Morphology, Plasmolysis, and Tetrazolium Bromide Stain as Criteria for Determining Viability of <i>Phytophthora</i> Oospores

Jiang, Jiping; Erwin, D. C.

doi:10.1080/00275514.1990.12025847

Cited by 28 publications

(29 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Since other organic salts were used to assess the viability of fungal spores [21][22][23][24][25][26][27] and the TTC has the ability to penetrate A.niger spores based on its molecular weight [28] , either the ethanol was not effective in extracting enough TF (to cause red color) or the ethanol extracted significant amount of black color from the spore which interfered with the measurement. Analysis of variance was performed on the formazan concentration (µg/mL) using SAS System (5th Ed, SAS Institute Inc., Cary, North Carolina) as shown in Table 2.…”

Section: Triphenyl Formazanyieldmentioning

confidence: 99%

Optimum Conditions for Measuring Dehydrogenase Activity of Aspergillus niger using TTC

Ghaly¹,

Mahmoud²

2006

American J. of Biochemistry and Biotechnology

View full text Add to dashboard Cite

ABSTRACT:The suitability of the triphenyl tetrazolium chloride (TTC) for dehydrogenase activity measurement of the vegetative cells of the fungal species Aspergillus niger was investigated. The triphenyl formazan (TF) yield increased with the increases in TTC concentration, pH, temperature and incubation time within the studied ranges. The effects of individual parameters, as well as the combined effects, on the TF yield were found to follow exponential expressions. The sensitivity analysis showed that the TF yield is more sensitive to changes in temperature followed by pH, TTC concentration and incubation time. Although the rate of increase in the enzyme activity decreased gradually with the increase in temperature, no enzyme denaturation was observer below 55 ºC. The optimum TTC-test conditions for measuring the dehydrogenase activity of A. niger vegetative cells are a TTC concentration of 15 g/L, a pH of 9, a temperature of 55 ºC and an incubation time of 4 h. Although the procedure has been successfully tested with mycelia of different stages of growth, it would be interesting to test the effects of different stages of growth on TTC reduction under both aerobic and anaerobic conditions.

show abstract

Section: Triphenyl Formazanyieldmentioning

confidence: 99%

Optimum Conditions for Measuring Dehydrogenase Activity of Aspergillus niger using TTC

Ghaly¹,

Mahmoud²

2006

American J. of Biochemistry and Biotechnology

View full text Add to dashboard Cite

show abstract

“…Filters containing oospores were introduced in 0.1% tetrazolium bromide (Sigma‐Aldrich) in 1 mmol l −1 potassium phosphate buffer (pH = 6.3) solution and incubated for 24 hr at 35°C in 0.1% of thiazolyl blue tetrazolium bromide (MTT) contained in a potassium phosphate buffer (1 mol l −1 , pH = 6.3; Jiang & Erwin, ). Filters were rinsed several times with deionized water, and then, samples were sonicated at 35 Hz for 90 s in 2 ml sterile water to completely detach oospores from the filter.…”

Section: Methodsmentioning

confidence: 99%

Testing of life history traits of a soilborne pathogen in vitro: Do characteristics of oospores change according the strains of Aphanomyces euteiches and the host plant infected by the pathogen?

Billard

Quillévéré‐Hamard

Lavaud

et al. 2019

Journal of Phytopathology

View full text Add to dashboard Cite

Aphanomyces euteiches is a polyphagous, homothallic soilborne pathogen producing asexual (zoospores) and sexual (oospores) spores. Even if oospores are essential for disease development and survival, to date, no study has focused on the production rates of oospores or the quality of the offspring produced by oospores. In this study, a nonabrasive oospore extraction method from infected roots of leguminous species (pea, faba bean and vetch) was developed. This methodology includes steps of grinding and filtration. The quality of oospores (viable, dormant and dead) was assessed with tetrazolium bromide staining, and germination of oospores was tested using exudates of peas, faba bean and vetch. The average yield of the extraction method was approximately 21%. Staining revealed some differences between strains and between leguminous species. The germination percentage of oospores extracted from pea, faba bean and vetch was 25%, 62% and 70%, respectively, and a significant difference was observed according to the origin of A. euteiches‐inoculated strains. Application of exudates seems to stimulate the germination of oospores (2% for the control, 18% for pea exudates and 1% for vetch exudates). Differences observed between A. euteiches strains and leguminous species indicate that more knowledge concerning the biology of oospores is needed. This will help to better estimate evolution process of the pathogen and manage resistance and crop successions.

