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Increased Free Radical Scavenging Activity and Consumer Preference in Garden Cress (Lepidium sativum L.) Subjected to Water Stress

Year 2023, Volume: 11 Issue: 2, 276 - 285, 28.12.2023
https://doi.org/10.33202/comuagri.1361583

Abstract

Tere (Lepidium sativum L.) bitkisinin çok çeşitli iklim koşullarında yetiştiği bilinmesine rağmen su stresinin bitkinin morfolojik, biyokimyasal ve duyusal özelliklerini nasıl etkileyebileceğine dair sınırlı sayıda çalışma bulunmaktadır. Kontrollü atmosfer koşullarında iki farklı su stresi seviyesi uygulanarak, dört tekrarlı gerçekleştirilen bu çalışmada; terenin bazı morfolojik parametreleri, biyokimyasal özellikleri ve duyusal özellikleri ölçülmüştür. Önceki çalışmalarda su stresi nedeniyle verim kaybı değerlendirilirken, bu çalışmalarda ürünün pazarlanabilirliği ve tüketicilerin tercihi dikkate alınmamıştır. Kontrol bitkilerine (%100 sulama) karşı iki farklı sulama (%50 ve %25 sulama) uygulanan çalışma sonucunda kontrol bitkilerinde prolin miktarı 5.98 mg-1100 g, olarak belirlenirken. %25 sulama suyu uygulanan bitkilerde 36.72 mg-1100g olarak tespit edilmiştir.. Toplam fenolik bileşikler ve toplam flavonoidler sırasıyla 61.26 ile 68.04 mg GA-1100g ve 8.35 ile 11.70 mg QE-1100g arasında değişirken, farklar istatistiksel olarak önem bulunmuştur. Ancak artan su stresine ile birlikte DPPH serbest radikal temizleme aktivitesinin artması istatistiksel olarak anlamlı bulunmuştur. Sonuç olarak %50 su stresi uygulanan Tere yapraklarının tüketiciler tarafından lezzet açısından en çok tercih edildiği ve kontrol bitkilerine göre daha yüksek radikal temizleme aktivitesine sahip olduğu tespit edilmiştir. Bu nedenle sulama yönetimi uygulamaları, küresel ısınma ve iklim değişikliğinin su kaynakları üzerindeki olası olumsuz etkileri altında Tere üretimi ile sınırlı su kullanımını sağlanabilir.

