Utilizing Plackett–Burman and Box–Behnken Designs for Plant Extract–Based AgNP Synthesis Optimization: Unveiling Antifungal Potential Against Phytophthora Species

Muharrem Türkkan

doi:10.18016/ksutarimdoga.vi.1523681

EN TR

Utilizing Plackett–Burman and Box–Behnken Designs for Plant Extract–Based AgNP Synthesis Optimization: Unveiling Antifungal Potential Against Phytophthora Species

Abstract

This study optimized a green synthesis method for silver nanoparticles (AgNPs) using aqueous extracts of black tea, linden, cherry laurel, kale, and melocan, employing a statistical design of experiments. The plant extracts acted as bio-reducing agents. Total and individual phenolic compounds in the extracts were quantified using ultraviolet-visible (UV–Vis) spectroscopy and ultra-high-performance liquid chromatography (UHPLC). AgNP yields were maximized through a combination of Plackett–Burman and Box–Behnken designs. The synthesized AgNPs were characterized by UV–Vis spectroscopy, Fourier transform infrared (FT–IR) spectroscopy, scanning electron microscopy (SEM) coupled with energy-dispersive X-ray spectroscopy (EDS), and transmission electron microscopy (TEM). Optimal AgNP production was achieved under the following conditions (determined by response surface methodology, RSM): 9.6 g of plant material, extraction heating at 80°C for 20 minutes, 10 mM AgNO3, 2.5 mL of extract, 800 W microwave irradiation, and a 90-second reaction time. FT–IR analysis confirmed the role of phenolic compounds in reducing and stabilizing AgNPs. The resulting AgNPs exhibited uniform spherical morphology, with average particle sizes of 5.30 nm (black tea), 8.74 nm (linden), 7.20 nm (cherry laurel), 6.32 nm (kale), and 9.44 nm (melocan). Antifungal assays against five Phytophthora species revealed that kale-derived AgNPs were most potent, with EC50, MIC, and MFC values ranging from 9.28–30.84 µg mL−1, 200–300 µg mL−1, and 200–400 µg mL−1, respectively. These results suggest that plant-extract-synthesized AgNPs offer a sustainable approach to managing Phytophthora diseases, warranting further research.

Keywords

Bitki Ekstraktı Bazlı AgNP Sentez Optimizasyonu için Plackett–Burman ve Box–Behnken Tasarımlarının Kullanımı: Phytophthora Türlerine Karşı Antifungal Potansiyelinin Ortaya Çıkarılması

Öz

Bu çalışma, siyah çay, ıhlamur, karayemiş, yaprak lahana ve melocan sulu ekstraktları kullanılarak gümüş nanoparçacıklar (AgNP’ler) için deneysel bir istatistiksel tasarım ile yeşil bir sentez yöntemini optimize etmiştir. Bu bitki ekstraktları biyo-indirgeyici ajanlar olarak işlev görmüştür. Ekstraktlardaki toplam ve bireysel fenolik bileşikler ultraviyole-görünür (UV–Vis) spektroskopi ve ultra yüksek performansli sivi kromatografisi (UHPLC) kullanılarak ölçülmüştür. AgNP verimleri, Plackett‒Burman ve Box‒Behnken tasarımlarının bir kombinasyonu ile maksimize edilmiştir. Sentezlenen AgNP’ler UV‒Vis spektroskopisi, Fourier dönüşümlü kizilötesi (FT–IR) spektroskopisi, taramali elektron mikroskobu (SEM)-enerji dağilimli X-ışını spektroskopisi (EDS) ve geçirimli elektron mikroskopisi (TEM) ile karakterize edilmiştir. Optimum AgNP üretimi aşağıdaki koşullar altında elde edilmiştir (yanıt yüzey metodolojisi, RSM ile belirlenmiştir): 9.6 g bitki materyali, 80°C’de 20 dakika ekstraksiyon ısıtma, 10 mM AgNO3, 2.5 mL ekstrakt, 800 W mikrodalga irritasyonu ve 90 saniyelik reaksiyon süresi. FT–IR analizi, fenolik bileşiklerin AgNP indirgenmesi ve stabilize edilmesindeki rolünü göstermiştir. Elde edilen AgNP’ler 5.30 nm (siyah çay), 8.74 nm (ıhlamur), 7.20 nm (karayemiş), 6.32 nm (yaprak lahana) ve 9.44 nm (melocan) ortalama partikül boyutları ile tek tip küresel morfoloji sergilemiştir. Beş Phytophthora türüne karşı yapılan antifungal deneyler, yaprak lahana türevi AgNP’lerin sırasıyla 9.28-30.84 µg mL−1, 200-300 µg mL−1 ve 200-400 µg mL−1 arasında değişen EC50, MIC ve MFC değerleri ile en güçlü olduğunu ortaya koymuştur. Bu sonuçlar, bitki ekstraktı ile sentezlenen AgNP’lerin Phytophthora hastalıkların yönetiminde sürdürülebilir bir yaklaşım sunduğunu ve daha fazla araştırılması gerektiğini göstermektedir.

