بررسی فعالیت ضدقارچی اسانس آویشن نانوکپسوله در برابر Botrytis cinerea در انگور قرمز شاهرودی

نوع مقاله : مقاله پژوهشی

نویسندگان

• حدیثه کریمی ¹
• حجت اله بداقی ¹
• احمد رجایی ²
• شیده موجرلو ¹

¹ گروه علوم باغبانی و گیاهپزشکی، دانشکده کشاورزی، دانشگاه صنعتی شاهرود، شاهرود، ایران.

² گروه علوم و صنایع غذایی، دانشکده کشاورزی، دانشگاه صنعتی شاهرود، شاهرود، ایران.

https://doi.org/10.22067/ifstrj.v16i4.76390

چکیده

پژوهش حاضر به‌منظور بررسی فعالیت ضدقارچی و پایداری اسانس آویشن درون‌پوشانی در نانوژل کیتوزان-مریستیک اسید در برابر قارچ Botrytis cinerea در انگور قرمز شاهرودی انجام گرفت. اثر نانوژل کیتوزان-مریستیک اسید در سه سطح صفر، 150 و 300 میکرولیتر در لیتر، اسانس آویشن خالص و اسانس آویشن درون‌پوشانی‌شده در نانوژل کیتوزان-مریستیک اسید در سه سطح 0، 75 و 150 میکرولیتر در لیتر در شرایط آزمایشگاهی و بر ماندگاری میوه‌های انگور در شرایط اتمسفر تعدیل‌یافته در طی 72 روز نگهداری در دمای 2 تا 4 درجه سانتی‌گراد موردبررسی قرار گرفت. نتایج طیف‌سنجی مادون قرمز (FTIR) اتصال موفقیت‌آمیز بین گروه‌های آمین کیتوزان و گروه‌های کربوکسیلیک اسید-مریستیک اسید را تأیید کرد و تصویر میکروسکوپ الکترونی روبشی اندازه ذرات نانوژل کیتوزان-مریستیک اسید حاوی اسانس را کمتر از 100 نانومتر نشان داد. تست رهایش نشان داد که انتشار اسانس آویشن از نانوژل کیتوزان-مریستیک اسید روندی دو مرحله‌ای دارد. نتایج حاصل از حبه‌های آلوده بسته‌بندی‌شده تأیید کرد با افزایش غلظت اسانس آویشن، اثر ضدقارچی نیز افزایش یافته است. کمترین میزان هدایت الکتریکی و تغییر رنگ، بالاترین مولفه L* و شاخص کروما در تیمار اسانس آویشن با غلظت 75 میکرولیتر در لیتر مشاهده گردید. نتایج نشان دادند که غلظت‌های پایین اسانس درون‎پوشانی‌شده در کنترل عامل قارچ بوتریتیس انگور بهتر از اسانس خالص با همان غلظت عمل کرد، درحالی‌که در آزمایش‌های مربوط به قابلیت انبارمانی میوه انگور تیمار اسانس آویشن با غلظت 75 میکرولیتر در لیتر مطلوبترین نتیجه را نشان داد.

کلیدواژه‌ها

• نانوکپسوله
• خاصیت ضد قارچی
• اسانس آویشن
• Botrytis cinerea
• انگور

عنوان مقاله [English]

Investigation of antifungal activity of nanoencapsulation of Thyme vulgaris essential oil against botrytis cinerea in red shahroodi grape (Vitis vinifera CV. Red)

نویسندگان [English]

• Hadise Karimi ¹
• Hojatollah Bodaghi ¹
• Ahmad Rajaei ²
• Shideh Mojerlou ¹

¹ Department of Horticulture and Plant Protection, Faculty of Agriculture, Shahroud University of Technology, Shahroud, Iran.

² Department of Food Science and Technology, Faculty of Agriculture, Shahroud University of Technology, Shahroud, Iran.

