Postharvest Biology and Technology 75 (2013) 2427

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Postharvest Biology and Technology

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Research note

Effectiveness of postharvest treatment with chitosan and other resistance inducers in the control of storage decay of strawberry
Gianfranco Romanazzi , Erica Feliziani, Marilla Santini, Lucia Landi
Department of Agricultural, Food and Environmental Sciences, Marche Polytechnic University, Via Brecce Bianche, 60131 Ancona, Italy

a r t i c l e

i n f o

a b s t r a c t
This study compared the effectiveness of practical grade chitosan when used in solution with acetic, glutamic, formic and hydrochloric acids, and a water-soluble commercial chitosan formulation, in controlling postharvest diseases of strawberry. The commercial chitosan formulation and other resistance inducers based on benzothiadiazole, oligosaccharides, soybean lecithin, calcium and organic acids, and Abies sibirica and Urtica dioica extracts were also tested. The commercial chitosan formulation was as effective as the practical grade chitosan solutions in the control of gray mold and Rhizopus rot of strawberries immersed in these solutions and kept for 4 days at 20 1 C. Moreover, the treatment with commercial and experimental resistance inducers reduced gray mold, Rhizopus rot and blue mold of strawberries stored 7 days at 0 1 C and then exposed to 3 days shelf-life. The highest disease reduction was obtained with the commercial chitosan formulation, followed by benzothiadiazole, calcium and organic acids. The compounds that provided the best results in postharvest applications to control storage decay of strawberries, should be tested in further trials through preharvest treatments, applied at owering and a few days before harvest. 2012 Elsevier B.V. All rights reserved.

Article history: Received 5 June 2012 Accepted 22 July 2012 Keywords: Botrytis cinerea Fragaria ananassa Penicillium spp. Postharvest diseases Resistance inducers Rhizopus stolonifer

1. Introduction Strawberries are a particularly perishable fruit during postharvest storage, susceptible to drying, mechanical injury, decay and physiological disorders. Gray mold and Rhizopus rot are caused by Botrytis cinerea (Pers.) and Rhizopus stolonifer (Ehrenb.), respectively, and they are the main causes of postharvest decay of strawberry (Fragaria ananassa Duch.) (Maas, 1998). The infection of the fruit by gray mold can be ascribed to an infection on the owers in the eld. The B. cinerea fungus remains latent underneath the sepals until fruit ripening, and then close to or after harvest it can turn from a saprophyte into a parasite (Powelson, 1960). The disease often starts close to the pedicel, and at times also in wounds on the fruit produced during harvest, resulting in its colonization. B. cinerea can also develop at low temperatures (even at 0 C), with the consequent shortening of the length of storage and marketing. Rhizopus rot can spread at temperatures greater than 46 C, and it is more common on fruit exposed to rain in the eld or grown under plastic tunnels in several rows, when located at their border. Both of these diseases spread quickly to other fruit, a phenomenon that is known as nesting. Infections from Penicillium spp. (blue mold) and Mucor spp. (Mucor rot) also occur occasionally (Maas, 1998).

Corresponding author. Tel.: +39 071 2204336; fax: +39 071 2204856. E-mail address: g.romanazzi@univpm.it (G. Romanazzi). 0925-5214/$ see front matter 2012 Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.postharvbio.2012.07.007

In conventional agriculture, these diseases are usually managed by fungicide treatments that are applied around owering, and are repeated up to harvest, depending on the disease pressure and the preharvest interval of the formulation. However, in organic agriculture and after harvest, the use of synthetic fungicides is not permitted, so there is a need for alternatives. Among these, the use of resistance inducers has the potential for large-scale application. Resistance inducers can increase plant defenses, and at times can also exploit their antimicrobial properties. Among the natural compounds, chitosan has received much interest for application in agriculture and for the food industry. Chitosan can decrease gray mold and Rhizopus rot of strawberries through the reduction of mycelial growth and spore germination, and induction of morphological alterations in the causal organisms (El Ghaouth et al., 1992). Moreover, chitosan acts as a potent elicitor, to enhance plant resistance against pathogens (Amborab et al., 2008). Chitosan needs to be dissolved in dilute acid solution to exploit its properties, and several acids can dissolve this biopolymer, the best of which are acetic, hydrochloric, glutamic and formic acids (Romanazzi et al., 2009). So far, there are no data on the effectiveness in the control of postharvest decay of strawberry of commercial chitosan formulations, either alone or compared with practical grade chitosan dissolved in dilute acids. A number of resistance inducers are available on the market today. Benzothiadiazole (BTH or acibenzolar-S-methyl) is an elicitor of systemic-acquired resistance in plants. It is a photostable

