Development of real-time PCR using Minor Groove Binding probe to monitor the biological control agent Candida oleophila (strain O)

Massart, Sébastien; Clercq, Deborah De; Salmon, Michel; Dickburt, C.; Jijakli, M. Haïssam

doi:10.1016/j.mimet.2004.08.012

Cited by 39 publications

(34 citation statements)

References 33 publications

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“…This experiment was repeated twice. The surface area of the apple was calculated as follows: [Area (cm 2 )=0.488×volume of displaced water (ml)+66.1] (De Clercq et al 2003;Massart et al 2005). Population sizes were expressed as cfu cm −2 .…”

Section: Organismsmentioning

confidence: 99%

Assessment of Pichia anomala (strain K) efficacy against blue mould of apples when applied pre- or post-harvest under laboratory conditions and in orchard trials

et al. 2008

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The yeast Pichia anomala strain K was selected in Belgium from the apple surface for its antagonistic activity against post-harvest diseases of apples. The efficacy of this strain against P. expansum was evaluated in the laboratory in three scenarios designed to mimic practical conditions, with different periods of incubation between biological treatment, wounding of fruit surface, and pathogen inoculation. Higher protection levels and higher final yeast densities were obtained when the applied initial concentration was 1×10 8 cfu ml −1 than when it was only 1×10 5 cfu ml −1 . The protection level correlated positively with the yeast density determined in wounds and was influenced by apple surface wetness. In orchard trials spanning two successive years, biological treatment against P. expansum, based on a powder of P. anomala strain K (1×10 7 cfu ml −1 ), β-1,3-glucans (YGT 2 g l −1 ), and CaCl 2 .2H 2 0 (20 g l −1 ), was applied to apples pre-or post-harvest under practical conditions and its effect compared with standard chemical treatments. The first year, the highest reduction (95.2%) against blue decay was obtained by means of four successive fungicide treatments and the next-highest level (87.6%) with pre-harvest high-volume spraying of the threecomponent mixture 12 days before harvest. The second year, the best results were obtained with post-harvest Sumico (carbendazim 25% and diethofencarb 25%) treatment and post-harvest biological treatment, both by dipping the apples, 88.3 and 56.3% respectively. A density threshold of 1×10 4 cfu cm −2 of strain K on the apple surface seemed to be required just after harvest for high protective activity, whatever the method and time of application. In the case of pre-harvest biological treatments, variations in meteorological conditions between the 2 years may have considerably affected strain K population density and its efficacies.

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Section: Organismsmentioning

confidence: 99%

Assessment of Pichia anomala (strain K) efficacy against blue mould of apples when applied pre- or post-harvest under laboratory conditions and in orchard trials

et al. 2008

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“…After 48 h of incubation, yeast cell recovery from intact fruit surfaces was performed as previously described (Massart et al, 2005). Briefly, apple fruits were introduced into 3000-ml plastic bags.…”

Section: Incubation Conditions and Recovery Of Yeast Cellsmentioning

confidence: 99%

“…Colony-forming units (CFUs) were counted out after a 72-h incubation at 25°C. The mean surface area of the apples was evaluated as previously described (De Clercq et al, 2003;Massart et al, 2005) according to the following linear regression curve: [Surface (in cm 2 ) = 0.488 × volume displaced water (ml) + 66.1 (r = 0.99)]. Three trials were carried out over time and each treatment contained 3 replicates per trial.…”

Section: Incubation Conditions and Recovery Of Yeast Cellsmentioning

confidence: 99%

A Box-Behnken design for predicting the combined effects of relative humidity and temperature on antagonistic yeast population density at the surface of apples

Lahlali

Massart

Serrhini

et al. 2008

International Journal of Food Microbiology

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The objective of this work was to develop models predicting the combined effects of relative humidity (RH, 75-98%), temperature (5-25°C), and initial applied yeast concentration (10 4 -10 8 CFU/ml) on the apple-surface population densities of two biocontrol agents fused against postharvest diseases; the antagonistic yeasts Pichia anomala strain K and Candida oleophila strain O. Experiments were carried out according to a Box-Behnken matrix. Multiple regression analyses showed that both models yielded a good prediction of yeast density. The effect of relative humidity appeared greater than that of temperature. The number of yeast colony-forming units per square centimeter of apple fruit surface increased with increasing relative humidity, temperature, and initial applied yeast concentration. The models predict that under optimal growth conditions (25°C, 98%), strains O and K should reach a density of 10 4 CFU/cm 2 when applied initially at 2 × 10 7 (strain O) or 10 7 CFU/ml (strain K). The model results suggest that rainfall was likely the principal cause of the variability of yeast efficacy reported for previous preharvest orchard trials spanning two successive years. Temperature may also contribute to this variation. The models developed here are important tools for predicting population densities of both strains on the apple surface within the experimental limits. The use of these results should contribute to achieving yeast densities of 10 4 CFU/cm 2 on apples by controlling yeast application and environmental factors such as relative humidity and temperature. The results of this study also confirm our previous in vitro findings that water activity has a greater effect than temperature on yeast population density.

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“…The characterization of these strains include also the development of a monitoring method (Massart et al, 2005) and the study of their ecological behavior (Lahlali et al, 2008). As strain K and strain O were already extensively characterized, it is easier to asses their efficacy against crown rot disease of banana than to select novel uncharacterized micro-organisms.…”

Section: Introductionmentioning

confidence: 99%

Biological control of crown rot of bananas with Pichia anomala strain K and Candida oleophila strain O

2008

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The antagonistic activity of two yeast strains (Pichia anomala (E.C. Hansen) Kurtzman, strain K and Candida oleophila Montrocher, strain O) against the parasitic complex responsible for banana crown rot was evaluated. The strains were applied at three different concentrations (10 6 , 10 7 , 10 8 cfu/ml) and their efficacy tested in vivo on three separate fungi (Colletotrichum musae (Berk. & Curt.) Arx, Fusarium moniliforme Sheldon, and Cephalosporium sp.) and on a parasitic complex formed by association of these three fungi. At the concentrations used C. musae appeared to be the most pathogenic. The complex showed intermediate aggressiveness between C. musae and both other fungi.Statistically significant antagonistic effects were observed on C. musae, F. moniliforme, and the fungal complex. The highest protection level (54.4%) was observed with strain O added at 10 8 cfu/ml on crowns previously inoculated with the fungal complex. The level was lower when the fungi were inoculated separately.Furthermore, the antagonistic effect was strongly reinforced when strain O at 10 8 cfu/ml was applied 24 h before fungal complex inoculation (59.9%), as compared to its application 15 min (24.3%) or 3 h (27.3%) after fungal complex inoculation. Bananas showed increased susceptibility to the fungal complex from March to June, and this influenced the level of protection by yeast, which decreased over the same period. A strict negative correlation (R 2 = 0.83) was highlighted between susceptibility of banana to crown rot and protection provided by yeast.

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Development of real-time PCR using Minor Groove Binding probe to monitor the biological control agent Candida oleophila (strain O)

Cited by 39 publications

References 33 publications

Assessment of Pichia anomala (strain K) efficacy against blue mould of apples when applied pre- or post-harvest under laboratory conditions and in orchard trials

Assessment of Pichia anomala (strain K) efficacy against blue mould of apples when applied pre- or post-harvest under laboratory conditions and in orchard trials

A Box-Behnken design for predicting the combined effects of relative humidity and temperature on antagonistic yeast population density at the surface of apples

Biological control of crown rot of bananas with Pichia anomala strain K and Candida oleophila strain O

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