International Journal of Agricultural

Science and Research (IJASR)

ISSN(P): 2250-0057; ISSN(E): 2321-0087
Vol. 6, Issue 2, Apr 2016, 195-200
TJPRC Pvt. Ltd.

ANTAGONIST YEAST: AN EFFICIENT BIOAGENT FOR

THE CONTROL OF POST HARVEST PATHOGENS OF FRUITS
J. STEN1, S. DIENGNGAN2 & S. DAS3
1,3
2

Department of Plant Pathology, BCKV, Mohanpur, Nadia, West Bengal, India

Department of Fruits and Orchard Management BCKV, Mohanpur, Nadia, West Bengal, India

ABSTRACT
Over the years, use of chemical fungicides has been employed as primary method in postharvest disease
management of fruits. Although effective, their long-term applications caused disruptions in equilibrium of ecosystems,
environmental problems, health hazards to human and animals. Thus, safety of synthetic fungicides in delicate foods
like fruits has alarmed legal enforcers and consumers to demand green technology and quality products. Antagonistic
organisms have enormous potential to influence agrochemical research. Among them, yeasts are found to be very
effective particularly against various post harvest diseases of fruits. Yeasts are a group of fungi which are very efficient
in colonizing the wound sites and hence provide protection against various postharvest pathogens through mechanisms

reactive oxygen species. Advantages of using of yeasts as biocontrol are controlling environmental conditions during
storage suitable for the bioagents as well as the fruits, easy application to the target site and economic viability
considering the high cost value of the fruits.
KEYWORDS: Antagonistic Organisms, Fruits, Post-harvest diseases, Yeasts

Original Article

like competition for nutrients and space, augmentation of various defense responses in the host and tolerance to

Received: Feb 24, 2016; Accepted: Mar 07, 2016; Published: Mar 15, 2016; Paper Id.: IJASRAPR201627

INTRODUCTION
Postharvest decay of fruits caused by pathogens account for a millions of dollars in losses of products
every year both in the industrialised as well as in developing countries. However, losses in the later is far more
extensive attributing to poor storage, processing and inefficient transportation system. The use of chemical
pesticides although effective, their indiscriminate usage has led to the disruption of the equilibrium of ecosystems,
that leads to dramatic disease outbreaks, wide spread development of pathogens resistant to one or more
chemicals, toxicity to non target organisms and environmental problems (Lee et al., 2009). These, ultimately have
alarmed legal enforcers and consumers to demand green technology and quality products from the food industry as
well as the scientific community and shift from the use of synthetic fungicides to a safer and environmental
friendly alternative for reducing the postharvest diseases in fruits (Mari et al., 2014). Researchers during the past
couple of decades show great interest in biological control, especially the application of microbial antagonists
against a wide variety of pathogens that appears to be promising as a viable supplement or alternative to chemical
control (Woodhead et al., 1990). Among the various biocontrol agents, several antagonist yeasts have been
developed in the recent years. Yeasts such as Pichia guilliermondii and Cryptoccocus laurentii, yeast that occur on
apple leaves, buds and fruits were the first to be applied for post harvest decay control on fruits. The yeast
Candida oleophila has been registered for control of post harvest decay of fruit crops (Kurtzman et al., 2001)