Anahtar Kelimeler

Supporting Institution

Ordu Üniversitesi BAP

Project Number

B2209

Thanks

The author is grateful to Dr. İlker Kurbetli for supplying the Phytophthora isolates, Dr. Umut Ateş for performing high-performance liquid chromatography (HPLC) analysis of the total phenolic content and individual phenolic compounds in the plant extracts, and Dr. Hamdi Güray Kutbay for confirming the identification of melocan (Smilax excelsa L.).

References

Adnan, M., Azad, M. O. K., Madhusudhan, A., Saravanakumar, K., Hu, X., Wang, M. H. & Ha, C. D. (2020). Simple and cleaner system of silver nanoparticle synthesis using kenaf seed and revealing its anticancer and antimicrobial potential. Nanotechnology, 31(26), 265101. https://doi.org/10.1088/1361-6528/ab7d72
Ahmad, N. & Sharma, S. (2012). Green Synthesis of silver nanoparticles using extracts of Ananas comosus. Green and Sustainable Chemistry, 2(4), 1-7. https://doi.org/10.4236/gsc.2012.24020
Akyuz, E., Şahin, H., Islamoglu, F., Kolayli, S. & Sandra, P. (2014). Evaluation of phenolic compounds in Tilia rubra subsp. caucasica by HPLC–UV and HPLC–UV–MS/MS. International Journal of Food Properties, 17, 331-343. https://doi.org/10.1080/10942912.2011.631252
Ali, M., Kim, B., Belfield, K. D., Norman, D., Brennan, M. & Ali, G. S. (2015). Inhibition of Phytophthora parasitica and P. capsici by silver nanoparticles synthesized using aqueous extract of Artemisia absinthium. Phytopathology, 105(9), 1183-1190. https://doi.org/10.1094/PHYTO-01-15-0006-R
Baytop, T. (1999). Therapy with medicinal plants in Turkey (past and present). Publication of the Istanbul University, 312.
Box, G. E. P. & Behnken, D. W. (1960). Simplex-sum designs: a class of second order rotatable designs derivable from those of first order. The Annals of Mathematical Statistics, 31(4), 838-864. https://doi.org/10.1214/aoms/1177705661
Buzea, C., Pacheco, I. I. & Robbie, K. (2007). Nanomaterials and nanoparticles: sources and toxicity. Biointerphases 2(4), MR17-MR71. https://doi.org/10.1116/1.2815690
Cai, Y., Piao, X., Gao, W., Zhang, Z., Nie, E. & Sun, Z. (2017). Large-scale and facile synthesis of silver nanoparticles via a microwave method for a conductive pen. RSC Advances, 7(54), 34041-34048. https://doi.org/10.1039/C7RA05125E