References

  • Abdel-Aty, A.M., Salama, W.H., Fahmy, A.S., Mohamed, S.A., 2019. Impact of germination on antioxidant capacity of garden cress: New calculation for determination of total antioxidant activity. Scientia Horticulturae. 246: 155-160.
  • Ahmadiani, N., Robbins, R.J., Collins, T.M., Giusti, M.M., 2014. Anthocyanins contents, profiles, and color characteristics of red cabbage extracts from different cultivars and maturity stages. Journal of agricultural and food chemistry. 62 (30): 7524-7531.
  • Ak I., Türker, G., 2018. Antioxidant activity of five seaweed extracts. New Knowl. J. Sci. 7: 149–155.
  • Al-Huqail, A., El-Dakak, R.M., Sanad, M.N., Badr, R.H., Ibrahim, M.M., Soliman, D., Khan, F., 2020. Effects of climate temperature and water stress on plant growth and accumulation of antioxidant compounds in sweet basil (Ocimum basilicum L.) leafy vegetable. Hindawi Scientifica. 3808909.
  • Al-Sammarraie, O.N., Alsharafa, K.Y., Al-Limoun, M.O., Khleifat, K.M., Al-Sarayreh, S.A., Al-Shuneigat, J.M., Kalaji, H.M., 2020. Effect of various abiotic stressors on some biochemical indices of Lepidium sativum plants. Scientific Reports. 10 (1): 1-10.
  • Arimi, K.S., 2021. Climate change adaptation and resilience among vegetable farmers. International Journal of Vegetable Science. 27(5): 496-504.
  • Aznar-Ramos, M.J., Razola-Díaz, M.D.C., Verardo, V., Gómez-Caravaca, A.M., 2022. Comparison between Ultrasonic Bath and Sonotrode Extraction of Phenolic Compounds from Mango Peel By-Products. Horticulturae 8: 1014.
  • Bates L.S. and R.P. Waldren. 1973. Teare ID, Plant & Soil. 39: 205.
  • Brand-Williams, W., Cuvelier, M.E., Berset, C., 1995. Use of a free radical method to evaluate antioxidant activity. LWT - Food Sci. Technol. 28: 25–30.
  • Cano-Lamadrid, M., Martínez-Zamora, L., Castillejo, N., Cattaneo, C., Pagliarini, E., Artés-Hernández, F., 2023. How does the phytochemical composition of sprouts and microgreens from Brassica vegetables affect the sensory profile and consumer acceptability? Postharvest Biology and Technology. 203: 112411.
  • Demir, H., Polat, E., 2017. Sera koşullarında domates-tere birlikte yetiştiriciliğinin verim ve kaliteye etkisi. Akademik Ziraat Dergisi, 6 51-62.
  • Diwakar, B.T., Dutta, P.K., Lokesh, B.R.,. Naidu, A.K., 2010. Physicochemical properties of garden cress (lepidium sativum l.) seed oil. J Am Oil Chem Soc 87: 539–548.
  • Ehsan, S., Ali, S., Noureen, S., Mahmood, K., Farid, M., Ishaque, W., Shakoor, M.B., Rizwan, M., 2014. Citric acid assisted phytoremediation of cadmium by Brassica napus L. Ecotox Environ Safe 106: 164–172.
  • El-Zaeddi, H., Calín-Sánchez, Á., Martínez-Tomé, J., Noguera-Artiaga, L., Burló, F., Barrachina, Á.A.C., 2016. Irrigation dose and plant density affect the essential oil content and sensory quality of parsley (Petroselinum sativum). Scientia horticulturae. 206: 1-6.
  • Erken, O., 2022. Some bioactive metabolites’ response to long-term water stress in red cabbage. Scientia Horticulturae. 293: 110731.
  • Escalante-Magaña, C., Aguilar-Caamal, L.F., Echevarría-Machado, I., Medina-Lara, F., Cach, L.S., Martínez-Estévez, M., 2019. Contribution of glycine betaine and proline to water deficit tolerance in pepper plants. HortScience. 54 (6): 1044-1054.
  • Giordano, M., Petropoulos, S.A., Rouphael, Y., 2021. Response and defence mechanisms of vegetable crops against drought, heat and salinity stress. Agriculture. 11 (5): 463.
  • Griñán, I., Galindo, A., Rodríguez, P., Morales, D., Corell, M., Centeno, A., Gonzales, J.C., Torrecillas, A., Barrachina, A.A., Hernandez, F., 2019. Volatile composition and sensory and quality attributes of quince (Cydonia oblonga Mill.) fruits as affected by water stress. Scientia Horticulturae. 244: 68-74.
  • Hayat, S., Hayat, Q., Alyemeni, M.N., Wani, A.S., Pichtel, J., Ahmad, A., 2012. Role of proline under changing environments: a review. Plant Signal Behav. 7 (11): 1456–1466.
  • Holden, M., 1976. Chlorophyll in chemistry and biochemistry of plant pigments. Vol. 2 (T. W. Goodwin, Ed.). Academic Press, London pp: 1 –37.
  • Jones, H.G., 2007. Monitoring plant and soil water status: established and novel methods revisited and their relevance to studies of drought tolerance. Journal of Experimental Botany, Integrated Approaches to Sustain and Improve Plant Production under Drought Stress Special Issue. 58 (2): 119-130.