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analog of salicylic acid, and it has proven to be effective in the management of gray mold of strawberry (Terry and Joyce, 2000; Munoz and Moret, 2010). Oligosaccharides can also elicit plant defenses, and their presence on host tissue can simulate the presence of pathogens and activate the plant responses (Shibuya and Minami, 2001). Abies sibirica and Urtica dioica extracts are available as commercial and experimental formulations, respectively, and as with some other plant extracts, they have recently gained popularity and scientic interest for their possible antimicrobial activities (Velzquez del Valle et al., 2008; Gatto et al., 2011). The objectives of this study were: (i) to compare the effectiveness in the control of postharvest diseases of strawberry of solutions obtained by dissolving practical grade chitosan in acetic, glutamic, formic and hydrochloric acids, and of the water-soluble commercial chitosan formulation and (ii) to evaluate the effectiveness of a commercial chitosan formulation, and benzothiadiazole, oligosaccharides, soybean lecithin, calcium and organic acids, and extracts of A. sibirica and U. dioica in the control of postharvest decay of strawberries.

2.3. Treatments Strawberries were pooled together and randomized, and then immersed for 10 s in a 5 L volume of the respective solutions. Strawberries immersed in deionised water were used as the control. After the treatments, the fruit were dried in air for 1 h, and then individually arranged in small plastic boxes. These were then placed in covered plastic boxes and stored for 7 days at 0 1 C, 9598% RH, and then exposed to 3 days shelf-life at 20 1 C, 9598% RH. Five replicates of 30 strawberries were used for each of the treatments. The infections which subsequently developed resulted from naturally occurring inoculum. 2.4. Data recording During the storage, the percentage of decayed strawberries was recorded. Disease severity was also recorded according to an empirical scale with six degrees: 0, healthy fruit; 1, 120% fruit surface infected; 2, 2140% fruit surface infected; 3, 4160% fruit surface infected; 4, 6180% fruit surface infected; 5, more than 81% of the strawberry surface infected and showing sporulation (Romanazzi et al., 2000). The empirical scale allowed the calculation of the McKinneys index, expressed as the weighted average of the disease as a percentage of the maximum possible level (McKinney, 1923). This parameter also includes information on both disease incidence and disease severity. 2.5. Experimental design and statistics The trials were arranged in a completely randomized design, and each experiment was repeated at least twice. Data from two or more experiments were pooled, as the statistical analysis to determine the homogeneity of variances was tested using Levenes test (SPSS Inc., Chicago, IL, USA). To normalize the data, the appropriate transformations were determined empirically using normal probability plots. The arcsin of the square root of the proportion was applied to the decay incidence data. The values were submitted to analysis of variance and the means were separated by Duncans Multiple Range Test (SuperANOVA, Abacus Concepts, Inc., Berkeley, CA, USA). Actual values are shown. 3. Results and discussion Research to reduce fungicide applications in agriculture through the discovery of new natural antimicrobials is needed to meet the growing consumer demand for food without chemical preservatives and to respond to the needs of sustainable farming. Due to the nontoxic and biocompatible properties of chitosan (Wu et al., 2005), it has been considered a candidate for substitution of fungi cides in horticultural cultivation (Bautista-Banos et al., 2006). The main difference between the practical grade chitosan solutions and the commercial chitosan formulation arises from the techniques of their preparation. Indeed, to dissolve the chitosan in the various acids, it is necessary to prepare the solutions 2 days in advance and to monitor and adjust the pH; in contrast, the commercial chitosan formulation can be prepared only 12 h before application, just by dissolving the powder in water. Moreover, the resulting solution with the commercial chitosan formulation has a lower viscosity compared to chitosan acetate, so it can be applied more easily in the eld using standard sprayers. These details are relevant when the practical application is proposed, as farmers can easily and quickly prepare and apply the compound at the eld scale. In our work, strawberries immersed in chitosan acetate, chloride, glutamate and formate, and in the commercial chitosan formulation, showed signicant reduction of gray mold and Rhizopus rot decay, as well as

