Production of monoterpenoids and aroma compounds from cell suspension cultures of Camellia sinensis

Abstract

Cell suspension cultures of Camellia sinensis were established in 250 ml shake flasks. Flasks contained 50 ml liquid medium of either Murashige and Skoog (MS), N/5 MS or Heller medium containing different levels of 6-benzyladenine (BA) (0.05–2 mg l−1), 2,4-dichlorophenoxyacetic acid (2,4-D) (1–10 mg l−1), and sucrose (10–50 g l−1). Moreover, the pH of the medium was varied from 5.2–6.2. In addition, cultures were subjected to light irradiation as well as to complete darkness. Following optimization of aroma and terpenoid extraction methods, cell cultures were analyzed for the volatile compounds using GC/MS. A total of 43 compounds were identified using the micro SDE apparatus. Among the major monoterpenoids obtained were α-terpineol and nerol. Moreover, other high aroma-value compounds, including 2-ethyl hexanol, benzyl alcohol, benzene acetaldehyde, nonanal and phenylethylalcohol were also detected. The highest levels of these compounds were obtained from cell suspension cultures grown in MS medium containing 5 mg l−1 2,4-D, 1 mg l−1 BA and 30 g l−1 sucrose at pH of 5.8 with incubation in complete darkness.