Postharvest Biology and Technology 78 (2013) 133138

Contents lists available at SciVerse ScienceDirect

Postharvest Biology and Technology

journal homepage: www.elsevier.com/locate/postharvbio

Pre- and postharvest treatment with alternatives to synthetic fungicides to control postharvest decay of sweet cherry
Erica Feliziani, Marilla Santini, Lucia Landi, Gianfranco Romanazzi
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
The effectiveness of alternatives to synthetic fungicides for the control of pathogens causing postharvest diseases of sweet cherry was tested in vitro and in vivo. When amended to potato dextrose-agar, oligosaccharides, benzothiadiazole, chitosan, calcium plus organic acids, and nettle macerate reduced the growth of Monilinia laxa, Botrytis cinerea and Rhizopus stolonifer. Treatment of sweet cherries three days before harvest or soon after harvest with oligosaccharides, benzothiadiazole, chitosan, calcium plus organic acids, nettle extract, r extract, laminarin, or potassium bicarbonate reduced brown rot, gray mold, Rhizopus rot, Alternaria rot, blue mold and green rot of cherries kept 10 d at 20 1 C, or 14 d at 0.5 1 C and then exposed to 7 d of shelf-life at 20 1 C. Among these resistance inducers, when applied either preharvest or postharvest, chitosan was one of the most effective in reducing storage decay of sweet cherry, and its antimicrobial activity in vitro and in eld trials was comparable to that of the fungicide fenhexamid. Benzothiadiazole was more effective when applied postharvest than with preharvest spraying. These resistance inducers could represent good options for organic growers and food companies, or they can complement the use of synthetic fungicides in an integrated disease management strategy. 2012 Elsevier B.V. All rights reserved.

Article history: Received 22 September 2012 Accepted 16 December 2012 Keywords: Botrytis cinerea Chitosan Monilinia spp. Prunus avium Resistance inducers

1. Introduction Sweet cherry (Prunus avium) is a perishable fruit, and during storage it can undergo postharvest decay. This is mainly caused by Monilinia spp. and Botrytis cinerea, and occasionally by Rhizopus stolonifer, Alternaria alternata, Penicillium expansum, and Cladosporium spp. (Romanazzi et al., 2001). At present, preharvest treatments with synthetic fungicides are the main means for postharvest disease control in stone fruit in general. However, alternatives to the use of synthetic fungicides are needed for the sweet cherry market, where no fungicides are registered for postharvest applications and none are allowed in organic agriculture. Compared to synthetic fungicides, alternative methods might also have the benets of lower risk of the development of fungal resistance, lower cost, and application close to the harvest. Moreover, they have the potential to reduce the impact of agriculture on the environment and on human health (Elmer and Reglinski, 2006; Mari et al., 2010). Natural compounds with antimicrobial activity and eliciting properties might represent alternatives to synthetic fungicides in the control of postharvest disease of fruit (Bautista-Banos et al., 2006). Resistance inducers are compounds that have a composition

Corresponding author. Fax: +39 071 2204336. E-mail addresses: g.romanazzi@univpm.it, drgiro@tin.it (G. Romanazzi). 0925-5214/$ see front matter 2012 Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.postharvbio.2012.12.004

based on pathogen or plant constituents, or their analogs, such that they can react with plant receptors and can activate plant defenses; this can then prevent pathogen infection (Terry and Joyce, 2004; Elmer and Reglinski, 2006). Benzothiadiazole is a synthetic analog of salicylic acid, and it has been reported to induce systemic acquired resistance in plants. Furthermore, it has been shown to be effective in the control of gray mold on strawberry (Terry and Joyce, 2000; Romanazzi et al., 2013). In the same way, some oligosaccharides that are derived from the degradation of fungal and plant cell wall polysaccharides represent a class of well-characterized elicitors that in some cases can induce plant defense responses at very low concentrations (Shibuya and Minami, 2001). Also, the natural polysaccharide chitosan has been reported to have antimicrobial activity against a long list of postharvest fungi and to be effective in inducing an array of responses in plant tissue (Bautista-Banos et al., 2006; Romanazzi et al., 2009; Reglinski et al., 2010; Feliziani et al., in press). Chitosan treatment can elicit plant defenses through the stimulation of enzymes related to pathogenesis and prolonged fruit and vegetable storage (Li and Yu, 2000; Meng et al., 2010; Romanazzi et al., 2012; Wang and Gao, in press). Additionally, chitosan treatment can form a coating on the surface of the fruit that slows down the respiration and ripening processes (Romanazzi et al., 2009; Dang et al., 2010). Recently, interest in the use of plant extracts and essential oils for their antimicrobial activity has increased, because these are considered to be safe for both the environment and human health.