2. Materials and methods 2.1. Fruit Trials were carried out on the strawberry cultivar Camarosa (Fragaria ananassa Duch) in commercial orchards located in the Marche region, central-eastern Italy, grown according to the standards of organic agriculture. The fruit were selected for the absence of defects, uniformity in size, and degree of ripening (2/3 red on the surface) (Rosati and Cantoni, 1993), and were used for the experiments on the day of harvest.

2.2. Resistance inducers The effectiveness in the control of postharvest strawberry diseases of chitosan dissolved in different acid solutions, and of the commercial chitosan formulation was tested. Crab shell chitosan (Sigma Chemical Co., St. Louis, MO, USA) was ground to a ne powder in a mortar, washed with distilled water, pelleted by lowspeed centrifugation, and air-dried. For experimental use, four different 1% solutions (w/v) of chitosan were prepared by dissolving the chitosan in 1% (v/v) acetic, hydrochloric, glutamic or formic acids under continuous stirring, to obtain chitosan acetate, chloride, glutamate and formate (Romanazzi et al., 2009). When dissolved, the pH of the chitosan solution was adjusted to 5.6 using 1 N NaOH, and 0.05% (w/v) Triton X-100 surfactant was added to improve the wetting properties of these solutions. A commercial chitosan-based formulation, known as Chito Plant (ChiPro GmbH, Bremen, Germany), was prepared by dissolving the powder (1%, w/v) directly in distilled water 2 h before use. Distilled water was used as the control. The effectiveness of different commercial resistance inducers in the control of postharvest strawberry diseases was compared. These were based on chitosan (Chito Plant, 1%, w/v), oligosaccharides (Algition, Socoa Trading, Bologna, Italy; 1%, v/v), benzothiadiazole (Bion, Syngenta Crop Protection, Switzerland; 0.2%, w/v), calcium and organic acids (Fitocalcio, Agrisystem, Lamezia Terme, CZ, Italy; 1%, v/v), soybean lecithin (Xedabio, Certis, Saronno, VA, Italy; 1%, v/v), an A. sibirica extract (Abies, Agritalia, Villa Saviola di Motteggiana, MN, Italy; 1%, v/v) and an experimental formulation based on a U. dioica extract (1%, w/v). This last compound was obtained by infusion of U. dioica leaves in water (10%, w/v) for one month, with the macerate ltered through a double layer of cheesecloth, and then diluted 1:10 in deionised water.

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Table 1 Decay, disease severity and McKinney index of gray mold and Rhizopus rot recorded on strawberries treated with solutions obtained by dissolving practical grade chitosan in acetic, glutamic, formic and hydrochloric acids, and with commercial chitosan formulation. The fruit were kept for 4 days at 20 1 C, 9598% RH. Treatment Decay (%) Gray mold Control Chitosan acetate Chitosan chloride Chitosan formate Chitosan glutamate Commercial chitosan 91.8 A 37.4 B 51.3 B 43.1 B 44.7 B 43.0 B Rhizopus rot 93.0 A 14.1 B 29.9 B 25.9 B 25.6 B 19.3 B Disease severity (15) Gray mold 4.6 A 3.1 B 3.2 B 3.4 B 3.4 B 3.5 B Rhizopus rot 4.8 A 3.7 B 3.4 B 3.4 B 3.6 B 3.6 B McKinney index (%) Gray mold 84.5 A 23.2 B 32.8 B 29.3 B 30.4 B 30.1 B Rhizopus rot 89.3 A 10.4 B 20.3 B 17.6 B 18.4 B 13.9 B

Values with the same letter are not statistically different according to Duncans Multiple Range Test at p < 0.01.

