Influence of Carbon Source, Nitrogen and PEG on Caffeic Acid Derivatives Production in Callus Cultures of Echinacea purpurea L.

Abstract

This study aimed to determine the effects of polyethylene glycol (PEG) as an abiotic elicitor and nutritional factors (different ammonium/nitrate ratios, carbon source and amount) in the culture medium on the production of Caffeic Acid Derivatives (CADs) in callus cultures of Echinacea purpurea L. Petiole and root explants were cultured on MS medium modified in terms of different types (sucrose and maltose) and amounts (sucrose 15, 45, 60 g l-1, and maltose 15, 30, 45, 60 g l-1) of carbon source, different concentrations (5, 10, 15 g l-1) of PEG and ammonium nitrate ratios (0:35, 5:25, 15:15, 35:0 mM). The amounts of CADs in the callus obtained at the end of the 10-week culture period were analysed. In both explant types, the highest amount of CADs were obtained from the medium containing 15 g l-1 sucrose and 15 or 30 g l-1 maltose applications, while the highest amount of CADs was obtained in the medium containing 0:35 mM ammonium/nitrate in nitrogen applications. While the highest amount of CADs in root explant was obtained from the medium containing 10 g l-1 PEG applications, CADs content could not be obtained in petiole explant. As a result, the highest amounts of caftaric, chlorogenic, caffeic, and chicoric acids (respectively, 9.38, 0.71, 0.29, and 34.77 mg g-1) were determined at callus obtained from root explant cultured on MS medium containing 30 g l-1 sucrose and 0:35 mM ammonium/nitrate. In conclusion, optimization of culture conditions and different elicitor applications were made to increase secondary metabolite content in E. purpurea L. under in vitro conditions and the results obtained were presented comparatively.

Keywords

• Sucrose
• Maltose
• Nitrogen
• PEG
• Caffeic acid derivatives
• Echinacea purpurea L.
• HPLC

Echinacea purpurea L. Kallus Kültürlerinde Karbon Kaynağı, Nitrojen ve PEG'in Kafeik Asit Türevlerinin Üretimine Etkisi

Abstract

Bu çalışmada, abiyotik bir elisitör olarak polietilen glikolün (PEG) ve besin ortamındaki farklı beslenme faktörlerinin (farklı amonyum/nitrat oranları, karbon kaynağı ve miktarı) Echinacea purpurea L. kallus kültürlerinde kafeik asit türevlerinin miktarlarına etkisinin belirlenmesi amaçlanmıştır. Petiyol ve kök eksplantları karbon kaynağının farklı türleri (sakkaroz ve maltoz) ve miktarları (sakkaroz 15, 45, 60 g l-1 ve maltoz 15, 30, 45, 60 g l-1), farklı amonyum nitrat oranları (0:35, 5:25, 15:15, 35:0 mM) ve farklı konsantrasyonlarda (5, 10, 15 g l-1) PEG içeren MS besin ortamlarında 10 hafta boyunca kültür alınmış ve kültür süresi sonunda elde edilen kalluslarda kaftarik, klorojenik, kafeik ve kikorik asit miktarları tespit edilmiştir. Her iki eksplant türünde de en yüksek kafeik asit türevlerinin miktarları, karbon kaynağı uygulamalarında, 15 g l-1 sakkaroz ile 15 ve 30 g l-1 maltoz; nitrojen uygulamalarında ise 0:35 mM amonyum/nitrat içeren besin ortamlarında uyarılan kalluslarda tespit edilmiştir. PEG uygulamalarında ise, en yüksek miktarlarda kafeik asit türevleri, 10 g l-1 PEG içeren besin ortamından kök eksplantından uyarılan kalluslarda elde edilirken, petiyol eksplantından elde edilen kallus dokularında kafeik asit türevlerinden hiçbiri tespit edilememiştir. E. purpurea L kallus kültürlerinde, kafeik asit türevlerinin miktarını artırmak için, in vitro şartlar altında, abiyotik elisitör olarak PEG ve farklı beslenme faktörlerinin optimizasyonu araştırıldığı bu çalışmada, elde edilen sonuçlar karşılaştırmalı olarak sunulmuştur. Tüm uygulamalar değerlendirildiğinde; sonuç olarak, en yüksek kaftarik, klorojenik, kafeik ve kikorik asit miktarları (sırasıyla 9.38, 0.71, 0.29 ve 34.77 mg g-1) 0:35 mM amonyum/nitrat ve 30 g l-1 sakkaroz içeren MS ortamında kültüre alınan kök eksplantından elde edilen kallus dokularında tespit edilmiştir.

Keywords

• Sakkaroz
• Maltoz
• Azot
• PEG
• Kafeik asit türevleri
• Echinacea purpurea L.
• HPLC

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