134

E. Feliziani et al. / Postharvest Biology and Technology 78 (2013) 133138

Indeed, some such preparations have shown a broad spectrum of activity against plant pathogens, and particularly those responsible for postharvest diseases of fruit and vegetables (Tripathi and Dubey, 2004; Gatto et al., 2011). Furthermore, inhibitory effects of inorganic salts against postharvest diseases have been reported on different commodities (Sanzani et al., 2009; Mari et al., 2010); among these, the control of postharvest rots on the sweet cherry by sodium bicarbonate and potassium sorbate has been demonstrated (Ippolito et al., 2005; Karabulut et al., 2001, 2005). The aims of the present study were to: (i) evaluate the in vitro ability of oligosaccharides, benzothiadiazole, chitosan, calcium plus organic acids, nettle extract, as alternatives to fungicides, to inhibit the growth of Monilinia laxa, B. cinerea, R. stolonifer and A. alternata, and (ii) evaluate the effectiveness of preharvest and postharvest applications of these and other resistance inducers, such as laminarin, potassium bicarbonate, r extract for the control of brown rot, gray mold, Rhizopus rot, Alternaria rot, blue mold and green rot during the storage of sweet cherries at room temperature (20 1 C) and at cold temperature (0.5 1 C). 2. Materials and methods 2.1. Antimicrobial activities of the resistance inducers in vitro The antimicrobial activities of a range of resistance inducers were tested for their ability to reduce mycelial growth of fungal colonies. Agar plugs, with a diameter of 6 mm, from M. laxa, B. cinerea, A. alternata or R. stolonifer colonies in active growth were placed in the centers of Petri dishes containing 10 mL potato dextroseagar in water without (control) or with additions, after autoclaving, of oligosaccharides (10 g/L, Algition, Socoa Trading, Bologna, Italy), benzothiadiazole (2 g/L, Bion, Syngenta, Basilea, Switzerland), chitosan (10 g/L, Chito Plant, ChiPro GmbH, Bremen, Germany), calcium plus organic acids (COA) (10 g/L, Fitocalcio, Agrisystem Srl, Lamezia Terme, Italy), extract from Urtica dioica (10 g/L), or fenhexamid (0.5 g/L, Teldor, Bayer CropScience, Monheim am Rhein, Germany). The U. dioica extract was prepared by macerating 10 kg of green leaves and stems of the nettle in 100 L water and leaving this for 1 month. The suspension thus obtained was ltered through a double layer of cheesecloth, and diluted 10fold. To determine the antimicrobial activities of the formulations used, the radial growth of the fungal colonies was measured daily, until one of the treatments reached the edge of the Petri dish. Seven replicates were used for each fungus and each treatment. This period corresponded approximately to 34 days for R. stolonifer colonies and to 1 week for B. cinerea, M. laxa and A. alternata. 2.2. Postharvest treatments Commercial sweet cherry Sweet Heart and Burlat were harvested from an organic orchard in Ancona, central-eastern Italy. The fruit were selected for uniformity in size and color, and absence of deformity or disease. The Sweet Heart cherries were treated with nettle extract (10 g/L), benzothiadiazole (2 g/L), chitosan (10 g/L), oligosaccharides (10 g/L), or COA (10 g/L). The Burlat cherries were treated with benzothiadiazole (2 g/L), chitosan (10 g/L), r extract from Abies sibirica (10 g/L, Abies, Agritalia, Villa Saviola di Motteggiana, Italy), laminarin (10 g/L, K&A Frontiere, BioAtlantis, Tralee, Ireland) or potassium bicarbonate at different concentrations (4, 9, 17, 26, 34 or 43 g/L) (Karma, Certis Europe, Saronno, Italy). Distilled water was used as the control. The cherries were randomized and immersed for 1 min in the tested solutions. Three replicates of thirty cherries per treatment were placed into small plastic boxes that were then placed into large boxes. To create humid condition of storage, a layer of wet paper was placed in the

