal centers. These data suggest that the strength of a germinal center reaction generated is Ag dependent. At least one regulatory parameter represents the quality of B cells that are initially primed.

Amino Acid Sequence↗

PubMed1984

Polyclonal activation of the murine immune system by an antibody to IgD V. Effect on germinal centers.

Heterologous anti-mouse delta chain has been shown to induce T-independent polyclonal B cell proliferation by 24 h after injection into mice and a T-dependent polyclonal increase in the number of IgG-secreting cells in spleen and lymph nodes 6 days after injection. The effect of anti-delta on germinal center cells has been difficult to determine, however, since anti-delta-induced blast transformation of mantle zone cells makes it difficult to distinguish these cells from germinal center cells. To clarify the effect of anti-delta on germinal centers, we stained lymph node sections from control and anti-delta-injected mice with peanut agglutinin conjugated to horseradish peroxidase which clearly differentiates the germinal center cells from other large and small lymphoid cells. For the first 5 days after i.v. injection of goat anti-mouse delta, the germinal center volumes did not differ significantly from those of mice injected with control antibodies. On days 6 and 7 after injection, the lymph node germinal center volumes of anti-delta-treated mice increased on average over 10-fold as compared to those of mice injected with an equal amount of control antibody. Thus, while anti-delta antibody directly induced blastogenesis of mantle layer B cells, it appears to have little direct effect on germinal center cells. In contrast, rapid germinal center development is seen in anti-delta-injected mice during the T-dependent phase of B cell activation and occurs simultaneously with the appearance of large numbers of IgG-secreting cells.

Animals↗

PubMed1998

The hybrid histidine kinase dhkB regulates spore germination in Dictyostelium discoideum.

Spore germination is a defined developmental process that marks a critical point in the life cycle of Dictyostelium discoideum. Upon germination the environmental conditions must be conducive to cell growth to ensure survival of emerged amoebae. However, the signal transduction pathways controlling the various aspects of spore germination in large part remain to be elucidated. We have used degenerate PCR to identify dhkB, a two-component histidine kinase, from D. discoideum. DhkB is predicted to be a transmembrane hybrid sensor kinase. The dhkB-null cells develop with normal timing to give what seem to be mature fruiting bodies by 22 to 24 h. However, over the next several hours, the ellipsoidal and encapsulated spores proceed to swell and germinate in situ within the sorus and thus do not respond to the normal inhibitors of germination present within the sorus. The emerged amoebae dehydrate due to the high osmolarity within the sorus, and by 72 h 4% or less of the amoebae remain as spores, while most cells are now nonviable. Precocious germination is suppressed by ectopic activation of or expression of cAMP-dependent protein kinase A. Additionally, at 24 h the intracellular concentration of cAMP of dhkB- spores is 40% that of dhkB+ spores. The results indicate that DHKB regulates spore germination, and a functional DHKB sensor kinase is required for the maintenance of spore dormancy. DHKB probably acts by maintaining an active PKA that in turn is inhibitory to germination.

8-Bromo Cyclic Adenosine Monophosphate↗

PubMed1994

Expression of desiccation-induced and lipoxygenase genes during the transition from the maturation to the germination phases in soybean somatic embryos.

Transitions between developmental programs during plant embryogenesis are usually characterized by differential gene expression. Soybean (Glycine max (L.) Merr.) somatic embryos from embryogenic suspension cultures germinate poorly on a germination medium even after being grown on medium containing abscisic acid until developmental changes are induced by desiccation. The objectives of this study were to investigate: (i) the relationship between water loss and germination of somatic embryos, (ii) changes in desiccation-induced polypeptides, and (iii) the expression of maturation- and germination-associated genes during the transition from maturation to germination of soybean somatic embryos. The results revealed that partial drying of somatic embryos at 76% relative humidity for 96 h resulted in more than 90% germination. Germination frequency was correlated with the percent fresh weight loss of the somatic embryos. Polypeptides induced by desiccation were classified into three groups based on the tissues in which they were expressed and the inducing agents to which they responded. Desiccation-induced polypeptides decreased or became undetectable upon inhibition. The expression of a maturation-associated gene, Mat1, was induced in both cotyledons and hypocotyl/radicle tissues of somatic embryos after 72 h desiccation. The transcript for this gene increased in concert with water loss from the embryos. During maturation of somatic embryos, the expression of embryonic lipoxygenase genes was turned on. Desiccation alone did not directly induce expression of germination-associated lipoxygenase genes SC514 and LOXB2 in somatic embryos.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Sequence↗

PubMed1983

Activation and germination characteristics observed in endospores of thermophilic strains of Bacillus.

