Integration of Biocontrol Agents and Food-Grade Additives for Enhancing Protection of Stored Apples from Penicillium expansum

Forty-nine compounds currently used as additives in foods were tested in combination with three biocontrol agents, the yeasts Rhodotorula glutinis, Cryptococcus laurentii, and the yeastlike fungus Aureobasidium pullulans, to increase their antagonistic activity against Penicillium expansum, the causal agent of blue mold on apples. Twelve additives dramatically improved the antagonistic activity of one or more of the tested biocontrol agents. In a two-way factorial experiment with these selected additives the percentage of P. expansum rots on apples was significantly influenced by the antagonist and the additive as well as by their interaction. The combination of the biocontrol agents and some additives resulted in a significantly higher activity with respect to the single treatments applied separately, producing additive or synergistic effects. Some of the selected additives combined with a low yeast concentration (106 cells per ml) had comparable or higher efficacy than the biocontrol agents applied alone at a 100-fold higher concentration (108 cells per ml). Some organic and inorganic calcium salts, natural gums, and some antioxidants displayed the best results. In general, the effect of each additive was specific to the biocontrol isolate used in the experiments. Possible mechanisms involved in the activity of these beneficial additives and their potential application in effective formulations of postharvest biofungicides are discussed.

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Control of Postharvest Decay in Pear by Four Laboratory-Grown Yeasts and Two Registered Biocontrol Products

Plant Disease ◽

10.1094/pdis.1999.83.2.155 ◽

1999 ◽

Vol 83 (2) ◽

pp. 155-158 ◽

Cited By ~ 55

Author(s):

David Sugar ◽

Robert A. Spotts

Keyword(s):

Pseudomonas Syringae ◽

Rhodotorula Glutinis ◽

Penicillium Expansum ◽

First Year ◽

Cryptococcus Laurentii ◽

Blue Mold ◽

Lesion Diameter ◽

Candida Oleophila ◽

Resistant Strains ◽

Second Year

Control of blue mold decay in Bosc pears was studied with the laboratory-grown yeasts Rhodotorula glutinis, Cryptococcus infirmo-miniatus, and two strains of Cryptococcus laurentii, as well as registered biocontrol products Aspire, containing the yeast Candida oleophila, and Bio-Save 11 (now Bio-Save 110), containing the bacterium Pseudomonas syringae. Both thiabendazole (TBZ)-sensitive and TBZ-resistant strains of Penicillium expansum were used. Aspire treatment reduced the average lesion diameter by approximately 65 and 45%, and reduced decay incidence by 27 and 9% with TBZ-resistant and TBZ-sensitive P. expansum, respectively, in the first year of the study, but did not result in significant decay control in the second year. Bio-Save 11 reduced decay lesion diameter by 32 to 72% and incidence by 21 to 40% over the 2 years. In both years, TBZ-sensitive P. expansum was completely controlled by the combination of either C. laurentii (both strains), R. glutinis, or C. infirmo-miniatus with 100 ppm TBZ. With TBZ-resistant P. expansum, control of wound infection with these yeasts alone or with 100 ppm TBZ ranged from 62.9 to 100%. In a packinghouse trial, control by Bio-Save 110 + 100 ppm TBZ and Aspire + 100 ppm TBZ was not different than control by TBZ at 569 ppm, the maximum label rate. The amount of decay following Aspire + 100 ppm TBZ treatment was significantly less than the amount of decay following Bio-Save 110 + 100 ppm TBZ treatment.

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Effect of the Biocontrol Yeast Rhodotorula glutinis Strain LS11 on Patulin Accumulation in Stored Apples

Phytopathology ◽

10.1094/phyto-95-1271 ◽

2005 ◽

Vol 95 (11) ◽

pp. 1271-1278 ◽

Cited By ~ 55

Author(s):

Raffaello Castoria ◽

Valeria Morena ◽

Leonardo Caputo ◽

Gianfranco Panfili ◽

Filippo De Curtis ◽

...