چکیده [English]

Introduction: Fresh grapes (Vitis vinifera L.) show severe lesions at the post-harvest stage and during the storage period. Decreasing the quality of grapes in the post-harvest stage limits its consumption and commercialization. Some methods such as using bio-control agents, natural antimicrobials, physical methods, disinfectants agents ,GRAS, (ozone, ethanol, acetic acid) have been used to control Botrytis cinerea after harvest. Encapsulation of the essential oils will increase their ability by increasing the effectiveness of the essential oils. Currently, chitosan has been interested for encapsulating bioactive compounds, as they are generally known to be safe and possess superior biological properties such as biodegradability, biocompatibility and non-toxicity. The aim of this study was to encapsulate thyme essential oil in chitosan nanogels to enhance and maintain its antifungal effect against B. cinerea in vitro and in vitro on Shahroodi red grape.
 
Materials and methods: For the purpose of this study, chitosan nanogels were first prepared and the infrared spectrum of chitosan-meric acid nanogels was measured using FT-IR430 infrared spectrophotometer at 20 ° C. The morphology of chitosan-meric acid nanogels and encapsulated essential oil was analyzed by SEM. Release test was then performed to determine the release rate of thyme essential oil encapsulated in chitosan-meric acid nanogels. Effect of chitosan-meristic acid nanogels at three levels of 0, 150 and 300 µl/L, pure thyme essential oil and thyme essential oil encapsulated in chitosan-meristic acid nanogels at three levels of 0, 75 and 150 µl / L in vitro and on the shelf life of grape fruits was studied under modified atmospheric conditions during 72 days storage at 2 to 4 °C. During storage, some traits such as firmness by manual penetrometer, electrical conductivity of fruit tissue, some components of fruit skin color and soluble solids were assessed by a refrectometer.
 
Results and discusion: Infrared spectroscopy (FTIR) results confirmed the successful coupling between chitosan amin groups and carboxylic acid-meristic acid groups and scanning electron microscopy image showed that the particle size of chitosan-meric acid nanogel was than 100 nm. The Comparison of the particle size in the present study with the previous studies on the size of chitosan-meric acid nanogels was smaller and more uniform. These differences could be related to several reasons, transform the long chitosan chain into smaller fragments by initial sonication, the important role of ultrasound in the reduction of the particle size and passing the nanogels through the filter. Release test showed that the diffusion of thyme essential oil from chitosan-meric acid nanogels has a two-step process. The chitosan-meric acid nanogels prepared in this study have hydrophilic (chitosan polymer) and hydrophilic (meristic acid fatty acid chain) regions, which led to the gradual release of thyme essential oil due to their hydrophobic nature. The results of the infected packaged berries confirmed that with increasing concentration of thyme essential oil, the antifungal effect was also increased. Evaluation of the effects of essential oil and nanogels - essential oils on grape berries infected with pathogenic spores showed the highest number of spores in control treatment (10.125 × 105 per ml) and the lowest number in NE2 (1.375 × 105 per ml) were observed. Coating treatments of chitosan-meric acid nanogel and thyme essential oil encapsulated in chitosan-meric acid nanogels showed better results at higher concentrations, but in the case of essential oil, lower spores were observed. The lowest electrical conductivity and discoloration, the highest L * component and chroma index were observed in thyme essential oil treatment with concentration of 75 µl/l. The results showed that the lower concentrations of the essential oil in the control of botrytis cinerea was better than the pure essential oils, whereas in the experiments on the storage of grape fruit, the treatment of thyme essential oil with a concentration of 75 µl/l It showed the most favorable result. Based on the results of the present study, the effect of essential oil nano-gel and essential oil alone on the shelf life after harvesting of grapes packed with polyethylene film confirmed that the essential oil performance was better than the essential oil nanogel. The lower performance of the essential oil nanogel than that of the essential oil alone can be due to inhibition of the essential oil diffusion through the packaging film while, the encapsulated essential oil release slower than the essential oil alone, it is likely to have an effect longer than expected in this study for the storage period of the grape. Considering this case and the antimicrobial capability of essential oil nanogel, it is necessary to investigate the effect of essential oil nanogel compared to essential oil on longevity of grapes for a longer of storage period.