The properties of endospores of some thermophilic strains of Bacillus were examined. Included were strains isolated from thermal pools and springs in Yellowstone National Park, a strain of B. thermodenitrificans and two strains of B. stearothermophilus, ATCC 7953 (smooth) and T-10. The spores of thermophilic strains of Bacillus contained relatively high levels of dipicolinic acid ranging from 11-14.8% of the spore dry weight, while the calcium levels were similar to those observed in other bacterial endospores including mesophilic bacilli and thermophilic actinomycetes. Spore populations of thermophilic bacilli could not be effectively germinated in solutions of sodium phosphate alone but germinated well in solutions supplemented with one of a variety of organic compounds. Solutions containing L-valine or L-leucine were particularly effective. A wide range of pH permitted the germination of fractions of spore populations, however, optimum germination was observed only at pH values of 6.0 and above. A range in incubation temperatures of less than 25 degrees C permitted 50% or more of the spores of each of the organisms to germinate. Freshly prepared spore did not germinate, but these spores germinated rapidly and completely if they were heated for 30 min at 100 degrees C just prior to germination testing, i.e., the spores were heat activatable. However, spores of thermophilic bacilli could also be activated by shifting them to and holding them at temperatures below their optimum growth temperature of 65 degrees C. Of the ten temperatures tested, ranging from 4 degrees C through 50 degrees C, the optimum reduced temperature for spore activation was 30 degrees C.

Bacillus↗

PubMed1980

Effect of volatile and gaseous metabolites of germinating pea seeds on Micromycetes.

Differences in the effect of volatile and gaseous metabolites of germinating pea seeds on the germination of spores of Mucor racemosus and macroconidia of Fusarium oxysporum are described. Germination of spores of M. racemosus was inhibited by seed metabolites whereas germination of macroconidia of F. oxysporum was stimulated during the first two days and inhibition occurred only after further two days of germination of the seeds. A pronounced inhibition of germination of spores of both micromycetes took place due to absorption of CO2 from volatile and gaseous metabolites. Absorption of some components of seed metabolites in a KMnO4 solution led to a decrease of the inhibitory effect on germination of spores of M. racemosus and stimulatory effect on germination of macroconidia of F. oxysporum.

Absorption↗

PubMed1999

Chloride dependence of endospore germination in halobacillus halophilus

The ion requirement for germination and outgrowth of endospores from the moderately halophilic salt marsh bacterium Halobacillus halophilus was studied. Germination and outgrowth of endospores plated onto nutrient broth was dependent on the salt concentration in the artificial seawater used as the source of ions. Maximal germination and outgrowth were observed when double-concentrated artificial seawater was used. Replacement of chloride salts in the artificial seawater by other salts resulted in a complete loss of germination and outgrowth that was restored upon addition of chloride. To analyze the role of chloride more directly and quantitatively, a defined growth medium was used in which the artificial seawater was substituted by a solution of magnesium sulfate and sodium chloride. Spore germination and outgrowth were strictly dependent on the chloride concentration; maximal germination and outgrowth were observed at approximately 1.3 M Cl(-). Chloride could be substituted by bromide, but not by sulfate or nitrate. Microscopic examinations of single spores clearly showed that germination is the chloride-dependent step. This first report on chloride dependence of spore germination in any endospore-forming bacterium adds another function to chloride in H. halophilus apart from its being essential for the physiology of the vegetative cell. http://link.springer. de/link/service/journals/00203/bibs/9172004/91720264.htm</HEA

Journal Article↗

PubMed2001

Gibberellins and Light-Stimulated Seed Germination.

Bioactive gibberellins (GAs) promote seed germination in a number of plant species. In dicots, such as tomato and Arabidopsis, de novo GA biosynthesis after seed imbibition is essential for germination. Light is a crucial environmental cue determining seed germination in some species. The red (R) and far-red light photoreceptor phytochrome regulates GA biosynthesis in germinating lettuce and Arabidopsis seeds. This effect of light is, at least in part, targeted to mRNA abundance of GA 3-oxidase, which catalyzes the final biosynthetic step to produce bioactive GAs. The R-inducible GA 3-oxidase genes are predominantly expressed in the hypocotyl of Arabidopsis embryos. This predicted location of GA biosynthesis appears to correlate with the photosensitive site determined by using R micro-beam in lettuce seeds. The GA-deficient non-germinating mutants have been useful for studying how GA stimulates seed germination. In tomato, GA promotes the growth potential of the embryo and weakens the structures surrounding the embryo. Endo-b-mannanase, which is produced specifically in the micropylar endosperm in a GA-dependent manner, may be responsible for breaking down the endosperm cell walls to assist germination. Recently, a role for GA in overcoming the resistance imposed by the seed coat was also suggested in Arabidopsis from work with a range of seed coat mutants. Towards understanding the GA signaling pathway, GA response mutants have been isolated and characterized, some of which are affected in GA-stimulated seed germination.