reduced severity and McKinney index, when compared to the control after 4 days shelf-life at 20 1 C (Table 1). The treatments with chitosan acetate, chitosan formate, the commercial chitosan, chitosan glutammate and chitosan chloride provided McKinney index reductions in gray mold of 73%, 65%, 64%, 64% and 61% respectively, and of Rhizopus rot of 88%, 80%, 84%, 79% and 77% respectively, as compared to the control. However, no signicant differences in disease control were observed for the solutions obtained starting from practical grade chitosan compared to the commercial chitosan formulation. In these trials, signicant infections of blue mold were not observed. The treatment of the strawberry slices with chitosan acetate signicantly decreased hydrogen peroxide production at 2, 4 and 6 h after treatment, as compared to the untreated control (data not shown). Chitosan solutions have antioxidant capacity, like hydrogen peroxide scavengers, and the use of chitosan as an antioxidant and anti-browning agent is widespread in the food industry (Devlieghere et al., 2004). The oxygen radicals scavenging capacities, the levels of phenylpropanoid compounds, and the antioxidant enzyme activity increased in strawberries after the treatment with chitosan (Wang and Gao, in press). On strawberries cold-stored 7 days (0 1 C) and then exposed to 3 days shelf-life (20 1 C), the reductions, as compared to the control, in the McKinney index for gray mold were 79%, 73%, 70%, 63%, 60%, 56% and 46% for the fruit treated with commercial chitosan, benzothiadiazole, calcium with organic acids, oligosaccharides, Abies extract, soybean lecithin, and Urtica extract, respectively and for blue mold were 90%, 84%, 71%, 61%, 59% and 31% for the fruit treated with commercial chitosan, benzothiadiazole, calcium with organic acids, Abies extract, Urtica extract and oligosaccharides, respectively. Only treatments with chitosan and calcium with organic acids reduced the McKinney Index of Rhizopus rot, respectively of 84% and 79%, as compared to the control (Table 2). Chitosan has a dual effect on hostpathogen interactions through its antifungal activity and its ability to induce plant defense responses (Romanazzi, 2010). Moreover, as chitosan can form an

edible lm when applied to the surface of fruit and vegetables, it is clearly effective in conferring a physical barrier to moisture loss, delaying dehydration and fruit shriveling. Therefore, its coating can prolong storage life, delay the drop in sensory quality, and control the decay of strawberry fruit (Han et al., 2004; Park et al., 2005; Chaiprasart et al., 2006; Hernndez-Munoz et al., 2006; Ribeiro et al., 2007). Chitosan coating can be used as a vehicle for incorporating functional ingredients, such as antimicrobials or nutraceutical compounds that could enhance the effects of chitosan coating or reinforce the nutritional value of the strawberries (Vargas et al., 2006; Vu et al., 2011; Perdones et al., 2012). Positive effects of treatment with practical grade chitosan coating on the decay of strawberries articially inoculated with B. cinerea and R. stolonifer and held at 13 C have been shown (El Ghaouth et al., 1992). Preharvest sprays of practical grade chitosan significantly reduced postharvest fungal rot of strawberries stored at 3 C and 13 C and maintained the quality of the fruit compared to the control (Reddy et al., 2000). In the same way, preharvest and postharvest treatments with practical grade chitosan on strawberries reduced the postharvest gray mold and Rhizopus rot after storage at 0 1 C followed by shelf-life at 20 1 C (Romanazzi et al., 2000). Benzothiadiazole is a functional analog of salicylic acid and an acquired systemic resistance activator that can elicit activation of genes involved in plant defense and pathogenesis-related proteins (Lawton et al., 1996; Vallad and Goodman, 2004). Our results are in agreement with Terry and Joyce (2000), who reported the possibility to delay the development of gray mold on strawberry fruit held at 5 C by about 1.2-fold, through single or multiple preharvest foliar treatments at anthesis with benzothiadiazole, with no phytotoxic effects seen for either fruit or plant. Postharvest treatment of strawberries with benzothiadiazole induced disease resistance by enhancing fruit antioxidant systems and free radical-scavenging capabilities (Cao et al., 2011). In the formulation where composition is based on calcium and organic acids, the calcium reinforces the structural composition of the plant cell wall through the binding of pectins with salts, and