www.tjprc.org

editor@tjprc.org

196

J. Sten, S Diengngan & S. Das

Yeasts Biocontrol
Yeasts are eukaryotic ascomycetes microorganisms belonging to the kingdom fungi. Yeasts are unicellular,
however, there are some yeasts species found to develop multicellular like characteristics by forming strings of connected
budding cells known as pseudohyphae (Kurtzman and Fell, 2005). Important characteristics of an ideal antagonist include
genetic stability, efficacy at low concentrations and against a wide range of pathogens on various fruit products, simple
nutritional requirements, survival in adverse environmental conditions, growth on cheap substrates in fermenters, lack of
pathogenicity for the host plant and no production of metabolites potentially toxic to humans, resistance to the most
frequently used pesticides and compatibility with other chemical and physical treatments (Wilson and Wisniewski 1994).
And as such, yeast seems to possess a good number of the above-mentioned features and, during the last several years,
research has been focused on the selection and study of yeast as biocontrol agent (Chalutz and Droby, 1998). Yeasts have
many properties that make them useful for biocontrol purposes. Yeasts have the ability to attach to hyphae or conidia of
phytopathogenic fungi. Attachment of phylloplane yeasts to Botrytis cinerea, Rhizoctonia solani, and Sclerotinia
homoeocarpa was determined using in vitro attachment assays. All non-attaching yeasts are found produced copious
extracellular polysaccharide (EPS) on PDA (Tom et al., 2004). Nutritional requirements of yeasts is quite simple and are
found to grow on inexpensive substrates in fermenters and are therefore easy to produce in large quantities (Druvefors,
2004). The suggested modes of action of biocontrol yeasts indicate less likelihood of any hazard to consumers (Arras et al.,
2007). Several reports show beneficial effects of yeast in food and feed where it is found to contain significantly good
amount of essential vitamins, minerals and amino acids. (Hussein et al., 1996)
Mechanism of Yeasts Antifungal Activities
At present information on the mechanisms of action for most of the antagonists is still incomplete because of
various factors such as difficulties encountered during the study of the complex interactions between host, pathogen,
antagonist and other microorganisms that might be present (Spadaro, 2003). However, it is important to understand the
mode of action before developing appropriate formulations and methods of application, and in order to obtain official
approval. Several possible biocontrol mechanisms have been suggested against post-harvest rots on fruit that includes
antibiosis, competition for nutrients and space, parasitism or direct interaction with the pathogen, and induction of
resistance in the host tissue (Droby and Chalutz, 1994).
Yeasts, act mainly competing for space or for the utilisation of some nutrients with the pathogen (Piano et al.,
1997). Yeast is found to successfully compete with the pathogen, inhibiting its growth but leaving it alive. In the
competition for space, yeasts are helped by the formation of an extracellular polysaccharide capsule that can promote
adhesion to the fruit surface (Andrews et al., 1994). Competition for nutrients was demonstrated for Pichia guilliermondii
against Penecillium digitatum co cultivated on synthetic media (Droby et al., 1989); the addition of exogenous nutrients
resulted in a reduced efficacy because the antagonists offered better results when nutrients were scarce. A rapid
multiplication and colonisation by antagonist cells in the wound was elucidated in various interactions (Piano et al., 1997).
Studies between the antagonistic yeast Cryptococcus laurentii and the pathogen Botrytis cinerea show a strong sugar
consumption by the biocontrol agents that blocks fungus conidial germination due to the deprivation of nutrients (Filonow,
1998). In fruit wounds, competition for nutrients is probably extended to other nutrients, such as nitrogen compounds
present in low concentration. Janisiewicz et al. (2000) have developed a non-destructive method using tissue culture plates
where in a defusing membrane at the lower end of cylindrical inserts was used to study the competition for nutrients
Impact Factor (JCC): 4.7987

NAAS Rating: 3.53

Antagonist Yeast: An Efficient Bioagent for the Control of Post Harvest Pathogens of Fruits

197

separated from the competition for space.

Direct parasitism can also be a possible yeast- pathogen inhibiting mechanism. A strong adhesion of Pichia
guilliermondii antagonist cells to B. cinerea mycelium was observed in vitro, perhaps due to a lectin link (Wisniewski et
al., 1991). P. Guilliermondii was found to shows high activity of -1,3-glucanase enzyme that could result in the
degradation of the fungal cell walls (Jijakli and Lepoivre, 1998). Some yeasts, such as Sporobolomyces roseus and
Cryptococcus laurentii, when applied are found to reduce the conidial adhesion and germination of B. cinerea on apples,
that is favoured by butyl acetate, a volatile aroma produced by the fruit (Filonow, 2001). These observations all suggest
that the antagonist stimulates some kind of host defence response.
Production of inhibitory substances was excluded, using agar diffusion test of filtrates from the culture plates. The
phenomenon of the killer yeast was first discovered by Makower and Beavan (1963) where they observed that certain
strains of Saccharomyces cerevisiae produced toxins that killed sensitive strains of the same species. Initially, it was
assumed that killer yeasts only killed yeasts belonging to the same or closelyrelated species. But later studies showed that
many killer toxins can affect other yeasts and evenbacteria and filamentous fungi (Polonelli et al., 1987; Polonelli and
Morace, 1986), and several applications within wine and beer fermentation (Passoth and Schnrer, 2003), food
preservation (Palpacelli et al., 1991) and antifungal therapy in humans and animals (Polonelli et al., 1986; Sguy et al.,
1994) have been suggested. Most killer proteins are only active within a very narrow temperature and pH range, which
complicates application in many environments (Suzuki and Nikkuni, 1989).
Antibiosis is the most common mode of action explored and this most likely reflects a chemical control. It is well
known that in vitro antibiosis activity does not often predict antibiosis in plant and many reports of antibiosis being
responsible for disease control are based only on in vitro observations. Antibiotic production in vitro is highly dependent
on the nutrients available in media that may or may not be available in planta. Pseudozyma flocculosa is found to produces
extracellular fatty acid that are act against Podosphaera xanthii (Avis et al., 2002). Antibiosis is an important mechanism
but not the only mode of action employed by this biocontrol agent.
The capacity of yeast cells to produce reactive oxygen species such as hydrogen peroxide (H2O2) and superoxide
-2