Chowdhury, S., Yusof, F., Faruck, M. O. & Sulaiman, N. (2016). Process optimization of silver nanoparticle synthesis using response surface methodology. Procedia Engineering, 148, 992-999. https://doi.org/10.1016/j.proeng.2016.06.552
Davis, P., Tan, K. & Mill, R.R. (1988). Flora of Turkey and the East Aegean Islands and Supplement I. Edinburgh University Press, Edinburgh.
El-Sawaf, A. K., El-Moslamy, S. H., Kamoun, E. A., & Hossain, K. (2024). Green synthesis of trimetallic CuO/Ag/ZnO nanocomposite using Ziziphus spina-christi plant extract: characterization, statistically experimental designs, and antimicrobial assessment. Scientific Reports, 14(1), 19718. https://doi.org/10.1038/s41598-024-67579-5
Encu, S. (2010). Lahana çeşitlerinin antioksidan kapasiteleri ve bileşenleri açısından değerlendirilmesi (Tez no 282642). [Yüksek Lisans Tezi, İstanbul Üniversitesi Fen Bilimleri Enstitüsü Kimya Ana Bilim Dalı]. Yükseköğretim Kurulu Ulusal Tez Merkezi.
Günal, N. (2013). Türkiye’de iklimin doğal bitki örtüsü üzerindeki etkileri. Acta Turcıca Çevrimiçi Tematik Türkoloji Dergisi, V(1), 1-22.
Fazil, M. M., Gul, A. & Jawed, H. (2024). Optimization of silver nanoparticles synthesis via Plackett–Burman experimental design: in vitro assessment of their efficacy against oxidative stress-induced disorders. RSC Advances, 14(29), 20809-20823. https://doi.org/10.1039/d4ra02774d
Firoozi, S., Jamzad, M. & Yari, M. (2016). Biologically synthesized silver nanoparticles by aqueous extract of Satureja intermedia C.A. Mey and the evaluation of total phenolic and flavonoid contents and antioxidant activity. Journal of Nanostructure in Chemistry, 6, 357-364. https://doi.org/10.1007/s40097-016-0207-0
Gevrek, C., Yiğit, U. & Türkkan, M. (2023). Optimization and antifungal activity of silver nanoparticles synthesized using the leaf extract of Corylus colurna L. (Turkish hazelnut). Akademik Ziraat Dergisi, 12(Özel Sayı), 159-172. https://doi.org/10.29278/azd.1335259
Halima, R., Narula, A. & Sravanthi, V. (2021). Optimization of process parameters for the green synthesis of silver nanoparticles using Plackett‒Burman and 3-level Box–Behnken Design. Journal of Huazhong University of Science and Technology, 50(3), 1-17.
İslam, A. & Deligöz, H. (2012). Ordu ilinde karayemiş (Laurocerasus officinalis L.) seleksiyonu. Akademik Ziraat Dergisi, 1(1), 37-44.
Jyoti, K., Baunthiyal, M. & Singh, A. (2016). Characterization of silver nanoparticles synthesized using Urtica dioica Linn. leaves and their synergistic effects with antibiotics. Journal of Radiation Research and Applied Sciences, 9(3), 217-227. https://doi.org/10.1016/j.jrras.2015.10.002
Kale, R., Barwar, S., Kane, P. & More, S. (2018). Green synthesis of silver nanoparticles using papaya seed and its characterization. International Journal for Research in Applied Science & Engineering Technology, 6, 168-174. https://doi.org/10.22214/ijraset.2018.2026
Karabegović, I.T., Stojicevi´c, S.S., Velickovic, D.T., Zoran B. Todorovic, Z.B., Nada C. Nikolic, N.C., Miodrag L. & Lazic, M.L. (2014). The effect of different extraction techniques on the compositionand antioxidant activity of cherry laurel (Prunus laurocerasus) leaf and fruit extracts. Industrial Crops and Products, 54, 142-148. https://doi.org/10.1016/j.indcrop.2013.12.047
