Integrated Analysis of the In Vitro Response of Melissa officinalis subsp. officinalis to Copper Oxide Nanoparticles

Year 2025, Volume: 13 Issue: 1, 179 - 191, 30.06.2025

Ersan Bektaş , Hüseyin Şahin , Halil İbrahim Güler , Kadriye İnan , Kaan Kaltalıoğlu , Nilhan Elif Uzun

https://doi.org/10.18586/msufbd.1706381

Abstract

This study explores the in vitro effects of copper oxide nanoparticles (CuO-NPs) on Melissa officinalis L. subsp. officinalis (lemon balm) by evaluating morphological, biochemical, and molecular responses. Seedlings were treated with CuO-NPs at 0.1, 1.0, and 10.0 mg/L to assess growth parameters, secondary metabolite production, antioxidant activity, enzyme inhibition, essential oil composition, and gene expression of PAL, TAT, and RAS. Low CuO-NP concentration (0.1 mg/L) significantly enhanced shoot elongation, while 10.0 mg/L induced strong root proliferation but inhibited shoot growth. Total phenolic (TPC) and flavonoid (TFC) contents peaked at 10.0 mg/L, correlating with increased antioxidant activity in CUPRAC, DPPH, and ABTS assays. HPLC analysis revealed elevated rosmarinic acid and reduced caffeic and p-coumaric acid levels, suggesting pathway redirection. Enzyme inhibition assays showed that 1.0 mg/L yielded the strongest AChE inhibition, while higher doses improved inhibition of MAO, urease, and HIV-1 RT. GC-MS analysis indicated altered essential oil profiles under CuO-NP exposure, including increased geranial and neral levels. qRT-PCR confirmed dose-dependent upregulation of PAL, TAT, and RAS genes, particularly at 10.0 mg/L, supporting enhanced phenylpropanoid biosynthesis. Collectively, CuO-NPs act as abiotic elicitors in a concentration-dependent manner, promoting bioactive compound production and antioxidant potential. These findings highlight the potential of CuO-NPs in improving the phytochemical quality of Melissa officinalis under controlled conditions.

Keywords

Antioxidant activity , CuO Nanoparticles , Gene expression , In vitro propagation , Melissa officinalis subsp. officinalis , Phenolic compounds

Supporting Institution

TUBITAK

Project Number

121Z818

Thanks

This study was financially supported by TUBITAK under Project No. 121Z818, and the authors gratefully acknowledge this support.

Melissa officinalis subsp. officinalis’in Bakır Oksit Nanoparçacıklarına Karşı İn Vitro Yanıtının Bütüncül Analizi

Abstract

Bu çalışma, bakır oksit nanoparçacıklarının (CuO-NP) Melissa officinalis L. subsp. officinalis (oğulotu) üzerindeki in vitro etkilerini morfolojik, biyokimyasal ve moleküler yanıtlar açısından değerlendirmektedir. Fideler 0.1, 1.0 ve 10.0 mg/L CuO-NP konsantrasyonlarıyla muamele edilerek büyüme parametreleri, sekonder metabolit üretimi, antioksidan aktivite, enzim inhibisyonu, uçucu yağ bileşimi ve PAL, TAT ve RAS genlerinin ekspresyonu analiz edilmiştir. Düşük CuO-NP konsantrasyonu (0.1 mg/L), sürgün uzamasını anlamlı şekilde artırırken, 10.0 mg/L kök gelişimini kuvvetli şekilde teşvik etmiş, ancak sürgün büyümesini baskılamıştır. Toplam fenolik (TPC) ve flavonoid (TFC) içerikleri 10.0 mg/L’de en yüksek değerlere ulaşmış; bu durum CUPRAC, DPPH ve ABTS testlerinde artan antioksidan aktivite ile ilişkilendirilmiştir. HPLC analizleri, rosmarinik asitte artış, kafeik ve p-kumarik asitlerde ise azalma göstermiş; bu da fenilpropanoid yolunun yön değiştirdiğine işaret etmiştir. Enzim inhibisyon analizlerinde, 1.0 mg/L konsantrasyonu AChE üzerinde en yüksek inhibisyon etkisi gösterirken, daha yüksek dozlar MAO, üreaz ve HIV-1 RT enzimlerinin inhibisyonunu artırmıştır. GC-MS analizleri, CuO-NP uygulaması altında uçucu yağ profillerinin değiştiğini ve özellikle geranial ve neral düzeylerinde artış olduğunu göstermiştir. qRT-PCR analizleri, özellikle 10.0 mg/L’de PAL, TAT ve RAS genlerinin doz bağımlı olarak yukarı regüle olduğunu doğrulamış ve fenilpropanoid biyosentezinin arttığını desteklemiştir. Genel olarak, CuO-NP’ler konsantrasyona bağlı şekilde abiyotik elisitör olarak işlev görmekte, biyolojik aktif bileşik üretimini ve antioksidan kapasiteyi artırmaktadır. Bu bulgular, kontrollü koşullarda Melissa officinalis bitkisinin fitokimyasal kalitesinin iyileştirilmesinde CuO-NP’lerin potansiyelini ortaya koymaktadır

Keywords

Antioksidan aktivite , CuO Nanopartikül , Fenolik bileşikler , Gen ekspresyonu , İn vitro propagasyon , Melissa officinalis subsp. officinalis

Supporting Institution

TÜBİTAK

Project Number

121Z818

Thanks

Bu çalışma TÜBİTAK tarafından 121Z818 numaralı proje kapsamında maddi olarak desteklenmiştir ve yazarlar bu destekten dolayı minnettardır.

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