  • Keutgen, N., Hausknecht, M., Tomaszewska-Sowa, M., Keutgen, A.J., 2021. Nutritional and sensory quality of two types of cress microgreens depending on the mineral nutrition. Agronomy. 11 (6): 1110.
  • Khalil, S.E., El-Noemani, A.A., 2012. Effect of irrigation intervals and exogenous proline application in improving tolerance of garden cress plant (Lepidium sativum L.) to water stress. Journal of Applied Sciences Research, (January), 157-167.
  • Kiremit, M.S., Osman, H.M., Arslan, H., 2023. Response of yield, growth traits, and leaf nutrients of garden cress to deficit saline irrigation waters. Journal of Plant Nutrition. 46 (6): 1050-1065.
  • Malar, J., Chairman, K., Singh, A.R.J., Vanmathi, J.S., Balasubramanian, A., Vasanthi, K., 2014. Antioxidative activity of different parts of the plant Lepidium sativum Linn. Biotechnol Rep (Amst). 3: 95-98.
  • Meilgaard, M.C., Carr B.T., Civille, G.V., 1999. Sensory evaluation techniques. CRC press.
  • Mohamed, E.A.A., Muddathir, A.M., Abdalla, A.H., 2020. Effects of organic and inorganic fertilization on growth, yield, seed fixed oil content, and fatty acids profile of garden cress (Lepidium sativum L.). SN Applied Sciences. 2 (10): 1-10.
  • Okunlola, G.O., Olatunji, O.A., Niewiadomska, E., Jimoh, M.A., Rufai, A.B., Ogunkunle, C.O., 2022. Foliar application of melatonin alleviates the deleterious effects of drought on the three most cultivated capsicum species in Africa. Gesunde Pflanzen, 1-11.
  • Painuli, S., Quispe, C., Herrera-Bravo, J., Semwal, P., Martorell, M., Almarhoon, Z.M., Cho, W.C., 2022. Nutraceutical profiling, bioactive composition, and biological applications of Lepidium sativum L. Oxidative Medicine and Cellular Longevity.
  • Rudzińska, A., Juchaniuk, P., Oberda, J., Wiśniewska, J., Wojdan, W., Szklener, K., Mańdziuk, S., 2023. Phytochemicals in cancer treatment and cancer prevention-review on epidemiological data and clinical trials. Nutrients. 15 (8): 1896.
  • Sat, I.G., Yildirim, E., Turan, M., Demirbas, M., 2013. Antioxidant and nutritional characteristics of garden cress (Lepidium sativum). Acta Sci. Polonorum-Hort. Cultus. 12: 173-179.
  • Selek, S., Koyuncu, I., Caglar, H.G., Bektas, I., Yilmaz, M.A., Gonel, A., Akyuz, E., 2018. The evaluation of antioxidant and anticancer effects of Lepidium Sativum Subsp Spinescens L. methanol extract on cancer cells. Cellular and Molecular Biology. 64 (3): 72-80.
  • Seyhan, S.A., 2019. DPPH antioksidan analizinin yeniden değerlendirilmesi. Batman Üniversitesi Yaşam Bilimleri Dergisi. 9 (2): 125-135.
  • Shraim, A.M., Ahmed, T.A., Rahman, M.M., Hijji, Y.M., 2021. Determination of total flavonoid content by aluminum chloride assay: A critical evaluation. LWT, 150.
  • Singh, S., Thakur, A., Puri, S., 2020. Effect of mannitol on lepidium sativum l. under abiotic stress (water stress) condition. Plant Archives. 20 (2): 8500-8510.
  • Singleton, V.L., Rossi, J.A., 1965. Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. American Journal of Enology and Viticulture. 16: 144-153.
  • Smolinska, B., Leszczynska, J., 2017. Photosynthetic pigments and peroxidase activity of Lepidium sativum L. during assisted Hg phytoextraction. Environmental Science and Pollution Research. 24 (15): 13384-13393.
  • Uyar, B.B., Gezmen-Karadağ, M., Şanlier, N., Günyel, S., 2013. Toplumumuzda sıklıkla kullanılan bazı bitkilerin toplam fenolik madde miktarlarının saptanması. Gıda. 38 (1): 23-29.
  • Yamasaki, S., Dillenburg, L.R., 1999. Measurements of leaf relative water content in araucaria angustifolia. Revista Brasilleira de Fisiologia Vegetal. 11 (2): 69-75.
  • Yanmaz, R., Yildirim, E., Koyuncu, D., 2010. Ülkemiz için yeni bir tere (lepidium sativum var. sativum) çeşit adayı: Dadaş. Journal of Agricultural Faculty of Atatürk University. 41 (2): 91-95.
  • Yildirim, E., Ekinci, M., Turan, M., Ağar, G., Ors, S., Dursun, A., Kul, R., Akgül, G., 2022. Physiological and biochemical changes of pepper cultivars under combined salt and drought stress. Gesunde Pflanzen, 1-9.
  • Zia-Ul-Haq, M., Ahmad, S., Calani, L., Mazzeo, T., Del Rio, D., Pellegrini, N., De Feo, V., 2012. Compositional study and antioxidant potential of Ipomoea hederacea Jacq and Lepidium sativum L. seeds. Molecules. 17 (9): 10306-10321.