Karakaş, İ., Hacıoğlu, N. & Özdemir, B. E. (2024). Green synthesis and antibiofilm activity of silver nanoparticles by Camellia sinensis L. (White tea leaf). Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi, 27(2), 285-292. https://doi.org/10.18016/ksutarimdoga.1297130
Kelly, K. L., Coronado, E., Zhao, L. L. & Schatz, G. C. (2003). The optical properties of metal nanoparticles: the influence of size, shape, and dielectric environment. The Journal of Physical Chemistry B, 107(3), 668-677. https://doi.org/10.1021/jp026731y
Kolaylı, S., Kucuk, M., Duran, C., Candan, F. & Dinçer, B. (2003). Chemical and antioxidant properties of Laurocerasus officinalis Roem. (Cherry laurel) fruit grown in the Black Sea Region. Journal of Agricultural and Food Chemistry, 51, 7489-94. https://doi.org/10.1021/jf0344486
Konwarh, R., Karak, N., Sawian, C. E., Baruah, S. & Mandal, M. (2011). Effect of sonication and aging on the templating attribute of starch for “green” silver nanoparticles and their interactions at biointerface. Carbohydrate Polymers, 83(3), 1245-1252. https://doi.org/10.1016/j.carbpol.2010.09.031
Kumar, K. M., Sinha, M., Mandal, B. K., Ghosh, A. R., Kumar, K. S. & Reddy, P. S. (2012). Green synthesis of silver nanoparticles using Terminalia chebula extract at room temperature and their antimicrobial studies. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 91, 228-233. https://doi.org/10.1016/j.saa.2012.02.001
Laime-Oviedo, L. A., Soncco-Ccahui, A. A., Peralta-Alarcon, G., Arenas-Chávez, C. A., Pineda-Tapia, J. L., Díaz-Rosado, J. C. & Vera-Gonzales, C. (2022). Optimization of synthesis of silver nanoparticles conjugated with Lepechinia meyenii (Salvia) using Plackett‒Burman Design and Response Surface Methodology—preliminary antibacterial activity. Processes, 10(9), 1727. https://doi.org/10.3390/pr10091727
Laime-Oviedo, L. A., Arenas-Chávez, C. A., Yáñez, J. A. & Vera-Gonzáles, C. A. (2023). Plackett‒Burman design in the biosynthesis of silver nanoparticles with Mutisia acuminatta (Chinchircoma) and preliminary evaluation of its antibacterial activity. F1000Research, 12. https://doi.org/10.12688/f1000research.140883.1
Magudapathy, P., Gangopadhyay, P., Panigrahi, B. K., Nair, K. G. M. & Dhara, S. (2001). Electrical transport studies of Ag nanoclusters embedded in glass matrix. Physica B: Condensed Matter, 299(1-2), 142-146. https://doi.org/10.1016/S0921-4526(00)00580-9
Naveed M., BiBi J., Kamboh A. A., Suheryani I., Kakar, I., Fazlani, S. A., FangFang, X., Kalhoro, S. A., Yunjuan L, Kakar, M. U, Abd El-Hack, M. E., Noreldin, A. E., Zhixiang, S., LiXia, C. & XiaoHui, Z. (2018). Pharmacological values and therapeutic properties of black tea (Camellia sinensis): A comprehensive overview. Biomed Pharmacotherapy, 100, 521-531. https://doi.org/10.1016/j.biopha.2018.02.048
Nikaeen, G., Yousefinejad, S., Rahmdel, S., Samari, F. & Mahdavinia, S. (2020). Central composite design for optimizing the biosynthesis of silver nanoparticles using Plantago major extract and investigating antibacterial, antifungal and antioxidant activity. Scientific Reports, 10(1), 9642. https://doi.org/10.1038/s41598-020-66357-3
Noroozi, M., Zakaria, A., Moksin, M. M., Wahab, Z. A. & Abedini, A. (2012). Green formation of spherical and dendritic silver nanostructures under microwave irradiation without reducing agent. International Journal of Molecular Sciences, 13(7), 8086-8096. https://doi.org/10.3390/ijms13078086
