Medium Optimization and Investigation of Phenolic Contents in Cell Suspension Culture of Hypericum retusum Auche

Abstract

Hypericum retusum has a potential value as a medicinal plant. In this study, we investigated the
effects of different combination of 6-benzyladenine (BAP) and kinetin (KIN) with 2,4-dichlorophenoxyacetic acid
(2,4-D), different sucrose concentrations (15, 30 and 50 g L-1) and different light conditions (continuous dark,
continuous light and control) and initial pH values (pH 4.5, 5.8 ve 6.5) on the accumulation of biomass and the
production of fve pharmacologically important phenolic constituents (hypericin, hyperoside, chlorogenic acid,
quercetin and pseudohypericin) in cell suspension culture of H. retusum. Our data suggested that the best biomass
accumulations obtained from on MS medium supplemented with 0.5 mg L-1 KİN+1.0 mg L-1 2,4-D, 30 g L-1
sucrose, 5.8 pH and under the darkness conditions in cell suspension culture of H. retusum. The highest phenolic
compound contents were obtained from on MS medium containing 0.1 mg L-1 BAP+ 0.5 mg L-1 2,4-D at and 15
g L-1 sucrose. The phenolic compound contents of suspension culture under the different light conditions varied
among the treatments. With this study, we have developed a novel cell suspension culture system for H. retusum,
suggesting a possible technology for large-scale production of phenolics.

Keywords

• Cell suspension culture,Hypericum retusum Aucher,Phenolic compounds,LC-MS/MS

Hypericum retusum Aucher’in Hücre Süspansiyon Kültürlerinin Optimizasyonu ve Fenolik Bileşen İçeriğinin İncelenmesi

Abstract

Hypericum retusum tıbbi bitki olarak potansiyel bir öneme sahiptir. Bu çalışmada, 6-benziladenin (BAP) ve
kinetin (KİN)’in 2,4-diklorofenoksiasetik asit (2,4-D) ile kombinasyonlarının, farklı sukroz konsantrasyonlarının
(15, 30 ve 50 g L-1), farklı ışık uygulamalarının (Sürekli karanlık, Sürekli aydınlık ve kontrol) ve farklı başlangıç pH
değerlerinin (pH 4.5, 5.8 ve 6.5), H. retusum’ un hücre süspansiyon kültürlerinde biyokütle birikimi ve farmakolojik
olarak önemli beş adet fenolik bileşiğin (hiperisin, hiperosid, klorogenik asit, kuersetin ve psödohiperisin) miktarları
üzerine etkisini araştırdık. Verilerimiz, H. retusum’ un hücre süspansiyon kültürlerinde en iyi biyokütle birikiminin,
0.5 mg L-1 KİN+ 1.0 mg L-1 2,4-D ve 30 g L-1 ilaveli MS besi ortamında, pH’sı 5.8 ve sürekli karanlık şartlar altında
olan ortamdan elde edildiğini ortaya koymuştur. En yüksek fenolik bileşik içeriği, 0.1 mg L-1 BAP+ 0.5 mg L-1
2,4-D ve 15 g L-1 sukroz ilaveli ve pH’sı 4.5 olan ortamdan elde edilmiştir. Farklı ışık uygulamalarına tabi tutulan
kültürlerdeki fenolik bileşen içeriği ise uygulamalar arasında farklılık göstermiştir. Bu çalışmayla, H. retusum
için, fenolik bileşiklerin geniş ölçekli üretimi için muhtemel bir teknoloji sunan yeni bir hücre süspansiyon kültür
sistemi geliştirdik.

Keywords

• Fenolik bileşik,Hücre süspansiyon kültürü,Hypericum retusum Aucher,LC-MS/MS

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