bottom of the large boxes. The Sweet Heart cherries were kept 10 d at 20 1 C, 95% to 98% relative humidity (RH), while the Burlat cherries were stored for 14 d at 0.5 1 C, and then exposed to 7 d of shelf-life at 20 1 C, 95% to 98% RH. 2.3. Preharvest treatments The trials were carried out in an experimental orchard located in a hilly area of the Ancona Province (43 31 60 N, 13 22 60 E; 203 m a.s.l.) in central-eastern Italy in 2009 and 2011. The trees were selected for uniformity of production and ripening. In 2009, the canopy of Sweet Heart trees was sprayed with a solution of chitosan (10 g/L), nettle extract (10 g/L), or fenhexamid (0.5 g/L), 3 days before the harvest. In 2011, Blaze Star trees were sprayed with a solution of chitosan (10 g/L), r extract (10 g/L), benzothiadiazole (2 g/L), or fenhexamid (0.5 g/L), 3 days before the harvest. The spraying used a back pump (model WJR2525, Honda, Tokyo, Japan) to deliver the equivalent volume of 1000 L/ha. Untreated trees were used as controls for both years. On the day of the harvest, the cherries were selected for uniformity in size and color, and absence of deformity or disease. Ten replicates of 750 g cherries per treatment were collected in plastic boxes that were then placed into large boxes. To create humid condition of storage, a layer of wet paper was placed in the bottom of the large boxes. The Sweet Heart and Blaze Star cherries were stored for 14 d at 0.5 1 C, and then exposed to 7 d of shelf-life at 20 1 C, 95% to 98% RH. In the present trials, we simulated real agricultural practices using preharvest applications of a commercial chitosan formulation. Commercial formulations for chitosan have the advantage of more practical use, as viscosity is lower than that of the biopolymer dissolved in acid solution, and it has the same effectiveness as chitosan dissolved in acetic acid (Romanazzi et al., 2009, 2013). 2.4. Data recording for the in vivo trials In the in vivo trials, at the end of the storage, the levels of decay due to each of the pathogens were assessed separately according to the symptoms. In any cases of doubt, isolations from rotted tissues were carried out on potato dextrose-agar, and the causal agent was identied according to the morphological properties. The diseases incidence was expressed as the percentage of infected fruit. The severity was assigned to ve classes according to the percentage of cherry surface covered by fungal mycelia: 0, uninfected cherry; 1, surface mycelia just visible to 25% of the cherry surface; 2, 26% to 50% of the cherry surface covered with mycelia; 3, 51% to 75% of the cherry surface covered with mycelia; and 4, >75% of the cherry surface covered with mycelia (Romanazzi et al., 2001). The infection index (or McKinney index), which incorporates both the incidence and severity of the disease, was expressed as the weighted means of the disease as a percentage of the maximum possible level (McKinney, 1923). Specically, it was calculated by the formula: I = [ (d f)/(N D)] 100, where d is the category of rot intensity scored on the sweet cherry and f its frequency; N the total number of sweet cherries examined (healthy and rotted) and D is the highest category of disease intensity occurring on the empirical scale (Romanazzi et al., 2001). 2.5. Statistical analysis The data were analyzed statistically by one-way ANOVA, followed by Tukeys HSD test, at P = 0.05 (Statsoft, USA). Percentage data were arcsine transformed before analysis to improve homogeneity of variance when the range of percentages was greater than 40. Actual values are shown.

E. Feliziani et al. / Postharvest Biology and Technology 78 (2013) 133138

135

Table 1 Radial mycelial growth (mm) of fungal colonies (Monilinia laxa, Botrytis cinerea, Alternaria alternata, Rhizopus stolonifer) on PDA amended with water (control), oligosaccharides, benzothiadiazole, chitosan, calcium plus organic acids, nettle extract, and fenhexamid. Treatment (g/L) Radial growth (mm) Monilinia laxa Control Oligosaccharides (10) Benzothiadiazole (2) Chitosan (10) Calcium plus organic acids (10) Nettle extract (10) Fenhexamid (0.5)
a

Botrytis cinerea 70 a 55 b 32 d 9e 49 c 32 d 0f

Alternaria alternata 80 a 70 b 37 d 31 e 71 b 57 c 18 f

Rhizopus stolonifer 25 a 21 ab 12 c 0d 22 ab 18 b 0d

29 aa 23 b 10 d 0e 11 d 18 c 0e

Values followed by different letters are signicantly different within columns, according to Tukeys HSD (P = 0.05).

Table 2 Effects of postharvest treatment with water (control), oligosaccharides, benzothiadiazole, chitosan, calcium plus organic acids, and nettle extract on McKinney infection index of postharvest diseases (brown rot, gray mold, Rhizopus rot, Alternaria rot, and total rot) of sweet cherries cv. Sweet Heart kept 10 d at 20 1 C, 95% to 98% RH. Treatment (g/L) McKinney infection index (%) Brown rot Control Oligosaccharides (10) Benzothiadiazole (2) Chitosan (10) Calcium plus organic acids (10) Nettle extract (10)
a b

Gray mold 21.3 a 12.2 b 13.9 b 11.5 b 16.4 b 6.5 b

Rhizopus rot 32.8 a 6.6 b 12.3 b 7.4 b 20.5 b 4.9 b

Alternaria rot 49.2 a 26.2 b 24.6 b 22.9 b 27.7 b 16.4 b

Total rota 67.2 a 36.1 b 44.2 b 29.5 b 49.3 b 23.0 b

24.6 a 11.5 b 9.8 b 6.6 b 9.8 b 4.9 b

Total rot includes brown rot, gray mold, Rhizopus rot, Alternaria rot, blue mold and green rot. Values followed by different letters are signicantly different within columns, according to Tukeys HSD (P = 0.05).