anion (O ) attributes as one of the suggested possible mode of action of some antagonist yeasts against post harvest wound
pathogens (Castoria et al., 2003).
Studies found that some antagonistic yeasts can be as effective as fungicides when they are applied before
pathogen inoculation. This suggested that yeast cell application might have induced resistance processes in the fruit skin.
Candida strains are able to cause chemical and osmotic changes in apple tissues, favouring antagonist settlement
(McLaughlin et al., 1990). Citrus fruits treated with yeast cells showed accumulation of phytoalexins (scoparon and
scopoletin) (Rodov et al., 1994).
Post Harvest Disease Management by Yeasts
At present, a class of products containing yeasts Candida oleophilaMontrocher, effective against Botrytis and
Penicillium spp. (Hofstein et al., 1994) and other yeasts species have been extensively tested and could be registered
relatively soon.

www.tjprc.org

editor@tjprc.org

198

J. Sten, S Diengngan & S. Das

Table 1: Biocontrol Yeasts

Name of the Antagonists
Metschnikowia pulcherrima (BIO
126)

Pathogen Controlled
Penicillium expansum and Botrytis cinerea
on apple

Saccharomyces cerevisiae

Fusarium oxysporum (sugar beet plants)

Pichia anomala J121

Pseudozyma flocculosa
Trichophyton sp. and Gliocladium
sp (filamentous)
Rhodotorula glutinis

Penicillium roqueforti (mould spoilage)

powdery mildews

Cryptococcus sp

Colletotrichum gloeosporioides (rubber)

Botrytis cinerea (geranium)
Botrytis cinerea, Rhizoctonia solani, and
Sclerotinia homoeocarpa

References
Spadaro et al., 2004
Shalaby and ElNady, 2008
Druvefors, 2004
Avis , 2002
Evueh and Ogbebor,
2008
Buck, 2004
Allen et al., 2004

Nevertheless with all the advantages of the potential biocontrol agents, certain significant limitations are also
found as such they have a narrow range of activity, because they act on specific hosts against well-defined pathogens under
particular environmental conditions (Lima et al., 1999). Methods to select antagonists with a broader spectrum of activity,
preferably for commercial development, includes efficacy tests for various pathogens and fruit species are still at its
infancy stage.
REFERENCES
1.

Allen, T.W, Burpee, L.L and Buck, J.W (2004). In vitro attachment of phylloplane yeasts to Botrytis cinerea, Rhizoctonia
solani, and Sclerotinia homoeocarpa. Can. J. of Microbiol. 50(12): 10411048

2.

Andrews, J.H., Harris, R.F., Spear, R.N., Lau, G.W., and Nordheim, E.V. (1994). Morphogenesis and adhesion of
Aureobasidium pullulans. Can. J. Microbiol. 40: 6-17.

3.

Arras, G., (2007). Enzymatic activity and sequencing of beta-1,3-glucanase gene in biocontrol yeasts. Novel approaches for
the control of post harvest diseases and disorders. Proceedings of the International Congress, Bologna, Italy,3-5 May,pp. 7277.

4.

Avis, T. J., Richard, R. and Belanger, R.R. (2002). Mechanism and means of detection of biological activity of Pseudozyma
yeasts against plant pathogenic fungi. Federation of European Microbiological Sciences: Yeast Research 2: 5-8.

5.

Buck, J.W. (2004). Combinations of Fungicides with Phylloplane Yeasts for Improved Control of Botrytis cinerea on
Geranium Seedlings. Phytopath. 94: 196-202.

6.

