Impaginato 3 1. Introduction Strawberry (Fragaria x annanassa Duch.) is one of the most important berries produced in the world. In Argentina, 33000 tons are produced, mainly cultivar Camarosa (Gómez Riera et al., 2013). It is highly accepted by consumers firstly because of its physical aspect and organoleptic and nutritional properties (Shin et al., 2008; Garriga et al., 2015). Nevertheless, its postharvest quality rapidly declines due to the soft texture, high metabolic activity and susceptibility to bacterial and fungal rots. Usually, there is quality decay during transport and commercialization (Dotto et al., 2011). Cell wall degradation is one of the principal causes of strawberry´s postharvest quality decay. As pectine synthesis occurs along the fruit maturation, they are less firmly attached to the cell wall. Additionally, mid- dle lamella´s debilitation and solubilization during fruit ripening diminishes cell cohesion (Lara et al., 2004). Calcium plays a preferential role on permeability and cell integrity and has a direct influence on fruit firmness and storage time (Fernández et al., 2006). Calcium functions as an intracellular cement because it forms calcium-pectine complexes that give firm- ness to vegetable tissues. Calcium´s presence also favours pectic material insolubilization and inhibits i t s d e g r a d a t i o n b y p o l y g a l a c t u r o n a s e e n z y m e (Alonso, 1995). An immersion in calcium at harvest could increase calcium´s content in strawberry, increasing fruit firm- ness (Galetto et al., 2010) and thereby storage peri- od. However, the key factor for quality maintenance is temperature - optimum for strawberry is 0°C (Mitcham et al., 2015). Could an immersion in calci- um allow an increase in storage temperature, main- taining fruit quality? Other factors, such as growing system and sub- s t r a t e , c a n i n f l u e n c e s t r a w b e r r y ´ s q u a l i t y . Greenhouse production improves organoleptic quali- ty of fruits and vegetables because they are not exposed directly to air conditions as in open field (Gruda, 2009). Soilless production, moreover, dimin- ishes incidence of diseases and pests (Urresterazu, 2004). Adv. Hort. Sci., 2017 31(1): 3-10 DOI: 10.13128/ahs-20719 A calcium lactate treatment at harvest, growing system and refrigerated modified atmosphere can affect strawberry’s ´Camarosa´ postharvest quality? M. Harris (*), M.C. Llorens, D. Frezza Universidad de Buenos Aires, Facultad de Agronomía, Departamento de Producción Vegetal, Cátedra de Horticultura, Avenida San Martin 4453 (1417) Ciudad Autónoma de Buenos Aires, Argentina. Key words: nutritional quality, organoleptic quality, perlite, postharvest behaviour, soil, storage temperature. Abstract: The aim of this work was to evaluate the effect of a calcium lactate treatment on postharvest behaviour, organoleptic and nutritional quality of strawberries (Fragaria x ananassa Duch., cv. Camarosa) grown in different grow- ing systems and stored in refrigerated modified atmosphere. Strawberry grown in perlite and soil in greenhouse and soil in open field was harvested and dipped in a calcium lactate 1% solution. Fruits were packed in modified atmosphere at 1°C and 8°C. At 1, 3 and 7 days of storage postharvest behaviour, organoleptic and nutritional quality was evaluated. Calcium had a positive effect on fruit firmness and no differences were observed between storage temperatures in calci- um treated fruits. Organoleptic quality (except visual quality) was better in fruits grown in open field soil, regardless cal- cium treatment and storage temperature. Nutritional quality was better in untreated fruits and stored at 1°C. (*) Corresponding author: mharris@agro.uba.ar Received for publication 6 June 2016 Accepted for publication 12 December 2016 Copyright: © 2017 Author(s). This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ Adv. Hort. Sci., 2017 31(1): 3-10 4 The aim of this work was to evaluate the effect of a calcium lactate treatment on postharvest behav- iour, organoleptic and nutritional quality of strawber- ries grown in three growing systems and stored at two temperatures in modified atmosphere during 7 days of storage. 