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Spore Germination (spore + germination)
Selected AbstractsUV-A/BLUE LIGHT,INDUCED REACTIVATION OF SPORE GERMINATION IN UV-B IRRADIATED ULVA PERTUSA (CHLOROPHYTA),JOURNAL OF PHYCOLOGY, Issue 2 2004Taejun Han Recent reduction in the ozone shield due to manufactured chlorofluorocarbons raised considerable interest in the ecological and physiological consequences of UV-B radiation (,=280,315 nm) in macroalgae. However, early life stages of macroalgae have received little attention in regard to their UV-B sensitivity and UV-B defensive mechanisms. Germination of UV-B irradiated spores of the intertidal green alga Ulva pertusa Kjellman was significantly lower than in unexposed controls, and the degree of reduction correlated with the UV doses. After exposure to moderate levels of UV-B irradiation, subsequent exposure to visible light caused differential germination in an irradiance- and wavelength-dependent manner. Significantly higher germination was found at higher photon irradiances and in blue light compared with white and red light. The action spectrum for photoreactivation of germination in UV-B irradiated U. pertusa spores shows a major peak at 435 nm with a smaller but significant peak at 385 nm. When exposed to December sunlight, the germination percentage of U. pertusa spores exposed to 1 h of solar radiation reached 100% regardless of the irradiation treatment conditions. After a 2-h exposure to sunlight, however, there was complete inhibition of germination in PAR+UV-A+UV-B in contrast to 100% germination in PAR or PAR+UV-A. In addition to mat-forming characteristics that would act as a selective UV-B filter for settled spores under the parental canopy, light-driven repair of germination after UV-B exposure could explain successful continuation of U. pertusa spore germination in intertidal settings possibly affected by intense solar UV-B radiation. [source] Influence of water activity and temperature on conidial germination and mycelial growth of ochratoxigenic isolates of Aspergillus ochraceus on grape juice synthetic medium.JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 10 2005Predictive models Abstract The first stages in the development of Aspergillus ochraceus, an ochratoxin A-producing fungus that infects grapes and may grow on them, have been studied on a synthetic nutrient medium similar to grape in composition. Spore germination and mycelial growth have been tested over a water activity (aw) and temperature range which could approximate to the real conditions of fungal development on grapes. Optimal germination and growth were observed at 30 °C for all three isolates tested. Maximal germination rates were detected at 0.96,0.99 aw at 20 °C, while at 10 and 30 °C the germination rates were significantly higher at 0.99 aw. Although this abiotic factor (aw) had no significant influence on mycelial growth, growth rates obtained at 0.98 aw were slight higher than those at other aw levels. Predictive models for the lag phase before spore germination as a function of water activity and temperature have been obtained by polynomial multiple linear regression, and the resulting response surface models have been plotted. Copyright © 2005 Society of Chemical Industry [source] Use of monoclonal antibodies to quantify the dynamics of ,-galactosidase and endo-1,4-,-glucanase production by Trichoderma hamatum during saprotrophic growth and sporulation in peatENVIRONMENTAL MICROBIOLOGY, Issue 5 2005Christopher R. Thornton Summary Trichoderma species are ubiquitous soil and peat-borne saprotrophs that have received enormous scientific interest as biocontrol agents of plant diseases caused by destructive root pathogens. Mechanisms of biocontrol such as antibiosis and hyperparasitism are well documented and the biochemistry and molecular genetics of these processes defined. An aspect of biocontrol that has received little attention is the ability of Trichoderma species to compete for nutrients in their natural environments. Trichoderma species are efficient producers of polysaccharide-degrading enzymes that enable them to colonize organic matter thereby preventing the saprotrophic spread of plant pathogens. This study details the use of monoclonal antibodies (mAbs) to quantify the production of two enzymes implicated in the saprotrophic growth of Trichoderma species in peat. Using mAbs specific to the hemicellulase enzyme ,-galactosidase (AGL) and the cellulase enzyme endo-1,4-,-glucanase (EG), the relationship between the saprotrophic growth dynamics of a biocontrol strain of Trichoderma hamatum and the concomitant production of these enzymes in peat-based microcosms was studied. Enzyme activity assays and enzyme protein concentrations derived by enzyme-linked immunosorbent assay (ELISA) established the precision and sensitivity of mAb-based assays in quantifying enzyme production during active growth of the fungus. Trends in enzyme activities and protein concentrations were similar for both enzymes, during a 21-day sampling period in which active growth and sporulation of the fungus in peat was quantified using an independent mAb-based assay. There was a sharp increase in active biomass of T. hamatum 3 days after inoculation of microcosms with phialoconidia. After 3 days there was a rapid decline in active biomass which coincided with sporulation of the fungus. A similar trend was witnessed with EG activities and concentrations. This showed that EG production related directly to active growth of the fungus. The trend was not found, however, with AGL. There was a rapid increase in enzyme activities and protein concentrations on day 3, after which they remained static. The reason for the maintenance of elevated AGL probably resulted from secretion of the enzyme from conidia and chlamydospores. ELISA, immunofluoresence and immunogold electron microscopy studies of these cells showed that the enzyme is localized within the cytoplasm and is secreted extracellularly into the surrounding environment. It is postulated that release of oligosaccharides from polymeric hemicellulose by the constitutive spore-bound enzyme leads to AGL induction and could act as an environmental cue for spore germination. [source] Adhesion and development of the root rot fungus (Heterobasidion annosum) on conifer tissues: effects of spore and host surface constituentsFEMS MICROBIOLOGY ECOLOGY, Issue 2 2000Frederick O Asiegbu Abstract The objective of this study was to correlate the occurrence of particular root and woody stump surface components with the ability of spores of the root rot fungus (Heterobasidion annosum) to adhere, germinate and establish