3. Results 3.1. Antimicrobial activities of resistance inducers in vitro When added to potato dextroseagar, all of the tested resistance inducers reduced the radial growth of M. laxa, B. cinerea and R. stolonifer, as compared to the controls. A. alternata growth was also inhibited except for the oligosaccharides and COA. Fenhexamid and chitosan had the highest ability of reducing the mycelial growth of all of the tested fungi. In particular, growth of M. laxa and A. alternata was completely inhibited with fenhexamid and chitosan, and B. cinerea did not grow with fenhexamid. R. stolonifer growth was inhibited by all of the resistance inducers, although none of them completely stopped its growth (Table 1). 3.2. Postharvest treatments The postharvest treatments with oligosaccharides, benzothiadiazole, chitosan, COA, and nettle extract all reduced the postharvest decay of the Sweet Heart cherries kept 10 d at 20 1 C, 95% to 98% RH (Table 2). There were no statistical differences among these treatments. Compared to the control, the application of nettle extract, chitosan, oligosaccharides, benzothiadiazole, and COA reduced the sweet cherry total rots of 66%, 56%, 46%, 34% and 27%, respectively. The infection index of the total rot, that included gray mold, brown rot, Rhizopus rot, Alternaria rot, blue mold and green rot, was lower than the sum of the single infection indices as multiple infections can occur on the same cherry. The postharvest treatment with benzothiadiazole, chitosan, r extract, the algal oligosaccharide laminarin, and potassium bicarbonate at different concentrations decreased the total rot of the Burlat cherries cold-stored for 14 d (0.5 1 C) and then exposed to 7 d of shelf-life (20 1 C, 9598% RH; Table 3). The most effective treatments in controlling postharvest total rots of sweet cherry were chitosan and potassium bicarbonate at concentrations ranging from 4 to 26 g/L. For brown rot, gray mold and total rot, chitosan reduced the infection indices by 67%, 88% and 75%, respectively, and 26 g/L potassium bicarbonate by 75%, 92% and 76%,

respectively. Gray mold infections were reduced by all the tested resistance inducers. While infection indices for brown rot were not decreased by potassium bicarbonate at 34 g/L and 43 g/L, but when it was used at lower concentrations. The treatment with potassium bicarbonate induced phytotoxic effects that were visible from concentrations >9 g/L, and that increased further with concentration (data not shown). These phytotoxic symptoms consisted of pedicel browning and the formation of dark spots on the cherry surface. Moreover, the pedicels of the sweet cherries treated with potassium bicarbonate dried earlier. 3.3. Preharvest treatments Preharvest treatments with chitosan, nettle macerate, and fenhexamid signicantly reduced brown rot, gray mold, and Rhizopus rot of the Sweet Heart cold-stored for 14 d (0.5 1 C) and then
Table 3 Effects of postharvest treatment with water (control), benzothiadiazole, chitosan, r extract, laminarin, and potassium bicarbonate at different concentrations on McKinney infection index of postharvest diseases (brown rot, gray mold, and total rots) of sweet cherries cv. Burlat stored 14 d at 0.5 1 C and then exposed to 7 d of shelf-life at 20 1 C, 95% to 98% RH. Treatment (g/L) McKinney infection index (%) Brown rot Control Benzothiadiazole (2) Chitosan (10) Fir extract (10) Laminarin (10) Potassium bicarbonate (4) Potassium bicarbonate (9) Potassium bicarbonate (17) Potassium bicarbonate (26) Potassium bicarbonate (34) Potassium bicarbonate (43) 44.9 a 8.7 b 14.9 b 14.4 b 13.1 b 12.7 b 15.8 b 12.4 b 11.1 b 23.6 ab 28.4 ab
b

Gray mold 61.3 a 33.8 b 7.6 c 30.7 b 25.6 bc 18.9 bc 14.4 bc 13.6 bc 5.1 c 14.9 bc 18.0 bc

Total rota 70.4 a 37.6 bc 17.3 de 35.6 bc 34.2 bcd 27.8 bcde 25.1 cde 22.7 cde 16.7 e 36.9 bc 44.2 b

a Total rot includes brown rot, gray mold, Rhizopus rot, Alternaria rot, blue mold and green rot. b Values followed by different letters are signicantly different within columns, according to Tukeys HSD (P = 0.05).

