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

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

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

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