The Effects of Seaweed and Humic Acid Seed Treatments on the Phytochemical Composition and Storage Stability of Wheatgrass Juice Derived from Different Triticum Species

Ayşe Gökdemir; Jale Bilgin; İmren Kutlu

doi:10.24925/turjaf.v14i8.2192-2201.8922

Authors

Ayşe Gökdemir Eskişehir Osmangazi University, Faculty of Agriculture, Department of Field Crops, 26100, Eskişehir, Türkiye https://orcid.org/0009-0005-1687-5877
Jale Bilgin Eskişehir Osmangazi University, Faculty of Agriculture, Department of Horticulture, 26100, Eskişehir, Türkiye https://orcid.org/0000-0003-1223-1087
İmren Kutlu Eskişehir Osmangazi University, Faculty of Agriculture, Department of Field Crops, 26100, Eskişehir, Türkiye https://orcid.org/0000-0002-3505-1479

DOI:

https://doi.org/10.24925/turjaf.v14i8.2192-2201.8922

Keywords:

Ancient wheat , Biostimulants , Antioxidant stability , Wheatgrass juice , Cold storage

Abstract

Wheatgrass juice is a prominent functional food; however, the comparative potential of ancient wheat species and the role of biostimulant seed priming in determining phytochemical stability during storage remain insufficiently defined. This study investigated the effects of seaweed extract and humic acid seed priming on the phytochemical composition and storage stability of wheatgrass juice derived from Triticum aestivum, Triticum sphaerococcum, Triticum macha, and Triticum spelta. Seeds were primed for 24 h with seaweed extract or humic acid (200 mg 100 g⁻¹ seed), grown for 10 days, and juice was obtained by cold pressing. Phytochemical attributes (total phenolic content, total flavonoid content, antioxidant capacity, vitamin C, and titratable acidity) were quantified in fresh samples and after 7 days of storage at 4°C and −20°C. Stability was defined as the percentage retention of bioactive compounds during storage. Data were analyzed using a completely randomized factorial design (three replicates) with analysis of variance, and mean comparisons were performed using Tukey’s test. Biostimulant treatments increased total phenolic content by up to 69% and antioxidant capacity by up to 7% compared to the control, with T. macha and T. sphaerococcum showing the strongest response, particularly under seaweed priming. During storage at 4°C, antioxidant capacity decreased by 49–87%, while total phenolic content declined by 84–94%. Whereas, flavonoids and vitamin C decreased by 12–21% and 25–34%, respectively. In contrast, storage at −20°C preserved flavonoids up to 99%, while mean retention rates of antioxidant capacity and total phenolic content were approximately 63% and 76%, respectively, across all genotypes and treatments. The improved retention of phytochemicals in primed samples may be associated with enhanced initial metabolite accumulation and reduced oxidative degradation. Among the evaluated genotypes, T. macha exhibited the highest phytochemical content and retention capacity. These findings demonstrate the interactive effects of genotype, biostimulant priming, and storage conditions on wheatgrass juice quality, and indicate that frozen storage is more effective than refrigeration for preserving bioactive compounds.

References

Ali, N., Popović, V., Koutchma, T., Warriner, K., & Zhu, Y. (2020). Effect of thermal, high hydrostatic pressure, and ultraviolet‐C processing on the microbial inactivation, vitamins, chlorophyll, antioxidants, enzyme activity, and color of wheatgrass juice. Journal of Food Process Engineering, 43(1), e13036. https://doi.org/10.1111/jfpe.13036

Ali, O., Ramsubhag, A., & Jayaraman, J. (2021). Biostimulant properties of seaweed extracts in plants: Implications towards sustainable crop production. Plants, 10(3), 531. https://doi.org/10.3390/plants10030531

Anjos Neto, A. P. D., Oliveira, G. R. F., Mello, S. D. C., Silva, M. S. D., Gomes-Junior, F. G., Novembre, A. D. D. L. C., & Azevedo, R. A. (2020). Seed priming with seaweed extract mitigate heat stress in spinach: effect on germination, seedling growth and antioxidant capacity. Bragantia, 79, 502-511. https://doi.org/10.1590/1678-4499.20200127

Aweya, J. J., Sharma, D., Bajwa, R. K., Earnest, B., Krache, H., & Moghadasian, M. H. (2025). Ancient grains as functional foods: integrating traditional knowledge with contemporary nutritional science. Foods, 14(14), 2529. https://doi.org/10.3390/foods14142529

Baltazar, M., Correia, S., Guinan, K. J., Sujeeth, N., Bragança, R., & Gonçalves, B. (2021). Recent advances in the molecular effects of biostimulants in plants: An overview. Biomolecules, 11(8), 1096. https://doi.org/10.3390/biom11081096