136

E. Feliziani et al. / Postharvest Biology and Technology 78 (2013) 133138

Table 4 Effect of treatments applied 3 days before the harvest with water (control), chitosan, nettle extract, and fenhexamid on McKinney infection index of postharvest diseases (brown rot, gray mold, Rhizopus rot, Alternaria rot, and total rots) of sweet cherries cv. Sweet Heart stored 14 d at 0.5 1 C and then exposed to 7 d of shelf-life at 20 1 C, 95% to 98% RH. Treatment (g/L) McKinney infection index (%) Brown rot Control Chitosan (10) Nettle extract (10) Fenhexamid (0.5)
a b

Gray mold 19.3 a 13.9 b 13.4 b 7.9 b

Rhizopus rot 18.6 a 12.9 b 13.6 b 10.7 b

Alternaria rot 33.6 a 17.9 b 28.6 ab 14.3 b

Total rota 74.3 a 42.9 b 48.6 b 31.4 c

13.6 a 5.0 b 5.0 b 2.9 b

Total rot includes brown rot, gray mold, Rhizopus rot, Alternaria rot, blue mold and green rot. Values followed by different letters are signicantly different within columns, according to Tukeys HSD (P = 0.05).

exposed to 7 d of shelf-life (20 1 C, 9598% RH, Table 4), and among these treatments, there were no statistical differences. Compared to the control, for brown rot, gray mold, Rhizopus rot and Alternaria rot, chitosan treatment reduced infection indices by 63%, 28%, 31% and 47%, respectively, while fenhexamid reduced them by 79%, 59%, 42% and 57%, respectively. Alternaria rot was not affected by the nettle extract, however it reduced the infection indices for brown rot, gray mold and Rhizopus rot by 63%, 31% and 27%, respectively. Fenhexamid provided the greatest reduction of the total rot, at 58%, a level signicantly greater than all of the other treatments. The preharvest treatments with chitosan, r extract, and fenhexamid reduced brown rot on the Blaze Star cherries cold-stored for 14 d (0.5 1 C) and then exposed to 7 d of shelf-life (20 1 C, 95% to 98% RH; Table 5). Compared to the control, the infection indices for brown rot (the most predominant decay that occurred in these trials) was reduced by 91%, 62% and 57% after treatments with fenhexamid, chitosan and r extract, respectively, and the effects of these treatments were not statistically different from each other. 4. Discussion The in vitro ability of chitosan to reduce mycelial growth of M. laxa, B. cinerea, R. stolonifer and A. alternata were comparable to those obtained with the synthetic fungicide fenhexamid. These data support previous studies that have reported that chitosan formulations reduced germination and radial growth of a list of decay-causing fungi, such as B. cinerea (El Ghaouth et al., 1992; Badawy and Rabea, 2009), A. alternata (Snchez-Domnguez et al., 2011), Rhizopus spp. (El Ghaouth et al., 1992; Garca-Rincn et al., 2010), and M. fructicola (Casals et al., 2012; Yang et al., 2012). Similarly, in the present study, benzothiadiazole and nettle extract had in vitro ability of reducing the mycelial growth of the tested fungi. A concentration of 2 g/L benzothiadiazole was sufcient to inhibit B. cinerea radial growth on potato dextroseagar media, and the fungus was progressively inhibited with increasing benzothiadiazole
Table 5 Effect of treatments applied 3 days before the harvest with water (control), chitosan, r extract, benzothiadiazole, and fenhexamid on McKinney infection index of postharvest diseases (brown rot, Alternaria rot, and total rots) of sweet cherries cv. Blaze Star stored 14 d at 0.5 1 C and then exposed to 7 d of shelf-life at 20 1 C, 95% to 98% RH. Treatment (g/L) McKinney infection index (%) Brown rot Control Chitosan (10) Fir extract (10) Benzothiadiazole (2) Fenhexamid (0.5) 25.0 a 9.4 bc 10.8 bc 17.0 ab 2.2 c
b

Total rota 25.2 a 9.5 bc 11.0 bc 17.3 ab 2.2 c

a Total rot includes brown rot, gray mold, Rhizopus rot, Alternaria rot, blue mold and green rot. b Values followed by different letters are signicantly different within columns, according to Tukeys HSD (P = 0.05).