Othman, L., Sleiman, A., & Abdel-Massih, R. M. (2019). Antimicrobial activity of polyphenols and alkaloids in Middle eastern plants. Frontiers in Microbiology,10, 911. https://doi.org/10.3389/fmicb.2019.00911.
Ovais, M., Khalil, A. T., Islam, N. U., Ahmad, I., Ayaz, M., Saravanan, M. & Mukherjee, S. (2018). Role of plant phytochemicals and microbial enzymes in biosynthesis of metallic nanoparticles. Applied Microbiology and Biotechnology, 102, 6799-6814. https://doi.org/10.1007/s00253-018-9146-7
Öz, M. (2022). Tilia rubra DC. subsp. caucasica V.Engler (Kafkas Ihlamuru) yaprak uçucu yağının kimyasal bileşimi. III. International Siirt Scientific Research Congress, Siirt, Türkiye, 18-19 November 2022, ss.14.
Özsoy, N., Can, A., Yanardag, R. & Akev, N. (2008). Antioxidant activity of Smilax excelsa L. leaf extracts. Food Chemistry, 110(3), 571-583. https://doi.org/10.1016/j.foodchem.2008.02.037
Özturk, B., Yıldız, K. & Küçüker, E. (2015). Effect of pre‐harvest methyl jasmonate treatments on ethylene production, water‐soluble phenolic compounds and fruit quality of Japanese plums. Journal of the Science of Food and Agriculture, 95(3), 583-591. https://doi.org/10.1002/jsfa.6787
Plackett, R. L. & Burman, J. P. (1946). The design of optimum multifactorial experiments. Biometrika, 33(4), 305-325. https://doi.org/10.1093/biomet/33.4.305
Reddy, L. V. A., Wee, Y. J., Yun, J. S. & Ryu, H. W. (2008). Optimization of alkaline protease production by batch culture of Bacillus sp. RKY3 through Plackett–Burman and response surface methodological approaches. Bioresource Technology, 99 (7), 2242-2249. https://doi.org/10.1016/j.biortech.2007.05.006
Sahan, Y. (2011). Effect of Prunus laurocerasus L. (Cherry laurel) leaf extracts on growth of bread spoilage fungi. Bulgarian Journal of Agricultural Science, 17 (1), 83-92.
Šamec, D., Urlić, B. & Salopek-Sondi, B. (2018). Kale (Brassica oleracea var. acephala) as a superfood: Review of the scientific evidence behind the statement. Critical Reviews in Food Science and Nutrition, 59(15), 2411-2422. https://doi.org/10.1080/10408398.2018.1454400
Shah, M., Fawcett, D., Sharma, S., Tripathy, S. K. & Poinern, G. E. J. (2015). Green synthesis of metallic nanoparticles via biological entities. Materials, 8(11), 7278-7308. https://doi.org/10.3390/ma8115377
Shameli, K., Bin Ahmad, M., Jaffar Al-Mulla, E. A., Ibrahim, N. A., Shabanzadeh, P., Rustaiyan, A. & Zidan, M. (2012). Green biosynthesis of silver nanoparticles using Callicarpa maingayi stem bark extraction. Molecules, 17(7), 8506-8517. https://doi.org/10.3390/molecules17078506
Sharma, N. K., Vishwakarma, J., Rai, S., Alomar, T. S., AlMasoud, N. & Bhattarai, A. (2022). Green route synthesis and characterization techniques of silver nanoparticles and their biological adeptness. ACS Omega, 7(31), 27004-27020. https://doi.org/10.1021/acsomega.2c01400
Siddiqi, K.S., Husen, A. & Rao, R.A.K. (2018). A review on biosynthesis of silver nanoparticles and their biocidal properties. Journal of Nanobiotechnology, 16, 14. https://doi.org/10.1186/s12951-018-0334-5
Siddiqui, A., Gul, A., Khan, H., Anjum, F. & Hussain, T. (2024). Bio-inspired synthesis of silver nanoparticles using Salsola imbricata and its application as antibacterial additive in glass ionomer cement. Nanotechnology, 35(35), 355101. https://doi.org/10.1088/1361-6528/ad50e4
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. https://doi.org/10.5344/ajev.1965.16.3.144
Singleton, V.L. (1999). Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. Methods in Enzymology, 299, 152-178.