کلیدواژه‌ها [English]

• Nanoencapsulation
• unti fungi property
• Thyme essential oil
• Botrytis cinerea
• Grape

مراجع

Abdolahi, A., Hassani, A., Ghuosta, Y., Bernousi, I., and Meshkatalsadat, M.H., 2010, In vitro efficacy of four plant essential oils against Botrytis cinerea Pers.: Fr. and Mucor piriformis A. Fischer. Journal of Essential Oil Bearing Plants, 13(1), 97-107.

-Arulmozhi, V., Pandian, K., and Mirunalini, S., 2013, Ellagic acid encapsulated chitosan nanoparticles for drug delivery system in human oral cancer cell line (KB). Colloids and Surfaces B: Biointerfaces, 110, 313-320.

-Artes-Hernandez, F., Aguayo, E., and Artes, F., 2004, Alternative atmosphere treatments for keeping quality of ‘autumn seedless’ table grapes during long-term cold storage. Postharvest Biology and Technology, 31(1), 59-67.

-Beyki, M., Zhaveh, S., Khalili, S. T., Rahmani-Cherati, T., Abdollahi, A., Bayat, M., Tabatabaei, M., and Mohsenifar, A., 2014, Encapsulation of Mentha piperita essential oil in chitosan-cinamic acid nanogel with enhanced antimicrobial activity against Aspergillus flavus. Industrial Crops products, 54, 310-319.

-Brummell, D.A., and Harpster, M.H., 2001, Cell wall metabolism in fruit softening and quality and its manipulation in transgenic plants. Plant molecular biology, 47(1-2), 311-339.

- Carvajal-Millan, E., Carvallo, T., Orozco, J.A., Martınez, M.A., Tapia, I., Guerrero, V.M., Chu, A.R., Llamas, J., Gardea, A.A.,2001, Polyphenol oxidase activity, color changes, and dehydration in table grape rachis during development and storage as affected by N-(2-chloro-4-pyridyl)-N-phenylurea. Jornal of Agricultural and Food Chemistry, 49, 946-951.

-Castillo, S., Navarro, D., Zapata, P.J., Guillen, F., Valero, D., Serrano, M., and Martinez-Romero, D., 2010, Antifungal efficacy of Aloevera in vitro and its use as a preharvest treatment to maintain postharvest table grape quality. Postharvest Biology and technology, 57(3), 183-188.

-Chauhan, S., Gupta, K.C., and Agrawal, M., 2014, Application of Biodegradable Aloe vera gel to control post harvest decay and longer the shelf life of Grapes. International Journal of Current Microbiology Applied Science, 3(3), 632-642.

- Crisosto, C.H., Garner, D., Crisosto, G., 2002, Carbon dioxide-enriched atmospheres during cold storage limit losses from Botrytis but accelerate rachis browning of ‘Redglobe’ table grapes. PostharVest Biology and Technology, 26, 181-189.

-Embuscado, M.E., and Huber, K.C., 2009, Edible films and coatings for food applications. New York, NY, USA: springer. pp. 213-214.

-Estevinho, B.M.A.N., Rocha, F.A.N., Santos, L.M.D.S., and Alves, M.A.C., 2013a, Using water-soluble chitosan for flavour microencapsulation in food industry. Journal of microencapsulation, 30(6), 571-579.

-Estevinho, B.N., Rocha, F., Santos, L., and Alves, A., 2013b, Microencapsulation with chitosan by spray drying for industry applications–A review. Trends in Food Science and Technology, 31(2), 138-155.

-Greay, S.J., and Hammer, K.A., 2015, Recent developments in the bioactivity of mono-and diterpenes: anticancer and antimicrobial activity. Phytochemistry reviews, 14(1), 1-6.

