PEER-REVIEW ARTICLE PEER-REVIEWED ARTICLE bioresources.cnr.ncsu.edu Almutairi et al. (2024). “Strawberry edible coatings,” BioResources 19(3), 6057-6075. 6057 Salicylic Acid and Chitosan Effects on Fruit Quality When Applied to Fresh Strawberry or During Different Periods of Cold Storage Khalid F. Almutairi,a,* Abdulaziz R. Alharbi,a Mohamed Ewis Abdelaziz,b,c and Walid F. A. Mosa d One of the biggest problems that threaten the production of strawberry in the world is the rapid damage and high rate of deterioration after harvest or during cold storage. Therefore, the current study was conducted to investigate the possibility of decreasing the post-harvest damage percentages and increasing the fruit quality of Fragaria x ananassa cv. ‘Estavana’ after harvest immediately or during the cold storage period. The strawberry plants were dipped for 3 to 5 minutes at 25 C in a solution of 10 L made from distilled water and containing salicylic acid (SA) at 0, 250, 500, and 1000 mg/L or chitosan (CHIT) at 0, 2.5, 5, and 10 mg/L during the period of 0, 3, 6, 9, 12, and 16 days after harvesting. The results showed that the fruit firmness was notably decreased, and the loss and decay percentages were increased by increasing the period of storage, but it could be decreased by using SA or CHIT. Fruit content from soluble solids, total sugars and anthocyanin was significantly increased in the 16 days stored fruits treated with 500 mg/L SA or 50mg/L CHIT. Treating the fresh harvested without or with SA or CHIT increased the fruit content from vitamin C. The highest fruit content from titratable acidity was in the fresh harvested fruits compared with treated fruits with SA or CHIT. DOI: 10.15376/biores.19.3.6057-6075 Keywords: Cold storage; Strawberry; Biostimulants; Fruit quality Contact information: a: Department of Plant Production, College of Food Science and Agriculture, King Saud University, P.O. Box 2460, Riyadh 11451, Saudi Arabia; b: The National Research and Development Center for Sustainable Agriculture (Estidamah), Riyadh, Kingdom of Saudi Arabia; c; Department of Vegetable Crops, Faculty of Agriculture, Cairo University, Giza, 12613, Egypt; d: Plant Production Department (Horticulture-Pomology), Faculty of Agriculture, Saba Basha, Alexandria University, Alexandria 21531, Egypt; *Corresponding author: almutairik@ksu.edu.sa INTRODUCTION Strawberry fruit is a popular food and is rich in many different bioactive compounds and antioxidants, including terpenoids, flavonoids, carotenoids, anthocyanins, and vitamin E (Pang et al. 2020; Kim et al. 2021). Strawberries are susceptibile to mechanical damage, dehydration, and microbial deterioration, and as a result, present a challenging post harvest protocol (Benavides and Franco 2023). Because strawberries are highly perishable and susceptible to fungal infections, their shelf life is usually fewer than five days at room temperature, and their quality properties deteriorate quickly after harvest (Cordenunsi et al. 2005; Feliziani and Romanazzi 2016; Barkaoui et al. 2021). It has been investigated whether bioactive edible coatings with CHIT can reduce or eliminate fungal and mechanical fruit damage (Benavides and Franco 2023). Edible coating treatments can provide a protective layer on the product's surface (Deng et al. 2017). When applied to mailto:almutairik@ksu.edu.sa PEER-REVIEWED ARTICLE bioresources.cnr.ncsu.edu Almutairi et al. (2024). “Strawberry edible coatings,” BioResources 19(3), 6057-6075. 6058 fresh fruit, these coatings can prevent microbial attack and help maintain the fruit’s desirable composition and marketability. Additionally, coatings can prevent moisture loss, create a modified internal atmosphere, and regulate the concentration of gases such as O2, CO2, and fragrance compounds to more desirable levels (Gutiérrez and Álvarez 2017). Edible coatings are a suitable replacement for preserving postharvest crops (No et al. 2007). Coatings have been demonstrated to slow down the rates of carbohydrate breakdown, which delays maturity (Yan et al. 2019), and to function as moisture and gas barriers, controlling microbial growth and preserving colour and texture (Chaudhary et al. 2020). CHIT, an environmentally safe and non-toxic edible polymer with antifungal properties, stimulates plant defense mechanisms. It is frequently used to preserve postharvest fruits and vegetables as a food additive and a successful replacement for synthetic fungicides (Romanazzi et al. 2017). Moreover, CHIT coating can alter the internal atmosphere of the fruit by lowering oxygen levels and/or increasing carbon dioxide levels. This leads to reduced respiration rates and metabolic activity, delays in sugar accumulation and starch degradation, and