on conifer tissues. With the aid of high performance liquid chromatography, several sugars (pinitol, xylitol, dulcitol, mannitol, D -glucose, mannose, fructose) were detected on both stump and fine root surfaces of Scots pine and Norway spruce. Of all the sugars observed, xylose and arabinose were poorly utilized for initiation of germ tube growth whereas spore germination was enhanced in the presence of D -glucose, mannose or fructose. Oxidation of these sugars by pretreatment of wood discs or roots with periodic acid abolished the ability of the spores to germinate. Non-sugar components such as long chain fatty acids on spores and root surfaces as detected with nuclear magnetic resonance were found to have a significant influence on adhesion and initiation of germ tube development. Removal of these aliphatic compounds from the root surface increased spore germination by 2-fold, whereas similar treatment on spores led to a 5-fold decrease in adhesiveness to root material. In vitro studies revealed that the di-ethyl ether extract from the roots had no long term adverse effect on spore germination which suggests that the fungus may possess the capability to detoxify this substance. Similarly, adhesion of spores was affected by low and freezing temperatures. The role of significant levels of mannitol and trehalose accumulated in spores and hyphae of the fungi on viability, survival and tolerance to adverse conditions such as oxidative stress, freezing and desiccation are discussed. [source] Analysis of the germination of spores of Bacillus subtilis with temperature sensitive spo mutations in the spoVA operonFEMS MICROBIOLOGY LETTERS, Issue 1 2004Venkata Ramana Vepachedu Abstract A Bacillus subtilis strain with a base substitution in the ribosome-binding site of spoVAC was temperature sensitive (ts) in sporulation and spores prepared at the permissive temperature were ts in l -alanine-triggered germination, but not in germination with Ca2+ -dipicolinic acid (DPA) or dodecylamine. Spores of a ts spo mutant with a missense mutation in the spoVAC coding region were not ts for germination with l -alanine, dodecylamine or Ca2+ -DPA. These findings are discussed in light of the proposal that SpoVA proteins are involved not only in DPA uptake during sporulation, but also in DPA release during nutrient-mediated spore germination. [source] Effects of modification of membrane lipid composition on Bacillus subtilis sporulation and spore propertiesJOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2009K.K. Griffiths Abstract Aims:, To determine effects of inner membrane lipid composition on Bacillus subtilis sporulation and spore properties. Methods and Results:, The absence of genes encoding lipid biosynthetic enzymes had no effect on B. subtilis sporulation, although the expected lipids were absent from spores' inner membrane. The rate of spore germination with nutrients was decreased c. 50% with mutants that lacked the major cardiolipin (CL) synthase and another enzyme for synthesis of a major phospholipid. Spores lacking the minor CL synthase or an enzyme essential for glycolipid synthesis exhibited 50,150% increases in rates of dodecylamine germination, while spores lacking enzymes for phosphatidylethanolamine (PE), phosphatidylserine (PS) and lysylphosphatidylglycerol (l-PG) synthesis exhibited a 30,50% decrease. Spore sensitivity to H2O2 and tert-butylhydroperoxide was increased 30,60% in the absence of the major CL synthase, but these spores' sensitivity to NaOCl or OxoneÔ was unaffected. Spores of lipid synthesis mutants were less resistant to wet heat, with spores lacking enzymes for PE, PS or l-PG synthesis exhibiting a two to threefold decrease and spores of other strains exhibiting a four to 10-fold decrease. The decrease in spore wet heat resistance correlated with an increase in core water content. Conclusions:, Changing the lipid composition of the B. subtilis inner membrane did not affect sporulation, although modest effects on spore germination and wet heat and oxidizing agent sensitivity were observed, especially when multiple lipids were absent. The increases in rates of dodecylamine germination were likely due to increased ability of this compound to interact with the spore's inner membrane in the absence of some CL and glycolipids. The effects on spore wet heat sensitivity are likely indirect, because they were correlated with changes in core water content. Significance and Impact of the Study:, The results of this study provide insight into roles of inner membrane lipids in spore properties. [source] Treatment with oxidizing agents damages the inner membrane of spores of Bacillus subtilis and sensitizes spores to subsequent stressJOURNAL OF APPLIED MICROBIOLOGY, Issue 4 2004D.E. Cortezzo Abstract Aims:, To determine if treatment of Bacillus subtilis spores with a variety of oxidizing agents causes damage to the spore's inner membrane. Methods and Results:, Spores of B. subtilis were killed 80,99% with wet heat or a variety of oxidizing agents, including betadine, chlorine dioxide, cumene hydroperoxide, hydrogen peroxide, OxoneTM, ozone, sodium hypochlorite and t-butylhydroperoxide, and the agents neutralized and/or removed. Survivors of spores pretreated with oxidizing agents exhibited increased sensitivity to killing by a normally minimal lethal heat treatment, while spores pretreated with wet heat did not. In addition, spores treated with wet heat or the oxidizing agents, except sodium hypochlorite, were more sensitive to high NaCl in plating media than were untreated spores. The core region of spores treated with at least two oxidizing agents was also penetrated much more readily by methylamine than was the core of untreated spores, and spores treated with oxidizing agents but not wet heat germinated faster with dodecylamine than did untreated spores. Spores of strains with very different levels of unsaturated fatty acids in their inner membrane exhibited essentially identical resistance to oxidizing agents. Conclusions:, Treatment of spores with oxidizing agents has been suggested to cause damage to the spore's inner membrane, a membrane whose integrity is essential for spore viability. The sensitization of spores to killing by heat and to high salt after pretreatment with oxidizing agents is consistent with and supports this suggestion. Presumably mild pretreatment with oxidizing agents causes some damage to the spore's inner membrane. While this damage may not be lethal under normal conditions, the damaged inner membrane may be less able to maintain its integrity, when dormant spores are exposed to high temperature or when germinated spores are faced with