doses (Terry and Joyce, 2000; Munoz and Moret, 2010). For the nettle extracts, there are no data in the literature on its effectiveness in the control of postharvest pathogens, although phenolic compounds derived from herb extracts are known to be effective against decay causing fungi of fruit and vegetables, such as B. cinerea, M. laxa, Penicillium spp. and Aspergillus spp. (Gatto et al., 2011). In the in vivo trials, the present study showed that preharvest and postharvest treatments with some of these tested resistance inducers can reduce the development of postharvest decay of sweet cherries. As previously reported, on sweet cherry, postharvest application of chitosan delayed their loss of water, maintained the quality attributes during storage, and induced peroxidase and catalase activity in the fruit (Dang et al., 2010). Indeed, the combination of hypobaric treatments and the practical grade chitosan coating applied either preharvest or postharvest had synergistic effects on the control of postharvest decay of sweet cherries cold-stored for 14 d (0 1 C) and then exposed to 7 d of shelf-life (Romanazzi et al., 2003). However, little information is available on the effects of preharvest or postharvest applications of the commercial chitosan formulation, which is easy for the growers to dissolve in water, on sweet cherry postharvest decay and the growth of M. laxa, which is one of the main cherry postharvest pathogens. Benzothiadiazole reduced the postharvest decay of sweet cherry when applied postharvest, although preharvest treatment with benzothiadiazole was not sufcient to control brown rot. In previous studies, benzothiadiazole treatments induced plant defense systems and protected strawberry from gray mold (Terry and Joyce, 2000; Romanazzi et al., 2013). Benzothiadiazole mimics the effects of salicylic acid, which is involved in plant signal transduction systems and is needed to activate the formation of defense compounds, such as polyphenols and pathogenesis-related proteins (Durang and Dong, 2004). On sweet cherry, preharvest treatments with salicylic acid signicantly reduced lesion diameters caused by M. fructicola, and induced phenylalanine ammonia-lyase, glucanase, and peroxidase activities during early storage of the fruit (Yao and Tian, 2005). In the present study, benzothiadiazole reduced the disease incidence when applied postharvest, and showed in vitro ability of reducing the mycelial growth of the tested fungi. This suggests that beside the well-known induced resistance of benzothiadiazole, it can also have a direct antimicrobial effect on several postharvest pathogens. These protective effects against plant pathogens can thus be ascribed to the combination of its defense-inducing activity on plants and its adverse effects on the growth and vigor of these pathogens. Postharvest application of potassium bicarbonate was effective in reducing postharvest brown rot and gray mold of these Burlat sweet cherries. Potassium bicarbonate has been shown to control Geotrichum candidum and P. expansum postharvest infections on peaches, nectarines and plums (Palou et al., 2009). In the present study, symptoms of potassium bicarbonate phytotoxicity were recorded only after applications at concentrations >9 g/L. Similarly, in a prior work, slight injury was seen to the stems of sweet cherries treated with 0.24 mol/L sodium bicarbonate (Karabulut

E. Feliziani et al. / Postharvest Biology and Technology 78 (2013) 133138

137

et al., 2005). As this was tested on just one cultivar here, it is not known whether this potassium bicarbonate phytotoxicity is cultivar dependent, and therefore further studies to understand the optimal dose and time of application of potassium bicarbonate are needed. We did not rinse the fruit after the potassium bicarbonate treatments, and this might be the reason for this phytotoxicity. In some packinghouses, rinsing is common practice, because the salt solutions must be washed off the fruit surface after treatment to prevent phytotoxic effects (Palou et al., 2009). Preharvest or postharvest applications of nettle and r extracts reduced these postharvest diseases of sweet cherry. Similarly, treatment with 10 g/L nettle or 10 g/L r extracts reduced postharvest decay of strawberries stored for 7 d at 0 C and then exposed to 3 d shelf-life at 20 C (Romanazzi et al., 2013). Applications of extracts from wild herbs reduced postharvest brown rot, gray mold and green mold on table grapes, apricots, nectarines and oranges (Gatto et al., 2011), and the application of a A. sibirica extract signicantly decreased the disease severity of downy mildew on grapevines under semi-controlled conditions, and its efcacy was equal to copper treatment (Dagostin et al., 2011). Postharvest applications of the experimental products containing oligosaccharides, or of the algal oligosaccharide laminarin, controlled these postharvest sweet cherry diseases. Oligosaccharides are signaling molecules that have been applied experimentally to activate plant defense responses (Esquerr-Tugay et al., 2000). The application of laminarin elicited defense responses and reduced disease infections of gray mold and powdery mildew on grapevine (Aziz et al., 2003). Thus, by simulating the presence of a pathogen, these oligosaccharides applied to the cherry tissue might have activated defense responses in advance, and avoided disease development. The commercial product based on COA decreased the development of sweet cherry decay, thus prolonging the shelf-life of the fruit. Indeed calcium ions strengthen the plant cell wall, to provide more resistance for the fruit during postharvest handling. Calcium improves fruit rmness by binding to the carboxyl groups of the pectic homogalacturonan backbones, and the reinforced cell wall would be a further barrier to pathogen penetration and infection, thus delaying disease development on fruit during storage (Ippolito et al., 2005). Immersion of strawberry fruit on a solution based on COA or on oligosaccharides reduced the infection index of postharvest gray mold and blue mold (Romanazzi et al., 2013). These alternative resistance inducers have here generally shown in vitro ability of reducing the mycelial growth of the tested fungi and the ability to reduce postharvest decay of sweet cherries, when applied either preharvest or postharvest. Compounds such as chitosan, algal extracts or potassium bicarbonate are substances dened as GRAS, Generally Recognized as Safe, by the United States Food and Drug Administration, and when applied to fruit, they are not expected to be harmful for humans or the environment. These therefore offer safer alternatives to synthetic fungicides, and have most of the requirements of an ideal means to control postharvest decay of fruit (Romanazzi et al., 2012). These resistance inducers could represent good options for organic growers and food companies, or they can complement the use of synthetic fungicides in an integrated disease management strategy. However, further studies on the impact of the treatments with these resistance inducers on the avor and quality characteristics of sweet cherries are needed.

