The indoline alkaloids accumulation by Rauwolfia serpentina cell lines upon surface and submerged maintenance

Abstract

The indoline al ka loids ac cu mu la tion by R. serpentina cell lines upon sur face and sub merged main te nanceThe In sti tute of Mo lec u lar Bi ol ogy and Ge net ics, NAS of Ukraine 150, Ac a de mi cian Zabolotny Str., Kyiv, 03143, Ukraine kunakh@imbg.org.ua Indoline al ka loids ac cu mu la tion, in clud ing ajmaline, in the bio mass of R. serpentina cell line cul ture on the ex am ple of hor mone in de pend ent highly pro duc tive strain K 27 at its sur face and sub merged main te nance in nu tri ent me dia with dif fer ing min eral com po si tion has been stud ied. Op ti mal nu tri ent me dia for both ways of main te nance have been iden ti fied. The con di tions for two step main te nance in sur face, fol lowed by sub merged cul tures, in compositionally sim ple nu tri ent me dia have been spec i fied. Two step main te nance in creases al ka loids ac cu mu la tion at the early stages of growth 3 4 times and al lows de creas ing the time of cal lus tis sue grow ing from 60 80 days to 20 40 days. Keywords: ajmaline, indoline al ka loids, plant tis sue cul ture, R. ser pen tine, cell lines -al ka loids producents. Primary CallusObtained by B. G. Butenko on MC medium according to [2], green body fragment of 5 year old plant, 1964 Callus Tissue MC medium, total alkaloid content 0.1-0.3 %, ajmaline 0.04 % (1964-1968) [3] Mutagen treatment by nitrogenous yperite in 1968 M1 Mutant Line MC medium, total alkaloid content 0.2-0.48 %, ajmaline app. 0.2 % [4]Selection on nutrition medium 5C, without phytohormones according to [5] (1972-1975) Cell Line A 5C medium without phytohormones, indoline alkaloid content 0.7-0.8 %, ajmaline app. 0.4 % Mutagen treatment by ethylenimine in 1981 and the adaptation to growth on 10C medium, without phytohormones according to [8] in the course of [1982][1983][1984][1985] K 27 Strain 10C medium, indoline alkaloid content 1.2-1.8 %, ajmaline 0.9-1.2 %

Comprehensive study of hormone-independent highly productive strain of Rauvolfia serpentina tissue culture as a source of indole alkaloids

“…The genealogy of the К-27 strain of R. serpentina, the details of its establishment and culturing along with the characteristics and peculiarities were reported earlier [3,[7][8][9]. Over more than 35 years, this strain was stable in terms of both productivity, genetic and cytological characteristics, being grown on both liquid and solid media of various composition with or without different organic additives [16,17,26,[18][19][20][21][22][23][24][25].…”

“…We have previously shown that the amount of DNA in cultured cells increases during a subculture cycle when they switch to alkaloid biosynthesis [29]. Obviously, in mixoploid cell populations of R. serpentina culture in vitro, there may be a dependence of strain productivity, in particular indoline alkaloid biosynthesis, on the ploidy of cells that formed the modal class [30]. However, other researchers found a negative correlation between the chromosome number and the accumulation of secondary metabolites in cultured R. serpentina cells [31].…”

“…Cell biomass of the K-27M strain R. serpentina tissue culture derived by the cell selection methods from the K-27 strain, which was maintained in vitro for more than 35 years, was studied. The details of establishment and charac teristics of the initial K-27 strain were published earlier [3,[16][17][18][19][20][21][22][23][24][25][26][27][28]. The newly established K-27M strain was cultured under thermostatic conditions at 27-28 °C in the dark, in 370 mL glass vessels.…”

Dynamics of changes in the genome of callus tissues of Rauwolfia serpentina upon a switch to conditions of submerged cultivation

“…Results of a study of the build-up of indole alkaloids and the overall productivity of the K-27 strain when grown under different conditions (surface and submerged cultivation) on different types of nutrient media have been published previously [5,6]. It was shown that accelerated buildup of alkaloids occurs under specially selected conditions of submerged cultivation; this, in turn, makes it possible to reduce the time needed to culture the tissues prior to harvesting.…”