Increased Free Radical Scavenging Activity and Consumer Preference in Garden Cress (Lepidium sativum L.) Subjected to Water Stress

Year 2023, Volume: 11 Issue: 2, 276 - 285, 28.12.2023
https://doi.org/10.33202/comuagri.1361583

Abstract

Although the Garden Cress is known to grow in a wide range of climatic conditions, there are limited studies on how water stress can affect their morphological, biochemical and sensory characteristics. In this research, these characteristics in the Garden Cress leaves were measured in response to water stress. In this study two different irrigation restrictions (50% and 25% irrigation) were applied against the control plants (100% irrigation) in four replications. As a result, the amount of proline was determined as 5.98 mg-1100g in control plants, and as 36.72 mg-1100g in 25% irrigated plants. While total phenolic compounds and total flavonoids varied from 61.26 to 68.04 mg of GA-1100gFW and 8.35 to 11.70 mg QE-1100gFW, respectively, the differences were statistically insignificant. The increase of DPPH free radical scavenging activity in line with increased water stress, however, was found to be statistically significant. In conclusion, the 50% water stress applied Garden Cress leaves were found to be most preferred by the consumers in terms of flavor and had higher radical scavenging activity than control plants. Irrigation management practices, therefore, could consider limited water use with Garden Cress production under possible negative effects of climate change on water resources.