potential initiation of fermentation processes (Silva et al. 2017). Furthermore, CHIT has a number of special advantages, including the capacity to form films, biocompatibility, biodegradability, antibacterial activity, and non- toxicity (Jiang et al. 2018a,b). The coating of fruit with CHIT has been demonstrated to have higher total soluble solids and antioxidant activity while decreasing hardness, decay, and weight loss (Adiletta et al. 2019). By minimizing losses in weight, soluble solids, vitamin C, titratable acidity, and firmness, CHIT coating preserves fruit quality during storage (Lin et al. 2020). By lowering the postharvest respiration rate of fruits, the coating’s use of polysaccharides such as CHIT acts as an effective oxygen and moisture barrier, delaying the deterioration of the product (Jung et al. 2020). An edible coating of CHIT can preserve the fruit freshness by minimizing microbial degradation, softening, and oxidative stress (Ghosh et al. 2021). Besides, CHIT works as an antimicrobial and antitranspirant agent, elicitor, and stimulator for both plant growth and beneficial microorganisms (Kipkoech et al. 2021). Additionally, the use of CHIT in coating mango fruit reduces the weight loss and the fruit content from TSS percentages, as well as preserves the peel firmness (Limon et al. 2021). Furthermore, CHIT is a great film-forming agent and biocompatible material for usage in the perishable fruit product sector. Fresh fruit coated with CHIT is thought to be safe for ingestion by humans because of its antibacterial and non-toxic qualities (Zhang et al. 2021). CHIT edible coating effectively reduces moisture loss, respiration rates, ethylene production, fruit ripening, softening, and decomposition, while preserving fruit quality and extending the postharvest life of various crops (Kumarihami et al. 2022). Salicylic acid (SA) is a relatively basic phenolic molecule that can be utilized as a safe and natural substitute chemical to regulate horticulture crops’ postharvest quality, and it is a naturally occurring substance that has the potential to significantly delay the fruit ripening (Mo et al. 2008). The application of SA effectively controls ethylene production, the respiration process, and enhances ascorbic acid content in fruits (Huang et al. 2008), lowering the post-harvest damages in horticultural crops (Asghari and Aghdam 2010), inducing the disease resistance (Shafiee et al. 2010), the preservation of post-harvest quality (Champa et al. 2015), and an extension of horticultural crop storage life. Additionally, it is frequently used to manage the firmness of crops to prolong their postharvest life (Kazemi et al. 2011). The preharvest treatment with SA significantly mitigates the harmful effects of heat and high light stress on photosystem, and it induces PEER-REVIEWED ARTICLE bioresources.cnr.ncsu.edu Almutairi et al. (2024). “Strawberry edible coatings,” BioResources 19(3), 6057-6075. 6059 the photosynthesis process (Zhao et al. 2011). Furthermore, Khan et al. (2012) documented that SA is essential for the organization of several plant physiological systems, including proline metabolism, antioxidant defense system, nitrogen absorption, photosynthesis, plant-water connections, and resistance to abiotic stressors. Besides, it was documented by many authors that SA is included in the improving resistances to various biotic and abiotic stresses, including heat, heavy metals toxicity, pathogens, and oxidative decomposition (Agnihotri et al. 2018; Ali et al. 2023), and cell wall enforcement (Jia et al. 2021). Treating peach cv. ‘Flordaking’ with SA at concentrations of 1, 2, and 3 mM improved fruit quality attributes such as flesh firmness, titratable acidity, and ascorbic acid content during storage. Conversely, these treatments reduced fruit weight loss, soluble solids content, membrane seepage, chilling injury, ethylene production, color deterioration, disease incidence, decomposition, and delayed the ethylene climacteric peak after six weeks of low- temperature storage (Ali et al. 2021). Besides, SA plays vital roles in raising the plant stress tolerance by activation the plant defense mechanisms (Alam et al. 2022), improvement of photosynthesis (George et al. 2022), the enhancing the plant tolerance to drought (Balfagón et al. 2022), cold (Raza et al. 2023), heat (Lafuente, and Romero 2022), and diseases (Khan et al. 2023). Besides, SA regulates the protein pattern, enhances the antioxidative system, growth and photosynthesis process, and Superoxide dismutase (SOD), Catalase (CAT), and Peroxidase (POD) activities under cold stress (Hernanz et al. 2023; Raza et al. 2023; Singh 2023). Therefore, the current study was conducted to increase