Kinetics of Germination of
            <i>Bacillus</i>
            Spores

Vary, James C.; Halvorson, Harlyn O.

doi:10.1128/jb.89.5.1340-1347.1965

Cited by 122 publications

(74 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Determination of the reduction in spore optical density has also been used to assess spore germination. The percent germination can be calculated from the absorbance decrease at 625 nm, a 60% decrease of initial absorbance for intact spores represents 100% germination of the spore population (Vary 1965;Parker 1969;Boschwitz et al 1983;Power et al 1989;Senesi et al 1992;Makino et al 1994). Other methods used to study spore germination include determining the loss of heat resistance during germination (Warth 1978b), or the amount of dipicolinic acid (DPA) released by germinating spores (Janssen et al 1958;Warth 1980).…”

Section: Introductionmentioning

confidence: 99%

A rapid staining technique for the detection of the initiation of germination of bacterial spores

Hamouda

Shih

Baker

2002

Lett Appl Microbiol

View full text Add to dashboard Cite

Aims: We propose to apply the Wirtz-Conklin staining technique to evaluate spore germination. Methods and Results: Spores at different stages of germination were stained with modified spore stain (Wirtz-Conklin) and evaluated for staining properties. Bacillus spores suspended in deionized water, which does not support germination, stained greenish-blue. Spores suspended in germination enhancers that did not form bacilli stained pink, indicating the initiation of germination. Spores suspended in culture media, which promotes bacterial outgrowth, formed bacilli and were also stained pink. Conclusions: Modified spore stain (Wirtz-Conklin) was found to be useful to detect the initiation of spore germination as early as 30 min following incubation in a germination environment. Significance and Impact of the Study: This simple staining procedure is useful in detecting the initiation of germination of bacterial spores.

show abstract

Section: Introductionmentioning

confidence: 99%

A rapid staining technique for the detection of the initiation of germination of bacterial spores

Hamouda

Shih

Baker

2002

Lett Appl Microbiol

View full text Add to dashboard Cite

show abstract

“…Many investigators have analyzed the effects of heat treatment on the germination response of the spores of various species and strains (2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17). Effects of heat (i) on the dormant state prior to germination, (ii) on the trigger reaction of germination, and (iii) on the later events during germination are discussed below.…”

mentioning

confidence: 99%

“…Heat also changes reversibly the tertiary structure of a protein responsible for the maintenance of the dormant state of the B. cereu s spore, resulting in a change in the permeability (8). Activation often reduces the microlag of germination as shown in B. cereus spores (3,17). The second point is concerned with the modification by heat of the site(s) for the germinant or related compounds.…”

mentioning

confidence: 99%

Disappearance of the Cooperative Effect of Glucose on L‐Alanine Binding during Heat Activation of Germination of Bacillus subtilis Spores

Yasuda

Tochikubo

1985

Microbiology and Immunology

View full text Add to dashboard Cite

Sublethal preheating of bacterial spores has been used to stimulate their germination. This treatment is known as "heat activation" or "heat shock." In our germination experiments of spores of Bacillus subtilis PCI219 (18-23), a sufficient and reproducible germination rate could be obtained with L-alanine and its analogs without preheating, and the heated spores required higher concentrations of Lalanine to germinate than the unheated spores in the presence of glucose. In the present study we examined the effect of heat treatment on the germination response, especially on the sites for the germinant and related compounds in the alanine receptor field of the spores (22). We found that heat treatment at 65 C for 30 min did not alter the binding sites for L-alanine, D-alanine, and the hydrophobic compounds, but it did affect the binding site for glucose. The results are described in some detail as follows.Actually, heat treatment of the spores of B. subtilis at various temperatures altered the germination rate (Fig. 1). At a sufficient concentration (1 mm) of L-alanine (21) the maximum germination rate was obtained by preheating at around 65 C for 30 min with or without glucose. The maximum germination rate of heated spores on L-alanine (Fig. 1A) or L-alanine plus glucose (Fig. 1B) was about 1.4 times more than that of the respective unheated ones, showing the heat activation. Heating at 70 C and higher temperatures caused a rapid decrease in germination rate. However, at lower concentrations of L-alanine only a little stimulation of the rate was observed between 50 and 60 C with L-alanine alone (Fig. 1A), and the germination rate decreased gradually as the preheating temperature in creased with L-alanine plus glucose (Fig. 1B). Germinability on glucose alone was not significant (Fig. 1B). These different responses of the germination rate to temperature indicated the complex effects of preheating.Preheating spores at 65 C for 30 min, a condition at which the maximum germination rate was obtained, was used in subsequent experiments. In the concentration-germination response curves the level of the maximum germination rate of heated spores in the absence of glucose was higher than that of unheated ones, but the apparent affinities of both spores for L-alanine, which were determined as the concentration of L-alanine showing 50 % of the maximum rate of each curve, were 101 1

