Control of Citrus Postharvest Penicillium Molds with Sodium Ethylparaben

Crop Protection 46 (2013) 44e51

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Crop Protection

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Control of citrus postharvest penicillium molds with sodium ethylparaben

Pedro A. Moscoso-Ramírez a,b, Clara Montesinos-Herrero a, Lluís Palou a,* a Laboratori de Patologia, Centre de Tecnologia Postcollita (CTP), Institut Valencià d’Investigacions Agràries (IVIA), 46113 Montcada, Valencia, Spain b Campus Tabasco, Colegio de Postgraduados, 86500 Tabasco, H. Cárdenas, Mexico article info abstract

Article history: The curative antifungal activity of postharvest sodium ethylparaben (SEP) treatments against citrus green Received 23 June 2012 mold (GM) and blue mold (BM) was determined on different citrus species and cultivars artificially Received in revised form inoculated with Penicillium digitatum or Penicillium italicum and incubated at 20 C and 90% RH for 7 d or 27 November 2012 stored at 5 C and 90% RH for 8 weeks plus 7 d of shelf-life at 20 C. The best concentration was selected Accepted 9 December 2012 in in vivo primary screenings with ‘Valencia’ oranges. SEP at 80 mM was tested at 20, 50 or 62 C for 30, 60 or 150 s in small-scale trials to determine the best dip treatment conditions. Dips of 80 mM SEP at Keywords: 20 C for 60 s were selected and applied alone or in combination with 25 mLL 1 of the conventional Penicillium digitatum þ m 1 Penicillium italicum fungicide imazalil (SEP IMZ 25). Imazalil at the very low concentrations of 25 (IMZ 25) or 50 LL fi Oranges (IMZ 50) was also tested. Effectiveness of SEP alone at 20 C for 60 s was signi cantly higher on oranges Disease incidence (cvs. ‘Valencia’ and ‘Lanelate’) than on mandarins (cvs. ‘Clemenules’, ‘Nadorcott’ and ‘Ortanique’), with Food additives GM and BM incidence reductions of up to 57e73% after 7 d at 20 C. SEP was compatible with IMZ 25 and Nonpollutant disease control consistently improved its performance, irrespective of citrus cultivars and storage conditions. All treatments were less effective on ‘Clemenules’ mandarins. On ‘Valencia’ oranges stored for 8 weeks at 5 C and 7 d at 20 C, the combined treatment was significantly more effective than the single treatments (reductions of GM and BM incidence of about 96e93% and 55e39%, respectively). In additional tests, SEP, IMZ 25 and the combination applied at 20 C for 60 s prevented GM on ‘Valencia’ oranges treated, inoculated with P. digitatum 24 h later and incubated at 20 C for 7 d. It can be concluded from these results that SEP might be an integrating nonpollutant control alternative to be included in citrus postharvest disease control programs in the future. Ó 2012 Elsevier Ltd. All rights reserved.

1. Introduction et al., 2002a), but the efﬁcacy of fungicides is decreasing due to development of resistance by pathogens, which is limiting fruit Postharvest green mold (GM), caused by Penicillium digitatum production worldwide (Brent and Hollomon, 2007). Furthermore, (Pers.:Fr.) Sacc., and blue mold (BM), caused by Penicillium italicum in the development and use of chemical fungicides for postharvest Wehmer, are the most economically important postharvest disease control, considerable attention must be given to the pres- diseases of citrus in Spain, California, and all citrus production areas ervation of the global environment. characterized by low summer rainfall (Eckert and Eaks, 1989; Palou Parabens are widely used as preservatives in food, cosmetic and et al., 2007). BM is especially important on citrus fruit kept under pharmaceutical products. Acute, subchronic and chronic studies in cold storage, but GM may cause 60e80% of decay under ambient rodents indicate that parabens can be considered as low-toxicity conditions (Brown and Eckert, 2000; Palou et al., 2002b). However, compounds. Parabens are rapidly absorbed, metabolized and the severity of losses due to these fungi varies depending upon excreted (Mills et al., 2004; Soni et al., 2005), and they are “generally production area, cultivar, climatic conditions and postharvest regarded as safe” (GRAS) compounds, approved for use in foods by the handling practices (Iqbal et al., 2012). Currently, these diseases are US Food and Drug Administration (FDA) and European Union (EU) primarily controlled by application of synthetic fungicides such as regulations (Mills et al., 2004). The antimicrobial activity of the par- imazalil (IMZ) or thiabendazole (Holmes and Eckert, 1999; Palou abens increases as the chain length of the alkyl group increases; however, their aqueous solubility decreases, and therefore, the

* Corresponding author. Tel.: þ34 963424117; fax: þ34 963424001. sodium salts of parabens are also frequently used in formulations E-mail address: [email protected] (L. Palou). (Giordano et al., 1999). It is generally believed that their inhibitory

