Optimization and Antifungal Activity of Silver Nanoparticles Synthesized Using the Leaf Extract of Corylus colurna L. (Turkish hazelnut)

Abstract

Objective: This study aims to optimize the production of microwave-synthesized silver nanoparticles (AgNPs) using Corylus colurna leaf extracts with response surface methodology based on the face-centered central composite design (FCCCD), characterize the synthesized nanoparticles by various spectroscopic and microscopic methods, and evaluate their antifungal effects on some Phytophthora species. Materials and Methods: In the study, the FCCCD of the response surface methodology was used to investigate the combined effect of three different synthesis variables (AgNO3 concentration, hazelnut leaf extract/AgNO3 ratio and reaction time) to achieve the maximum amount of AgNP at the spectral wavelength of 350-420 nm. The estimated area under the spectral curve was calculated with the help of Microsoft Office Excel program using a simple midpoint rule. Ultraviolet Visible (UV-Vis) Spectroscopy, Fourier Transform Infrared (FT-IR) Spectroscopy, and Transmission Electron Microscope (TEM) were used to characterize hazelnut leaf extract-AgNPs synthesized under optimum conditions. The antifungal activity of AgNPs was tested against six Phytophthora species (P. cactorum, P. capsici, P. cinnamomi, P. citrophthora, P. nicotianae and P. palmivora) under in vitro conditions, and the experiment was carried out with 6 replications. Results: UV-Vis spectroscopy revealed that typical surface plasmon resonance values of AgNPs synthesized under different conditions were in the wavelength range of 396 to 411 nm. Optimal AgNP production was achieved within the investigated range when the AgNO3 concentration, plant leaf extract/AgNO3 ratio, and reaction time were 5 mM, 0.1, and 90 seconds, respectively. FT-IR spectrum showed that AgNPs contain O–H, N–H, C=C, C–N, and C–O groups, and various compounds in hazelnut leaf extract play an important role in the synthesis of AgNPs. TEM analysis results revealed that AgNPs were in spherical form with an average size of 17.48 nm (Gaussian fit). Green synthesized AgNPs decreased mycelial growth of P. cactorum, P. capsici, P. cinnamomi, P. citrophthora, P. palmivora and P. nicotianae by 81.67%, 74.80%, 73.54%, 81.01%, 74.50%, and 62.39%, respectively. In addition, it was determined that the EC50 values of AgNPs varied between 118.58-292.56 µg ml-1, and the MIC values were above 340 µg ml-1. Conclusion: This study suggests that AgNPs synthesized by hazelnut leaf extract should be further investigated for use in combating diseases caused by Phytophthora species.

Keywords

• Corylus colurna
• silver nanoparticles
• face-centered central composite design
• Phytophthora
• toxicity

Corylus colurna L. (Türk Fındığı)’nin yaprak ekstraktı kullanılarak sentezlenen gümüş nanopartiküllerin optimizasyonu ve antifungal aktivitesi

Öz

Amaç: Bu çalışma, yüz merkezli merkezi kompozit tasarım (FCCCD)’a dayalı yanıt yüzey yöntemi (RSM) ile Corylus colurna yaprak ekstraktı kullanılarak mikrodalgada sentez edilen gümüş nanopartiküllerin (AgNP’lerin) üretimini optimize etmeyi, sentezlenen nanopartikülleri çeşitli spektroskopik ve mikroskobik yöntemlerle karakterize etmeyi ve bazı Phytophthora türleri üzerindeki antifungal etkilerini değerlendirmeyi amaçlamaktadır. Materyal ve Yöntem: Çalışmada, yanıt yüzey yönteminin FCCCD’i, 350-420 nm spektral dalga aralığında maksimum AgNP miktarını elde etmek için üç farklı sentez değişkeni (AgNO3 konsantrasyonu, fındık yaprak ekstraktı/AgNO3 oranı ve reaksiyon süresi)’nin birleşik etkisini araştırmak için kullanılmıştır. Spektral eğri altındaki tahmini alan basit bir orta nokta kuralı kullanılarak Microsoft Office Excel programı yardımı ile hesaplanmıştır. Optimum koşullar altında sentezlenen fındık yaprak ekstraktı-AgNP’leri karakterize etmek için Ultraviyole Görünür (UV-Vis) Spektroskopisi, Fourier Dönüşümlü Kızılötesi (FT-IR) Spektroskopisi ve Transmisyon Elektron Mikroskobu (TEM) kullanılmıştır. AgNP’lerin antifungal etkinliği, altı Phytophthora türü (P. cactorum, P. capsici, P. cinnamomi, P. citrophthora, P. nicotianae ve P. palmivora)’ne karşı in vitro koşullarda denenmiş olup, deneme 6 tekerrürlü olarak yürütülmüştür. Araştırma Bulguları: UV-Vis spektroskopisi, farklı koşullar altında sentezlenen AgNP’lerin tipik yüzey plazmon rezonans değerlerinin 396 ile 411 nm dalga boyu aralığında değiştiğini ortaya koymuştur. AgNO3 konsantrasyonu, bitki yaprak ekstraktı/AgNO3 oranı ve reaksiyon süresi sırasıyla 5 mM, 0.1 ve 90 saniye olduğunda, araştırılan aralıkta optimum AgNP üretimi elde edilmiştir. FT-IR spektrumu, AgNP’lerin O–H, N–H, C=C, C–N ve C–O gruplarını içerdiğini ve fındık yaprak ekstraktındaki çeşitli bileşiklerin AgNP’lerin sentezinde önemli bir rol oynadığını göstermiştir. TEM analiz sonuçları, AgNP’lerin ortalama 17.48 nm (Gauss uyumu) büyüklüğe sahip küresel formda olduğunu ortaya koymuştur. Yeşil sentezlenen AgNP’lerin P. cactorum, P. capsici, P. cinnamomi, P. citrophthora, P. palmivora ve P. nicotianae’nın misel gelişimini sırasıyla %81.67, %74.80, %73.54, %81.01, %74.50 ve %62.39’a kadar azaltmıştır. Ayrıca AgNP’lerin EC50 değerlerinin 118.58-292.56 µg ml-1 arasında değiştiği ve MIC değerlerinin ise 340 µg ml-1’in üzerinde olduğu belirlenmiştir. Sonuç: Bu çalışma, fındık yaprak ekstraktı ile sentezlenen AgNP’lerin, Phytophthora türlerinin neden olduğu hastalıkların mücadelesinde kullanılmak üzere daha fazla araştırılması gerektiğini önermektedir.

Anahtar Kelimeler

• Corylus colurna
• Gümüş nanopartikül
• Yüz Merkezli Merkezi Kompozit Tasarım
• Phytophthora
• toksisite

Kaynakça

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