Keyword(s):

Rhodotorula Glutinis ◽

Biocontrol Agents ◽

Yeast Cells ◽

Penicillium Expansum ◽

Cryptococcus Laurentii ◽

Fungicide Residues ◽

Tlc Plates ◽

Layer Chromatography

Contamination of apples (Malus domestica) and derived juices with fungicide residues and patulin produced by Penicillium expansum are major issues of food safety. Biocontrol agents represent an alternative or supplement to chemicals for disease control. Our data show that these microbes could also contribute to actively decreasing patulin accumulation in apples. Three biocontrol agents, Rhodotorula glutinis LS11, Cryptococcus laurentii LS28, and Aureobasidium pullulans LS30, were examined for their in vitro growth in the presence of patulin and for their capability to decrease mycotoxin recovery from the medium. Strain LS11 yielded the highest growth rates and the greatest decrease of toxin recoveries. Further, it caused the appearance of two major spots on thin-layer chromatography (TLC) plates, suggesting possible metabolization of the mycotoxin. In vivo, i.e., in the low percentage of LS11-pretreated apples infected by P. expansum, patulin accumulation was significantly lower than in nontreated infected fruits. Yeast cells survived and increased in infected apples and, in a model system emulating decaying apple, resulted in accelerated breakdown of patulin and the production of the same TLC spots as those detected in vitro. These data suggest that biocontrol yeast cells surviving in decaying apples could metabolize patulin and/or negatively affect its accumulation or synthesis. To our knowledge, this is the first report describing the effect of a biocontrol agent on patulin accumulation in vivo.

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Resistance of Postharvest Biocontrol Yeasts to Oxidative Stress: A Possible New Mechanism of Action

Phytopathology ◽

10.1094/phyto.2003.93.5.564 ◽

2003 ◽

Vol 93 (5) ◽

pp. 564-572 ◽

Cited By ~ 117

Author(s):

Raffaello Castoria ◽

Leonardo Caputo ◽

Filippo De Curtis ◽

Vincenzo De Cicco

Keyword(s):

Oxidative Stress ◽

Superoxide Anion ◽

Rhodotorula Glutinis ◽

Antagonistic Activity ◽

Model System ◽

Penicillium Expansum ◽

Cryptococcus Laurentii ◽

Oxygen Species ◽

Combined Application

We detected the generation of the reactive oxygen species (ROS) superoxide anion ( O·- 2) and hydrogen peroxide (H2O2) in apple wounds 2 immediately after wounding, and assessed the relationships between (i) timely colonization of apple wounds by biocontrol yeasts, (ii) resistance of these microorganisms to oxidative stress caused by ROS, and (iii) their antagonism against postharvest wound pathogens. We analyzed a model system consisting of two yeasts with higher (Cryptococcus laurentii LS-28) or lower (Rhodotorula glutinis LS-11) antagonistic activity against the postharvest pathogens Botrytis cinerea and Penicillium expansum. LS-28 exhibited faster and greater colonization of wounds than LS-11. In contrast to LS-28, the number of LS-11 cells dropped 1 and 2 h after application, and then increased only later. In vitro, LS-28 was more resistant to ROS-generated oxidative stress. The combined application of biocontrol yeasts and ROS-deactivating enzymes in apple wounds prevented the decrease in number of LS-11 cells mentioned above, and enhanced colonization and antagonistic activity of both biocontrol yeasts against B. cinerea and P. expansum. Polar lipids of LS-11 contained the more unsaturated and oxidizable α-linolenic acid, which was absent in LS-28. Resistance to oxidative stress could be a key mechanism of biocontrol yeasts antagonism against postharvest wound pathogens.