Journal Article↗

PubMed2004

Lipids stimulate spore germination in the entomopathogenic ascomycete Ascosphaera aggregata.

The alfalfa leafcutting bee (Megachile rotundata) is solitary and managed on a large scale for pollination of alfalfa seed crops. The bees nest in holes drilled in wood or polystyrene blocks, and their larvae are highly prone to a fungal disease called chalkbrood. The most prevalent form of chalkbrood is caused by Ascosphaera aggregata, but this ascomycete is difficult to culture. Hyphae will grow on standard fungal media, but spore germination is difficult to achieve and highly variable. We found that germination can be enhanced with oils. Lipids derived from plants and bee larvae increased germination from 50% (without oil) to 75-85% (with oil). Percent germination was significantly greater in the presence of lipids but germination was not significantly different when different oils, including mineral oil, were used. A. aggregata spores oriented along the oil-aqueous interface in the broth in a polar fashion, with swelling and germ tube formation always occurring into the aqueous portion of the broth. The other half of the spore tended to attach to a lipid droplet, where it remained, without swelling, during germ tube formation. The physical attachment of spores to the oil-aqueous interface is what most probably stimulates spore germination, as opposed to some nutritional stimulation. However, further research is needed to determine if and where the spores encounter such an interface when germinating in the host gut, where germination normally occurs.

Animals↗

PubMed1987

Effects of sawdust pollution on the germination of fungal spores in Lagos Lagoon.

Four fungi, Aspergillus flavus, Aspergillus giganteus, Cladosporium oxysporum a and Trichoderma aureoviride, isolated from Lagos Lagoon, were tested for spore germination in aqueous sawdust extract of different hardwood species (Khaya ivorensis, Mitragyna ciliata and Triplochiton scleroxylon). Extracts of M. ciliata were inhibitory to spore germination especially at higher concentrations. Germ tubes of spores decreased in length with increase in extract concentration for C. oxysporum and T. aureoviride, while Aspergillus species showed increase in length up to a peak at 3% sawdust extract concentration, and thereafter germ tube lengths decreased with increasing concentration. Extracts of T. scleroxylon stimulated the spores and percentage germination increased at higher extract concentrations with no significant difference in germ tube length. Similarly, K. ivorensis had a stimulatory effect on spore germination and length of germ tube, especially at higher extract concentrations. Addition of soluble exogenous carbon and nitrogen sources to sawdust extract enhanced spore germination. Apart from A. flavus, which had only 9% germination, all the spores failed to germinate in lagoon water having 21% salinity. The spores were also inhibited by in lagoon water having 21 per thousand salinity. The spores were also inhibited by various inorganic salts, i.e. CaCl(2), KH(2)PO(4), MgSO(4) and NaCl, present in Lagos Lagoon. The results suggest that sawdust pollution causes enrichment of the lagoon, thereby enhancing spore germination.

Journal Article↗

PubMed2004

Esterase activity as a novel parameter of spore germination in Bacillus anthracis.

Spores of Bacillus anthracis were shown to produce esterase activity about 4 min after exposure to conventional germinants such as combinations of amino acids and purine ribosides. Neither amino acids nor ribosides alone induce germination and esterase activity. Expression of esterase activity was chloramphenicol resistant, and correlated with loss of spore refractivity, a traditional parameter of early germination. Based on these observations, we hypothesized that esterase activity could be used as a novel parameter for quantifying early events during spore germination. To test this hypothesis, we measured expression of esterase activity under a variety of germinating conditions. Using diacetyl fluorescein as fluorogenic substrate of esterases, we demonstrated that esterase activity was invariably induced whenever spores were triggered by known germinants. Moreover, D-alanine, an inhibitor of L-alanine-mediated germination, was found to significantly inhibit expression of esterase activity. In terms of molecular mechanisms, esterase expression could represent activation of proteases at the onset of spore germination.

Amino Acids↗

PubMed2003

Cuticular lipids and silverleaf whitefly stage affect conidial germination of Beauveria bassiana and Paecilomyces fumosoroseus.