osmotic stress. Triggering of spore germination by dodecylamine likely involves action by this agent on the spore's inner membrane allowing release of the spore core's depot of dipicolinic acid. Presumably dodecylamine more readily alters the permeability of a damaged inner membrane and thus more readily triggers germination of spores pretreated with oxidizing agents. Damage to the inner spore membrane by oxidizing agents is also consistent with the more rapid penetration of methylamine into the core of treated spores, as the inner membrane is likely the crucial permeability barrier to methylamine entry into the spore core. As spores of strains with very different levels of unsaturated fatty acids in their inner membrane exhibited essentially identical resistance to oxidizing agents, it is not through oxidation of unsaturated fatty acids that oxidizing agents kill and/or damage spores. Perhaps these agents work by causing oxidative damage to key proteins in the spore's inner membrane. Significance and Impact of the Study:, The more rapid heat killing and germination with dodecylamine, the greater permeability of the spore core and the osmotic stress sensitivity in outgrowth of spores pretreated with oxidizing agents is consistent with such agents causing damage to the spore's inner membrane, even if this damage is not lethal under normal conditions. It may be possible to take advantage of this phenomenon to devise improved, less costly regimens for spore inactivation. [source] Control of Aspergillus growth and aflatoxin production using antioxidants at different conditions of water activity and pHJOURNAL OF APPLIED MICROBIOLOGY, Issue 2 2003A. Nesci Abstract Aims: The effect of butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), trihydroxybutyrophenone (THB) and propyl paraben (PP) (at concentrations of 1, 10 and 20 mmol l,1) on germination, growth and aflatoxin B1 production by Aspergillus section Flavi was evaluated. Methods and Results: Studies on the percentage of spore germination, elongation rate, growth rate and aflatoxin B1 production were carried out in vitro in relation to water activity (aw) at 0·982, 0·937, 0·809 and 0·747 values. At 0·809 and 0·747aw values none of the isolates was able to germinate. Overall, PP and BHA were the antioxidants most effective at inhibiting germination of both species. In the presence of the lowest concentration of BHA and PP (1 mmol l,1) the conidial germination percentage ranged from 2 to 19% after 15 h of incubation at the highest water activity tested. BHA and PP at 10,20 mmol l,1 completely inhibited conidial germination. The antioxidants more efficient in controlling Aspergillus elongation rate were PP, BHT and BHA. All strains were much more sensitive to all antioxidants tested on the percentage of spore germination and growth rate at 0·937aw. The antioxidants PP and BHA completely inhibited aflatoxin B1 production by all strains when added at 1 mmol l,1. Decreased aflatoxin B1 levels in comparison with the control, were observed with BHT at 1, 10 and 20 mmol,1 with the strain T20 at 0·982aw. In contrast, stimulation was observed with the antioxidant THB at 10 and 20 mmol l,1 at 0·937aw with the strains T20 and T23. The effect of BHA and PP at 1 mmol l,1 on lag phase and growth rate was maintained in the pH range between 6 and 8. At all pH values the inhibitory effect of BHA was higher than PP. No aflatoxin B1 was detected at all pH values. Conclusions: The data show that BHA and PP could be considered as effective fungitoxicants for A. flavus and A. parasiticus. Significance and Impact of the Study: The information obtained show promise for controlling growth and aflatoxin B1 in stored maize. Futher studies should be carried out to examine the potential for antioxidants, such as BHA and PP to effectively control both growth and aflatoxin production. [source] An antifungal compound produced by Bacillus subtilis YM 10,20 inhibits germination of Penicillium roqueforti conidiosporesJOURNAL OF APPLIED MICROBIOLOGY, Issue 2 2003G.S. Chitarra Abstract Aims: To identify and characterize an antifungal compound produced by Bacillus subtilis YM 10-20 which prevents spore germination of Penicillium roqueforti. Methods and Results: The antifungal compound was isolated by acid precipitation with HCl. This compound inhibited fungal germination and growth. Identification by HPLC and mass spectrometry analysis showed high similarity to iturin A. Permeabilization and morphological changes in P. roqueforti conidia in the presence of the inhibitor were revealed by fluorescence staining and SEM, respectively. Conclusions: The iturin-like compound produced by B. subtilis YM 10-20 permeabilizes fungal spores and blocks germination. Significance and Impact of the Study: Fluorescence staining in combination with flow cytometry and scanning electron microscopy are efficient tools for assessing the action of antifungal compounds against spores. Iturin-like compounds may permeabilize fungal spores and inhibit their germination. [source] Bacillus megaterium spore germination is influenced by inoculum sizeJOURNAL OF APPLIED MICROBIOLOGY, Issue 5 2002M.L. Caipo Aims:,The effect of spore density on the germination (time-to-germination, percent germination) of Bacillus megaterium spores on tryptic soy agar was determined using direct microscopic observation. Methods and Results:,Inoculum size varied from approximately 103 to 108 cfu ml,1 in a medium where pH=7 and the sodium chloride concentration was 0·5% w/v. Inoculum size was measured by global inoculum size (the concentration of spores on a microscope slide) and local inoculum size (the number of spores observed in a given microscope field of observation). Both global and local inoculum sizes had a significant effect on time-to-germination (TTG), but only the global inoculum size influenced the percentage germination of the observed spores. Conclusions:,These results show that higher concentrations of Bacillus megaterium spores encourage more rapid germination and more spores to germinate, indicating that low spore populations do not behave similarly to high spore populations. Significance and Impact of the Study:,A likely explanation for the inoculum size-dependency of germination would be chemical signalling or quorum sensing between Bacillus spores. [source] Effect of sporulation and recovery medium on the heat resistance and amount of injury of spores from spoilage bacilliJOURNAL OF APPLIED MICROBIOLOGY, Issue 5 2001A.E. Cazemier Aims:,To assess the influence of sporulation media on heat resistance, and the use of stress recovery media to measure preservation injury of spores of five representative