to Dr. Alberto Belleggia for the assistance in the rst year eld trials, and to Dr. Giorgio Murri of the Experimental farm Pasquale Rosati, Marche Polytechnic University for the help in the second year eld trials. References
Aziz, A., Poinssot, B., Daire, X., Adrian, M., Bzier, A., Lambert, B., Jean-Marie, J., Pugin, A., 2003. Laminarin elicits defense responses in grapevine and induces protection against Botrytis cinerea and Plasmopara viticola. Mol. Plant Microbe Interact. 16, 11181128. Bautista-Banos, S., Hernndez-Lauzardo, A.N., Velzquez-del Valle, M.G., Hernndez-Lpez, 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 Prot. 25, 108118. Badawy, M.E.I., Rabea, E.I., 2009. Potential of the biopolymer chitosan with different molecular weights to control postharvest gray mold of tomato fruit. Postharvest Biol. Technol. 51, 110117. Casals, C., Elmer, P.A.G., Vinas, I., Teixid, N., Sisquella, M., Usall, J., 2012. The combination of curing with either chitosan or Bacillus subtilis CPA-8 to control brown rot infections caused by Monilinia fructicola. Postharvest Biol. Technol. 64, 126132. Dagostin, S., Schrer, H.-J., Pertot, I., Tamm, L., 2011. Are there alternatives to copper for controlling grapevine downy mildew in organic viticulture? Crop Prot. 30, 776788. Dang, Q.F., Yan, J.Q., Li, Y., Cheng, X.J., Liu, S.C., Chen, X.G., 2010. Chitosan acetate as an active coating material and its effect on the storing of Prunus avium L. J. Food Sci. 75, 125131. Durang, W.E., Dong, X., 2004. Systemic acquired resistance. Annu. Rev. Phytopathol. 42, 185209. El Ghaouth, A., Arul, J., Grenier, J., Asselin, A., 1992. Antifungal activity of chitosan on two postharvest pathogens of strawberry fruits. Phytopathology 82, 398402. Elmer, P.A.G., Reglinski, T., 2006. Biosuppression of Botrytis cinerea in grapes. Plant Pathology 55, 155177. Esquerr-Tugay, M.T., Bourdart, G., Dumas, B., 2000. Cell wall degrading enzymes, inhibitory proteins, and oligosaccharides participate in the molecular dialogue between plants and pathogens. Plant Physiol. Biochem. 38, 157163. Feliziani, E., Smilanick, J.L., Margosan, D.A., Mansour, M.F., Romanazzi, G., Gu, S., Gohil, H. L., Rubio Ames, Z. Preharvest fungicide, potassium sorbate, or chitosan use on quality and storage decay of table grapes. Plant Dis., in press. Garca-Rincn, J., Vega-Prez, J., Guerra-Snchez, M.G., Hernndez-Lauzardo, A.N., Pena Daz, A., Velzquez-Del Valle, M.G., 2010. Effect of chitosan on growth and plasma membrana properties of Rhizopus stolonifer. Pestic. Biochem. Physiol. 97, 275278. 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 Biol. Technol. 61, 7282. Ippolito, A., Schena, L., Pentimone, I., Nigro, F., 2005. Control of postharvest rots of sweet cherries by pre- and postharvest applications of Aureobasidium pullulans in combination with calcium chloride or sodium bicarbonate. Postharvest Biol. Technol. 36, 245252. Karabulut, O.A., Lurie, S., Droby, S., 2001. Evaluation of the use of sodium bicarbonate, potassium sorbate and yeast antagonists for decreasing postharvest decay of sweet cherries. Postharvest Biol. Technol. 23, 233236. Karabulut, O.A., Arslan, U., Ilhan, K., Kuruoglu, G., 2005. Integrated control of postharvest disease of sweet cherry with yeast antagonists and sodium bicarbonate applications within a hydrocooler. Postharvest Biol. Technol. 37, 135141. Li, H., Yu, T., 2000. Effect of chitosan on incidence of brown rot, quality and physiological attributes of postharvest peach fruit. J. Sci. Food Agric. 81, 269274. Mari, M., Neri, F., Bertolini, P., 2010. New approaches for postharvest disease control in Europe. In: Prusky, D., Gullino, M.L. (Eds.), Postharvest Pathology. Springer, Heidelberg, D, pp. 119136. McKinney, H.H., 1923. Inuence of soil temperature and moisture on infection of wheat seedlings by Helmintosporium sativum. J. Agric. Resour. 26, 195218. Meng, X.-H., Qin, G.-Z., Tian, S.-P., 2010. Inuences of preharvest spraying Cryptococcus laurentii combined with postharvest chitosan coating on postharvest diseases and quality of table grapes in storage. LWT-Food Sci. Technol. 43, 596601. Munoz, Z., Moret, A., 2010. Sensitivity of Botrytis cinerea to chitosan and acibenzolarS-methyl. Pest Manage. Sci. 66, 974979. Palou, L., Smilanick, J.L., Crisosto, C., 2009. Evaluation of food additives as alternative or complementary chemicals to conventional fungicides for the control of major postharvest diseases of stone fruit. J. Food Prot. 72, 10371046. Reglinski, T., Elmer, P.A.G., Taylor, J.T., Wood, P.N., Hoyte, S.M., 2010. Inhibition of Botrytis cinerea growth and suppression of botrytis bunch rot in grapes using chitosan. Plant Pathology 59, 882890. Romanazzi, G., Nigro, F., Ippolito, A., Salerno, M., 2001. Effect of short hypobaric treatments on postharvest rots of sweet cherries, strawberries and table grapes. Postharvest Biol. Technol. 22, 16. Romanazzi, G., Nigro, F., Ippolito, A., 2003. Short hypobaric treatments potentiate the effect of chitosan in reduction storage decay of sweet cherries. Postharvest Biol. Technol. 29, 7380.