Table 2 Decay, severity and McKinney index of gray mold, Rhizopus rot and blue mold recorded on strawberries treated with commercial and experimental resistance inducers. The fruit were stored for 7 days at 0 1 C, 9598% RH, followed by 3 days of shelf life at 20 1 C, 9598% RH. Treatment Decay (%) Gray mold Control Abies extract Oligosaccharides Benzothiadiazole Chitosan Ca-organic acids Urtica extract Soybean lecithin 63.5 a 29.8 bc 29.0 bc 25.1 c 20.4 c 23.5 c 44.6 b 36.8 bc Rhizopus rot 48.9 a 36.2 ab 36.3 ab 20.8 ab 8.6 b 12.7 ab 24.2 ab n.d.a Blue mold 56.9 a 28.3 bc 40.4 ab 12.6 cd 4.8 d 28.5 bc 28.2 bc n.d.a Disease severity (15) Gray mold 4.2 a 2.2 c 3.4 ab 2.9 bc 2.7 bc 3.4 b 2.9 bc 3.2 b Rhizopus rot 3.8 a 3.8 a 2.5 a 2.2 a 1.8 a 1.6 a 2.3 a n.d.a Blue mold 3.8 a 2.9 abc 3.4 ab 1.6 bc 1.0 c 2.2 abc 1.9 abc n.d.a McKinney index (%) Gray mold 53.3 a 21.2 bc 19.7 bc 14.6 bc 11.1 c 16.0 bc 27.9 b 23.6 bc Rhizopus rot 44.8 a 29.6 ab 32.8 ab 15.2 ab 7.2 b 9.6 b 13.6 ab n.d.a Blue mold 40.8 a 16.0 bc 28.0 b 6.4 c 4.0 c 12.0 c 16.8 bc n.d.a

Values with the same letter are not statistically different according to Duncans Multiple Range Test at p < 0.05. a Disease not developed in the trials in which the compound was used.

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therefore provides more resistance during the manipulation and transport of fruit. Calcium is one of the most widely used treatment alternatives to fungicides with table grapes, with the aim to protect the berries from preharvest and postharvest gray mold, and it is used in both organic and conventional agriculture (Romanazzi et al., 2012). When oligosaccharides are applied to plants, these can simulate the presence of a pathogen and thus induce plant defense responses (Chisholm et al., 2006). These compounds derived from the degradation of plant cell-wall polysaccharides are one class of well characterized elicitors that, in some cases, can induce defense responses at very low concentrations (Shibuya and Minami, 2001). In the present study, the application of soybean lecithin was tested as a resistance inducer. Hoa and Ducamp (2008) reported that treatments with soybean lecithin delayed mango ripening during storage at ambient temperatures, thus slowing the changes of the biochemical ripening indicators. Lecithin could also work as an antioxidant, since hydrogen peroxide content in strawberry tissues treated with lecithin was reduced compared to the control (data not shown). In the food industry, soybean lecithin is normally used as a natural and non-toxic compound with antioxidant properties, and it is approved by the United States Food and Drug Administration for human consumption, with the status of Generally Recognized As Safe. Over the last few years, there has been increasing interest for research into plant extracts for their antimicrobial actions and their safe application (Gatto et al., 2011). The activity of the A. sibirica extract could be due to its triterpene acids, which act as plant growth regulators and facilitate cell division and shoot regeneration (Korolev et al., 2003). From the present study, the A. sibirica extract appears to have good antimicrobial activity. The U. dioica extract has also been shown to have antimicrobial and antioxidant effects (Glcin et al., 2004), and here we show its antimicrobial properties, as it reduced gray and blue molds. Moreover, this U. dioica extract is used by organic farmers, who claim that they can achieve a reduction in aphid numbers. Among these formulations tested, the commercial chitosan formulation and benzothiadiazole provided the highest disease reduction, which indicates their possible application in IPM. Resistance inducers also have the advantage of triggering widespectrum resistance, for activity against several classes of plant pathogen and pest (Inbar et al., 1998). However, further studies are needed to better understand the mechanisms of action of these resistance inducers, and the appreciation from the consumers of treated fruit. Acknowledgement The research was granted by EUBerry Project: EU FP7 KBBE 2010-4, Grant Agreement No. 265942. References
Amborab, B.E., Bonmort, J., Fleurat-Lessart, P., Roblin, G., 2008. Early events induced by chitosan on plant cells. Journal of Experimental Botany 59, 23172324. Bautista-Banos, S., Hernndez-Lauzardo, A.N., Velzquez-del Valle, M.G., Hernndez-Lopez, M., Ait Barka, E., Bosquez-Molina, E., Wilson, C.L., 2006. Chitosan as a potential natural compound to control pre- and postharvest diseases of horticultural commodities. Crop Protection 25, 108118. Cao, S., Hu, Z., Zheng, Y., Yang, Z., Lu, B., 2011. Effect of BTH on antioxidant enzymes radical-scavenging activity and decay in strawberry fruit. Food Chemistry 125, 145149. Chaiprasart, P., Hansawasdi, C., Pipattanawong, N., 2006. The effect of chitosan coating and calcium chloride treatment on postharvest qualities of strawberry fruit (Fragaria ananassa). Acta Horticulture 708, 337342. Chisholm, S.T., Coaker, G., Day, B., Staskawicz, B.J., 2006. Hostmicrobe interactions: shaping the evolution of the plant immune response. Cell 124, 803814. Devlieghere, F., Vermeulen, A., Debevere, J., 2004. Chitosan: antimicrobial activity interactions with food components, and applicability as a coating on fruit and vegetables. Food Microbiology 21, 703714.