Castoria, R., Caputo, L., De Curtis, F. and De Cicco, V. (2003). Biological Control: Resistance of Postharvest Biocontrol
Yeasts to Oxidative Stress: A Possible New Mechanism of Action. Phytopathology 93(5): 564-572.

7.

Chalutz, E., and Droby, S. (1998). Biological control of postharvest disease. In: Boland, GJ, Kuykendall, LD (eds.), PlantMicrobe Interactions and Biological Control. Marcel Dekker, New York, USA. 157-170.

8.

Droby, S., and Chalutz, E. (1994).Mode of action of biocontrol agents of postharvest diseases. In: Wilson, C.L., Wisniewski,
M.E. (Eds.), Biological Control of Postharvest Diseases. Theory and Practice. CRC Press, Boca Raton, USA.63-75

9.

Droby, S., Chalutz, E. and Wilson, C. L. (1991).Antagonistic microorganisms as biological control agents of postharvest
diseases of fruits and vegetables.Postharvest News and Information 2, 169-73.

10. Droby, S., Chalutz, E., Wilson, C.L. and Wisniewski, M. (1989).Characterization of the biocontrol activity of
Debaryomyceshanseniiin the control of Penicilliumdigitatumon grapefruit.Can. J. of Microbiol.35, 794-800.

Impact Factor (JCC): 4.7987

NAAS Rating: 3.53

Antagonist Yeast: An Efficient Bioagent for the Control of Post Harvest Pathogens of Fruits

199

11. Druvefors, U.A. (2004). Yeast biocontrol of grain spoilage moulds Mode of action of Pichiaanomola. Doctoral thesis
Swedish University of Agri. Sciences. Sweden.
12. Evueh, G. A. and Ogbebor, N. O. (2008). Use of phylloplane fungi as biocontrol agent against Colletotrichum leaf disease of
rubber (Hevea brasiliensis Muell. Arg.). African J. Biotechnol. 7 : 2569-2572.
13. Filonow, A. (2001). Butyl acetate and yeasts interact in adhesion and germination of Botrytis cinerea conidia in vitro and in
fungal decay of golden delicious apple. J. Chem. Ecol. 27: 831-844
14. Filonow, A. (1998). Role of competition for sugars by yeasts in the biocontrol of graymold of apple.BiocontrolSc.and Tech.8,
243-56.
15. Hofstein, R., Friedlender, B., Chalutz, E. and Droby, S. (1994). Large scale production and pilot testing of biocontrol agents
of postharvest diseases. In: Wilson, C.L., Wisniewski, M. (Eds.), Biological Control of Postharvest Diseases Theory and
Practice. CRC Press Inc., Boca Raton, FL, USA. pp. 89-100.
16. Hussein, H.S.,Mackie, R.I., Merchen, N.R., Baker, D.H. and Parson , C.M. (1996). Effects of oleaginous yeast on growth
performance, fatty acid composition of muscles and energy utilization by poultry.Bioresources Tech. 55:125-130.
17. Janisiewicz, W.J., Tworkoski, T.J. and Sharer, C. (2000). Characterizing the mechanism of biological control of postharvest
diseases on fruits with a simple method to study competition for nutrients. Phytopathol. 90: 1196-1200
18. Jijakli, H.M.,and Lepoivre, P. (1998). Characterization of an Exo--1,3-Glucanase produced by Pichiaanomalastrain K,
antagonist of Botrytis cineraon apples. Phytopathol.88, 335-343.
19. Kurtzman C.P., Fell, J. W. (2005). Biodiversity and Ecophysiology of Yeasts (in: The Yeast Handbook, Gbor P, de la Rosa
CL, eds.). Berlin: Springer. pp. 1130.
20. Kurtzman, C.P. and Droby, S. (2001) :Metschnikowia fructicola, A New Ascosporic Yeast With Potential For Biocontrol Of
Post Harvest Fruit Rots. System. Appl. Microliol. 24:395-399.
21. Lee Y. S., Kim J; Lee S. G; Oh E; Lee S, C., and Park I. K. (2009). Effects of plant essential oils and components from oriental
sweetgum (Liquiddambar orientalis) on growth and morphogenesis of three phytopathogenic fungi.Pestic.Biochem. Phis; 93:
138-43
22. Lima, G., Arru, S., De Curtis, F., and Arras, G. (1999). Influence of antagonist, host fruit and pathogen on the biological
control of postharvest fungal diseases by yeasts. J. Ind. Microbiol. Biot. 23: 223-229
23. Makower, M.,and Bevan, E.A. (1963).In 11th International Congress in Genetics 1, 202.
24. Mari M, Francesco A.D., and Bertolini P. (2014).Control of fruit postharvest diseases: old issues and innovative approaches.
Stewart Postharvest Review, 1:1.
25. Mclaughlin, R.J., Wisniewski, M.E., Wilson, C.L. and Chalutz, E. (1990). Effects of inoculum concentration and salt solutions
on biological control of postharvest diseases of apple with Candida sp.. Phytopathol. 80 : 456-461.
26. Palpacelli, V., Ciani, M.,andRosini, G. (1991). Activity of different killer yeasts on strains of yeast species undesirable in the
food-industry. FEMSMicrobiology Letters 84, 75-78.
27. Passoth, V.,andSchnrer, J. (2003). In Functional genetics of industrial yeasts(Ed, de Winde, H.) Springer Verlag Berlin,
Heidelberg, pp. 297-330.
28. Piano, S., Neyrotti, V., Migheli, Q.,andGullino, M.L. (1997).Biocontrol capability of Metschnikowiapulcherrimaagainst
Botrytis postharvest rot of apple.Postharvest Biol. and Technol. 11, 131-140.