show abstract

“…One to two hours were needed for the majority of TrigoCor spores to germinate (Figs 1, 2, 4 and 5), which is longer than typical times reported for heat-activated Bacillus spores (Vary and Halvorson 1965;Hashimoto et al 1969;Atluri et al 2006), and similar or shorter than times commonly reported for non-heat-activated spores (Evans and Curran 1943;Kong et al 2011). The time required for Bacillus spore germination is dependent on the lag time between addition of germinant and release of DPA.…”

Section: Discussionmentioning

confidence: 63%

“…The time required for Bacillus spore germination is dependent on the lag time between addition of germinant and release of DPA. This lag time is variable between species, strains and even individual spores (Chen et al 2006;Zhang et al 2010;Setlow 2013) and is influenced by factors such as activation via heat treatment, concentration of germinant, type of germinant, temperature, number of germinant receptors present in spore membranes and storage conditions (Keynan et al 1964;Vary and Halvorson 1965;Zhang et al 2010Zhang et al , 2013Setlow 2013). Whereas the absence of a heat treatment likely lengthened the lag time of TrigoCor spores, it is possible that the lag time was also somewhat shortened due to the use of high germinant concentrations and/or germinants such as A-GFK which act on multiple germinant receptors.…”

Section: Discussionmentioning

confidence: 99%

Nutrient-induced spore germination of aBacillus amyloliquefaciensbiocontrol agent on wheat spikes

2014

View full text Add to dashboard Cite

Aims: In this study, we investigated the feasibility of applying nutrient germinants to plant surfaces to stimulate the spore germination of the plant disease biocontrol agent Bacillus amyloliquefaciens strain TrigoCor. Methods and Results: Using the terbium chloride assay and phase-contrast microscopy, we screened potential germinants of TrigoCor spores and found that a combination of D-glucose, D-fructose and potassium chloride (GFK), in addition to either L-asparagine (Asn-GFK) or L-alanine (Ala-GFK), induced maximal levels of TrigoCor spore germination in vitro. The germinant mixture Asn-GFK was also able to significantly stimulate Bacillus spore germination on wheat surfaces. Conclusions: The successful in vivo stimulation of Bacillus spore germination suggests that nutrient-induced spore germination on plant surfaces is a feasible strategy for improving Bacillus biocontrol. Significance and Impact of the Study: One of the challenges of applying Bacillus biological control agents to aboveground plant parts is that Bacillus cells transition to a metabolically dormant spore state on plant surfaces, making them unable to prevent subsequent pathogen attacks. This study demonstrates that using nutrients to stimulate Bacillus spore germination in vivo is a promising option for improving disease control and should be pursued further.

show abstract

Kinetics of Germination of Bacillus Spores

Cited by 122 publications

References 10 publications

A rapid staining technique for the detection of the initiation of germination of bacterial spores

A rapid staining technique for the detection of the initiation of germination of bacterial spores

Disappearance of the Cooperative Effect of Glucose on L‐Alanine Binding during Heat Activation of Germination of Bacillus subtilis Spores

Nutrient-induced spore germination of aBacillus amyloliquefaciensbiocontrol agent on wheat spikes

Contact Info

Product

Resources

About