El Ghaouth, A., Arul, J., Grenier, J., Asselin, A., 1992. Antifungal activity of chitosan on two post-harvest pathogens of strawberry fruits. Phytopathology 82, 398402. Gatto, M.A., Ippolito, A., Linsalata, V., Cascarano, N.A., Nigro, F., Vanadia, S., Di Venere, D., 2011. Activity of extracts from wild edible herbs against postharvest fungal diseases of fruit and vegetables. Postharvest Biology and Technology 61, 7282. Glcin, I., Kfrevio lu, .I., Oktay, M., Bykokuro lu, M.E., 2004. Antioxidant, g g antimicrobial, antiulcer and analgesic activities of nettle (Urtica dioica L.). Journal of Ethnopharmacology 90, 205215. Han, C., Zhao, Y., Leonard, S.W., Traber, M.G., 2004. Edible coatings to improve storability and enhance nutritional value of fresh and frozen strawberries (Fragaria ananassa) and raspberries (Rubus ideaus). Postharvest Biology and Technology 33, 6778. Hernndez-Munoz, P., Almenar, E., Ocio, M.J., Gavara, R., 2006. Effect of calcium dips and chitosan coatings on postharvest life of strawberries (Fragaria ananassa). Postharvest Biology and Technology 39, 247253. Hoa, T.T., Ducamp, M.N., 2008. Effects of different coatings on biochemical changes of cat Hoa loc mangoes in storage. Postharvest Biology and Technology 48, 150152. Inbar, M., Doostdar, H., Sonoda, R.M., Leibee, G.L., Mayer, R.T., 1998. Elicitors of plant defensive systems reduce insect densities and disease incidence. Journal of Chemical Ecology 24, 135149. Korolev, K.G., Lomovskii, O.I., Rozhanskaya, O.A., Vasilev, V.G., 2003. Mechanochemical preparation of water-soluble forms of triterpene acids. Chemistry of Natural Compounds 39, 366372. Lawton, K., Friedrich, L., Hunt, M., Weymann, K., Kessmann, H., Staub, T., Ryals, J., 1996. Benzothiadiazole induces disease resistance in Arabidopsis by activation of the systemic acquired resistance signal transduction pathway. Plant Journal 10, 7182. Maas, J.L., 1998. Compendium of Strawberry Diseases, 2nd ed. APS Press, St. Paul. McKinney, H.H., 1923. Inuence of soil temperature and moisture on infection of wheat seedlings by Helmintosporium sativum. Journal of Agricultural Research 26, 195218. Munoz, Z., Moret, A., 2010. Sensitivity of Botrytis cinerea to chitosan and acibenzolarS-methyl. Pest Management Science 66, 974979. Park, S.I., Stan, S.D., Daeschel, M.A., Zhao, Y., 2005. Antifungal coatings on fresh strawberries (Fragaria ananassa) to control mold growth during cold storage. Journal of Food Science 70, 202207. Perdones, A., Snchez-Gonzlez, L., Chiralt, A., Vargas, M., 2012. Effect of chitosan-lemon essential oil coatings on storage-keeping quality of strawberry. Postharvest Biology and Technology 70, 3241. Powelson, R.L., 1960. Initiation of strawberry fruit rot caused by Botrytis cinerea. Phytopathology 50, 491494. Reddy, M.V.B., Belkacemi, K., Corcuff, R., Castaigne, F., Arul, J., 2000. Effect of preharvest chitosan sprays on post-harvest