Stepanov, A. L. (2004). Optical properties of metal nanoparticles synthesized in a polymer by ion implantation: a review. Technical Physics, 49, 143-153. https://doi.org/10.1134/1.1648948
Trivedi, P., Khandelwal, M. & Srivastava, P. (2014). Statistically optimized synthesis of silver nanocubes from peel extracts of Citrus limetta and potential application in waste water treatment. Journal of Microbial & Biochemical Technology, 4(004). https://doi.org/10.4172/1948-5948.S4-004
Tuttu, G., Ursavaş, S. & Söyler, R. (2017). Ihlamur çiçeğinin Türkiye’deki hasat miktarları ve etnobotanik kullanımı. Anadolu Orman Araştırmaları Dergisi, 3(1), 60-66
Türkkan, M. & Gürel, Y. (2024). Plackett‒Burman and Box‒Behnken Designs for optimizing Polygonum cognatum Meissn-mediated AgNP synthesis: Antifungal activity against diverse Phytophthora spp. Akademik Ziraat Dergisi, 13(2), 272-286. https://doi.org/10.29278/azd.1522321
Vijayaraghavan, K., Nalini, S. K., Prakash, N. U. & Madhankumar, D. J. M. L. (2012). Biomimetic synthesis of silver nanoparticles by aqueous extract of Syzygium aromaticum. Materials Letters, 75, 33-35. https://doi.org/10.1016/j.matlet.2012.01.083
Ye, M., Yang, W., Zhang, M., Huang, H., Huang, A. & Qiu, B. (2023). Biosynthesis, characterization, and antifungal activity of plant-mediated silver nanoparticles using Cnidium monnieri fruit extract. Frontiers in Microbiology, 14, 1291030. https://doi.org/10.3389/fmicb.2023.1291030
Yeşilada, E., Sezik, E., Honda, G., Takasihi, Y., Takeda, Y. & Tanaka, T. (1999). Traditional medicine in Turkey IX. Folk Medicine in Noth-West Anatolia. Journal of Ethnopharmacology, 64, 195- 210
Yiğit, U. & Türkkan, M. (2023). Antifungal activity and optimization procedure of microwave-synthesized silver nanoparticles using linden (Tilia rubra subsp. caucasica) flower extract. International Journal of Chemistry and Technology, 7(1), 25-37. https://doi.org/10.32571/ijct.1194356
Yi̇ği̇t, U., Gürel, Y., Ilhan, H. & Türkkan, M. (2023). Antifungal activity and optimization procedure of silver nanoparticles green synthesized with Prunus laurocerasus L. (Cherry laurel) leaf extract. International Journal of Life Sciences and Biotechnology, 6 (1), 1–20. https://doi.org/10.38001/ijlsb.1168628
Yilmaz, M., Yilmaz, A., Karaman, A., Aysin, F. & Aksakal, O. (2021). Monitoring chemically and green-synthesized silver nanoparticles in maize seedlings via surface-enhanced Raman spectroscopy (SERS) and their phytotoxicity evaluation. Talanta, 225, 121952. https://doi.org/10.1016/j.talanta.2020.121952

Details

Primary Language

English

Subjects

Phytopathology

Journal Section

Research Article

Authors

Muharrem Türkkan ^*
0000-0001-7779-9365
Türkiye

Early Pub Date

March 20, 2025

Publication Date

March 27, 2025

Submission Date

July 28, 2024

Acceptance Date

February 17, 2025

Published in Issue

Year 1970 Volume: 28 Number: 2

DOI

https://doi.org/10.18016/ksutarimdoga.vi.1523681

IZ

https://izlik.org/JA83KT66KB

Cite

RIS / Bibtex

APA

Türkkan, M. (2025). Utilizing Plackett–Burman and Box–Behnken Designs for Plant Extract–Based AgNP Synthesis Optimization: Unveiling Antifungal Potential Against Phytophthora Species. Kahramanmaraş Sütçü İmam Üniversitesi Tarım Ve Doğa Dergisi, 28(2), 516-534. https://doi.org/10.18016/ksutarimdoga.vi.1523681

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