References

  • Abdel-Aty, A.M., Salama, W.H., Fahmy, A.S., Mohamed, S.A., 2019. Impact of germination on antioxidant capacity of garden cress: New calculation for determination of total antioxidant activity. Scientia Horticulturae. 246: 155-160.
  • Ahmadiani, N., Robbins, R.J., Collins, T.M., Giusti, M.M., 2014. Anthocyanins contents, profiles, and color characteristics of red cabbage extracts from different cultivars and maturity stages. Journal of agricultural and food chemistry. 62 (30): 7524-7531.
  • Ak I., Türker, G., 2018. Antioxidant activity of five seaweed extracts. New Knowl. J. Sci. 7: 149–155.
  • Al-Huqail, A., El-Dakak, R.M., Sanad, M.N., Badr, R.H., Ibrahim, M.M., Soliman, D., Khan, F., 2020. Effects of climate temperature and water stress on plant growth and accumulation of antioxidant compounds in sweet basil (Ocimum basilicum L.) leafy vegetable. Hindawi Scientifica. 3808909.
  • Al-Sammarraie, O.N., Alsharafa, K.Y., Al-Limoun, M.O., Khleifat, K.M., Al-Sarayreh, S.A., Al-Shuneigat, J.M., Kalaji, H.M., 2020. Effect of various abiotic stressors on some biochemical indices of Lepidium sativum plants. Scientific Reports. 10 (1): 1-10.
  • Arimi, K.S., 2021. Climate change adaptation and resilience among vegetable farmers. International Journal of Vegetable Science. 27(5): 496-504.
  • Aznar-Ramos, M.J., Razola-Díaz, M.D.C., Verardo, V., Gómez-Caravaca, A.M., 2022. Comparison between Ultrasonic Bath and Sonotrode Extraction of Phenolic Compounds from Mango Peel By-Products. Horticulturae 8: 1014.
  • Bates L.S. and R.P. Waldren. 1973. Teare ID, Plant & Soil. 39: 205.
  • Brand-Williams, W., Cuvelier, M.E., Berset, C., 1995. Use of a free radical method to evaluate antioxidant activity. LWT - Food Sci. Technol. 28: 25–30.
  • Cano-Lamadrid, M., Martínez-Zamora, L., Castillejo, N., Cattaneo, C., Pagliarini, E., Artés-Hernández, F., 2023. How does the phytochemical composition of sprouts and microgreens from Brassica vegetables affect the sensory profile and consumer acceptability? Postharvest Biology and Technology. 203: 112411.
  • Demir, H., Polat, E., 2017. Sera koşullarında domates-tere birlikte yetiştiriciliğinin verim ve kaliteye etkisi. Akademik Ziraat Dergisi, 6 51-62.
  • Diwakar, B.T., Dutta, P.K., Lokesh, B.R.,. Naidu, A.K., 2010. Physicochemical properties of garden cress (lepidium sativum l.) seed oil. J Am Oil Chem Soc 87: 539–548.
  • Ehsan, S., Ali, S., Noureen, S., Mahmood, K., Farid, M., Ishaque, W., Shakoor, M.B., Rizwan, M., 2014. Citric acid assisted phytoremediation of cadmium by Brassica napus L. Ecotox Environ Safe 106: 164–172.
  • El-Zaeddi, H., Calín-Sánchez, Á., Martínez-Tomé, J., Noguera-Artiaga, L., Burló, F., Barrachina, Á.A.C., 2016. Irrigation dose and plant density affect the essential oil content and sensory quality of parsley (Petroselinum sativum). Scientia horticulturae. 206: 1-6.
  • Erken, O., 2022. Some bioactive metabolites’ response to long-term water stress in red cabbage. Scientia Horticulturae. 293: 110731.
  • Escalante-Magaña, C., Aguilar-Caamal, L.F., Echevarría-Machado, I., Medina-Lara, F., Cach, L.S., Martínez-Estévez, M., 2019. Contribution of glycine betaine and proline to water deficit tolerance in pepper plants. HortScience. 54 (6): 1044-1054.
  • Giordano, M., Petropoulos, S.A., Rouphael, Y., 2021. Response and defence mechanisms of vegetable crops against drought, heat and salinity stress. Agriculture. 11 (5): 463.
  • Griñán, I., Galindo, A., Rodríguez, P., Morales, D., Corell, M., Centeno, A., Gonzales, J.C., Torrecillas, A., Barrachina, A.A., Hernandez, F., 2019. Volatile composition and sensory and quality attributes of quince (Cydonia oblonga Mill.) fruits as affected by water stress. Scientia Horticulturae. 244: 68-74.
  • Hayat, S., Hayat, Q., Alyemeni, M.N., Wani, A.S., Pichtel, J., Ahmad, A., 2012. Role of proline under changing environments: a review. Plant Signal Behav. 7 (11): 1456–1466.
  • Holden, M., 1976. Chlorophyll in chemistry and biochemistry of plant pigments. Vol. 2 (T. W. Goodwin, Ed.). Academic Press, London pp: 1 –37.
  • Jones, H.G., 2007. Monitoring plant and soil water status: established and novel methods revisited and their relevance to studies of drought tolerance. Journal of Experimental Botany, Integrated Approaches to Sustain and Improve Plant Production under Drought Stress Special Issue. 58 (2): 119-130.
  • Keutgen, N., Hausknecht, M., Tomaszewska-Sowa, M., Keutgen, A.J., 2021. Nutritional and sensory quality of two types of cress microgreens depending on the mineral nutrition. Agronomy. 11 (6): 1110.
