Aronya ve asma in vitro kültürlerinde elisitör kaynaklı biyoaktif metabolit üretimi ve bu ekstraktların fonksiyonel yoğurt zenginleştirilmesinde kullanımı

Öz

Bu çalışma, aronya (Aronia melanocarpa L.) ve asma (Vitis vinifera L.) in vitro kültürlerinde elisitör uygulamalarının biyoaktif metabolit birikimi üzerine etkilerini ve elde edilen ekstraktların fonksiyonel yoğurt zenginleştirilmesindeki potansiyelini belirlemek amacıyla yürütülmüştür. In vitro koşullarda oluşturulan aronya (‘Nero’) ve asma (‘Yalova İncisi’) kültürleri Murashige ve Skoog (MS) besi ortamında yetiştirilmiş ve 14 gün süreyle salisilik asit (100-200 µM), metil jasmonat (50-100 µM), PEG (%5 w/v) ve UV-B (10 dk/gün) uygulamalarına tabi tutulmuştur. Toplam fenolik, flavonoid içerikleri ve antioksidan aktiviteleri (DPPH, ABTS, FRAP) spektrofotometrik olarak belirlenmiştir. En etkili elisitör uygulamalarından elde edilen ekstraktlar (aronya için 100 µM MeJA, asma için 200 µM SA), %1 (w/v) oranında set tipi yoğurtlara ilave edilmiştir. Yoğurt üretimi 90 °C’de 10 dakika pastörizasyon, 43 °C’de inokülasyon ve pH 4.6’ya ulaşana kadar fermentasyon koşullarında gerçekleştirilmiştir. Kontrol ve zenginleştirilmiş örnekler 4 °C’de depolanarak 0., 7. ve 14. günlerde analiz edilmiştir. MeJA ve UV-B uygulamaları aronya kültürlerinde en yüksek fenolik (75.8 mg GAE g-1 DW) ve flavonoid (37.5 mg QE g-1 DW) düzeylerini sağlamıştır. SA, asma kültürlerinde fenolik içeriği 57.4 mg GAE g-1 DW’ye yükseltmiştir. Zenginleştirilmiş yoğurtlar daha yüksek antioksidan aktivite (ABTS: 1.92; FRAP: 1.48 mmol g-1 DW), daha düşük sinerezis (12.9%) ve maya-küf gelişiminde azalma (1.10 log CFU g-1) göstermiştir. Elde edilen bulgular, elisitörle zenginleştirilen kültürlerin fonksiyonel süt ürünlerinde sürdürülebilir biyoaktif bileşik kaynakları olarak değerlendirilebileceğini ortaya koymaktadır.

Anahtar Kelimeler

• Aronya (Aronia melanocarpa L.)
• Asma (Vitis vinifera L.)
• Elisitör uygulamaları
• Biyoaktif bileşikler
• Gıda zenginleştirme

Elicitor-induced production of bioactive metabolites in aronia and grapevine in vitro cultures and their application in functional yogurt fortification

Abstract

This study investigated the effects of elicitor applications on bioactive metabolite accumulation in aronia (Aronia melanocarpa L.) and grapevine (Vitis vinifera L.) in vitro cultures and evaluated the potential of the obtained extracts for functional yogurt fortification. In vitro cultures of aronia (‘Nero’) and grapevine (‘Yalova İncisi’) were established on Murashige and Skoog (MS) medium and treated with salicylic acid (100-200 µM), methyl jasmonate (50-100 µM), polyethylene glycol (5% w/v), and UV-B (10 min/day) for 14 days. Total phenolic, flavonoid, and antioxidant capacities (DPPH, ABTS, FRAP) were determined spectrophotometrically. The most effective extracts (100 µM MeJA for aronia and 200 µM SA for grapevine) were incorporated into set-type yogurts at 1% (w/v) following pasteurization (90 °C, 10 min), inoculation with starter culture at 43 °C, and fermentation to pH 4.6. Control and fortified yogurts were stored at 4 °C and analyzed on days 0, 7, and 14. MeJA and UV-B treatments yielded the highest phenolic (75.8 mg GAE g-1 DW) and flavonoid (37.5 mg QE g-1 DW) levels in aronia, while SA increased phenolics in grapevine to 57.4 mg GAE g-1 DW. Fortified yogurts showed higher antioxidant activity (ABTS: 1.92; FRAP: 1.48 mmol g-1 DW), lower syneresis (12.9%), greater firmness (0.41 N), and reduced yeast-mold growth (1.10 log CFU g-1). These findings confirm that elicitor-enhanced cultures provide sustainable bioactive sources for functional dairy applications.