-Guo, P., Anderson, J.D., Bozell, J.J., and Zivanovic, S., 2016, The effect of solvent composition on grafting gallic acid onto chitosan via carbodiimide. Carbohydrate polymers, 140, 171-180.

-Hadian, M., Rajaei, A., Mohsenifar, A., and Tabatabaei, M., 2017, Encapsulation of Rosmarinus officinalis essential oils in chitosan-benzoic acid nanogel with enhanced antibacterial activity in beef cutlet against Salmonella typhimurium during refrigerated storage. LWT-Food Science and Technology, 84, 394-401.

-Hsieh, W.C., Chang, C.P., and Gao, Y.L., 2006, Controlled release properties of chitosan encapsulated volatile citronella oil microcapsules by thermal treatments. Colloids and Surfaces B: Biointerfaces, 53(2), 209-214.

-Keawchaoon, L., and Yoksan, R., 2011, Preparation, characterization and in vitro release study of carvacrol-loaded chitosan nanoparticles. Colloids and surfaces B: Biointerfaces, 84(1), 163-171.

-Khalili, S.T., Mohsenifar, A., Beyki, M., Zhaveh, S., Rahmani-Cherati, T., Abdollahi, A., and Tabatabaei, M., 2015, Encapsulation of Thyme essential oils in chitosan-benzoic acid nanogel with enhanced antimicrobial activity against Aspergillus flavus. LWT-Food Science and Technology, 60(1), 502-508.

-Lee, J., Lee, C., Kim, T.H., Lee, E.S., Shin, B.S., Chi, S.C., Park, E.S., Lee, K.C., Youn, Y.S., 2012, Self-assembled glycol chitosan nanogels containing palmityl-acylated exendin-4 peptide as a long-acting anti-diabetic inhalation system. Journal of controlled release 161(3), 728-734.

-Lopez-Reyes, JG., Spadaro, D., Gullinoa, Ml., Garibaldia, A., 2010, Efficacy of plant essential oils on postharvest control of rot caused by fungi on four cultivars of apples in vivo. Flavour Fragr Journal, 25, 171-177.

-Mahdavi, S.A., Jafari, S.M., Ghorbani, M., and Assadpoor, E., 2014, Spray-drying microencapsulation of anthocyanins by natural biopolymers: A review. Drying technology, 32(5), 509-518.

-Martin, A., Varona, S., Navarrete, A., and Cocero, M.J., 2010, Encapsulation and co-precipitation processes with supercritical fluids: applications with essential oils. Open Chemical Engineering Journal, 4(1), 31-41.

-Mohammadi, A., Hashemi, M., and Hosseini, S.M., 2015, Nanoencapsulation of Zataria multiflora essential oil preparation and characterization with enhanced antifungal activity for controlling Botrytis cinerea, the causal agent of gray mould disease. Innovative Food Science and Emerging Technologies, 28, 73-80.

-Mohammadrezakhani, S., and Pakkish, Z., 2015, Reduction of chilling injury and peroxide hydrogen accumulation in Thompson grape (Vitis vinifera L.) fruit by proline and ascorbic acid during storage. Journal of Plant Physiology & Breeding, 5(1), 1-12.

-Muftuoglu, F., Ayhan, Z., and Esturk, O., 2010, Modified atmosphere packaging of kabaasi apricot (Prunus armeniaca L.Kabaasi): Effect of atmosphere, packaging material type and coating on the physicochemical properties and sensory quality. Food and Bioprocess Tecnology , 12.

-Munoz, Z., and Moret, A., 2010, Sensitivity of Botrytis cinerea to chitosan and acibenzolar‐S‐methyl. Pest management science, 66(9), 974-979.

-Nasti, A., Zaki, N.M., de Leonardis, P., Ungphaiboon, S., Sansongsak, P., Rimoli, M. G., and Tirelli, N., 2009, Chitosan/TPP and chitosan/TPP-hyaluronic acid nanoparticles: systematic optimisation of the preparative process and preliminary biological evaluation. Pharmaceutical research, 26(8), 1918-1930.