infection by Botrytis cinerea and quality of strawberry fruit. Postharvest Biology and Technology 20, 3951. Ribeiro, C., Vicente, A.A., Teixeira, J.A., Miranda, C., 2007. Optimization of edible coating composition to retard strawberry fruit senescence. Postharvest Biology and Technology 44, 6370. Romanazzi, G., Nigro, F., Ippolito, A., 2000. Effectiveness of pre and postharvest chitosan treatments on storage decay of strawberries. Frutticoltura 62, 7175. Romanazzi, G., Mlikota Gabler, F., Margosan, D., Mackey, B.E., Smilanick, J.L., 2009. Effect of chitosan dissolved in different acids on its ability to control postharvest gray mold of table grapes. Phytopathology 99, 10281036. Romanazzi, G., 2010. Chitosan treatment for the control of postharvest decay of table grapes, strawberries and sweet cherries. In: Sivakumar, D. (Ed.), Fresh Produce Special Issues: New Trends in Postharvest Management of Fresh Produce, vol. 4. Global Science Books, Ltd., UK, pp. 111115. Romanazzi, G., Lichter, A., Mlikota Gabler, F., Smilanick, J., 2012. Natural and safe alternatives to conventional methods to control postharvest gray mold of table grapes. Postharvest Biology and Technology 63, 141147. Rosati, P., Cantoni, L., 1993. Fragola: tecnologia e ricerche avanzate nel mondo. Frutticoltura 55, 927. Shibuya, N., Minami, E., 2001. Oligosaccharide signalling for defence responses in plants. Physiological and Molecular Plant Pathology 59, 223233. Terry, L.A., Joyce, D.C., 2000. Suppression of grey mould on strawberry fruit with the chemical plant activator acibenzolar. Pest Management Science 56, 989992. Vallad, G.E., Goodman, R.M., 2004. Systemic acquired resistance and induced systemic resistance in conventional agriculture. Crop Science 44, 19201934. Vargas, M., Albors, A., Chiralt, A., Gonzlez-Martnez, C., 2006. Quality of cold-stored strawberries as affected by chitosan-oleic acid edible coatings. Postharvest Biology and Technology 41, 164171. Velzquez del Valle, M.G., Bautista Banos, S., Hernndez Lauzardo, A.N., Guerra Snchez, M.G., Amora Lazcano, E., 2008. Estrategias de control de Rhizopus stolonifer Ehrenb. (Ex Fr.) lind agente causal de pudriciones postcosecha en productos agrcolas. Revista Mexicana de Fitopatologa 26, 4955. Vu, K.D., Hollingsworth, R.G., Leroux, E., Salmieri, S., Lacroix, M., 2011. Development of edible bioactive coating based on modied chitosan for increasing the shelf life of strawberries. Food Research International 44, 198203. Wang, S.Y., Gao, H., 2012. Effect of chitosan-based edible coating on antioxidants, antioxidant enzyme system, and postharvest fruit quality of strawberries (Fragaria ananassa Duch.). LWT-Food Science and Technology, in press. Wu, T., Zivanovic, S., Draughon, F.A., Conway, W.S., Sams, C.E., 2005. Physiochemical properties and bioactivity of fungal chitin and chitosan. Journal of Agricultural and Food Chemistry 53, 38883894.