spoilage bacilli. Methods and Results:,Bacillus spores prepared on nutrient agar supplemented with Ca2+, Mg2+, Mn2+, Fe2+ and K+ were more heat-resistant than spores obtained from nutrient agar with Mn2+. This increased heat resistance correlated with a decrease in the protoplast water content as determined by buoyant density sedimentation. The degree of preservation injury severity could be assessed on media containing NaCl at moderate pH and organic acids at acid pH. Ca-DPA, K+ or proline were added to the recovery media to demonstrate that heat probably caused injury to both spore germination and the outgrowth system. Significance and Impact of the Study:,The metal content of sporulation media can strongly effect the validity of preservation resistance studies. The distinctive recovery media developed here can be relevant for assessing and comparing new preservation technologies. [source] Chitosan Protects Cooked Ground Beef and Turkey Against Clostridium perfringens Spores During ChillingJOURNAL OF FOOD SCIENCE, Issue 6 2006Vijay K. Juneja ABSTRACT:, We investigated the inhibition of Clostridium perfringens spore germination and outgrowth by the biopolymer chitosan during abusive chilling of cooked ground beef (25% fat) and turkey (7% fat) obtained from a retail store. Chitosan was mixed into the thawed beef or turkey at concentrations of 0.5%, 1.0%, 2.0%, or 3.0% (w/w) along with a heat-activated 3-strain spore cocktail to obtain a final spore concentration of 2 to 3 log10 CFU/g. Samples (5 g) of the ground beef or turkey mixtures were then vacuum-packaged and cooked to 60 °C in 1 h in a temperature-controlled water bath. Thereafter, the products were cooled from 54.4 to 7.2 °C in 12, 15, 18, or 21 h, resulting in 4.21, 4.51, 5.03, and 4.70 log10 CFU/g increases, respectively, in C. perfringens populations in the ground beef control samples without chitosan. The corresponding increases for ground turkey were 5.27, 4.52, 5.11, and 5.38 log10 CFU/g. Addition of chitosan to beef or turkey resulted in concentration- and time-dependent inhibition in the C. perfringens spore germination and outgrowth. At 3%, chitosan reduced by 4 to 5 log10 CFU/g C. perfringens spore germination and outgrowth (P, 0.05) during exponential cooling of the cooked beef or turkey in 12, 15, or 18 h. The reduction was significantly lower (P < 0.05) at a chilling time of 21 h, about 2 log10 CFU/g, that is, 7.56 log10 CFU/g (unsupplemented) compared with 5.59 log10 CFU/g (3% chitosan). The results suggest that incorporation of 3% chitosan into ground beef or turkey may reduce the potential risk of C. perfringens spore germination and outgrowth during abusive cooling from 54.4 to 7.2 °C in 12, 15, or 18 h. [source] UV-A/BLUE LIGHT,INDUCED REACTIVATION OF SPORE GERMINATION IN UV-B IRRADIATED ULVA PERTUSA (CHLOROPHYTA),JOURNAL OF PHYCOLOGY, Issue 2 2004Taejun Han Recent reduction in the ozone shield due to manufactured chlorofluorocarbons raised considerable interest in the ecological and physiological consequences of UV-B radiation (,=280,315 nm) in macroalgae. However, early life stages of macroalgae have received little attention in regard to their UV-B sensitivity and UV-B defensive mechanisms. Germination of UV-B irradiated spores of the intertidal green alga Ulva pertusa Kjellman was significantly lower than in unexposed controls, and the degree of reduction correlated with the UV doses. After exposure to moderate levels of UV-B irradiation, subsequent exposure to visible light caused differential germination in an irradiance- and wavelength-dependent manner. Significantly higher germination was found at higher photon irradiances and in blue light compared with white and red light. The action spectrum for photoreactivation of germination in UV-B irradiated U. pertusa spores shows a major peak at 435 nm with a smaller but significant peak at 385 nm. When exposed to December sunlight, the germination percentage of U. pertusa spores exposed to 1 h of solar radiation reached 100% regardless of the irradiation treatment conditions. After a 2-h exposure to sunlight, however, there was complete inhibition of germination in PAR+UV-A+UV-B in contrast to 100% germination in PAR or PAR+UV-A. In addition to mat-forming characteristics that would act as a selective UV-B filter for settled spores under the parental canopy, light-driven repair of germination after UV-B exposure could explain successful continuation of U. pertusa spore germination in intertidal settings possibly affected by intense solar UV-B radiation. [source] COALESCENCE VERSUS COMPETITION: FIELD AND LABORATORY STUDIES OF INTRA- AND INTERSPECIFIC ENCOUNTERS AMONG COALESCING SEAWEEDSJOURNAL OF PHYCOLOGY, Issue 2000B. Santelices Classical ecological theory predicts that whenever growing individuals share a common and limiting resource, such as substratum in mid-intertidal and shallow subtidal habitats, preemptive competition will occur determining species abundance and distribution patterns. However, conspecificity of several ecologically dominant Rhodophyta may coalesce when grown in laboratory cultures. The extent at which intraspecific coalescence occurs in the field and whether the process may also happens during interspecific encounters remain to be determined. If intra- and interspecific coalescence effectively occurs, then coexistence through coalescence rises as an alternative to competition among red-algal dominated intertidal and shallow subtidal communities. Populations of Mazzaella laminarioides and Nothogenia fastigiata living in mid-intertidal, semi-exposed rocky habitats in Central Chile are being used to test the above ideas. Intra- and interspecific encounters occur in the field throughout the year. Coalescence does occur among conspecific partners but it has not been detected in interspecific encounters. Rather, a thick interface of compressed cells, necrotic tissues and cyanobacterial nodules is formed between the two contacting partners. In addition, observations of laboratory cultures indicate that spore germination, germling survival and differentiation of erect axes in bispecific cultures may be reduced when compared to single-species controls. Interspecific differences in growth and differentiation rates appear as the mechanisms explaining a lack of coalescence and negative effects during interspecific contacts. On the other hand, the existence of conspecific coalescence in the field suggests this process should be considered as a real alternative to intraspecific competition among coalescing Rhodophyta. [source] Rapid Screening Method of Cassava Cultivars for Resistance to Colletotrichum gloeosporioides f.sp. manihotisJOURNAL