Keywords

• Aronia (Aronia melanocarpa L.)
• Grapevine (Vitis vinifera L.)
• Elicitor treatments
• Bioactive compounds
• Food fortification

References

1. Bayram, O.Y., Kinik, O., & Büyükkileci, C. (2025). Functional and phenolic characterization of medicinal plant-enriched strained yogurt: bioactivity and storage stability. Journal of Food Measurement and Characterization, 19 (10), 7557-7570.
2. Benzie, I.F., & Strain, J.J. (1996). The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: the FRAP assay. Analytical Biochemistry, 239 (1), 70-76.
3. Bourne, M. (2002). Food texture and viscosity: concept and measurement. Elsevier.
4. Brand-Williams, W., Cuvelier, M.E., & Berset, C.L.W.T. (1995). Use of a free radical method to evaluate antioxidant activity. LWT-Food science and Technology, 28 (1), 25-30.
5. Bustos, A.Y., Taranto, M.P., Gerez, C.L., Agriopoulou, S., Smaoui, S., Varzakas, T., & Enshasy, H.A.E. (2025). Recent advances in the understanding of stress resistance mechanisms in probiotics: Relevance for the design of functional food systems. Probiotics and Antimicrobial Proteins, 17 (1), 138-158.
6. Cao, W., Li, L., Wang, J., Guo, W., Chen, W., Pan, L., & Li, D. (2025). Effects of Black Garlic Polyphenols on the Physicochemical Characteristics, Antioxidant Activity, and Sensory Evaluation of Yogurt. Gels, 11 (7), 561.
7. Chang, C.C., Yang, M.H., Wen, H.M., & Chern, J.C. (2002). Estimation of total flavonoid content in propolis by two complementary colorimetric methods. Journal of Food and Drug Analysis, 10 (3).
8. Cunniff, P., & Washington, D. (1997). Official methods of analysis of AOAC International. J. AOAC Int, 80 (6), 127A.