the shelf life and maintain the strawberry fruits during storage and reduce their decay and fungal attack by the application of CHIT and SA. EXPERIMENTAL The current work was performed to study the effect of the dipping of strawberry fruits (Fragaria x ananassa cv. ‘Estavana’) for 3 to 5 min at 25 C with SA (LHCHEM company, Jinan City, Shandong Province, China) at 0, 250, 500, and 1000 mg/L and CHIT (Realfine Chemicals (Shanghai) Co., Ltd., Wuxi, Jiangsu, China) at 0, 2.5, 5, and 10 mg/L during the period of 0, 3, 6, 9, 12 and 16 days after harvesting. The volume of the used solution was 10 L from distilled water. Split-Plot design was used to perform this experiment where the time was the main factor, and SA or CHIT was the sub main factor in eight replicates were used as blocks. During the storage periods, several parameters were measured. Physical Fruit Characteristics A total of 40 fruits for each treatment (10 fruits as a replicate) were chosen to perform the study during the experiment times and the fruit quality was measured as follows: Fruit firmness (lb/inch2) was assessed by using a Magness and Taylor pressure tester (mod. FT 02 (0-2 lb, Alfonsine, Italy). The number of strawberry fruits that had rotted and showed signs of pitted peel or pathogen incidence relative to the overall quantity of strawberries was used to calculate the percentage of fruit decay, which was then represented as a percentage following each storage period (Huang et al. 2023). By comparing the difference between the fruits’ original and final weights using an electronic balance, the percentage of fruit weight loss was calculated (PA4102 OHAUS Corporation, USA) and expressed in percentage (%). https://realfinesh.en.alibaba.com/index.html?from=detail&productId=1601151623002 PEER-REVIEWED ARTICLE bioresources.cnr.ncsu.edu Almutairi et al. (2024). “Strawberry edible coatings,” BioResources 19(3), 6057-6075. 6060 Fruit Chemical Characteristics Total soluble solids (TSS) were assessed by triplicate with a digital refractometer (Atago N1; Atago Co. Ltd., Tokyo, Japan) at 20 °C and expressed as %. Total Titratable Acidity (TA) The TA was determined according to an AOAC method (AOAC 2005) by triplicate using an automatic titration device (877 Titrino plus, Metrohm ion analyses CH9101, Herisau, Switzerland) with 0.1 N NaOH up to pH 8.1, using 1 mL diluted juice in 25 mL distilled H2O, and the results were expressed as g malic acid per 100 g fw (Celikel et al. 2008). Malondialdehyde (MDA) content was assayed using the TBA method, where 2.0 g of frozen juice sac was extracted with 5 mL of 10% (m/v) TCA solution and centrifuged (10000 × g at 4 °C for 20 min). Afterwards, 2.0 mL of the supernatant was mixed by adding the same volume of 0.67% TBA (dissolved in 50 mM NaOH) solution, followed by a boiling water bath for 20 min, and then quickly cooled in an ice bath. Finally, the absorbance of the supernatant was recorded at three specific wavelengths (450 nm, 532 nm, and 600 nm) using a UV-Vis spectrophotometer (model: TU-1950, Persee General Instrument Co., Ltd., Beijing, China), with the results were reported as millimole per gram fruit weight (Bakpa and Zhang 2022). Fruit Total Sugars The quantity was determined by using phenol sulfuric acid, and fruit reducing sugars content was determined calorimetrically (Nielsen 2010). The content of Vitamin C mg/100 mL juice was determined by the titration of 2,6-dichlorophenol-indo-phenol (Huang et al. 2023). Antioxidant Enzyme Activity Assays A 10 g fruit sample was chosen and homogenized using a Kinematica tissue processor (Kinematica AG Werkstrasse 7 c-d Switzerland) in 25 mL of ice-cold extraction buffer and 0.5 g of polyvinyl-polypyrrolidone (PVPP) (Crl-6010, Kriens-LU, Switzerland). For the catalase (CAT) and superoxide dismutase (SOD) estimation, the extraction buffer used was 50 mM sodium phosphate with a pH of 7.8. The homogenized mixture was centrifuged at 27,000 g for 50 min at 4 °C, and the resulting supernatants were used to measure CAT and SOD activity (Wang et al. 2005). The reaction combination for CAT analysis consisted of 2.8 mL H2O2 (40 mM in 50 mM sodium phosphate buffer, pH 7.0) and 0.2 mL enzyme extract. The decrease of H2O2 (substrate) was determined by measuring the decrease in absorbance at 240 nm during a 120-s period using a spectrophotometer (Model UH4150AD UV-Vis-NIR Spectrophotometer Hitachi, Ltd., Tokyo, Japan). The activity was expressed as unit g−1 FW, where one unit of catalase equals one molecule of H2O2 per mass of fruit per minute at 30 °C. For the SOD activity assay, a volume of approximately 3 mL in the SOD activity experiment comprised 65 mM sodium phosphate cradle (pH 7.8). Finally, 13 mM methionine, 75 mM nitro-blue tetrazolium (NBT), 10 mM EDTA, and 2 mM riboflavin were mixed, along with 0.1 mL of the enzyme extract. After illumination of the combinations for 10 minutes with light (60 Mol m−2 s−1), the absorbance at 560 nm was evaluated using a spectrophotometer. The reaction solution took 3 min to settle. The reaction was measured in units of g−1 FW, where one unit represented the amount of chemical that caused a 50% reduction in the SOD-inhibitable NBT per mass per hour. PEER-REVIEWED ARTICLE bioresources.cnr.ncsu.edu Almutairi et al. (2024). “Strawberry edible coatings,” BioResources 19(3), 6057-6075. 