Beauveria bassiana and Paecilomyces fumosoroseus are generalist entomopathogenic fungi that infect the silverleaf whitefly (Bemisia argentifolii). We found second and third instar whiteflies to be the most susceptible larval stage to both fungi. Conidia of B. bassiana germinated most readily on the cuticle of second instars (54% germinated) and P. fumosoroseus germination was highest on third instar cuticle (45%). Fourth instars (the ultimate instar) had low susceptibility to these pathogens, and spore germination on the cuticle of fourth instars was very low for B. bassiana (7%) and intermediate for P. fumosoroseus (33%). Cuticular lipids were found to have toxic or inhibitory effects on conidia of B. bassiana and P. fumosoroseus when the spores were germinated on nutrient agar in the presence of the lipids. In the absence of added nutrients, P. fumosoroseus conidial germination increased in the presence of the lipids. To test if the inhibitory effects of the lipids were due solely to hydrophobicity (preventing water from coming into contact with the conidia) we tested the effects of synthetic long-chain wax esters. The synthetic wax esters inhibited germination of P. fumosoroseus to a degree that was similar to the effect of the cuticular lipid extracts, but the synthetic lipids did not have a significant effect on B. bassiana. Thus, the thick coating of long-chain wax esters produced by whitefly nymphs affect spore germination of fungal pathogens, but whether they play a significant role in defense against disease is not clear.

Animals↗

PubMed2006

Impact of moisture on in vitro germination of Metarhizium anisopliae and Beauveria bassiana and their activity on Triatoma infestans.

The in vitro germination of 11 Metarhizium anisopliae and 11 Beauveria bassiana isolates originating from substrates collected in rural peridomestic areas in Central Brazil where triatomines are common was tested. Conidia completed germination up to 24 h after exposure to water activity of > 0.99 aw in all isolates tested. At lower 0.93 aw germination was delayed but conidia of most isolates germinated at high rates (> 98%) within 216 h of incubation. Activities of 2 M. anisopliae and 2 B. bassiana isolates with different patterns of germination at 0.93 aw were tested in Triatoma infestans third instar nymphs. There was no relationship between germination kinetics in vitro at 0.93 aw and their activity in vivo at 98, 75 and 43% relative humidity (rh). Isolates with accelerated germination at 0.93 aw were not more virulent at 75 and 43% rh compared with isolates with retarded or no germination. Highest mortalities were observed at 98% rh, and they did not exceed 25% after 25 d incubation at lower 75 and 43% rh. Isolates that originated from a region with an extensive annual arid period showed no adaptation to lower humidity in their activity against T. infestans.

Animals↗

PubMed2001

Heat shock, mass-dependent germination, and seed yield as related components of fitness in Cistus ladanifer.

The different weight-number strategies of seed production displayed by individuals of a Mediterranean fire-prone plant species (Cistus ladanifer) were investigated in relation to seed germination responses to pre-germination heating. A control (no heating), a high temperature during a short exposure time (100 degrees C during 5 min) and a high temperature during a long exposure time (100 degrees C during 15 min) were applied to seeds from different individual plants with different mean seed weight. These pre-germination treatments resemble natural germination scenarios for the studied species, absence of fire, typical Mediterranean shrub fire, and severe fire with high fuel load. Seed germination was related to heat treatments and seed mass. Seed heating increased the proportion of seeds germinating compared with the control treatment. Mean seed weight was positively correlated to the proportion of germinated seeds but only within heat treatments. These results suggest that in periods without fire, the relative contributions to the population dynamics are equal for all seeds, regardless of their mass, whereas heavier seeds would be the main contribution after wildfire events. Since lighter seeds can be produced in higher quantities than heavier ones within a given fruit, the number of seedlings produced per fruit depended strongly on the germination conditions. In the absence of wildfire, fruits producing lighter seeds gave rise to more seedlings; nevertheless, they were numerically exceeded by those producing heavy seeds after a wildfire. The implications of these results are discussed in relation to their consequences on the population dynamics of this species, considering also additional information on stand flammability and changes in seed mass with plant age.

Journal Article↗

PubMed1997

Effect of water activity, modified atmosphere packaging and storage temperature on spore germination of moulds isolated from prunes.