2. Materials and Methods Plant material and growth conditions The experiment was conducted in the Horticulture experimental field, School of Agriculture, University of Buenos Aires (latitude 45° S, longitude 58° 31´ W, altitude 26 m asl). Strawberry seedlings (commercial variety ‘Camarosa’) were planted in three growing systems: soil in open field, soil and perlite in green- house. Plant density was 7 plants m-2. Soil treatments were covered with a black plastic mulching. All treat- ments were fertirrigated: nutrient solutions were for- mulated according to strawberry´s requirements (Table 1). Hourly intervals of temperature (°C), rela- tive humidity (%) and radiation (W·m-2) were mea- sured in open field and greenhouse with datalogger HOBO. Harvest Mature strawberry fruits (at least 75% red colour) were harvested and immediately submerged in cold water to decrease fruit temperature. Half of the fruits of each growing system were treated with a calcium lactate 1% solution during one minute. Although calcium chloride is most commonly used, calcium lactate was used as a firming agent (Codex Alimentarius - World Health Organization, 2015) because chloride can give a bitter taste to fruits (Oms-Oliu et al., 2010). Sixty-five grams (g) of fruit were packed in a modified atmosphere (medium density polyethylene semi rigid container). Modified atmosphere increases postharvest life in fruits and vegetables by reducing respiratory rate (Sandhya, 2010). Containers were stored at storage chambers at 1°C and 8°C. Postharvest At 1, 3 and 7 days of storage, quality characteris- tics of fruits were evaluated as follows: Postharvest behaviour - Oxygen and carbon dioxide concentration in the container was measured with Dansensor gas analyzer and expressed as percentage. - Fresh weight loss. Fruits were weighted and weight loss was expressed as percentage relative to the initial value. Organoleptic quality - Visual quality. The overall visual and sanitary qual- ity was determined by scoring each strawberry using a 1-10 hedonic scale, being 10 excellent and 6 the commercialization limit. - Colour was measured with Minolta Chromameter CR300. L, a, b, c, h parameters were determined in four spots in equatorial zone of three fruits per treat- ment. - Firmness was measured in equatorial zone with Ludwig penetrometer fitted with a 3 mm diameter round probe. - Total soluble solids were determined with Atago refractometer and expressed as °Brix. Nutritional quality - Ascorbic and dehidroascorbic acid was deter- mined by liquid chromatography and expressed as mg of ascorbic acid 100 g-1 of fresh weight. - Antioxidant capacity. Antioxidant capacity was estimated by determining the free-radical scavenging capacity evaluated with the stable radical DPPH (adapted from Brand Williams et al., 1995 and Leong and Shui, 2002). Two g of edible portion of the fruit was homogenized using a blender and inserted into a 50 ml centrifuge tube. Twenty ml of 50% aqueous ethanol was added (1:10 w/v) and mixed in a vortex mixer for 15-30 seconds. The extract was centrifuged at 2000 g for 5 min al 4°C. The supernatant was fil- tered before using. A 25 mg solution of DPPH (1,1 diphenyl-2-picrylhydrazyl) was prepared in methanol. For calibration curve, aliquots of ascorbic acid (0, 25, 50, 75, and 100 μl) solved in aqueous ethanol 50% (0.1 ml ml-1) were placed in tubes with in 3 ml DPPH. Absorbance at 517 was measured at 1, 10, 30, 60, 90 and 120 minutes. An aliquot of 50 ml of an antioxi- dant/fruit extract solution was added to 3 ml of the DPPH solution. The decrease in absorbance at 517 Macroelements Microelements Element Concentration (mg l-1) Element Concentration (mg l-1) Nitrogen 64 Iron 2.8 Phosphorus 31 Sodium 1.2 Potassium 200 Manganese 0.5 Sulfur 64 Boron 0.5 Magnesium 48 Copper 0.02 -- Zinc 0.05 Molybdenum 0.01 Table 1 - Nutrient solution formulation Harris et al. - Calcium lactate treatments effects on postharvest of strawberry cultivar Camarosa 5 nm was measured at 0, 1, 5 and then every 10 min- utes until the reaction reached a