Barabaschi, D., Volante, A., Faccioli, P., Povesi, A., Tagliaferri, I., Mazzucotelli, E., & Cattivelli, L. (2025). Ancient diversity of Triticum aestivum subspecies as source of novel loci for bread wheat improvement. Frontiers in Plant Science, 16, 1536991. https://doi.org/10.3389/fpls.2025.1536991

Canellas, L. P., Canellas, N. O., da S. Irineu, L. E. S., Olivares, F. L., & Piccolo, A. (2020). Plant chemical priming by humic acids. Chemical and Biological Technologies in Agriculture, 7(1), 12. https://doi.org/10.1186/s40538-020-00178-4

Chen, B., Fu, J., Zheng, Y., Liu, G., Fu, L., & Li, Y. (2024). Biostimulants in agriculture: innovations in seed treatment and coating technologies. Seed Science and Technology, 52(3), 227-249. https://doi.org/10.15258/sst.2024.52.3.02

Deyalage, S. T. (2025). Comparative study on nutritional characteristics and physicochemical properties of ancient wheat species. Msc. Thesis, Department of Food and Human Nutritional Sciences University of Manitoba Winnipeg, 188 p. http://hdl.handle.net/1993/38852

Djordjevic, T. M., Šiler-Marinkovic, S. S., & Dimitrijevic-Brankovic, S. I. (2011). Antioxidant Activity and Total Phenolic Content in Some Cereals and Legumes. International Journal of Food Properties, 14(1), 175–184. https://doi.org/10.1080/10942910903160364

Drygaś, B., Piechowiak, T., Balawejder, M., Matłok, N., Kreczko, J., & Puchalski, C. (2024). The eliciting effect of aqueous extracts from Ascophyllum nodosum algae on the cultivation of arugula (Eruca sativa Mill.) microgreens. Sustainability, 16(17), 7436. https://doi.org/10.3390/su16177436

Feszterová, M., Mišiaková, M., & Kowalska, M. (2023a). Bioactive vitamin C content from natural selected fruit juices. Applied Sciences, 13(6), 3624. https://doi.org/10.3390/app13063624

Feszterová, M., Kowalska, M., & Mišiaková, M. (2023b). Stability of vitamin C content in plant and vegetable juices under different storing conditions. Applied Sciences, 13(19), 10640. https://doi.org/10.3390/app131910640

Gao, F., Wang, S., Wang, Z., Li, Z., & Liu, J. (2025). Phenolic Compounds in Cereals: Advances and Challenges in the Extraction, Detection, and Health Implications. Food Reviews International, 1-39. https://doi.org/10.1080/87559129.2025.2581051

Ghumman, A., Singh, N., & Kaur, A. (2017). Chemical, nutritional and phenolic composition of wheatgrass and pulse shoots. International Journal of Food Science & Technology, 52(10), 2191–2200. https://doi.org/10.1111/ijfs.13498

Gogoi, P., Sharma, P., Goudar, G., Mahajan, A., & Banjara, S. K. (2025). Phenolic composition and antioxidant potential of Indian pigmented rice landraces cultivated under identical agro-climatic conditions. Journal of Cereal Science, 122, 104120. https://doi.org/10.1016/j.jcs.2025.104120

Gonçalves, B., Santos, M., Silva, V., Rodrigues, A., Oliveira, I., Lopes, T., ... & Guinan, K. J. (2025). Biostimulants in Fruit Crop Production: Impacts on Growth, Yield, and Fruit Quality. Horticulturae, 11(12), 1452. https://doi.org/10.3390/horticulturae11121452

Grubišić, S., Kristić, M., Lisjak, M., Mišković Špoljarić, K., Petrović, S., Vila, S., & Rebekić, A. (2022). Effect of Wheatgrass Juice on Nutritional Quality of Apple, Carrot, Beet, Orange and Lemon Juice. Foods, 11(3), 445. https://doi.org/10.3390/foods11030445

Gunjal, M., Kaur, J., Rasane, P., Singh, J., Kaur, S., Bakshi, M., ... & Ercisli, S. (2024). Nutritional significance of wheatgrass: cultivation practices and opportunities for its processing and preservation. Recent Advances in Food, Nutrition & Agriculture, 15(3), 163-177. https://doi.org/10.2174/012772574X275920231214053122

Hait, R., Das, A., & Mitra, R. D. (2023). Exploring the effect of wheat grass juice on thalassaemia: A comprehensive review. Journal of Advanced Zoology, 44(S6), 1636-1639. https://doi.org/10.17762/jaz.v44iS6.2583

Kaplan Evlice, A., Pehlivan, A., Keçeli, A., Şanal, T., Karabak, S., Zencirci, N., & Yaman, H. M. (2022). Nutritional and technological aspects of ancient wheat. In Ancient Wheats (pp. 147-179). Cham: Springer International Publishing. https://doi.org/10.1007/978-3-031-07285-7_7