6061 Total Phenols Content Phenol content was measured by the reduction of phosphotungstic-phospho- molybdic acid to blue pigments in an alkaline solution, following the Folin method as described by Salem et al. (2018). A 100 μL aliquot of the diluted sample (1/100) in ethanol was mixed with 400 μL of 1/10 diluted Folin-Ciocalteu reagent. After 5 minutes, 500 μL of a 10% (w/v) sodium carbonate solution was added. Following a 1-hour incubation at room temperature, the absorbance was measured at 765 nm in triplicate. The total polyphenol content was expressed as mg gallic acid equivalents per g dry weight of fruit (mg GAE/g DW). Total Anthocyanin Content (TAC) One mL of each fruit extract sample was separately added to 980 μL of KCl buffer (pH 1.0) and NaOAc buffer (pH 4.5). After a 15-min incubation at room temperature, the absorbance was measured at 510 nm and 700 nm using a spectrophotometer, with 50% ethanol as a blank. The total anthocyanin content (TAC) was calculated using Eq. 1, and the results were expressed as mg of cyanidin-3-glucoside equivalents per 100 g of dry weight. TAC = (A*MW*DF *1000/Ɛ*L) (1) where, A: Absorbance = [(A 510 nm to A 700 nm)] pH 1.0 – [(A 510 nm to A 700 nm)] pH 4.5; MW: molecular weight (449.2 g mol-1); DF: dilution factor; L: is the cell path length (1 cm); Ɛ: molar absorptivity coefficient of cyanidin- 3-glucoside (26900 L mol-1 cm-1) (Jakobek et al. 2007). Statistical Analysis The obtained results were statistical analysis using Split-Plot Design by using CoHort Software (Pacific Grove, CA, USA), and the Least Significant Difference (LSD) at 0.05% was used to compare the means of treatments (Mishra et al. 2019). RESULTS AND DISCUSSION The data in Table 1 show that the fruit firmness was significantly increased in the fresh harvested fruits, especially when they were treated with 1000 SA. On the contrary, the fruit firmness was decreased by increasing the period of the storage. For 9 days stored fruits, the the statistical analysis indicated that there are no significant differences between the treatments. Additionally, the treating the stored fruits for 3 or 12 days with CHIT show that there are no significant differences between the different concentrations. The results in Table 2 show that fruit decay percentages in the fresh fruits were zero. However, the decay percentages were significantly increased by increasing the time of storage, where the period of 16 or 12 days registered the highest percentages. The usage of SA or CHIT decreased the decay percentages in the stored fruits. Moreover, the percentage of decay was also minimized by raising the used concentration from SA or CHIT. PEER-REVIEWED ARTICLE bioresources.cnr.ncsu.edu Almutairi et al. (2024). “Strawberry edible coatings,” BioResources 19(3), 6057-6075. 6062 Table 1. Effect of Storage Time and Postharvest Treatment of SA and CHIT on Fruit Firmness Fruit Firmness (Ib/inch2) Treatment Concentration (mg/L) Time of the storage (days) 0 3 6 9 12 16 SA 0 2.47ab 2.27a-e 2.00a-e 1.77a-e 1.60de 1.67b-e 250 2.40a-d 2.32a-e 2.00a-e 1.75a-e 1.72a-e 1.80a-e 500 2.35a-e 1.75a-e 1.85a-e 2.37a-e 1.77a-e 1.82a-e 1000 2.52a 1.92a-e 2.20a-e 2.45a-c 1.77a-e 1.77a-e CHIT 0 2.32a-e 2.35a-e 2.07a-e 1.57de 2.25a-e 1.55e 2.5 2.32a-e 2.10a-e 1.75a-e 1.75a-e 2.22a-e 1.62c-e 5 2.37a-e 2.20a-e 2.00a-e 2.37a-e 2.52a 1.60de 10 2.37a-e 2.37a-e 2.07a-e 2.20a-e 2.05a-e 1.92a-e LSD0.05 0.41 The treatments that have the same letters mean that there are no significant differences between them. Table 2. Effect of Storage Time and Postharvest Treatment of SA and CHIT on the Fruit Decay Percentage Decay % Treatment Concentration (mg/L) Time of the storage (days) 0 3 6 9 12 16 SA 0 0 x 4.57t 8.57o 14.34k 25.15c 33.83a 250 0 x 3.75u 5.99r 11.78m 20.65f 27.37b 500 0 x 3.19uv 5.93r 10.02n 17.57h 23.29d 1000 0 x 2.76vw 5.12st 8.65o 13.35 l 20.10g CHIT 0 0 x 4.58t 8.65o 14.36k 25.20c 34.13a 2.5 0 x 3.23uv 6.97q 10.12n 17.75h 23.52d 5 0 x 2.90vw 5.39rs 9.11o 15.98i 21.17e 10 0 x 2.43w 4.50t 7.61p 15.17j 17.69h LSD0.05 0.50 The treatments that have the same letters mean that there are no significant differences between