Acknowledgements The research was carried out within the project Pre and postharvest treatments to control storage decay of sweet cherries granted by Marche Polytechnic University. Thanks are expressed

138

E. Feliziani et al. / Postharvest Biology and Technology 78 (2013) 133138 Terry, L.A., Joyce, D.C., 2000. Suppression of grey mold on strawberry fruit with the chemical plant activator acibenzolar. Pest Manage. Sci. 56, 989992. Terry, L.A., Joyce, D.C., 2004. Elicitors of induced disease resistance in postharvest horticulture crops: a brief review. Postharvest Biol. Technol. 32, 113. Tripathi, P., Dubey, N.K., 2004. Exploitation of natural products as an alternative strategy to control postharvest fungal rotting of fruit and vegetables. Postharvest Biol. Technol. 32, 235245. Wang, S.Y., Gao, H. Effect of chitosan-based edible coating on antioxidants, antioxidant enzyme system, and postharvest fruit quality of strawberries (Fragaria ananassa Duch.). LWT-Food Sci. Technol., in press Yang, L.Y., Zhang, J.-L., Bassett, C.L., Meng, X.-H., 2012. Differences between chitosan and oligochitosan in growth of Monilia fructicola and control of brown rot in peach fruit. LWT-Food Sci. Technol. 46, 254259. Yao, H., Tian, S., 2005. Effects of pre- and post-harvest application of salicylic acid or methyl jasmonate on inducing disease resistance of sweet cherry fruit in storage. Postharvest Biol. Technol. 35, 253262.

Romanazzi, G., Gabler, F.M., 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., Lichter, A., Mlikota Gabler, F., Smilanick, J., 2012. Natural and safe alternatives to conventional methods to control postharvest gray mold of table grapes. Postharvest Biol. Technol. 63, 141147. Romanazzi, G., Feliziani, E., Santini, M., Landi, L., 2013. Effectiveness of postharvest treatment with chitosan and other resistance inducers in the control of storage decay of strawberry. Postharvest Biol. Technol. 75, 2427. Snchez-Domnguez, D., Ros, M.Y., Castillo-Ocampo, P., Zavala-Padilla, G., Ramos Garca, M., Buatista-Banos, S., 2011. Cytological and biochemical changes induces by chitosan in the pathosystem Alternaria alternata-tomato. Pesticide Biochem. Physiol. 99, 250255. Sanzani, S.M., Nigro, F., Mari, M., Ippolito, A., 2009. Innovations in the control of postharvest diseases of fresh fruits and vegetables. Arab J. Plant Prot. 27, 240244. Shibuya, N., Minami, E., 2001. Oligosaccharide signalling for defence responses in plant. Physiol. Mol. Plant Pathol. 59, 223233.