Karakaş, F. P. (2026). Evaluation of harvest timing and genotypic effect on bioactive composition and antioxidant activity of wheatgrass from traditional and modern wheat cultivars. International Journal of Secondary Metabolite, (Advanced Online Publication), 144-159. https://doi.org/10.21448/ijsm.1700714

Karaşahin, M. (2017). The effects of different seaweed doses on yield and nutritional values of hydroponic wheatgrass juice. Turkish Journal of Agriculture-Food Science and Technology, 5(3), 226-230. https://doi.org/10.24925/turjaf.v5i3.226-230.994

Kaur, B., Kumar, B., Sirhindi, G., Guleria, N., & Kaur, J. (2023). Phenolic biotransformations in wheatgrass juice after primary and secondary fermentation. Foods, 12(8), 1624. https://doi.org/10.3390/foods12081624

Kaya, C., Ugurlar, F. (2025). Non-microbial Biostimulants for Quality Improvement in Fruit and Leafy Vegetables. In: Ahammed, G.J., Zhou, J. (eds) Growth Regulation and Quality Improvement of Vegetable Crops. Springer, Singapore. https://doi.org/10.1007/978-981-96-0169-1_16

Khalid, A., Hameed, A., & Tahir, M. F. (2023). Wheat quality: A review on chemical composition, nutritional attributes, grain anatomy, types, classification, and function of seed storage proteins in bread making quality. Frontiers in Nutrition, 10, 1053196. https://doi.org/10.3389/fnut.2023.1053196

Khan, M. K., Pandey, A., Hamurcu, M., & Gezgin, S. (Eds.). (2025). Wheat Wild Relatives: Developing Abiotic Stress Tolerance under Climate Change. Elsevier.

Lorenc-Kukuła, K., Zuk, M., Kulma, A., Czemplik, M., Kostyn, K., Skala, J., ... & Szopa, J. (2009). Engineering flax with the GT family 1 Solanum sogarandinum glycosyltransferase SsGT1 confers increased resistance to Fusarium infection. Journal of agricultural and food chemistry, 57(15), 6698-6705. https://doi.org/10.1021/jf900833k

Lumactud, R. A., Gorim, L. Y., & Thilakarathna, M. S. (2022). Impacts of humic-based products on the microbial community structure and functions toward sustainable agriculture. Frontiers in Sustainable Food Systems, 6, 977121. https://doi.org/10.3389/fsufs.2022.977121

Melini, F., Fasano, S., & Melini, V. (2026). Role of Solvent and Citric Acid-Mediated Solvent Acidification in Enhancing the Recovery of Phenolics, Flavonoids, and Anthocyanins from Apple Peels. Applied Sciences, 16(2), 671. https://doi.org/10.3390/app16020671

Monteiro, E., De Lorenzis, G., Ricciardi, V., Baltazar, M., Pereira, S., Correia, S., ... & Castro, I. (2024). Exploring seaweed and glycine betaine biostimulants for enhanced phenolic content, antioxidant properties, and gene expression of Vitis vinifera cv.“Touriga Franca” berries. International Journal of Molecular Sciences, 25(10), 5335. https://doi.org/10.3390/ijms25105335

Nabi, F., Sarfaraz, A., Kama, R., Kanwal, R., & Li, H. (2025). Structure-based function of humic acid in abiotic stress alleviation in plants: a review. Plants, 14(13), 1916. https://doi.org/10.3390/plants14131916

Parit, S. B., Dawkar, V. V., Tanpure, R. S., Pai, S. R., & Chougale, A. D. (2018). Nutritional Quality and Antioxidant Activity of Wheatgrass (Triticum aestivum) Unwrap by Proteome Profiling and DPPH and FRAP assays. Journal of Food Science, 83(8), 2127–2139. https://doi.org/10.1111/1750-3841.14224

Rana, S., Kamboj, J. K., & Gandhi, V. (2011). Living life the natural way – Wheatgrass and Health. Functional Foods in Health and Disease, 1(11), 444. https://doi.org/10.31989/ffhd.v1i11.112

Rodríguez, F. C., Gallagher, E., Rai, D. K., & Burgess, C. M. (2022). Nutritional and physiochemical properties of wheatgrass juice and preservation strategies. Food Chemistry Advances, 1, 100136. https://doi.org/10.1016/j.focha.2022.100136

Roumia, H., Kókai, Z., Mihály-Langó, B., Csobod, É. C., & Benedek, C. (2023). Ancient Wheats-A Nutritional and Sensory Analysis Review. Foods, 12(12), 2411. https://doi.org/10.3390/foods12122411