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Synergistic Effects of Combining Biocontrol Agents with Silicon against Postharvest Diseases of Jujube Fruit

Journal of Food Protection ◽

10.4315/0362-028x-68.3.544 ◽

2005 ◽

Vol 68 (3) ◽

pp. 544-550 ◽

Cited By ~ 41

Author(s):

SHIPING TIAN ◽

GUOZHENG QIN ◽

YONG XU

Keyword(s):

Alternaria Alternata ◽

Synergistic Effects ◽

Rhodotorula Glutinis ◽

Penicillium Expansum ◽

Cryptococcus Laurentii ◽

Postharvest Diseases ◽

Jujube Fruit ◽

Zizyphus Jujuba ◽

Fungistatic Effect ◽

Chinese Date

The synergistic effects of biocontrol yeasts Cryptococcus laurentii and Rhodotorula glutinis combined with silicon (Si) against Alternaria alternata and Penicillium expansum molds were investigated in jujube fruit (Chinese date, Zizyphus jujuba) stored at 20 and 0°C, respectively. Combinations of C. laurentii and R. glutinis at 5 × 107 cells/ml with 2% Si was most effective in controlling the diseases caused by A. alternata and P. expansum on jujube fruit stored at 20°C. When fruits were stored at 0°C, combining C. laurentii and R. glutinis with Si was as effective against P. expansum as was Si or the yeasts applied alone and was more effective in controlling A. alternata. Si may have a fungistatic effect by directly inhibiting pathogen growth, but it did not greatly influence the growth of the antagonists.

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Effect of pyrimethanil on Cryptococcus laurentii, Rhodosporidium paludigenum, and Rhodotorula glutinis biocontrol of Penicillium expansum infection in pear fruit

International Journal of Food Microbiology ◽

10.1016/j.ijfoodmicro.2013.04.012 ◽

2013 ◽

Vol 164 (2-3) ◽

pp. 155-160 ◽

Cited By ~ 20

Author(s):

Chen Yu ◽

Tao Zhou ◽

Kuang Sheng ◽

Lizhen Zeng ◽

Changzhou Ye ◽

...

Keyword(s):

Rhodotorula Glutinis ◽

Penicillium Expansum ◽

Cryptococcus Laurentii ◽

Pear Fruit

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Indole-3-Acetic Acid Improves Postharvest Biological Control of Blue Mold Rot of Apple by Cryptococcus laurentii

Phytopathology ◽

10.1094/phyto-99-3-0258 ◽

2009 ◽

Vol 99 (3) ◽

pp. 258-264 ◽

Cited By ~ 22

Author(s):

Ting Yu ◽

Jishuang Chen ◽

Huangping Lu ◽

Xiaodong Zheng

Keyword(s):

Acetic Acid ◽

Apple Fruit ◽

Natural Resistance ◽

Penicillium Expansum ◽

Cryptococcus Laurentii ◽

Blue Mold ◽

Biocontrol Efficacy ◽

Indole 3 Acetic Acid

Cryptococcus laurentii is a well-known postharvest biocontrol yeast; however, it cannot provide satisfactory levels of decay control when used alone. Here, we evaluated the effects of indole-3-acetic acid (IAA), a plant growth regulator, on the biocontrol efficacy of the yeast antagonist C. laurentii against blue mold rot caused by Penicillium expansum in apple fruit. Results showed that the addition of IAA at 20 μg/ml to suspensions of C. laurentii greatly enhanced inhibition of mold rot in apple wounds compared with that observed with C. laurentii alone. The addition of IAA at 20 μg/ml or lower did not influence the population growth of C. laurentii in wounds, but adverse effects were seen on C. laurentii when the concentration of IAA was increased to 200 μg/ml or above in vitro and in vivo. P. expansum infection in apple wounds was not inhibited when the pathogen was inoculated into the fruit wounds within 2 h after application of IAA; however, infection was reduced when inoculated more than 12 h after IAA application. Treatment of wounds with IAA at 20 μg/ml 24 h before pathogen inoculation resulted in significant inhibition of P. expansum spore germination and host infection. Application of IAA at 20 μg/ml also reduced P. expansum infection when it was applied 48 h before pathogen inoculation in the intact fruit. Thus, IAA could reinforce the biocontrol efficacy of C. laurentii in inhibiting blue mold of apple fruit by induction of the natural resistance of the fruit.

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Production, formulation and antagonistic activity of the biocontrol like-yeast Aureobasidium pullulans against Penicillium expansum

Biotechnology Letters ◽

10.1007/s10529-006-9269-2 ◽

2007 ◽

Vol 29 (4) ◽

pp. 553-559 ◽

Cited By ~ 23

Author(s):

R. Mounir ◽

A. Durieux ◽

E. Bodo ◽

C. Allard ◽

J.-P. Simon ◽

...