them. . Table 3. Effect of Storage Time and Postharvest Treatment of SA and CHIT on the Fruit Loss Percentage Loss % Treatment Concentration (mg/L) Time of the storage (days) 0 3 6 9 12 16 SA 0 0v 1.15rs 1.52op 2.08n 6.93ef 8.91a 250 0v 1.26qr 1.71o 2.31m 6.73f 7.69d 500 0v 1.167rs 1.58op 2.16n 6.18g 8.19b 1000 0v 1.13rs 1.53op 2.10n 5.92h 7.84cd CHIT 0 0v 1.13rs 1.53op 2.09n 7.03e 7.90c 2.5 0v 0.94stu 1.27qr 1.74o 5.09j 6.74f 5 0v 0.86tu 1.17rs 1.60op 4.57k 6.05gh 10 0v 0.77u 1.04rst 1.42pq 4.16l 5.52i LSD0.05 0.17 The treatments that have the same letters mean that there are no significant differences between them. PEER-REVIEWED ARTICLE bioresources.cnr.ncsu.edu Almutairi et al. (2024). “Strawberry edible coatings,” BioResources 19(3), 6057-6075. 6063 Table 3 shows that the loss percentages were significantly enhanced by the storage of the strawberry fruits. The highest percentage was observed in 16 days of stored fruits. The percentage of fruit loss was increased by increasing the period of the storage and using SA or CHIT played a good role in decreasing the fruit loss percentage. The efficacy of SA or CHIT was increased in parallel to increasing the used concentration. As shown in Table 4, the storage of strawberry fruits increased the fruit content from TSS % where the period of 16 days registered the highest percentage from TSS in particularly when 500 mg/L SA or 10 mg/L CHIT. Increasing the used concentration from SA or CHIT was more effective in improving the fruit content from soluble solids during the storage period. In the 16 days stored fruits, the differences between the effect of 250 and 1000 mg/L SA or between 2.5 and 5 mg/L CHIT were not significant. Table 4. Effect of Storage Time and Postharvest Treatment of SA and CHIT on TSS % TSS% Treatment Concentration (mg/L) Time of the storage (days) 0 3 6 9 12 16 SA 0 8.07r 8.40o-r 8.55n-r 9.35jk 10.45e-h 11.57bc 250 8.15qr 8.82k-q 9.00j-o 9.87hi 10.75d-g 11.72ab 500 8.12r 8.65m-r 8.72l-r 9.55ij 10.72d-g 12.12a 1000 8.22qr 8.67m-r 9.32j-l 10.15gh 10.77d-g 11.50bc CHIT 0 8.10r 8.07r 8.20qr 8.92k-p 10.05h 10.95de 2.5 8.07r 8.45o-r 8.65m-r 9.27j-m 10.42e-h 11.47bc 5 8.27p-r 8.30p-r 8.42o-r 9.12j-n 10.30f-h 11.15cd 10 8.20qr 8.75k-r 8.90k-p 10.20h 10.82d-f 12.12a LSD0.05 0.38 The treatments that have the same letters mean that there are no significant differences between them. Table 5 shows that when the strawberry fruits were stored, anthocyanin was increased in parallel to increasing the period of the storage, where the highest concentration of anthocyanin was in the 16 days stored fruits. The SA and CHIT helped to improve the fruit coloring by increasing the content of anthocyanin, where the 250, 500, and 1000 SA or 2.5, 5, and 10 mg/L CHIT in 16 days stored fruits gave the highest concentrations. Table 5. Effect of Storage Time and Postharvest Treatment of SA and CHIT on Anthocyanin Content Anthocyanin (mg/100g) Treatment Concentration (mg/L) Time of the storage (days) 0 3 6 9 12 16 SA 0 30.72p 35.32m-o 37.675l-n 42.17h-j 48.82de 51.95a-c 250 31.37p 35.67m-o 39.52kl 42.92h-j 49.82cd 51.90a-c 500 31.75p 34.77no 38.10lm 43.12hi 52.10a-c 52.25a-c 1000 31.45p 36.20m-o 41.80h-k 46.72ef 47.65d-f 52.77ab CHIT 0 30.62p 34.50o 36.67m-o 40.40jk 45.75fg 50.07b-d 2.5 31.70p 35.47m-o 37.40l-o 40.60i-k 46.05fg 51.45a-c 5 31.55p 34.87no 36.95m-o 40.72i-k 46.50e-g 50.15b-d 10 31.15p 36.67m-o 44.17gh 47.17ef 48.70de 53.62a LSD0.05 1.80 Treatments that take the same letters mean that there are no significant differences between them. PEER-REVIEWED ARTICLE bioresources.cnr.ncsu.edu Almutairi et al. (2024). “Strawberry edible coatings,” BioResources 19(3), 6057-6075. 6064 The results in Table 6 show that the storage of the strawberry fruits gradually increased the fruit content from the total sugars percentages and the highest percentages were noticed when the fruits were treated with SA or CHIT. The differences between the effects of the different concentrations of SA or CHIT were not significant. The fruit content from total sugars was significantly low in fresh harvested fruits and when the fruits were stored for 3, 6 and 9 days after harvested. Table 6. Effect of Storage Time and Postharvest Treatment of SA and CHIT on Total Sugars Content % Total sugars % Treatment Concentration (mg/L) Time of the storage (days) 0 3 6 9 12 16 SA 0 6.62l-n 6.77j-n 7.22i-l 7.6f-i 8.40a-d 8.71ab 250 6.55l-n 6.82j-n 6.82j-n 7.45g-j 8.35b-d 8.99a 500 6.52l-n 6.57l-n 6.70k-n 7.37h-k 8.13b-f 9.00a 1000 6.27n 6.42mn 6.50l-n 7.227i-l 7.93c-h 8.70ab CHIT 0 6.50l-n 6.80j-n 6.87j-n 7.65f-i 8.30b-e 8.53a-c 2.5 6.55l-n 6.52l-n 6.87j-n 7.20i-l 8.01c-g 8.77ab 5 6.45mn 6.55l-n 6.50l-n 7.147i-m 7.80d-i 9.01a 10 6.45mn 6.27n 6.30n 6.85j-n 7.72e-i 8.44a-c LSD0.05 