0261-2194/$ e see front matter Ó 2012 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.cropro.2012.12.007 P.A. Moscoso-Ramírez et al. / Crop Protection 46 (2013) 44e51 45 effects on membrane transport and mitochondrial function processes Table 1 are key for their mode of action (Soni et al., 2005). Therefore, the use Physicochemical properties of sodium ethylparaben. of parabens might be considered as one of the nonpolluting alterna- Property Description tive methods to conventional fungicides for the control of postharvest Formula C9H9NaO3 (Fig. 1) diseases of horticultural products (Palou et al., 2008; Montesinos- Molecular 188.1631 Herrero et al., 2009; Valencia-Chamorro et al., 2009). weight (g/mol) One of the paraben sodium salts, sodium ethylparaben (SEP), Synonyms p-hydroxybenzoic acid ethyl ester sodium salt Sodium ethyl-p-hydroxybenzoate has demonstrated to be useful to control postharvest pathogens on Sodium 4-ethoxycarbonylphenoxide citrus fruit (Valencia-Chamorro et al., 2008, 2009). However, in Ethyl-4-hydroxy benzoic acid sodium salt these studies, SEP was tested in in vitro conditions or in applications Sodium 4-(ethoxycarbonyl) phenolate benzoic acid incorporated to edible coatings. Thus, little information is available 4-hydroxy-, ethyl ester, sodium salt E number E215 about the use of aqueous solutions of SEP applied as potential Physical state White hygroscopic crystalline powder postharvest antifungal treatments, particularly against penicillium Melting point (C) 115e118 molds of citrus fruit. The present research had the following aims: Boiling point (C) 297e298 (1) to preliminarily evaluate the curative activity of SEP at different Solubility in water Soluble e concentrations against GM and BM, (2) to optimize the SEP dip pH 9.5 10.5 (10% w/v aqueous solution) treatment conditions, (3) to determine the SEP compatibility with Source: Merck KgaA, Darmstadt, Germany. low doses of imazalil under selected dip conditions, (4) to determine the effectiveness of SEP treatments on economically impor- control citrus postharvest GM and BM on fruit previously inoculated tant citrus species and cultivars, (5) to evaluate the curative activity with the pathogens. A sterile mother solution of SEP was prepared at of SEP treatments on long-term cold-stored citrus fruit and (6) to a concentration of 100 Mm. Sterile solutions at concentrations of 0.1, evaluate the preventive activity of SEP treatments against citrus 1, 10, 20, 30, 40, 70 and 100 mM SEP were prepared by diluting with postharvest GM and BM. sterile water. Inoculum preparation was carried out following the procedure described above. In this case, 30 mL of conidial suspension 2. Materials and methods of P. digitatum or P. italicum were placed, using a micropipette, on rind wounds made with the stainless steel rod described above. 2.1. Fruit Different sets of fruit were used for each pathogen. About 24 h after the inoculation of the pathogen, 30 mL of SEP solution at the above ‘ ’ ‘ ’ The experiments were conducted with Valencia and Lanelate mentioned concentrations were placed, using a micropipette, in the ‘ ’ oranges (Citrus sinensis (L.) Osbeck), Clemenules (synonyms: same inoculation rind wound. Control fruit were treated with 30 mL ‘ ’ ‘ ’ Nules , Clementina de Nules ) clementine mandarins (Citrus retic- of sterile distilled water. For each combination of concentration of ‘ ’ ulata Blanco) and Nadorcott (C. reticulata C. sinensis; synonyms: SEP and pathogen, four replicates of five ‘Valencia’ oranges each ‘ ’ ‘ ’ ‘ ’ Afourer , W. Murcott ) and Ortanique [C. reticulata x(Citrus were used. Treated fruit were incubated at 20 C and 90% RH for 6 d, ‘ ’ sinensis C. reticulata); synonym: Topaz ] hybrid mandarins. Fruit at which time disease incidence (% of infected fruit) and severity were collected from commercial orchards in the Valencia area (lesion diameter) and pathogen sporulation (% of fruit showing (Spain) and used the same day or stored up to 1 week at 5 C and spores) were determined. Severity and sporulation were assessed 90% relative humidity (RH) before use. Before each experiment, over the entire fruit sample, not only over infected and symptomatic fruit were selected, randomized, washed with tap water and fruit. allowed to air dry at room temperature. 2.3.2. Dip treatment conditions 2.2. Fungal inoculation Small-scale trials were conducted using ‘Valencia’ oranges to establish the best dip treatment conditions. Fungal inoculation with Penicillium digitatum and P. italicum, isolates NAV-7 and MAV-1, a concentration of 106 spores mL 1 was carried out following the respectively, from the fungal culture collection of the IVIA CTP, procedure mentioned above. e were cultured on potato dextrose agar (PDA, Sigma Aldrich Chem- Stainless steel buckets containing 10 L of aqueous solution of ical Co., St. Louis, MA, USA) plates at 25 C. Conidia of each fungus 80 mM (13.29 g L 1; 1.3% w/v) SEP were used. This concentration of from 7- to 14-day-old cultures were taken from the agar surface with SEP was selected according to previous results obtained in the a sterile rod and transferred to a sterile aqueous solution of 0.05% Ò in vivo primary screening tests. When needed, solutions were Tween 80 (Panreac, S.A.U., Barcelona, Spain). Conidial suspensions heated by placing the buckets in a 250-L stainless steel water tank were filtered through two layers of cheesecloth to separate hyphal fi 5 6 1 tted with two electrical resistances (4.5 kW each), a thermostat, fragments and adjusted to a concentration of 10 or 10 spores mL and an automatic water-recirculating system. Fruit were placed using a hemocytometer. With the exceptions of in vivo primary into 18 L multi-perforated wall stainless steel containers, exactly screenings and preventive activity tests, the fruit inoculation fitting in the above mentioned buckets, and completely immersed procedure was as follows: the tip of a stainless steel rod, 1 mm wide in the treatment solution for 30, 60 or 150 s at 20, 50 or 62 C. After and 2 mm in length, was immersed in the conidial suspension and inserted in the fruit rind afterward. Fruit were inoculated at two opposite points in the fruit equatorial zone, one with P. digitatum and the opposite with P. italicum. Inoculated fruit were kept in a temperature-controlled room at 20 C for 24 h, until treatment.

2.3. Curative activity

2.3.1. In vivo primary screenings SEP (Ethyl 4-hydroxybenzoate sodium salt; Merck KgaA, Darm- stadt, Germany; Table 1, Fig. 1) was tested at eight concentrations to Fig. 1. Chemical structure of sodium ethylparaben. Download English Version: https://daneshyari.com/en/article/4506241

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