-Nopwinyuwong, A., Trevanich, S., and Suppakul, P., 2010, Development of a novel colorimetric indicator label for monitoring freshness of intermediate-moisture dessert spoilage. Talanta, 81(3), 1126-1132.

-Nuisin, R., Krongsin, J., Noppakundilograt, S., and Kiatkamjornwong, S., 2013, Microencapsulation of menthol by crosslinked chitosan via porous glass membrane emulsification technique and their controlled release properties. Journal of microencapsulation, 30(5), 498-509.

-Peng, H., Xiong, H., Li, J., Chen, L., Zhao, Q., 2010a, Methoxy poly (ethylene glycol) -grafted-chitosan based microcapsules: Synthesis, characterization and properties as a potential hydrophilic wall material for stabilization and controlled release of algal oil. Journal of Food Engineering, 101, 113-119.

-Peng, H., Xiong, H., Li, J., Xie, M., Liu, Y., Bai, C., and Chen, L., 2010b, Vanillin cross-linked chitosan microspheres for controlled release of resveratrol. Food Chemistry, 121(1), 23-28.

-Perrico, M., Arace, E., Corbe, M.R., Sinigagila, M., and Bevilacqua, A., 2015, Bioactivity of essential oils. A review on their interaction with food components. Frontiers in Microbiology, 1-7.

-Romanazzi, G., Karabulut, O.A., and Smilanick, J.L., 2007, Combination of chitosan and ethanol to control postharvest gray mold of table grapes. Postharvest biology and technology, 45(1), 134-140.

-Romanazzi, G., Lichter, A., Gabler, F.M., and Smilanick, J.L., 2012, Recent advances on the use of natural and safe alternatives to conventional methods to control postharvest gray mold of table grapes. Postharvest Biology and Technology, 63(1), 141-147.

-Sabounchi, S., and Massoud, R., 2016, The effects of Zataria multiflora essential oil on some characteristics of sultana table grapes contaminated with Botrytis cinerea. Journal of Food Biosciences and Technology, 6(1), 49-54.

-Shin, M.H., Kim, J.H., Choi, H.W., Keum, Y.S., and Chun, S.C., 2014, Effect of thymol and linalool fumigation on postharvest diseases of table grapes. Mycobiology, 42(3), 262-268.

-Silva, P.S., Viccini, L.F., Singulani, J.L., de Siqueira, E.P., Zani, C.L., Alves, T.M.A., 2010, Chemical composition of the essential oil and hexanic fraction of Lippia and Lantana species. Revista Brasileira Farmacognosia, 20(6), 843-849.

-Supapvanich, S., and Tucker, G.A., 2011, Physicochemical changes in fresh-cut Honeydew melon fruit during storage. African Journal of Agricultural Research, 6(12), 2737-2742.

-Topalovic, A., and Milukovic-Petkovsek, M., 2010, Changes in sugars, organic acids and phenolics of grape berries of cultivar Cardinal during ripening. Journal Food Agriculture Environ, 8(3&4), 223-7.

-Valderruten, N.E., Valverde, J.D., Zuluaga, F., and Ruiz-Durantez, E., 2014, Synthesis and characterization of chitosan hydrogels cross-linked with dicarboxylic acids. Reactive and Functional Polymers, 84, 21-28.

-Valero, D., Valverde, J.M., MartInez-Romero, D., Guillen, F., Castillo, S., and Serrano, M., 2006, The combination of modified atmosphere packaging with eugenol or thymol to maintain quality, safety and functional properties of table grapes. Postharvest Biology and Technology, 41(3), 317-327.

-Wang, X.H., Li, D.P., Wang, W.J., Feng, Q.L., Cui, F.Z., Xu, Y.X., and van der Werf, M., 2003, Crosslinked collagen/chitosan matrix for artificial livers. Biomaterials, 24(19), 3213-3220.