0.40 The treatments that take the same letters mean that there are no significant differences between them. The data in Table 7 show that treating fresh fruits with 500 mg/L SA significantly increased their Vitamin C (VC) content. Additionally, treating fruits with 500 or 1000 mg/L SA and also by 2.5, 5, or 10 mg/L CHIT also greatly improved the VC. The results showed that the storage of strawberry fruits for 3, 6, 12, and 16 days solely or by treating them with 250, 500, and 1000 SA or by 2.5, 5, and 10 mg/L decreased VC. Additionally, there is an inverse relationship between the vitamin C content in the fruit and the storage duration, with the lowest values observed after 12 or 16 days of storage. This trend was noted whether the fruit was untreated or treated with 250, 500, and 1000 mg/L SA or 2.5, 5, and 10 mg/L CHIT. Table 7. Effect of Storage Time and Postharvest Treatment of SA and CHIT on Vitamin C Levels Vitamin C (mg/100 mL) Treatment Concentration (mg/L) Time of the storage (days) 0 3 6 9 12 16 SA 0 58.20b 51.42f 45.02k 42.00n 35.32s 36.47qr 250 57.90b 50.65fg 46.57i 42.60mn 36.32qr 36.32qr 500 59.27a 52.37e 48.20h 44.20l 37.87p 36.70qr 1000 58.15b 53.00e 48.90h 44.90kl 38.32p 36.90q CHIT 0 57.97b 51.40f 46.62i 42.85m 35.80rs 36.47qr 2.5 58.27b 52.60e 48.45h 44.12l 37.9p 36.62qr 5 58.27b 54.47d 50.12g 45.72j 39.37o 37.57p 10 58.00b 55.22c 50.70fg 46.25ij 39.65o 37.97p LSD0.05 0.67 The treatments that take the same letters mean that there are no significant differences between them. PEER-REVIEWED ARTICLE bioresources.cnr.ncsu.edu Almutairi et al. (2024). “Strawberry edible coatings,” BioResources 19(3), 6057-6075. 6065 The results in Table 8 show that the fresh harvested fruits were characterized by a high content of titratable acidity. Fresh fruit treated with 250, 500, and 1000 SA or with 2.5, 5, and 10 mg/L contained high quantities of titratable acidity. Additionally, the 3 and 6 days stored fruit contained higher titratable acidity than fruits stored for 9, 12, and 16 days, only or after treatment with 250, 500, and 1000 mg/L SA and 2.5, 5, and 10 mg/L CHIT. Additionally, the results showed that as the period of storage increased, the percentage of titratable acidity decreased. Table 8. Effect of Storage Time and Postharvest Treatment of SA and CHIT on the Fruit Titratable Acidity Titratable acidity % Treatment Concentration (mg/L) Time of the storage (days) 0 3 6 9 12 16 SA 0 0.87a 0.83e 0.82ef 0.79ij 0.77l-n 0.71q 250 0.85cd 0.83e 0.81e-g 0.78jk 0.76m-o 0.70qr 500 0.86a-c 0.82ef 0.81f-h 0.78j-l 0.76n-p 0.70qr 1000 0.87ab 0.81e-g 0.80gh 0.77k-n 0.75op 0.69r CHIT 0 0.87a 0.83e 0.81e-g 0.78jk 0.77m-o 0.70qr 2.5 0.86b-d 0.82ef 0.81f-h 0.78j-l 0.76n-p 0.70qr 5 0.85d 0.82e-g 0.80gh 0.77l-n 0.75op 0.70r 10 0.85d 0.81f-h 0.80hi 0.77k-m 0.75p 0.69r LSD0.05 0.01 The treatments that take the same letters mean that there are no significant differences between them. The results in Table 9 show that the fruit content from phenol was significantly increased in the 3 days of stored fruits. Additionally, fruits stored for 3 days after treatment with 250, 500, and 1000 SA or with 2.5, 5, and 10 mg/L CHIT also contained a high quantity of phenols. Moreover, the 6 or 9 days of stored fruits solely or after treatment with SA or CHIT are characterized by a high phenol content. By increasing the storage period, the fruit content from phenols was decreased even with using SA or CHIT. Table 9. Effect of Storage Time and Postharvest Treatment of SA and CHIT on Phenols Phenols (mg/g dry weight) Treatment Concentration (mg/L) Time of the storage (days) 0 3 6 9 12 16 SA 0 106.00op 165.50a 153.00b 141.25e 121.50l 108.00op 250 106.75op 156.00b 145.00cd 138.00f 125.00k 112.50n 500 105.75op 153.00b 141.75de 137.00fg 127.00jk 115.75m 1000 104.75p 148.00c 136.00fg 131.00hi 130.75hi 116.00m CHIT 0 105.5op 164.25a 147.75c 142.50de 121.50l 108.75o 2.5 108.00op 153.75b 142.50de 135.75fg 127.00jk 115.25m 5 106.00op 147.50c 136.50fg 131.00hi 132.00h 116.25m 10 105.25op 142.50de 133.75gh 128.50ij 134.25gh 116.50m LSD0.05 2.44 The treatments that take the same letters mean that there are no significant differences between them. The data in Table 10 show that a high significant concentration of MDA was in fruits stored for 16 days. Moreover, fruit stored for 16 days and treated with 250, 500, and PEER-REVIEWED ARTICLE bioresources.cnr.ncsu.edu Almutairi et al. (2024). “Strawberry edible coatings,” BioResources 19(3), 6057-6075. 