9. Da Silva, N., Taniwaki, M.H., Junqueira, V.C., Silveira, N., Okazaki, M.M., & Gomes, R.A.R. (2018). Microbiological examination methods of food and water: a laboratory manual. CRC Press.
10. Frumuzachi, O., Mocan, A., Rohn, S., & Gavrilaș, L. (2025). Impact of a Chokeberry (Aronia melanocarpa (Michx.) Elliott) Supplementation on Cardiometabolic Outcomes: A Critical Systematic Review and Meta-Analysis of Randomized Controlled Trials. Nutrients, 17 (9), 1488.
11. Georgiev, V., Ananga, A., & Tsolova, V. (2014). Recent advances and uses of grape flavonoids as nutraceuticals. Nutrients, 6 (1), 391-415.
12. Giri, C.C., & Zaheer, M. (2016). Chemical elicitors versus secondary metabolite production in vitro using plant cell, tissue and organ cultures: recent trends and a sky eye view appraisal. Plant Cell, Tissue and Organ Culture (PCTOC), 126 (1), 1-18.
13. Go, M.Y., Kim, J., Jeon, C.Y., & Shin, D.W. (2024). Functional activities and mechanisms of Aronia melanocarpa in our health. Current Issues in Molecular Biology, 46 (8), 8071-8087.
14. González-Martınez, C., Becerra, M., Cháfer, M., Albors, A., Carot, J.M., & Chiralt, A. (2002). Influence of substituting milk powder for whey powder on yoghurt quality. Trends in Food Science & Technology, 13 (9-10), 334-340.
15. Gülhan, A. (2024). Evaluation of some physicochemical, nutritional, and microbiological properties and sensory profiles of green tea and almond milk kombucha beverages. Journal of Food Measurement and Characterization, 18 (12), 9879-9889.
16. Hamidi Moghaddam, R., Mansouripour, S., & Soltani, M. (2024). Functional set-type yogurt prepared using turnip extract (Brassica rapa L.): physicochemical, antioxidant, microbiological, rheological, microstructural, and sensory characteristics. Journal of Food Measurement and Characterization, 18 (2), 1204-1217.
17. Harrigan, W.F. (1998). Laboratory methods in food microbiology 3rd ed. Academic Pres, London, UK.
18. Hu, X., Ma, W., Zhang, D., Tian, Z., Yang, Y., Huang, Y., & Hong, Y. (2025). Application of natural antioxidants as feed additives in aquaculture: A review. Biology, 14 (1), 87.
19. Irondi, E.A., Bankole, A.O., Awoyale, W., Ajani, E.O., & Alamu, E.O. (2024). Antioxidant, enzymes inhibitory, physicochemical and sensory properties of instant bio-yoghurts containing multi-purpose natural additives. Frontiers in Nutrition, 10, 1340679.
20. Ivanov, Y., Atanasova, M., & Godjevargova, T. (2025). Nutritional and functional values of grape seed flour and extract for production of antioxidative dietary supplements and functional foods. Molecules, 30 (9), 2029.
21. Jain, D., Bisht, S., Parvez, A., Singh, K., Bhaskar, P., & Koubouris, G. (2024). Effective biotic elicitors for augmentation of secondary metabolite production in medicinal plants. Agriculture, 14 (6), 796.
22. Jiang, J., & Xiong, Y.L. (2016). Natural antioxidants as food and feed additives to promote health benefits and quality of meat products: A review. Meat Science, 120, 107-117.
23. Karastergiou, A., Gancel, A.L., Jourdes, M., & Teissedre, P.L. (2024). Valorization of grape pomace: A review of phenolic composition, bioactivity, and therapeutic potential. Antioxidants, 13 (9), 1131.
24. Kaya, Ç. (2025). Süper meyve aronya (Aronia melanocarpa L.)’nın biyokimyasal bileşimi, sağlık yararları ve gıda sanayinde kullanım potansiyeli. Nevşehir Bilim ve Teknoloji Dergisi, 14 (1), 1-15. https://doi.org/10.17100/nevbiltek.1626095
25. Kaya, Ç., & Özatay, Ş. (2024). Development of an effective sterilization protocol for plant tissue culture studies in superfruit aronia [Aronia melanocarpa (Michaux) Elliot]. Black Sea Journal of Agriculture, 7 (6), 679-685. https://doi.org/10.47115/bsagriculture.1548432
26. Kaya, Ç., & Sarıyer, T. (2024). Comparative biplot analysis of micropropagation of viking aronia cultivar in different plant tissue culture media. Anadolu Ege Tarımsal Araştırma Enstitüsü Dergisi, 34 (1), 90-96. https://doi.org/10.18615/anadolu.1494423
27. Khoo, H.E., Azlan, A., Tang, S.T., & Lim, S.M. (2017). Anthocyanidins and anthocyanins: Colored pigments as food, pharmaceutical ingredients, and the potential health benefits. Food & Nutrition Research, 61 (1), 1361779.
28. Kulling, S.E., & Rawel, H.M. (2008). Chokeberry (Aronia melanocarpa)–A review on the characteristic components and potential health effects. Planta Medica, 74 (13), 1625-1634.
29. Manoharan, S., Dubey, P.K., & Sharma, M. (2024). Recent advances in deep‐fat frying through pretreatments and edible coating to reduce oil absorption. Journal of Food Process Engineering, 47 (8), e14706.