www.tjprc.org

editor@tjprc.org

200

J. Sten, S Diengngan & S. Das

29. Polonelli, L.,andMorace, G. (1986).Reevaluation of the yeast killer phenomenon.J.of Clinical Microbiol. 24, 866-869.
30. Polonelli, L., Dettori, G., Cattel, C.,and Morace, G. (1987).Biotyping of micelial fungus cultures by the killer system.
European J. of Epidemiol. 3, 23
31. Rodov, V., Ben-Yehoshua, S., Dhallewin, G., Castina, T. and Farg, D. (1994). Accumulation of the phytoalexins, scoparone
and scopoletine in citrus fruit subjected to various postharvest treatments. Int. Symp. Natural Phenols in Plant Resistance,
Acta Hortic. 3 : 517
32. Sguy, N., Aliquat, E.M., Dei-Cas, E., Polonelli, L., Camus, D. andCailliez, J.C. (1994) In Workshops on opportunistic
protists, pp. S109.
33. Shalaby, M.E. and El- Nady, M.F. (2008). Application of Saccharomyces cerevisiae as a biological control agent against
Fusarium infection of sugar beet plants. Acta Biologica Szegediensis 52:271-275
34. Sparado, D., Garibaldi, A., Gullino, M.L. (2004). Control of Penicillium expansum and Botrytis cinerea on apple combining a
biocontrol agent with hot water dipping and acibenzolar-S-methyl, baking soda, or ethanol application. Postharvest Biol.
Technol. 33: 141-151.
35. Sparado, D. (2003). Biological control of post harvest diseases of pome fruits using yeast antagonists. Dottorato Di Ricerca
In Scienze Agrarie, Forestali Ed Agroalimentari .Curriculum Difesa Integrata E Biologica Delle Colture XVI CICLO.
Chapter1. pp 1-22.
36. Suzuki, C. and Nikkuni, S. (1989). Purification and properties of the killer toxin produced by a halotolerant yeast,
Pichiafarinosa. Agri. andBiol. Chem.53, 2599-2604.
37. Tom W., Leon L. and James W. (2004).In vitro attachment of phylloplane yeasts to Botrytis cinerea, Rhizoctoniasolani, and
Sclerotiniahomoeocarpa. Can. J. Microbiol. 50: 10411048
38. Wilson, C.L. and Wisniewski, M.E. (1994).Biological Control of Postharvest Diseases Theory and Practice.CRC Press, Boca
Raton, Florida.Pp 182.
39. Wisniewski, M.E., Biles, C., Droby, S., Mclaughlin, R., Wilson, C.L. and Chalutz, E., (1991). Mode of action of the postharvest
biocontrol yeast, Pichiaguilliermondii.I. Characterization of attachment to Botrytis cinerea. Physiol. Mol. Plant P. 39: 245-58.
40. Woodhead, S.H., D.J. O'Leary & S.C. Rabatin. (1990). Discovery, development, and registration of a biocontrol agent from an
industrial perspective. Can. J. Plant Pathol. 12: 328-331

Impact Factor (JCC): 4.7987

NAAS Rating: 3.53