show abstract

“…Jiang and D.C. Erwin 1990), (ii) a tetrazolium bromide stain (0.1% MTT [Sigma, St. Louis, Missouri]), pH 6.5, 35 C for 48 h (Van der Gaag 1994) and (iii) microscopic examination at 400 X magnification. Two replicated experiments were conducted, the first with 8-wk-old oospores and the second with 10-wk-old oospores.…”

Section: Methodsmentioning

confidence: 99%

“…Oospores were presumed viable if they stained a rose or lavender color with tetrazolium bromide (Sutherland and Cohen 1983, Jiang and Erwin 1990, Van der Gaag 1994 or if plasmolysis occurred in 4 M NaCl (Mayton et al 2000, Jiang and Erwin 1990, Pittis and Shattock 1994. Oospores were presumed viable if they stained a rose or lavender color with tetrazolium bromide (Sutherland and Cohen 1983, Jiang and Erwin 1990, Van der Gaag 1994 or if plasmolysis occurred in 4 M NaCl (Mayton et al 2000, Jiang and Erwin 1990, Pittis and Shattock 1994.…”

Section: Introductionmentioning

confidence: 99%

Viability and Maturation of Aphanomyces cochlioides Oospores

Dyer¹,

Windels

2003

Mycologia

View full text Add to dashboard Cite

JSTOR is a not-for-profit service that helps scholars, researchers, and students discover, use, and build upon a wide range of content in a trusted digital archive. We use information technology and tools to increase productivity and facilitate new forms of scholarship. For more information about JSTOR, please contact support@jstor.org. This content downloaded from 155.247.166.234 on Fri, 01 Jan 2016 07:14:02 UTC All use subject to JSTOR Terms and Conditions Mycologia, 95(2), 2003, pp. 321-326.Abstract: Plasmolysis, tetrazolium bromide staining and microscopic appearance were tested for their usefulness in determining viability of oospores of Aphanomyces cochlioides. For comparison, three lethal treatments were employed to contrast the reaction of dead oospores and untreated, presumably viable oospores. Few oospores stained with tetrazolium bromide, even though plasmolysis and microscopic appearance indicated that 85% were viable. Cytoplasm of viable oospores was densely organized and uniformly granular (DOUG), whereas cytoplasm of oospores exposed to lethal treatments was loosely organized and non-uniformly granular (LONG). Doseresponse bioassay experiments were conducted with untreated oospores of varying inoculum densities or with mixtures of untreated DOUG and heat-treated LONG oospores in varying proportions. The number of DOUG oospores was correlated (R2 = 0.62, P < 0.001) with severity of damping-off of sugar beet seedlings caused by A. cochlioides. Thus, the granular appearance of cytoplasm offered a fast, easy and reliable indicator of viability of A. cochlioides oospores. Tests with newly formed oospores/oogonia showed that >80% harvested at 3-4 d after inoculation of hypocotyls stained with tetrazolium, but by 8-9 d <10% stained, apparently because of declining permeability of the spore wall to tetrazolium as oospores matured.

show abstract

Morphology, Plasmolysis, and Tetrazolium Bromide Stain as Criteria for Determining Viability of Phytophthora Oospores

Cited by 28 publications

References 10 publications

Optimum Conditions for Measuring Dehydrogenase Activity of Aspergillus niger using TTC

Optimum Conditions for Measuring Dehydrogenase Activity of Aspergillus niger using TTC

Testing of life history traits of a soilborne pathogen in vitro: Do characteristics of oospores change according the strains of Aphanomyces euteiches and the host plant infected by the pathogen?

Viability and Maturation of Aphanomyces cochlioides Oospores

Contact Info

Product

Resources

About