Asmada Hücre Süspansiyon Kültürlerine Elisitör Uygulamaları ile Fenolik Bileşiklerin Üretilmesi

Bu araştırmada asmada hücre süspansiyon kültürlerinde fenolik bileşiklerin üretiminde kadmiyum sülfat CdSO4 floresan radyasyonu, metil jasmonat MeJA ve sukroz uygulamalarının etkileri incelenmiştir. Gamay, Kalecik karası ve Öküzgözü üzüm çeşitlerine ait yaprak saplarından elde edilen kallus hücre süspansiyon kültürlerine CdSO4 0, 1 ve 1.5 mM , MeJA 0 ve 10 µM ve sukroz 0, 0.20 ve 0.25 M uygulanmıştır. Floresan radyasyonu için hücre süspansiyonları tümüyle karanlıkta ya da 10000 lux ışık altında tutulmuştur. Toplam fenolik, toplam flavanol, toplam flavonol ve antosiyanin içerikleri spektrofotometrik olarak; trans-resveratrol içeriği ise HPLC ile belirlenmiştir. 1.5 mM konsantrasyonundaki CdSO4 ve 10 µM konsantrasyonundaki MeJA bütün çeşitlerde en yüksek fenolik bileşik üretimini sağlamıştır. Özellikle 1.5 mM CdSO4 uygulanmış Kalecik karası en yüksek toplam fenolik madde 3.144 mg g-1 , antosiyanin 1.672 CV g-1 ve trans-resveratrol 3.650 µg g-1 içeriğine sahip olmuştur. 10 µM konsantrasyonundaki MeJA uygulaması ise Öküzgözü çeşidinde trans-resveratrol miktarının 11.681 µg g-1 gibi yüksek bir değere çıkmasını sağlamıştır. Sukroz uygulamaları içinde 0.20 M dozu en yüksek toplam fenolik 4.215 mg g-1 ve trans-resveratrol 7.550 µg g-1 miktarını Kalecik karasında, en yüksek antosiyanin birikimini ise Gamay çeşidinde sağlamıştır. Karanlık uygulaması bütün çeşitlerde trans-resveratrol birikimini kuvvetli bir şekilde artırmıştır. Toplam fenolik ve antosiyanin sentezinin ışık tarafından uyarıldığı belirlenmiştir. Sonuçlar CdSO4 , MeJA, sukroz ve floresan radyasyonu gibi elisitör uygulamalarının üzümde fenolik bileşiklerin üretiminde önemli bir yaklaşım olabileceğini göstermiştir

Anahtar Kelimeler:

Asma, Hücre süspansiyonu, Kadmiyum, Metil jasmonat, Antosiyanin, trans-resveratrol

Elicitor Applications to Cell Suspension Culture for Production of Phenolic Compounds in Grapevine

In this study, the effects of cadmium sulphate CdSO4 , fleuresans irradiation, methyl jasmonate MeJA and sucrose treatments on the production of phenolic compounds in grapevine cell suspension cultures initiated from callus from petiole tissues of Vitis vinifera L. cvs. Gamay, Kalecik karası and Öküzgözü were investigated. As the elicitors of CdSO4 0, 1 and 1.5 mM , MeJA 0 and 10 µM and sucrose 0, 0.20 and 0.25 M were applied. Cell suspensions were exposed to visible light 10,000 lux for fleuresans irradiation or cultured in dark constantly. Total phenolics, total flavanols, total flavonols and anthocyanin were determined spectrophotometrically while trans-resveratrol was quantified by HPLC. CdSO4 at 1.5 mM concentration and MeJA at 10 µM concentration yielded the highest phenolic productions in all cultivars. Especially, Kalecik Karası treated with CdSO4 at 1.5 mM had the highest total phenolic 3.144 mg g-1 , anthocyanin 1.672 CV g-1 and trans-resveratrol 3.650 µg g-1 contents. MeJA application at 10 µM provided the transresveratrol accumulation as high as 11.681 µg g-1 in Öküzgözü. 0.20 M sucrose concentration resulted in the highest total phenolics 4.215 mg g-1 and trans-resveratrol 7.550 µg g-1 in Kalecik Karası cultures while the most anthocyanin accumulation 2.024 CV g-1 was achieved from Gamay. Darkness had strongly increased trans-resveratrol content in all cultivars, whereas total phenolics and anthocyanin synthesis were induced by light. Elicitor applications of CdSO4, MeJA, sucrose and fleuresans irradiation can be an efficient approach for the production of phenolics in grapevines

Keywords:

Grapevine, Cell suspension, Cadmium, Methyl jasmonate, Anthocyanin, trans-resveratrol,

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