30. Marant, B., Flourat, A. L., Lanoue, A., Magot, F., Parent, L., Leclère, V., Guihard, G., Aggad, D., Jacquard, C., Courot, E., Aziz, A., & Crouzet, J. (2025). Peroxidase 4-based enzymatic synthesis of stilbene oligomers in methyl jasmonate-elicited grapevine cell suspensions. Journal of Agricultural and Food Chemistry, 73 (3), 1929-1939.
31. Mendoza, D., Cuaspud, O., Arias, J.P., Ruiz, O., & Arias, M. (2018). Effect of salicylic acid and methyl jasmonate in the production of phenolic compounds in plant cell suspension cultures of Thevetia peruviana. Biotechnology Reports, 19, e00273.
32. Murashige, T., & Skoog, F. (1962). A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum, 15 (3).
33. Murthy, H.N., Joseph, K.S., Paek, K.Y., & Park, S.Y. (2024). Bioreactor configurations for adventitious root culture: recent advances toward the commercial production of specialized metabolites. Critical Reviews in Biotechnology, 44 (5), 837-859.
34. Naeem, S., Ihsan, A., & Ajaz, H. (2024). Enhancing antioxidant content and consumer appeal in greek yogurt fortified with grape seed and strawberry pulp. International Journal of Social Sciences Bulletin, 2 (4), 861-870.
35. Nallakaruppan, N., & Thiagarajan, K. (2024). In vitro elicitation of anthraquinones—A review. Plant Cell, Tissue and Organ Culture (PCTOC), 156 (3), 70.
36. Namdeo, A.G. (2007). Plant cell elicitation for production of secondary metabolites: a review. Pharmacogn Rev, 1 (1), 69-79.
37. Prashant, S.P., & Bhawana, M. (2024). An update on biotechnological intervention mediated by plant tissue culture to boost secondary metabolite production in medicinal and aromatic plants. Physiologia Plantarum, 176 (4), e14400.
38. Quideau, S., Deffieux, D., Douat‐Casassus, C., & Pouységu, L. (2011). Plant polyphenols: Chemical properties, biological activities, and synthesis. Angewandte Chemie International Edition, 50 (3), 586-621.
39. Pourghorban, S., Yadegarian, L., Jalili, M., & Rashidi, L. (2022). Comparative physicochemical, microbiological, antioxidant, and sensory properties of pre‐and post‐fermented yoghurt enriched with olive leaf and its extract. Food Science & Nutrition, 10 (3), 751-762.
40. Rao, M.J., Duan, M., Zhou, C., Jiao, J., Cheng, P., Yang, L., Wei, W., Shen, Q., Ji, P., Yang, Y., Conteh, O., Yan, D., Yuan, H., Rauf, A., Ai, J., & Zheng, B. (2025). Antioxidant defense system in plants: Reactive oxygen species production, signaling, and scavenging during abiotic stress-induced oxidative damage. Horticulturae, 11 (5), 477.
41. Rao, S.R., & Ravishankar, G.A. (2002). Plant cell cultures: Chemical factories of secondary metabolites. Biotechnology Advances, 20 (2), 101-153.
42. Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M., & Rice-Evans, C. (1999). Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology and Medicine, 26 (9-10), 1231-1237.
43. Salama, M., Li, X., Sun, H., Khalifa, I., Zahran, H.A., Sheng, L., & Cai, Z. (2025). Animal protein-based fat replacers: new prospective and applications in dairy products and their analogues. Food Reviews International, 41 (3), 857-882.
44. Šedík, P., Horská, E., Predanócyová, K., Grunert, K.G., Hudecová, M., & Nagyová, Ľ. (2025). Consumption trends and perspectives for milk and dairy products in Slovakia. Journal of International Food & Agribusiness Marketing, 37 (1), 72-94.
45. Selwal, N., Goutam, U., Akhtar, N., Sood, M., & Kukreja, S. (2024). Elicitation:“A Trump Card” for enhancing secondary metabolites in plants. Journal of Plant Growth Regulation, 43 (9), 3027-3047.
46. Shahidi, F., & Ambigaipalan, P. (2015). Phenolics and polyphenolics in foods, beverages and spices: Antioxidant activity and health effects–A review. Journal of Functional Foods, 18, 820-897.
47. Shori, A.B. (2024). Comparative analysis of Lactobacillus starter cultures in fermented camel milk: Effects on viability, antioxidant properties, and sensory characteristics. Foods, 13 (22), 3711.
48. Singleton, V.L., & Rossi, J.A. (1965). Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. American journal of Enology and Viticulture, 16 (3), 144-158.
49. Sivaprakasam, S., Chinnaiyan, U., Varatharajan, S., Uthayasuryan, E., Chandrasekaran, M., Paramasivam, S., & Ramasamy, P. (2025). Unveiling the physicochemical attributes and antioxidant potential of cashew apple juice varieties. Applied Fruit Science, 67 (2), 74.
50. Terpou, A., Papadaki, A., Lappa, I.K., Kachrimanidou, V., Bosnea, L.A., & Kopsahelis, N. (2019). Probiotics in food systems: Significance and emerging strategies towards improved viability and delivery of enhanced beneficial value. Nutrients, 11 (7), 1591.
51. Walker, K.A., Wendeln, M.L., & Jaworski, E.G. (1979). Organogenesis in callus tissue of Medicago sativa. The temporal separation of induction processes from differentiation processes. Plant Science Letters, 16 (1), 23-30.