OF PHYTOPATHOLOGY, Issue 1 2002C. N. FOKUNANG An in vitro method for assessing cassava anthracnose disease (CAD) resistance was developed as a preliminary screen to a CAD-resistant breeding programme. Potato dextrose agar (PDA) media was amended by extracts from the stem cortex of 10 cassava cultivars (30001; 30572, 30211, 88/02549, 88/00695, 88/01336, 91/00344, 91/00313, 91/00684 and 91/00475), and assayed for efficacy of inhibition of the growth of Colletotrichum gloeosporioides f. sp. manihotis isolates (05FCN, 10FCN, 12FCN, and 18FCN). Morphological and physiological data indicated that there was a significant difference (P , 0.05), in mycelial growth, spore germination and sporulation among the four isolates on PDA amended with cassava stem extracts. Extracts from cassava cultivars 30211, 91/00684 and 91/00313 showed higher inhibition of germ tube development, mycelial growth and sporulation of the fungal isolates, whereas cultivars 88/02549 and 88/01336 showed the least inhibition. The 10 cultivars were further tested in both greenhouse and field conditions, under disease pressure for two planting seasons, to corroborate resistance to the fungus as observed in vitro. Greenhouse and field trials with the 10 cassava cultivars showed a significant difference (P , 0.05) in CAD resistance. Cultivars 88/02549 and 88/01336 were highly CAD-susceptible, as shown in the in vitro assays and confirmed in the greenhouse and field tests. The other eight cultivars were either resistant (30211, 91/00684), or moderately resistant (30572, 88/00695, 91/00475, 91/00344, 30001 and 91/00313) to CAD. The study shows that an in vitro screening assay of cassava for resistance to CAD could serve as a convenient preliminary screening technique to discriminate CAD-resistant from CAD-susceptible cassava cultivars. The in vitro screening method considerably reduces time and labour in comparison with the current screening techniques of cassava, which involve field planting, inoculation and evaluation. [source] Antifungal Activity of a Bowman,Birk-type Trypsin Inhibitor from Wheat KernelJOURNAL OF PHYTOPATHOLOGY, Issue 7-8 2000G. Chilosi A trypsin inhibitor from wheat kernel (WTI) was found to have a strong antifungal activity against a number of pathogenic fungi and to inhibit fungal trypsin-like activity. WTI inhibited in vitro spore germination and hyphal growth of pathogens, with protein concentration required for 50% growth inhibition (IC50) values ranging from 111.7 to above 500 ,g/ml. As observed by electron microscopy, WTI determined morphological alterations represented by hyphal growth inhibition and branching. One of the fungal species tested, Botrytis cinerea produced a trypsin-like protease, which was inhibited by the trypsin inhibitor. WTI, as well as other seed defence proteins, appear to be an important resistance factor in wheat kernels during rest and early germination when plants are particularly exposed to attack by potential soil-borne pathogens. Zusammenfassung Ein Trypsinhemmer aus Weizenkörnern (WTI) zeigte eine starke antifungale Aktivität gegenüber verschiedenen pathogenen Pilzen und hemmte deren trypsinähnliche Aktivität. WTI hemmte in vitro die Sporenkeimung und das Hyphenwachstum der Pathogene, wobei die IC50 -Werte zwischen 111,7 und mehr als 500 ,g/ml lagen. Elektronenmikroskopische Untersuchungen zeigten, dai WTI morphologische Veränderungen bewirkte, die aus einer Hemmung des Hyphenwachstums und einer veränderten Verzweigung bestanden. Eine der untersuchten Pilzarten, Botrytis cinerea, bildete eine trypsinähnliche Protease, die durch den Trypsininhibitor gehemmt wurde. Ebenso wie andere sameneigene Abwehrproteine scheint WTI während der Keimruhe und in den frühen Stadien der Keimung, wenn die Pflanzen gegenüber möglichen bodenbürtigen Pathogenen besonders exponiert sind, ein wichtiger Resistenzfaktor in Weizenkörnern zu sein. [source] Technique for visual demonstration of germinating arbuscular mycorrhizal spores and their multiplication in potsJOURNAL OF PLANT NUTRITION AND SOIL SCIENCE, Issue 5 2007Jitendra Panwar Abstract We describe a simple technique for the germination of arbuscular mycorrhizal (AM),fungal spores and their multiplication in pots. Glomus fasciculatum, G. mosseae, and Gigaspora margarita were used. A single wheat seedling was tied to a glass slide, previously covered with filter paper with the help of thread. One single surface-sterilized AM-fungal spore was placed on the middle portion of the root of the wheat seedling using a sterilized syringe. The slide was placed vertically in a 100,mL glass beaker filled with 25,mL of root exudates,water (1:4, v/v) solution, which was collected by growing twenty wheat seedlings in a 150,mL beaker filled with 100,mL sterilized distilled water for 7 d. The slide was observed daily using a compound microscope to follow the time course of germination. In this technique, the spore is directly in contact with the host root, and a visualization of spore germination, hyphal development, and appressorium formation is possible without disrupting fungal growth or the establishment of the symbiosis. The method allows to document the germination events and to assess hyphal-elongation rates by photographing the same spore on consecutive days. The inoculated seedling was used to initiate single-spore multiplication in a sterilized (autoclave on 3 alternate days at 120°C for 120,min at 1.05,kg,cm,2 pressure) potted sandy soil (150,mL volume) into which the slide with the inoculated seedling was inserted carefully through a previously made slit. The wheat seedlings in all pots (4 treatments and 15 replications) became colonized by mycorrhiza, confirming that the establishment of the AM-fungal symbiosis is highly reproducible. Our technique permits the relatively undisturbed growth of the symbiotic partners, the visualization of germinating AM-fungal spores, and their multiplication in pots. This simple and low-cost method facilitates the production of pure lines of AM fungi from single spores, allowing for the study of intraspecific variation and potentiality for cytological, biochemical, physiological, and taxonomical studies. [source] Synergistic effect of oligochitosan and silicon on inhibition of Monilinia fructicola infectionsJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 4 2010Lingyu Yang Abstract BACKGROUND: Oligochitosan has broad-spectrum antimicrobial activity and shows an obvious inhibitory effect on phytopathogens. In addition, as an exogenous elicitor, it can induce various defence responses, including affecting the activities of several defence-related enzymes and substances in some plants. Owing