Document Type : Original Article
Authors
1
Plant Biotechnology Department, Biotechnology Research Institute, National Research Centre (NRC), Dokki, Giza 12622, Egypt.
2
Department of Plant Biotechnology, Biotechnology Research Institute, National Research Centre
3
Plant Biotechnology Department, National Research Centre, Cairo, Egypt
4
Plant biotechnology Department, Biotechnology Research Institute, National Research Centre
5
Department of Plant Biotechnology, National Research Centre
Abstract
Catharanthus roseus is a perennial small herb that produces terpenoid indole alkaloids (TIAs) of high medicinal importance. Carbon source is an absolute requirement for growth, development, and secondary metabolite production in plant tissue culture systems. TIAs are found in limited quantities within the plant, and their commercial demand is rapidly increasing. This study aims to evaluate the effects of different types and concentrations of carbon sources on alkaloid accumulation in hairy root cultures of C. roseus. Leaf explants were infected with Agrobacterium rhizogenes, strain ATTCC 15834, and several parameters were investigated for the maximum efficiency of the transformation protocol. In addition, the effects of sucrose, glucose and fructose on the accumulation of vincristine, vinblastine, ajmalicine and catharanthine in hairy root cultures were analyzed by HPLC. The results indicated 30-min period as the best time for inoculating the explants with A. rhizogenes. Maximum transformation efficiency was obtained with the supplementation of acetosyringone at a concentration of 100 µM into the co-cultivation media. Successful transformation was confirmed by polymerase chain reaction (PCR) with the RolA gene serving as an indicator of effective integration. While fructose at a concentration of 30 g/L significantly enhanced the levels of vinblastine, ajmalicine and catharanthine on hairy root cultures of C. roseus, sucrose at a concentration of 30 g/L was superior over other types of carbon sources for the maximum production of vincristine. These results offer insights into the optimal growth conditions, particularly the appropriate carbon source and provide a basis for further research aiming at enhancing commercial production of TIAs.
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