6066 1000 mg/L SA and also with 2.5, 5, and 10 mg/L CHIT were characterized by a high content of MDA. Its content was also significantly increased in the 12 days solely or after the treatment with 250, 500, and 1000 SA or with 2.5, 5, and 10 mg/L CHIT. In the fresh harvest fruits, MDA concentration was less than that after the storage, whereas when the storage time increased, the concentration of MDA in the fruit increased. Table 10. Effect of Storage Time and Postharvest Treatment of SA and CHIT on MDA Enzyme MDA ((µmol/g fruit weight) Treatment Concentration (mg/L) Time of the storage (days) 0 3 6 9 12 16 SA 0 40.40z 51.90n 55.60j 61.5f 65.70b 68.70a 250 40.57yz 46.72s 52.45m 56.67i 62.47e 65.62b 500 40.60yz 44.55u 50.55p 55.42j 60.62g 64.27c 1000 41.00y 43.95v 48.85q 53.40l 63.57d 63.57d CHIT 0 40.70yz 51.50o 55.50j 61.55f 65.60b 68.52a 2.5 40.32z 44.92t 50.50p 55.60j 61.57f 65.40b 5 40.62yz 43.47w 48.87q 54.57k 60.45g 64.45c 10 40.67yz 42.72x 47.55r 53.72l 58.57h 63.57d LSD0.05 0.34 The treatments that take the same letters mean that there are no significant differences between them. At 6 days, stored fruits were characterized by the high significant content of catalase enzyme (CAT), followed by 3 or 9 days stored fruits solely or after the usage of SA or CHIT (Table 11). The lowest concentrations of CAT enzyme were noticed in the 16 days stored fruits or even after the usage of SA or CHIT. The amount of CAT enzyme was significantly different in the fresh fruits compared to fruits treated with SA or CHIT. Table 11. Effect of Storage Time and Postharvest Treatment of SA and CHIT on CAT Enzyme CAT (U/Fruit weight) Treatment Concentration (mg/L) Time of the storage (days) 0 3 6 9 12 16 SA 0 48.60i 55.50b 58.55a 54.67bc 40.45k 30.62r 250 48.57i 53.67cd 55.60b 52.70d-f 41.65k 33.65op 500 48.52i 52.70d-f 54.62bc 50.5gh 38.60l 32.70pq 1000 48.50i 51.42fg 53.72cd 52.02e-g 36.62m 31.42qr CHIT 0 48.62i 55.65b 58.67a 54.62bc 40.55k 30.57r 2.5 48.77i 52.65d-f 55.45b 51.65fg 43.60j 35.50n 5 48.57i 50.62gh 53.47c-e 50.57gh 41.55k 34.60no 10 48.52i 49.67hi 51.70fg 48.67i 38.55l 31.62qr LSD0.05 1.11 The treatments that take the same letters mean that there are no significant differences between them. Treating the 3, 6 and 9 days stored fruits with 250, 500, and 1000 mg/L SA or with 2.5, 5, and 10 mg/L CHIT increased the superoxide dismutase (SOD) enzyme (Table 12). The superior concentration of SOD was noticed in the 6 days stored fruits after treatment of 1000 mg/L SA and in the 3, 6 and 9 days stored fruits treated with 10 mg/L CHIT. It was noticed also that SOD decreased gradually by increasing the storage period where its PEER-REVIEWED ARTICLE bioresources.cnr.ncsu.edu Almutairi et al. (2024). “Strawberry edible coatings,” BioResources 19(3), 6057-6075. 6067 concentration in the 12 and 16 stored fruits was significantly decreased particularly when the fruits were not treated with SA or CHIT. Table 12. Effect of Storage Time and Postharvest Treatment of SA and CHIT on SOD Enzyme SOD (U/Fruit weight) Treatment Concentration (mg/L) Time of the storage (days) 0 3 6 9 12 16 SA 0 10.77ef 9.65g 8.55i 7.65j 7.60j 6.90k 250 10.42f 11.57d 12.67b 11.65d 10.42f 8.60i 500 10.55ef 12.62b 12.87b 12.02cd 10.95ef 9.12h 1000 10.52ef 12.75b 13.65a 12.72b 11.55d 9.62g CHIT 0 10.70ef 9.57g 8.65i 7.65j 7.35j 7.40j 2.5 10.50ef 11.57d 12.50b 12.35bc 10.67ef 9.57g 5 10.45f 12.62b 12.72b 12.77b 11.02e 10.50ef 10 10.57ef 13.47a 13.42a 13.42a 11.60d 10.80ef LSD0.05 0.34 The treatments that take the same letters mean that there are no significant differences between them. DISCUSSION The results showed that the post-implementation of CHIT on strawberry increased their fruit shelf life, and decreased the percentage of decay and fungal diseases. These results were in agreement with Wang and Gao (2013); they reported that untreated strawberry fruits with CHIT may have low anthocyanin concentrations due to pigment degradation and accelerated fruit senescence. Because of its propensity to produce semi- permeable membranes, the CHIT-coated film on the fruit surface can block epidermal stomata and lenticels (Hosseinnejad and Jafari 2016), which decreases water and nutrient loss from the fruit and prevents respiration (Xu et al. 2018). Treating kiwifruit with CHIT after harvesting increased the fruit firmness as a result of the creation of a semipermeable layer on the fruit surface, which functions as a protective barrier to minimise the respiration rate, hence reducing metabolic activity and textural changes (Drevinskas et al. 2017; Zheng et al. 2017). Besides, CHIT may create an altered environment surrounding the fruit surface, which inhibits the breakdown of pectin and postpones the fruit’s loss of firmness (He et al. 2018). CHIT-based coatings have been effectively applied to a range of fresh fruits and vegetables. They can function as barriers, slow down senescence and maturation, lessen dehydration, and postpone microbiological and fungal decomposition (Pagno et