to this dual function of oligochitosan, it can be used to control postharvest diseases of fruits. Silicon, like oligochitosan, also has a dual function. In this study the synergistic effect of oligochitosan and silicon on the decay control of apple fruit was investigated. RESULTS:In vitro, both oligochitosan and silicon significantly inhibited spore germination, germ tube elongation and mycelial growth of Monilinia fructicola, with higher concentrations having a greater effect. The synergistic effect of oligochitosan and silicon at half-maximal inhibitory concentration on disease control at 25 °C was much better than the effect of oligochitosan or silicon alone, not only in vitro but also in vivo. CONCLUSION: The results showed that a combination of oligochitosan and silicon had a synergistic effect on the control of disease caused by M. fructicola in apple fruit at 25 °C. Copyright © 2010 Society of Chemical Industry [source] Influence of water activity and temperature on conidial germination and mycelial growth of ochratoxigenic isolates of Aspergillus ochraceus on grape juice synthetic medium.JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 10 2005Predictive models Abstract The first stages in the development of Aspergillus ochraceus, an ochratoxin A-producing fungus that infects grapes and may grow on them, have been studied on a synthetic nutrient medium similar to grape in composition. Spore germination and mycelial growth have been tested over a water activity (aw) and temperature range which could approximate to the real conditions of fungal development on grapes. Optimal germination and growth were observed at 30 °C for all three isolates tested. Maximal germination rates were detected at 0.96,0.99 aw at 20 °C, while at 10 and 30 °C the germination rates were significantly higher at 0.99 aw. Although this abiotic factor (aw) had no significant influence on mycelial growth, growth rates obtained at 0.98 aw were slight higher than those at other aw levels. Predictive models for the lag phase before spore germination as a function of water activity and temperature have been obtained by polynomial multiple linear regression, and the resulting response surface models have been plotted. Copyright © 2005 Society of Chemical Industry [source] Inhibitory activity of tea polyphenol and Hanseniaspora uvarum against Botrytis cinerea infectionsLETTERS IN APPLIED MICROBIOLOGY, Issue 3 2010H.M. Liu Abstract Aims:, To investigate the effect of tea polyphenol (TP) and Hanseniaspora uvarum alone or in combination against Botrytis cinerea in grapes and to evaluate the possible mechanisms involved. Methods and Results:, TP alone was effective in controlling grey mould in grape at all concentrations. TP at 0·5 and 1·0% in combination with H. uvarum (1 × 106 CFU ml,1) showed a lower infection rate of grey mould. TP at 0·01% or above significantly inhibited the spore germination of B. cinerea. TP at 0·1% showed inhibition ability on mycelium growth of B. cinerea. The addition of TP did not affect the growth of H. uvarum in vitro and significantly increased the population of H. uvarum in vivo. Conclusions:, TP exhibited an inhibitory effect against B. cinerea and improved the biocontrol efficacy of H. uvarum. The inhibitory effects of spore germination and mycelial growth of B. cinerea and the increased populations of H. uvarum in vivo may be some of the important mechanisms of TP. Significance and Impact of the Study:, The results suggested that TP alone or in combination with biocontrol agents has great potential in the commercial management of postharvest diseases of fruits. [source] Influence of baking enzymes on antimicrobial activity of five bacteriocin-like inhibitory substances produced by lactic acid bacteria isolated from Lithuanian sourdoughsLETTERS IN APPLIED MICROBIOLOGY, Issue 6 2008V. Narbutaite Abstract Aim:, To evaluate the effect of four different baking enzymes on the inhibitory activity of five bacteriocin-like inhibitory substances (BLIS) produced by lactic acid bacteria (LAB) isolated from Lithuanian sourdoughs. Methods and Results:, The overlay assay and the Bioscreen methods revealed that the five BLIS exhibited an inhibitory effect against spore germination and vegetative outgrowth of Bacillus subtilis, the predominant species causing ropiness in bread. The possibility that the observed antibacterial activity of BLIS might be lost after treatment with enzymes used for baking purposes was also examined. Conclusions:, The enzymes tested; hemicellulase, lipase, amyloglucosidase and amylase had little or no effect on the majority of the antimicrobial activities associated with the five BLIS studied. Significance and Impact of the Study:, This study suggests a potential application in the sourdough baking industry for these antimicrobial producing LAB strains in the control of B. subtilis spore germination and vegetative outgrowth. [source] Evidence for quorum sensing in Clostridium botulinum 56ALETTERS IN APPLIED MICROBIOLOGY, Issue 1 2006L. Zhao Abstract Aims:, Experiments were designed to detect quorum-sensing signals produced by Clostridium botulinum. Methods and Results:,Clostridium botulinum 56A cell-free supernatants obtained at the end of lag phase, the mid-exponential phase and early stationary phase of growth were assayed for bioluminescence in the Vibrio harveyi quorum-sensing assay system. Twelve and 16-h culture supernatants induced bioluminescence in the auto-inducer 2 (AI-2) but not the auto-inducer 1 (AI-1) assay. Intra-species quorum sensing was also assayed as the ability of the supernatants to promote spore germination and outgrowth in a microtitre plate system. Spore populations exposed to C. botulinum supernatant from the end of lag phase became positive for growth sooner than controls. Conclusions:, The influence of cell-free supernatant on ungerminated spores and detection of bioluminescence in the AI-2 assay are evidence for a signalling molecule(s) and provide a first step in characterizing C. botulinum quorum sensing. Significance and Impact of the Study:, This study suggests that spores do not behave independently of each other and may explain the inocula size effects observed in challenge studies. Whether AI-2 production in C. botulinum serves as an inter-species signal or as a detoxification mechanism remains to be determined. [source] Non-uniform assembly of the Bacillus anthracis exosporium and a bottle cap model for spore germination and outgrowthMOLECULAR MICROBIOLOGY, Issue 2 2007Christopher T. Steichen Summary Spores of Bacillus anthracis are enclosed by an exosporium composed of a basal layer and an external hair-like nap. The