The combined effect of medium water activity (aw) modified atmosphere packaging (MAP) on the conidial germination of Aspergillus niger, Eurotium amstelodami, Penicillium chrysogenum and Fusarium oxysporum was studied by applying response surface methodology (RSM) for 45 days incubation at 20, 30 and 40 degrees C. Oxytetracycline glucose yeast extract (OGYE) agar media were prepared over a wide range of aw (0.80-0.95), controlled by glucose. The headspace atmosphere composition varied from 0-20% for O2 and from 0-80% for CO2 with N2 as balance in the ratio medium/gas mixture of 1/3 (v/v). The results of this experiment were confirmed by repeating some combinations (aw-MAP) in the zone where the inhibition was effective. In contrast to A. niger which germinated and grew better at 30 and 40 degrees C, P. chrysogenum and F. oxysporum were capable of germinating and growing at 20 degrees C but not at 40 degrees C. E. amstelodami was less influenced by the incubation temperature than were the other species. Under anaerobic atmospheres, germination and growth of all tested species were completely inhibited at low aw, while at high aw because of residual O2, conidia germinated but failed to grow. At any aw value between 0.88 and 0.92, conidia failed to germinate when 100% CO2 was applied. Similar results were obtained for E. amstelodami and F. oxysporum in 80% CO2-20% N2 and 60% CO2-40%N2. Under aerobic conditions, at 5% O2 germination and growth occurred only at high aw, while 10 or 20% O2 combined with either 80 or 60% CO2 conidial germination and mould growth were only delayed comparing to the control (air).

Culture Media↗

PubMed1978

Developmental regulation in cotton seed germination: polyadenylation of stored messenger RNA.

Evidence that RNA preexisting in the cotyledons of mature cotton seed (stored mRNA) is polyadenylated during the first day of germination is presented, based on three different experimental data sets. First, actinomycin D is found to inhibit 32PO4 incorporation into mRNA-poly(A) by 62%, into mRNA by 70%, but into poly(A) only 30%. Second, far more 32PO4 and [2-3H]adenosine are incorporated into the poly(A) portion of mRNA-poly(A) than into the mRNA portion as would be expected from their relative sizes and base composition. This underlabeling of the mRNA moiety is enhanced when cotyledons are germinated in actinomycin D. However, an expected distribution of the isotopes between the mRNA and poly(A) moieties is found in cotyledons labeled later in germination. Third, spectral measurements of the absolute amount of mRNA-poly(A) accumulated during the first day of germination in cotyledons germinated in actinomycin D are larger than would be expected from the 70% inhibition of mRNA labeling caused by the drug. The three sets of data suggest that over 50% of the total mass of mRNA polyadenylated during early germination exists in the mature seed. Its complexity, however, has not been measured. These data may explain the sensitivity of much of germination enzyme synthesis to inhibition by 3'dAdo during early germination and its insensitivity to actinomycin D during this period.

Adenosine Monophosphate↗

PubMed1999

Regulation by biotic and abiotic stress of a wheat germin gene encoding oxalate oxidase, a H2O2-producing enzyme.

Germins and germin-like proteins (GLPs) constitute a ubiquitous family of plant proteins that seem to be involved in many developmental and stress-related processes. Wheat germin has been extensively studied at the biochemical level: it is found in the apoplast and the cytoplasm of germinating embryo cells and it has oxalate oxidase activity (EC 1.2.3.4). Germin synthesis can also be induced in adult wheat leaves by auxins and by a fungal pathogen but it remains to be determined whether the same gene is involved in developmental, hormonal and stress response. In this work, we have studied the expression of one of the wheat germin genes, named gf-2.8, in wheat as well as in transgenic tobacco plants transformed with either this intact gene or constructs with GUS driven by its promoter. This has allowed us to demonstrate that expression of this single gene is both developmentally and pathogen-regulated. In addition, we show that expression of the wheat gf-2.8 germin gene is also stimulated by some abiotic stresses, especially the heavy metal ions Cd2+, Cu2+ and Co2+. Several chemicals involved in stress signal transduction pathways were also tested: only polyamines were shown to stimulate expression of this gene. Because regulation of the wheat gf-2.8 germin gene is complex and because its product results in developmental and stress-related release of hydrogen peroxide in the apoplast, it is likely that it plays an important role in several aspects of plant growth and defence mechanisms.

Antioxidants↗

PubMed1999

Water activity and temperature effects on germination and growth of Eurotium amstelodami, E. chevalieri and E. herbariorum isolates from bakery products.

This study investigated the effect of temperature (5-30 degrees C), water activity (0.775-0.90 aw) and their interactions on the temporal rates of germination and mycelial growth of three species of Eurotium on flour wheat sucrose medium. Germination was quite rapid at aw >0.85, with an almost linear increase with time for all isolates. However, under more extreme water stress, germination was slower. The aw minima for germination were usually lower than those 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 using the Gompertz model modified by Zwietering. Eurotium spp. had shown short lag times at 0.90 aw over a wide range of temperatures. At marginal temperatures, lag phases were significantly longer, especially at >15 degrees C. The temperature x aw profiles for mycelial growth varied between species in terms of rates (mm d(-1)). Predictions of the effect 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 predict fungal spoilage in food products.

Ascomycota↗