plateau. The decreased absorbance of DPPH remaining at the steady-state was calculated and expressed as mg of a s c o r b i c a c i d ( A A ) e q u i v a l e n t s p e r 1 0 0 g o f homogenate (AEAC). The AEAC was calculated using the following equation: AEAC=Δ A x f x V x 100 x 1/W where Δ A is the change of absorbance after addition of fruit extract, f is the inverse of the calibration curve slope, V is the volume of filtrate (ml) and W is the weight of homogenate used for extraction (g). Statistical analysis A completely randomized factorial design with 3 replicates per treatment was used. The results were analyzed by multivariate analysis of variance repeat- ed in time with a 5% significance level. Tukey test to compare means was used (Kuehl, 2001). Infostat software was used (Di Rienzo et al., 2015). 3. Results and Discussion Oxygen and carbon dioxide concentration in the con- tainer Temperature, as expected, generated the most significant differences: strawberries stored at 1°C respired less than those stored at 8°C (Fig. 1). Nevertheless, in all cases, at equal temperatures, strawberries treated with calcium decreased the res- piration. On the other hand, an interaction between grow- ing systems and calcium treatment was observed (p<0,0001) both in oxygen and carbon dioxide levels: strawberries grown in soil systems (both open field and greenhouse) and without calcium treatment showed larger differences between storage tempera- tures: those stored at 8°C showed a high oxygen decrease and carbon dioxide increase, especially at the 7th day of storage. Treated fruits, instead, showed similar behaviour during the seven days of storage. Thus, calcium treatment could be considered as a regulator because differences between tempera- t u r e s i n t r e a t e d s t r a w b e r r i e s w e r e s m a l l e r . Waghmare and Annapure (2013) observed that a combination of modified atmosphere, calcium chlo- ride and nitric acid treatment in chopped papayas stored at 5°C had a significant decrease of oxygen and increase of carbon dioxide compared to fruits only stored in modified atmosphere. Equilibrium modified atmosphere was not estab- lished, especially those stored at low temperatures. This could be because medium density polyethylene was used and this material has low permeability to gases: 2600 cm 3/m 2.d.atm for oxygen and 7600 cm3/m2.d.atm for carbon dioxide (Sandhya, 2010). Additionally, as temperature increases, material per- meability increases as well. Thus, containers stored at 8°C had a higher permeability (Oliveira et al., 2015). Fig. 1 - Oxygen and carbon dioxide concentration (%) in containers at 1, 3 and 7 days of postharvest of strawberries grown in different growing systems, calcium lactate treatment and two storage temperatures. Adv. Hort. Sci., 2017 31(1): 3-10 6 Fresh weight loss Maximum allowable weight loss in strawberry is 6% (Laurin et al., 2003). All treatments, except one case, were below those values (Fig. 2). The smaller w e i g h t l o s s w a s o b s e r v e d i n o p e n f i e l d f r u i t s (p<0.0001), independently storage temperature and calcium treatment. Preharvest temperatures can affect postharvest shelf life. For example, fruits grown at high temperatures can exhibit water soak- ing (Benkeblia et al., 2011). In this work, differences between greenhouse and open field temperatures reached 7°C. Fruits harvested in greenhouse soil and soilless systems had surely higher temperature, what determined higher weight loss in those systems. Visual quality Fruits grown in perlite had better quality: 7.7% more than open field soil and 20% compared to greenhouse soil fruits (Fig. 3). At the 7th day of postharvest, all the strawberries stored at 1°C pre- Fig. 2 - Fresh weight loss (%) at 1, 3 and 7 days of postharvest of strawberries grown in different growing systems, calcium lactate treat- ment and two storage temperatures. Maximum allowable weight loss (6%) is indicated. Fig. 3 - Visual quality at 1, 3 and 7 days of postharvest of strawberries grown in different growing systems, calcium lactate treatment and two storage temperatures. Each