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Thymol production in hairy root culture of Sahendian savory (Satureja sahendica Bornm)
Bahmani Helia,Maroufi Asad,Majdi Mohammad,Fakheri Barat Ali 한국식물생명공학회 2021 Plant biotechnology reports Vol.15 No.2
Sahendian savory (Satureja sahendica Bornm) is a medicinal plant species which produces valuable secondary metabolites including thymol, p-cymene and γ-terpinene. In this study, optimization of hairy roots culture in S. sahendica using Agrobac�terium rhizogenes was efciently established. High frequency of genetic transformation was obtained using node and inter�node explants and three bacterial strains (ATCC 15834, A4, and LBA 9402). However, leaf explants showed no susceptibility to induce hairy roots with the A. rhizogenes strains. The node explants and LBA 9402 strain were the best combination for root induction. Infection time of 20 min caused greater transformation occurrence compared to 10 and 30 min, respectively. Node explants infected with LBA 9402 were found to produce more transformants upon co-cultivation for 2 days. The maxi�mum induction of hairy roots was obtained from bacterial infection at a density of OD600=0.8. Polymerase chain reaction by gene-specifc primers from rol B gene confrmed the positive transformation events. S. sahendica efciently provided and produced a large number of hairy roots which could be used for the production of secondary metabolites or other molecular and biotechnological applications. Production of thymol an important secondary metabolite of S. sahendica from hairy root cultures was confrmed by TLC and GC analysis. Elicitor-treated hairy root cultures with methyl jasmonate and ammonium nitrate produced more amount of thymol compared to untreated roots. These results demonstrate that secondary metabolite production such as thymol in hairy root culture of Sahendian savory could be used to increase low secondary metabolite production in this valuable medicinal plant.