Keyword(s):

Aureobasidium Pullulans ◽

Antagonistic Activity ◽

Penicillium Expansum

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Control of Postharvest Pathogens and Colonization of the Apple Surface by Antagonistic Microorganisms in the Field

Phytopathology ◽

10.1094/phyto.1997.87.11.1103 ◽

1997 ◽

Vol 87 (11) ◽

pp. 1103-1110 ◽

Cited By ~ 117

Author(s):

Wolfgang Leibinger ◽

Barbara Breuker ◽

Matthias Hahn ◽

Kurt Mendgen

Keyword(s):

Aureobasidium Pullulans ◽

Random Amplified Polymorphic Dna ◽

Rhodotorula Glutinis ◽

Growing Season ◽

Penicillium Expansum ◽

Promising Alternative ◽

Antagonistic Microorganisms ◽

Fruit Decay ◽

Population Sizes ◽

Control Post

Selected isolates of Aureobasidium pullulans, Rhodotorula glutinis, and Bacillus subtilis reduced the size and number of lesions on wounded apples caused by the postharvest pathogens Penicillium expansum, Botrytis cinerea, and Pezicula malicorticis. Combinations of the antagonistic microorganisms were applied to apple trees in the field late in the growing season of two consecutive years. The population dynamics of the introduced microorganisms and the incidence of fruit decay were determined. Population sizes of introduced antagonists on apple surfaces increased in the field following application of treatments until harvest. After transfer of the fruit from the field into cold storage, the populations of the introduced antagonists remained higher than in the control treatments. Identification of the applied isolates of A. pullulans and R. glutinis during the experiments was achieved by isolate-specific DNA probes generated from random amplified polymorphic DNA. A combination of two strains of A. pullulans and one strain of R. glutinis suppressed rotting of apple to the same extent as the commonly used fungicide Euparen. Our data demonstrate that the application of antagonistic microorganisms in the field represents a promising alternative to fungicide treatments to control post-harvest diseases of apple.

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Laboratory effect of Boni Protect containing Aureobasidium pullulans (de Bary) Arnoud in the control of some fungal diseases of apple fruit

Acta Agrobotanica ◽

10.5586/aa.2013.009 ◽

2013 ◽

Vol 66 (1) ◽

pp. 77-88 ◽

Cited By ~ 5

Author(s):

Anna Wagner ◽

Beata Hetman ◽

Marek Kopacki ◽

Agnieszka Jamiołkowska ◽

Paweł Krawiec ◽

...

Keyword(s):

Aureobasidium Pullulans ◽

Disease Incidence ◽

Brown Rot ◽

Apple Fruit ◽

Penicillium Expansum ◽

Blue Mold ◽

Monilinia Fructigena ◽

Control Product ◽

Grey Mold ◽

Apple Cultivars

The efficacy of Aureobasidium pullulans (in the biopreparation Boni Protect) against different pathogens of apples (Botrytis cinerea, Monilinia fructigena, Penicillium expansum, and Pezicula malicorticis) was evaluated under laboratory con- ditions. The biocontrol product was applied at concentrations of 0.05%, 0.1%, and 0.5%. Fruits of apple cultivars 'Jonagold Decosta' and 'Pinova' were used. Boni Protect was very effective against B. cinerea on cv. 'Jonagold Decosta', reducing disease incidence by 55–83.8%. On 'Pinova' apples, this biological control product was the most efficient at earlier stages of the experiment. It inhibited grey mold by 65% after 5 days from inoculation and only by 14% after 20 days. On cv. 'Jonagold Decosta', Boni Protect at a concentration of 0.1% was also effective against M. fructigena, reducing brown rot by 31.4–74.5%, but its efficiency on cv. 'Pinova' was not significant. Blue mold caused by P. expansum was inhibited only slightly by the biocontrol product, while P. malicorticis proved to be the most resistant to its antagonistic abilities.

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