graph represents a combination of growing system and storage temperature. Commercialization limit (6) is indicated. Harris et al. - Calcium lactate treatments effects on postharvest of strawberry cultivar Camarosa 7 sented visual and sanitary quality above commercial- ization level. Fruits stored at 8°C, instead, were below that level and in most cases they had Botrytis cinerea symptoms. Temperature management is a key factor to minimize postharvest deterioration in strawberry. At high storage temperatures, fruits have higher respiration rates, and consequently, shorter postharvest shelf life (Shin et al., 2008). Colour Significant differences were not observed for all colour parameters. Shin et al. (2008) did not find colour changes during 7 days of storage of L c and h° values. Firmness An interaction was observed between growing system and calcium treatment (p=0,0203): treated strawberries grown in soil systems, both open field and greenhouse had 20% more firmness than others. In perlite, difference between treated and untreated fruit was not significant (Fig. 4). Firmness increased in fruits stored at 1°C along the storage time. Shin et al. (2007) also observed an increase in firmness along 4 days of storage at high (10.5°C ) and low (0.5°C) temperatures. The same authors investigated firmness during 12 days of stor- age and observed a positive tendency in fruits stored at 3°C and a decrease in those stored at 10°C (Shin et al., 2008). Firmness increase in low storage tempera- tures is due to physical changes in cell wall: cold pro- duces an increase in pectin viscosity, which impacts positively in fruit firmness (Lara et al., 2004). Total soluble solids Total soluble solid content (Table 2) in all cases was above the minimum content recommended (7°Brix) for the postharvest quality maintenance (Mitcham et al., 2015). A significant interaction was found between temperature and growing system (p<0,0001): fruits stored at 8°C and grown in open Table 2 - Total soluble solids (°Brix) at 1, 3 and 7 days of postharvest of strawberries grown in different growing systems, calcium lactate treatment and two storage temperatures Growing system Storage 1°C Storage 8°C 1 day 3 days 7 days 1 day 3 days 7 days Ca with Ca without Ca with Ca without Ca with Ca without Ca with Ca without Ca with Ca without Ca with Ca without Soil open field 8 A bc 10 A d 7.63 AB ab 7.83 AB bc 8 A bc 6.33 A a 8.50 A bc 7.17 A a 9.17 AB cd 12 A e 8 A ab 9 AB cd Soil greenhouse 8 A bc 7.33 B a 7.75 AB ab 8.67 BC cd 8.17 AB bc 9.33 AB de 8.33 A cd 8.17 BC bc 8.33 B cd 7.33 BC ab 6 B a 8.83 B e Perlite greenhouse 8 A a 9.5 A bc 8 A a 9.17 CD ab 8 A a 8.83 BC ab 9 AB cd 6.5 AB a 8 BC ab 8.33 CD bc 10.5 C de 7.67 BC ab Different capital letters indicate differences between row and different small letters indicate differences between columns (p<0.05). Fig. 4 - Firmness (kPa) at 1, 3 and 7 days of postharvest of straw- berries grown in different growing systems, calcium lac- tate treatment and two storage temperatures. Adv. Hort. Sci., 2017 31(1): 3-10 8 field had higher content of soluble solids. It was only in the open field system that a difference between storage temperatures was found: those stored at 8°C had a 12.5% higher content of total soluble solids. Calcium lactate immersion affected negatively the total soluble solid content. Similar results were found by other authors with different calcium sources. Singh et al. (2007) observed that weekly foliar appli- cations of calcium chloride since flowering decreased total soluble solid content in 10% at harvest. Dunn and Able (2006), as well, found that a calcium defi- ciency during growth stage increased significantly soluble solids content in fruits. Ascorbic acid content In almost all cases, ascorbic acid content (Fig. 5) decreased during storage period. Phillips et al. (2016) also observed a decrease in strawberries stored at - 1.5°C during 7 days. Only the fruits stored at freezers (-10 to -20°C) and ultra freezers (-55°C) maintained ascorbic acid contents similar to those observed at harvest time. Low temperature storage is a key factor to