nap is formed by a collagen-like glycoprotein called BclA, while the basal layer contains many different proteins, one of which is a spore-specific alanine racemase (Alr). In this study, we employed fluorescence microscopy and a fluorescently labelled anti-Alr monoclonal antibody (mAb) to examine the distribution of Alr within the exosporium. Binding of the mAb occurred over approximately three-quarters of the exosporium but not in a cap-like region at one end of the spore, indicating the absence or inaccessibility of Alr in this region. We also determined that the cap-like region, or cap, corresponds to the first part of the exosporium assembled within the mother cell during sporulation and the only part of the exosporium assembled in a ,exsY mutant strain of B. anthracis. Our results provide the first direct evidence that exosporium assembly is a non-uniform process and suggest that exosporium formation is discontinuous. Finally, we demonstrated that during spore germination and outgrowth, the outgrowing cell always escapes from its exosporium shell by popping through the cap, suggesting that the cap is designed to facilitate the emergence of the outgrowing cell. [source] Functional analysis of the Alternaria brassicicola non-ribosomal peptide synthetase gene AbNPS2 reveals a role in conidial cell wall constructionMOLECULAR PLANT PATHOLOGY, Issue 1 2007KWANG-HYUNG KIM SUMMARY Alternaria brassicicola is a necrotrophic pathogen causing black spot disease on virtually all cultivated Brassica crops worldwide. In many plant pathosystems fungal secondary metabolites derived from non-ribosomal peptide synthetases (NPSs) are phytotoxic virulence factors or are antibiotics thought to be important for niche competition with other micro-organisms. However, many of the functions of NPS genes and their products are largely unknown. In this study, we investigated the function of one of the A. brassicicola NPS genes, AbNPS2. The predicted amino acid sequence of AbNPS2 showed high sequence similarity with A. brassicae, AbrePsy1, Cochliobolus heterostrophus, NPS4 and a Stagonospora nodorum NPS. The AbNPS2 open reading frame was predicted to be 22 kb in length and encodes a large protein (7195 amino acids) showing typical NPS modular organization. Gene expression analysis of AbNPS2 in wild-type fungus indicated that it is expressed almost exclusively in conidia and conidiophores, broadly in the reproductive developmental phase. AbNPS2 gene disruption mutants showed abnormal spore cell wall morphology and a decreased hydrophobicity phenotype. Conidia of abnps2 mutants displayed an aberrantly inflated cell wall and an increase in lipid bodies compared with wild-type. Further phenotypic analyses of abnps2 mutants showed decreased spore germination rates both in vitro and in vivo, and a marked reduction in sporulation in vivo compared with wild-type fungus. Moreover, virulence tests on Brassicas with abnps2 mutants revealed a significant reduction in lesion size compared with wild-type but only when aged spores were used in experiments. Collectively, these results indicate that AbNPS2 plays an important role in development and virulence. [source] Microscopy reveals disease control through novel effects on fungal development: a case study with an early-generation benzophenone fungicide,PEST MANAGEMENT SCIENCE (FORMERLY: PESTICIDE SCIENCE), Issue 5 2006Mark R Schmitt Abstract The benzophenones are a new class of agricultural fungicides that demonstrate protectant, curative and eradicative/antisporulant activity against powdery mildews. The chemistry is represented in the marketplace by the fungicide metrafenone, recently introduced by BASF and discussed in the following paper. The benzophenones show no evidence of acting by previously identified biochemical mechanisms, nor do they show cross-resistance with existing fungicides. The value of microscopy in elucidating fungicide mode of action is demonstrated through identification of the effects of an early benzophenone, eBZO, on mildew development. eBZO caused profound alterations in the morphology of powdery mildews of both monocotyledons and dicotyledons, affecting multiple stages of fungal development, including spore germination, appressorial formation, penetration, surface hyphal morphology and sporogenesis. Identification of analogous effects of eBZO on sporulation in the model organism Aspergillus nidulans (Eidam) Winter provides a unique opportunity to elucidate important morphogenetic regulatory sites in the economically important obligate pathogens, the powdery mildews. Benzophenones provide a further example of the benefits of whole-organism testing in the search for novel fungicide modes of action. Copyright © 2006 Society of Chemical Industry [source] Control of post-harvest decay of apples by pre-harvest and post-harvest application of ammonium molybdatePEST MANAGEMENT SCIENCE (FORMERLY: PESTICIDE SCIENCE), Issue 12 2001Carla Nunes Abstract Ammonium molybdate was tested as a potential fungicide for use in apples (cv Golden Delicious) against blue and grey mould, important post-harvest diseases of pome fruits. In tests in vivo at 20,°C, ammonium molybdate (15,mM) reduced lesion diameters of Penicillium expansum, Botrytis cinerea and Rhizopus stolonifer by 84%, 88% and 100% respectively. When apples treated with ammonium molybdate were stored at 1,°C for three months, a significant reduction in severity and incidence of P expansum and B cinerea was observed in both years of study (1998 and 1999). In the second year of the experiment the reduction in disease severity was greater than 88% for both pathogens, and the level of control was similar to, or greater than, that observed with the fungicide imazalil. When ammonium molybdate was applied as a pre-harvest treatment, a significant reduction in blue mould decay was observed after three months in cold storage. In vitro, ammonium molybdate greatly inhibited spore germination of P expansum and B cinerea, although better inhibition was obtained against grey mould. Ammonium dimolybdate, sodium molybdate and potassium molybdate were also tested in vitro in comparison with ammonium molybdate as inhibitors of spore germination, but only ammonium molybdate inhibited spore germination by more than 50%. © 2001 Society of Chemical Industry [source] Sensitivity of the Early Life Stages of Macroalgae from the Northern Hemisphere to Ultraviolet Radiation,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 4 2007Michael Y. Roleda The reproductive cells of macroalgae are regarded as the life history stages most susceptible to various environmental stresses, including UV radiation (UVR). UVR is proposed