Quinoline Alkaloid Production by Transformed Cultures ofCinchona ledgeriana

Abstract

A culture of CINCHONA LEDGERIANA, transformed with AGROBACTERLUM TUMEFACIENS A6 capable of growing and producing quinoline alkaloids in medium free of exogenous phytohormones has been obtained. Unlike the untransformed culture of this species, addition of ZR in combination with either IAA or IBA to medium did not affect alkaloid production. Growing the transformed culture in the dark, however, produced a marked enhancement of alkaloid accumulation, up to 50 times that of cultures grown in the light. This dark-dependent accumulation was not confined to any particular time in the growth cycle, although the extent of the stimulatory effect increased the longer cultures were kept in darkness. Blue light was detrimental to alkaloid accumulation but in red or green light the level of accumulation was equivalent to that in the dark. Alternating cultures for several 28-day periods between light and dark conditions resulted in alternate periods of low and higfralkaloid productivity, indicating this was not an adaption effect. These findings correlate to previously reported differences in key enzymes from the biosynthetic pathway. Increasing the phosphate or nitrate concentration above that of Gamborg B5, increasing sucrose above 2% (w/v), replacing sucrose with glucose or adding tryptophan or casamino acids to the medium resuited in lower alkaloid yields by dark-grown cultures of the transformed line.

Production of Alkaloids in Plant Cell Culture

“…ledgeriana cultures transformed with Agrobacterium tumefaciens (Payne et al, 1987b) were also capable of producing these quinoline alkaloids without the addition of phytohormones; however, the type and amount of alkaloid did not seem to differ from those reported for the untransfonned lines of C. ledgeriana. Quinine and quinidine are of commercial importance: quinine is used as an antimalarial drug and as a bitter flavoring in the soft drink industry, while quinidine is used to treat cardiac arrhythmias.…”

“…where alkaloid production is associated with the chloroplast (Wink et al, 1983). However, quinoline alkaloid production, particularly quinine in Cinchona cells transformed by Agrobacterium tumefaciens, is increased 10-to 12-fold if cells are grown in the dark (Payne et al, 1987b).…”

Enhanced Production Level of Quinine in Cell Suspension Culture of Cinchona ledgeriana Moens by Paclobutrazol

“…could solve the above problem. Several groups of researchers have attempted enhancement of cinchona alkaloid production in cell suspension cultures by using stressing agents, precursors, elicitors, auxins, and enzymes (Harkes et al 1986, Payne et al 1987, Toruan-Mathius et al 2006. Previous research on C. ledgeriana clone QRC 313 increased quinine content to 0.12%, which was 11-fold more than that in control cells (Ratnadewi & Sumaryono 2010).…”

Assessment of inter- and intra-population variation in quinoline alkaloid profile of juvenile shoot cultures of Cinchona ledgeriana: effect of method of nutrient delivery on alkaloid profile

“…In recent years, various studies have focussed on the potential of technologies in vitro such as callus (Mulder-Krieger et al 1982a,b;Scragg et al 1986), suspension (Anderson et al 1982;Rhodes et al 1986;Robins et al 1986;Wijnsma et al 1986), whole-tissue (Staba & Chung 1981;Anderson et al 1982;Chung & Staba 1987) and transformed-organ (Payne et al 1987) culture to produce Cinchona -derived quinoline alkaloids. The results of these and related studies (see below) clearly indicate that alkaloid production by Cinchona spp.…”