Sakr, M. T., Ibrahim, H. M., ElAwady, A. E., & Abo ElMakarem, A. A. (2019). Effect of humic acid, seaweed extract and essential oils as antioxidants on pre-and post-harvest quality of red radish plants. Horticult Int J, 3(3), 129-138. https://doi.org/10.15406/hij.2019.03.00120

Shahrajabian, M. H., & Sun, W. (2024). The importance of salicylic acid, humic acid and fulvic acid on crop production. Letters in Drug Design & Discovery, 21(9), 1465-1480. https://doi.org/10.2174/1570180820666230411102209

Sidiqui, B., Sidiqi, I. (2025). Sustainable fruit cultivation using biostimulants: Improving nutrient efficiency, defense, and productivity. Journal of Pharmacognosy and Phytochemistry, 14(5), 378-386. https://www.doi.org/10.22271/phyto.2025.v14.i5e.15595

Singh, R. P., Chidambara Murthy, K. N., & Jayaprakasha, G. K. (2002). Studies on the antioxidant activity of pomegranate (Punica granatum) peel and seed extracts using in vitro models. Journal of agricultural and food chemistry, 50(1), 81-86. https://doi.org/10.1021/jf010865b

Singleton, V. L., Orthofer, R., & Lamuela-Raventós, R. M. (1999). [14] Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. In Methods in enzymology (Vol. 299, pp. 152-178). Academic press. https://doi.org/10.1016/S0076-6879(99)99017-1

Skoczylas, Ł., Korus, A., Tabaszewska, M., Gędoś, K., & Szczepańska, E. (2018). Evaluation of the quality of fresh and frozen wheatgrass juices depending on the time of grass harvest. Journal of Food Processing and Preservation, 42(1), e13401. https://doi.org/10.1111/jfpp.13401

Souza, A. C., Olivares, F. L., Peres, L. E. P., Piccolo, A., & Canellas, L. P. (2022). Plant hormone crosstalk mediated by humic acids. Chemical and Biological Technologies in Agriculture, 9(1), 29. https://doi.org/10.1186/s40538-022-00295-2

Stirk, W. A., Rengasamy, K. R., Kulkarni, M. G., & van Staden, J. (2020). Plant biostimulants from seaweed: An overview. The chemical biology of plant biostimulants, 31-55. https://doi.org/10.1002/9781119357254.ch2

Stoica, R., Senin, R., Radu, E., Manolache, M., Oancea, F., & Ceausu, M. (2016). Analysis of the storage stability of the wheatgrass (Triticum aestivum L.) Juice in terms of antioxidant activity and Their polyphenols content. Rev. De Chim, 67, 1669-1672. https://scholar.google.com/scholar_lookup?title=Analysis+of+the+storage+stability+of+the+wheatgrass+(Triticum+aestivum+L.)+Juice+in+terms+of+antioxidant+activity+and+Their+polyphenols+content&author=Stoica,+R.&author=Senin,+R.&author=Radu,+E.&author=Manolache,+M.&author=Oancea,+F.&author=Ceausu,+M.&publication_year=2016&journal=Rev.+De+Chim.&volume=67&pages=1669%E2%80%931672

Tariq, H., Awan, S. I., Sabir, S. M., & Ilyas, M. (2020). Hexaploid-tetraploid landraces and wild species of wheat revealed diversity for antioxidants and total phenolics. Philipp. Agric. Sci, 103, 29-37. https://www.ukdr.uplb.edu.ph/journal-articles/3974

Vu, N. D., Nhi Tran, T. Y., Tan Nguyen, T. X., Nhu Hang Nguyen, T. T., Huong Pham, T. T., Long, H. B., & Pham, B. A. (2025). Physicochemical and biological properties of cashew apples in the form of juice, concentrated juice, and fresh under different storage conditions. Journal of Food Quality, 2025(1), 4458814. https://doi.org/10.1155/jfq/4458814

Zhang, Y., Li, Y., Ren, X., Zhang, X., Wu, Z., & Liu, L. (2023). The positive correlation of antioxidant activity and prebiotic effect about oat phenolic compounds. Food Chemistry, 402, 134231. https://doi.org/10.1016/j.foodchem.2022.134231

Zhang, C. L., Li, L., Wang, Q. C., Hong, L. Z., Zhang, P., Zhang, X. S., ... & Qin, P. (2025). Integrative physicochemical and metabolic profiling of grain components related to nutritional quality in Yunnan hulled wheat and common wheat. Journal of the Science of Food and Agriculture, 105(14), 7686-7709. https://doi.org/10.1002/jsfa.70063

Zhishen, J., Mengcheng, T., & Jianming, W. (1999). The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food chemistry, 64(4), 555-559. https://doi.org/10.1016/S0308-8146(98)00102-2