Salvia officinalis L.'nin İn Vitro Kallus Kültürü ve Bazı Amino Asitlerin Rosmarinik Asit Birikimi Üzerine Etkisi

Öz

Bu çalışmada Salvia officinalis (adaçayı) bitkisinin çeşitli eksplantlarının in vitro kallus oluşturma yeteneği ve oluşan kalluslardaki rosmarinik asit üretimi araştırılmıştır. Kallus oluşumu için genç yapraklar (<1-1.5 cm), yapraklar (>1-1.5 cm), nodlar ve gövde parçaları olmak üzere 4 çeşit eksplant 0.8 mg/L 2.4-D, 0.5 mg/L NAA ve 2.0 mg/L BAP içeren MS ortamında kültüre alınmıştır. En yüksek kallus oluşum oranı (%100) genç yaprak eksplantlarından elde edilmiştir. 4 hafta sonra oluşan kalluslar 0.1 mg/L NAA+1.0 mg/L BAP içeren MS ortamına aktarılmış, ayrıca in vitro rosmarinik asit üretimini artırmak için besin ortamına L-tirozin (10 mg/L) ve L-fenilalanin (10 mg/L) eklenmiştir. Kalluslar bu ortamlarda 1 ve 2 ay boyunca kültüre alınmış ve adaçayı kalluslarındaki rosmarinik asit üretimi HPLC ile analiz edilmiştir. Amino asitlerin ortama eklenmesi kalluslarda rosmarinik asit üretimini önemli ölçüde arttırmıştır. Ayrıca sonuçlar amino asitler ve kallus kültür süreleri bakımından önemli ölçüde farklılıklar göstermiştir. Tirozinin adaçayı kalluslarında rosmarinik asit üretimini artırmada daha etkili olduğu gözlenmiştir. Hem tirozin hem de fenilalanin ile desteklenmiş ortamlarda 2 ay boyunca kültüre alınan kallusların, yalnızca 1 ay boyunca kültüre alınan gruba göre daha yüksek miktarda rosmarinik asit ürettiği belirlenmiştir.

Anahtar Kelimeler

• Adaçayı
• Kallus
• Rosmarinik asit
• L-tirozin
• L-fenilalanin

In Vitro Callus Culture of Salvia Officinalis L. and the Effect of Some Amino Acids on Rosmarinic Acid Accumulation

Abstract

This study investigated the in vitro callus formation ability of various explants obtained from the Salvia officinalis (common sage) and the production of rosmarinic acid in the resulting calli. Four types of explants, including young leaves (<1-1.5 cm), older leaves (>1-1.5 cm), nodes, and stem pieces, were cultured on MS medium containing 0.8 mg/l 2.4-D, 0.5 mg/l NAA, and 2.0 mg/l BAP to induce callus formation. The highest callus formation (100%) was achieved from young leaf explants. After 4 weeks, the formed calli were transferred to an MS medium containing 0.1 mg/l NAA+1.0 mg/l BAP, and in vitro rosmarinic acid production was enhanced by adding L-tyrosine (10 mg/l) and L-phenylalanine (10 mg/l) to the nutrient medium. The calli were cultured in these media for 1 and 2 months, and the production of rosmarinic acid in sage calli was analyzed using HPLC. The addition of amino acids to the medium significantly increased rosmarinic acid production in the calli. Furthermore, the results varied significantly with different amino acids and callus culture durations. Tyrosine was observed to be more effective in increasing rosmarinic acid production in sage calli. Moreover, calli cultured for 2 months in both tyrosine- and phenylalanine-supplemented media exhibited higher rosmarinic acid production compared to the control group cultured for only 1 month.

Keywords

• Common sage
• Callus
• Rosmarinic acid
• L-tyrosine
• L-phenylalanine

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