Selection of protoplasts-derived cell lines for high yields indole alkaloid from suspension cultures of Vinca(Catharanthus roseus) = 일일초(Catharanthus roseus) 현탁배양으로부터 원형질체유래 indole

Abstract

To produce economically important indole alkaloids by cell cultures, we selected protoplasts--derived clc nes (protoclones) of vinca (Catharanthus roseus、for high yields of catharanthine and ajmalicine. Protoplasts were enzymatically isolated from suspension - cultured cells. The highest plating effi iency (1%) was obtained when protoplasts were plated at a density of 1X105 protoplasts / m in a culture medium solidified with 0.4% Seaplaque agarose. The growth rates of 40 protoclc les subcultured on a solid medium varied over a wide range. Protoclone VPC-6, which had she highest growth rate, was observed to produce relatively high yields of catharanthine and ajmalicine when cultured in a liquid medium. Although the original cell line did not produce cat laranthine at a detectable level by HPLC, protoclone VPC-10 produced it at a level of 5.9서g / g fresh weight of cells for 10 days of culture. Under the same conditions, protoclone VPC-15 pr< duced ajmalicine at a level of 133.6a授 /g, of which productivity was improved about 3 times th; n that of the original cell line. The results indicate that differences in the growth rate and ir dole alkaloid yield among the protoclones reflect the somaclonal variation in suspe-nsion-cultu ed cells.