al. 2018; Kabanov and Novinyuk 2020). CHIT is a good edible coating material due to its outstanding film-forming properties, high mechanical strength, and antibacterial qualities (Jiao et al. 2019). CHIT treatment inhibited fungal and bacterial growth, implying that pathogens grew more slowly in these settings (Youssef and Hashim 2020). Wantat et al. (2022) described a CHIT-montmorillonite nanocomposites coating that might lower ethylene generation and respiration rates, hence preserving banana storage quality. The results of this experiment showed that the usage of SA on strawberry increased their fruit shelf life and decreased decay percentages and fungal diseases. These results are in the same line with those documented by Barman et al. (2016). They reported that SA plays a significant function in different physiological processes such as fruit ripening, PEER-REVIEWED ARTICLE bioresources.cnr.ncsu.edu Almutairi et al. (2024). “Strawberry edible coatings,” BioResources 19(3), 6057-6075. 6068 minimizing fruit damage by increasing ethylene production, and preserving the fruit firmness and color. Additionally, SA has been reported to enhance disease resistance (Zhang et al. 2010), reduce chilling (Luo et al. 2011), increase the storability of horticultural crops (Valero et al. 2011), and have antisenescence properties, delaying the postharvest ripening process (Gimenez et al. 2017). These effects lead to improving the fruit content from TSS percentage (Ahmad et al. 2013). According to García-Pastor et al. (2020), the application of SA resulted in a more intense red hue by raising the concentration of anthocyanins, hence boosting pomegranate profits on the global market. SA can reduce the metabolic activity and transpiration of fruit, thereby inhibiting weight loss (Amiri et al. 2021), which is a crucial quality parameter for commercial fruit, as it directly impacts visual quality and freshness (Koyuncu et al. 2019; Madhav et al. 2021). Batool et al. (2022) reported that postharvest treatments with SA significantly preserved total soluble solids, titratable acidity, color profile, ascorbic acid content, and total phenolic content in apricot varieties. These treatments also enhanced antioxidant activity and texture, maintained the visual color of apricots compared to the control, and reduced chilling injury index, weight loss, and decay percentage. The use of SA has been shown to improve storage quality by lowering respiration rates and ethylene production, preventing changes in fruit color and softening, preserving sugars and organic acids, reducing chilling injury, and boosting both pathogen resistance and the antioxidant system (Chen et al. 2023). CONCLUSIONS 1. The percentages of fruit loss and decay were significantly increased in parallel to increasing the period of the storage while, the fruit firmness was high in the fresh harvested fruits particularly, the fruits treated with 1000 mg/L salicylic acid (SA). 2. Treating strawberry fruits with salicylic acid (SA) or chitosan (CHIT) increased the total soluble solids (TSS), total sugars, vitamin C, and anthocyanin content. It was noticed that after 3 days storage, the phenol content was increased especially when they were not treated with SA or CHIT. 3. The CAT and SOD enzyme contents were decreased during the storage period, and the treatment of SA and CHIT increased their content, while malondialdehyde (MDA) was increased by increasing the time of the storage while its concentration was decreased after treatment of SA or CHIT. ACKNOWLEDGMENTS The authors would like to extend their gratitude to the Saudi Arabia Ministry of Environment, Water and Agriculture for providing financial, technical, and administrative support to fund this work through the initiative of encouraging the agricultural “Applied research project in the field of Promising Crops no. 220011487”. The authors also extend their thanks and appreciation to The National Research and Development Center for Sustainable Agriculture (Estidamah) for overcoming the difficulties and facilitating the tasks to achieve this study. PEER-REVIEWED ARTICLE bioresources.cnr.ncsu.edu Almutairi et al. (2024). “Strawberry edible coatings,” BioResources 19(3), 6057-6075. 6069 REFERENCES CITED Adiletta, G., Zampella, L., Coletta, C., and Petriccione, M. (2019). “Chitosan coating to preserve the qualitative traits and improve antioxidant system in fresh figs (Ficus carica L.),” Agriculture 9(4), article 84. 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DOI: 10.1007/s11947-017-1957-5 Article submitted: April 4, 2024; Peer review completed: June 8, 2024; Revised version received and accepted: July 7, 2024; Published: July 17, 2024. DOI: 10.15376/biores.19.3.6057-6075