maintain ascorbic acid content during postharvest (Lee and Kader, 2000). An interaction was observed between calcium treatment and storage temperature (p=0.0026). Untreated fruits stored at 1°C had 26% more ascorbic acid compared to the others. Many authors recog- nize that a calcium treatment at harvest enhances ascorbic acid content in fruit mainly due to an increase in fruit firmness (Aghdam et al., 2013). Nevertheless, other authors did not find differences between calcium treatment and the control (Shaffie et al., 2010). Regardless the calcium treatment, tem- perature was the key factor to maintain ascorbic acid content in strawberries. Fruits grown in perlite had a 49% higher content compared to those grown at soil (both open field and greenhouse). These results are in agreement with de data reported by Treftz and Omaye (2015), who observed that soilless grown strawberries had a 74% higher content of ascorbic acid compared to soil grown fruits. For other fruits, the results were con- tradictory: Isabelle et al. (2010) observed higher ascorbic acid content in pepper and Özcelik and Akilli (1999) in tomato. However, Gruda (2005) didn´t find differences in strawberry. Antioxidant capacity Several interactions were observed for antioxi- dant capacity. Untreated and greenhouse soil grown fruits presented 13% higher antioxidant capacity ( p = 0 , 0 0 0 3 ) ( T a b l e 3 ) . W a n g a n d Z h e n g ( 2 0 0 1 ) observed that strawberries grown under higher tem- perature (day and night) had higher antioxidant capacity, as it was observed in this work, where dif- ferences of temperatures between greenhouse and Fig. 5 - Ascorbic acid content (mg 100 g-1) at 1, 3 and 7 days of postharvest of strawberries grown in different growing systems, calcium lactate treatment and two storage temperatures. Harris et al. - Calcium lactate treatments effects on postharvest of strawberry cultivar Camarosa 9 open field reached 7°C. As well, untreated fruits stored at 1°C had 10% h i g h e r a n t i o x i d a n t c a p a c i t y ( p = 0 . 0 2 9 5 ) . M a n y authors, as described in 3.7, explain that a calcium treatment enhances ascorbic acid content (and con- sequently antioxidant capacity) due to an increase in cell wall firmness. Nevertheless, other authors express that a disruption in cell wall composition increases antioxidant capacity, but this increase is different depending the product (Reyes et al., 2006). With respect to storage temperature, Shin et al. (2008) found significant differences since the 12th day. Different capital letters indicate differences between columns and different small letters indicate differences between rows (p< 0.05). Yield in the growing systems Although yield was not an objective of this work, we observed that the perlite system had 35% and 17% higher yield than soil in greenhouse and soil in open field systems, respectively. 4. Conclusions Strawberry is highly accepted by consumers but its postharvest quality rapidly declines. A calcium lac- tate treatment can increase principally strawberry’s firmness. This, in addition to storage temperature and growing system can increase postharvest shelf life of fruits. In this research, the calcium lactate treatment had a positive effect on fruit firmness, especially in those grown in open field and greenhouse soil. Furthermore, treated fruits had less respiration dur- ing storage time and there was no significant differ- ence between storage temperatures in fruits treated with calcium lactate. Equilibrium modified atmos- phere was not established. The other variables were not affected or had a negative response to calcium lactate treatment. Weight loss and organoleptic quality (except visual quality) were better in fruits grown in open field soil, regardless calcium treatment and storage tempera- ture. On the other hand, nutritional quality was bet- ter in untreated fruits and stored at 1°C. In conclusion, even though temperature is sub- stantial to maintain fruit quality at postharvest, a cal- cium lactate treatment could be useful to improve strawberry (cv. 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