  • Khalil, S.E., El-Noemani, A.A., 2012. Effect of irrigation intervals and exogenous proline application in improving tolerance of garden cress plant (Lepidium sativum L.) to water stress. Journal of Applied Sciences Research, (January), 157-167.
  • Kiremit, M.S., Osman, H.M., Arslan, H., 2023. Response of yield, growth traits, and leaf nutrients of garden cress to deficit saline irrigation waters. Journal of Plant Nutrition. 46 (6): 1050-1065.
  • Malar, J., Chairman, K., Singh, A.R.J., Vanmathi, J.S., Balasubramanian, A., Vasanthi, K., 2014. Antioxidative activity of different parts of the plant Lepidium sativum Linn. Biotechnol Rep (Amst). 3: 95-98.
  • Meilgaard, M.C., Carr B.T., Civille, G.V., 1999. Sensory evaluation techniques. CRC press.
  • Mohamed, E.A.A., Muddathir, A.M., Abdalla, A.H., 2020. Effects of organic and inorganic fertilization on growth, yield, seed fixed oil content, and fatty acids profile of garden cress (Lepidium sativum L.). SN Applied Sciences. 2 (10): 1-10.
  • Okunlola, G.O., Olatunji, O.A., Niewiadomska, E., Jimoh, M.A., Rufai, A.B., Ogunkunle, C.O., 2022. Foliar application of melatonin alleviates the deleterious effects of drought on the three most cultivated capsicum species in Africa. Gesunde Pflanzen, 1-11.
  • Painuli, S., Quispe, C., Herrera-Bravo, J., Semwal, P., Martorell, M., Almarhoon, Z.M., Cho, W.C., 2022. Nutraceutical profiling, bioactive composition, and biological applications of Lepidium sativum L. Oxidative Medicine and Cellular Longevity.
  • Rudzińska, A., Juchaniuk, P., Oberda, J., Wiśniewska, J., Wojdan, W., Szklener, K., Mańdziuk, S., 2023. Phytochemicals in cancer treatment and cancer prevention-review on epidemiological data and clinical trials. Nutrients. 15 (8): 1896.
  • Sat, I.G., Yildirim, E., Turan, M., Demirbas, M., 2013. Antioxidant and nutritional characteristics of garden cress (Lepidium sativum). Acta Sci. Polonorum-Hort. Cultus. 12: 173-179.
  • Selek, S., Koyuncu, I., Caglar, H.G., Bektas, I., Yilmaz, M.A., Gonel, A., Akyuz, E., 2018. The evaluation of antioxidant and anticancer effects of Lepidium Sativum Subsp Spinescens L. methanol extract on cancer cells. Cellular and Molecular Biology. 64 (3): 72-80.
  • Seyhan, S.A., 2019. DPPH antioksidan analizinin yeniden değerlendirilmesi. Batman Üniversitesi Yaşam Bilimleri Dergisi. 9 (2): 125-135.
  • Shraim, A.M., Ahmed, T.A., Rahman, M.M., Hijji, Y.M., 2021. Determination of total flavonoid content by aluminum chloride assay: A critical evaluation. LWT, 150.
  • Singh, S., Thakur, A., Puri, S., 2020. Effect of mannitol on lepidium sativum l. under abiotic stress (water stress) condition. Plant Archives. 20 (2): 8500-8510.
  • Singleton, V.L., Rossi, J.A., 1965. Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. American Journal of Enology and Viticulture. 16: 144-153.
  • Smolinska, B., Leszczynska, J., 2017. Photosynthetic pigments and peroxidase activity of Lepidium sativum L. during assisted Hg phytoextraction. Environmental Science and Pollution Research. 24 (15): 13384-13393.
  • Uyar, B.B., Gezmen-Karadağ, M., Şanlier, N., Günyel, S., 2013. Toplumumuzda sıklıkla kullanılan bazı bitkilerin toplam fenolik madde miktarlarının saptanması. Gıda. 38 (1): 23-29.
  • Yamasaki, S., Dillenburg, L.R., 1999. Measurements of leaf relative water content in araucaria angustifolia. Revista Brasilleira de Fisiologia Vegetal. 11 (2): 69-75.
  • Yanmaz, R., Yildirim, E., Koyuncu, D., 2010. Ülkemiz için yeni bir tere (lepidium sativum var. sativum) çeşit adayı: Dadaş. Journal of Agricultural Faculty of Atatürk University. 41 (2): 91-95.
  • Yildirim, E., Ekinci, M., Turan, M., Ağar, G., Ors, S., Dursun, A., Kul, R., Akgül, G., 2022. Physiological and biochemical changes of pepper cultivars under combined salt and drought stress. Gesunde Pflanzen, 1-9.
  • Zia-Ul-Haq, M., Ahmad, S., Calani, L., Mazzeo, T., Del Rio, D., Pellegrini, N., De Feo, V., 2012. Compositional study and antioxidant potential of Ipomoea hederacea Jacq and Lepidium sativum L. seeds. Molecules. 17 (9): 10306-10321.
There are 42 citations in total.

Details

Primary Language English
Subjects Ecology (Other)
Journal Section Articles
Authors

Okan Erken 0000-0001-5177-7432

Çiğdem Pala 0000-0003-1784-9134

Bayram Kızılkaya 0000-0003-4508-2516

Publication Date December 28, 2023
Published in Issue Year 2023 Volume: 11 Issue: 2

Cite

APA Erken, O., Pala, Ç., & Kızılkaya, B. (2023). Increased Free Radical Scavenging Activity and Consumer Preference in Garden Cress (Lepidium sativum L.) Subjected to Water Stress. ÇOMÜ Ziraat Fakültesi Dergisi, 11(2), 276-285. https://doi.org/10.33202/comuagri.1361583