to determine the upper depth distribution limit of macroalgae on the shore. These hypotheses were tested by UV-exposure experiments, using spores and young thalli of the eulittoral Rhodophyceae Mastocarpus stellatus and Chondrus crispus and various sublittoral brown macroalgae (Phaeophyceae) with different depth distribution from Helgoland (German Bight) and Spitsbergen (Arctic). In spores, the degree of UV-induced inhibition of photosynthesis is lower in eulittoral species and higher in sublittoral species. After UV stress, recovery of photosynthetic capacity is faster in eulittoral compared to sublittoral species. DNA damage is lowest while repair of DNA damage is highest in eulittoral compared to sublittoral species. When the negative impact of UVR prevails, spore germination is inhibited. This is observed in deep water kelp species whereas the same UVR doses do not inhibit germination of shallow water kelp species. A potential acclimation mechanism to increase UV tolerance of brown algal spores is the species-specific ability to increase the content of UV-absorbing phlorotannins in response to UV-exposure. Growth rates of young Mastocarpus and Chondrus gametophytes exposed to experimental doses of UVR are not affected while growth rates of all young kelp sporophytes exposed to UVR are significantly lowered. Furthermore, morphological UV damage in Laminaria ochroleuca includes tissue deformation, lesion, blistering and thickening of the meristematic part of the lamina. The sensitivity of young sporophytes to DNA damage is correlated with thallus thickness and their optical characteristics. Growth rate is an integrative parameter of all physiological processes in juvenile plants. UV inhibition of growth may affect the upper distribution depth limit of adult life history stages. Juveniles possess several mechanisms to minimize UVR damage and, hence, are less sensitive but at the expense of growth. The species-specific susceptibility of the early life stages of macroalgae to UVR plays an important role for the determination of zonation patterns and probably also for shaping up community structure. [source] Induction of resistance to downy mildew (Peronospora parasitica) in cauliflower by DL-,-amino- n -butanoic acid (BABA)PLANT PATHOLOGY, Issue 1 2002D. Silué Of the three isomers of aminobutyric acid, only the , isomer (BABA) was effective in inducing resistance against Peronospora parasitica, the causal agent of downy mildew, in cauliflower (Brassica oleracea var. botrytis). A single foliar spray applied to 7-day-old seedlings protected the plants against Peronospora parasitica for at least 15 days. Of the enantiomers (R and S), only the R was effective. Resistance was accompanied by a hypersensitive-like reaction (necrotic spots) which was evident before inoculation. BABA was systemically effective when applied to the roots, but failed to protect cotyledons adjacent to treated ones. Unlike other chemical inducers, BABA was effective when applied several hours postinoculation. It had no effect on P. parasitica spore germination. In cauliflower seedlings, BABA did not induce the accumulation of the pathogenesis-related protein PR-1, PR-2, PR-3, PR-5 and PR-9. Only treated and challenged seedlings accumulated PR-2. [source] Early events of Bacillus anthracis germination identified by time-course quantitative proteomicsPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 19 2006Pratik Jagtap Abstract Germination of Bacillus anthracis spores involves rehydration of the spore interior and rapid degradation of several of the protective layers, including the spore coat. Here, we examine the temporal changes that occur during B. anthracis spore germination using an isobaric tagging system. Over the course of 17,min from the onset of germination, the levels of at least 19 spore proteins significantly decrease. Included are acid-soluble proteins, several known and predicted coat proteins, and proteins of unknown function. Over half of these proteins are small (less than 100 amino acids) and would have been undetectable by conventional gel-based analysis. We also identified 20 proteins, whose levels modestly increased at the later time points when metabolism has likely resumed. Taken together, our data show that isobaric labeling of complex mixtures is particularly effective for temporal studies. Furthermore, we describe a rigorous statistical approach to define relevant changes that takes into account the nature of data obtained from multidimensional protein identification technology coupled with the use of isobaric tags. This study provides an expanded list of the proteins that may be involved in germination of the B. anthracis spore and their relative levels during germination. [source] Enhanced resistance to foliar fungal pathogens in carrot by application of elicitorsANNALS OF APPLIED BIOLOGY, Issue 1 2009J. Jayaraj Abstract Treatment of greenhouse-grown carrot plants with salicylic acid (SA) (100 ,m), chitosan (0.02%) and the nutrient-chelate product Alexin (1%) followed 10 h later by inoculation with the necrotrophic fungal pathogens Alternaria radicina and Botrytis cinerea significantly reduced disease development 10 days after inoculation (d.a.i.) compared with control plants sprayed with water. The most effective treatment was chitosan, followed by Alexin and SA. Additional sprays of elicitors resulted in significantly lower disease development 25 d.a.i. Treated plants had elevated transcript levels of pathogenesis-related protein 1 (PR1), chitinase, lipid transfer protein (LTP), chalcone synthase, nonexpressor of PR1 and pathogenesis-related protein 5 (PR5) genes compared with control plants when assayed 10,70 h after treatment. The activity of peroxidase, polyphenoloxidase, phenylalanine ammonia-lyase, chitinase, ,-1,3-glucanase and lipoxygenase was significantly increased in elicitor-treated plants compared with control plants 12,72 h after treatment. Microscopic examination of treated leaves revealed reduced fungal growth and colonisation, 48 h after treatment, accompanied by fewer lesions at 120 h, compared with the control. Protein extracts from elicitor-treated plants reduced spore germination and germ tube elongation of the pathogens in vitro by 30,45%. Elicitor-treated plants accumulated higher amounts of total phenolics, 6-methoxymellin and H2O2 compared with the control. Both chitosan and Alexin induced responses similar to that of SA, suggesting that these elicitors may activate the salicylate pathway, leading to induction of defence genes, enzymes, phytoalexin and phenolics, which collectively reduced fungal colonisation. [source] | ||||||||||||||||||||||||||||