Colchicine Mutagenesis from Long-term Cultured Adventitious Roots Increases Biomass and Ginsenoside Production in Wild Ginseng (Panax ginseng Mayer)

Abstract

Panax ginseng Mayer is a perennial herb that has been used as a medicinal plant in Eastern Asia for thousands of years. The aim of this study was to enhance root biomass and ginsenoside content in cultured adventitious roots by colchicine mutagenesis. Adventitious P. ginseng roots were treated with colchicine at different concentrations (100, 200, and 300 mg·L−1) and for different durations (1, 2, and 3 days). Genetic variability of mutant lines was assessed using random amplification of polymorphic DNA (RAPD) analysis. Ginsenoside biosynthesis gene expression, ginsenoside content, enzyme activities, and performance in bioreactor culture were assessed in four mutant lines (100–1-2, 100–1-18, 300–1-16, and 300–2-8). The results showed that ginsenoside productivity was enhanced in all mutant lines, with mutant 100–1-18 exhibiting the most pronounced increase (4.8-fold higher than the control). Expression of some ginsenoside biosynthetic enzymes was elevated in mutant lines. Enzyme activities varied among lines, and lipid peroxidation activity correlated with root biomass. All four lines were suitable for bioreactor cultivation, with mutant 100–1-18 exhibiting the highest biomass after culture scale-up. The results indicated that colchicine mutagenesis of P. ginseng roots increased biomass and ginsenosides production. This technique, and the root lines produced in this study, may be used to increase industrial yields of P. ginseng biomass and ginsenosides.

Polyploidy in Industrial Crops: Applications and Perspectives in Plant Breeding

“…Polyploids of Panax ginseng Mayer formed thicker roots with more abundant and shorter lateral roots than the control plants' roots. The root biomass obtained after six weeks of culture in the bioreactor was about 1.5 times higher for mutant roots than for the control roots [114]. Tetraploids of E. purpurea also produced fewer, thicker roots with more lateral roots compared to diploids [76,115].…”

“…To date, many studies have been conducted on the effects of polyploidisation on the production by plants of desirable compounds used in the pharmaceutical industry [3,90,105,[110][111][112]115,127,129] and/or the cosmetic industry [3,109,114,130] or as natural insecticides [79].…”

Induced autopolyploidy—a promising approach for enhanced biosynthesis of plant secondary metabolites: an insight

“…However, low doses of colchicine were added to the media so as to prevent long time exposure-related injury to the tissues. Le et al [ 66 ] have reported initial inoculation in colchicine supplemented basal media for the respective treatment durations and subsequent culturing in fresh colchicine free basal media. The most promising treatment duration was 24 h (Table 1 ), wherein maximum yield of secondary metabolites was recovered.…”

“…Improvements in secondary biometabolites productions could be due to morphological, physiological as well as genetic manifestations engendered upon by induced autoploids. Some of such illustrative causes of autoploidy-mediated secondary metabolite augmentation are increase in stomatal sizes and densities leading to increased photosynthates output [ 67 ], heterosis and gene redundancy imparting transgressive meliorism [ 56 , 66 , 68 ]. Shmeit et al [ 62 ] reckoned that improvised primary metabolism could be the presumptive cause of ameliorated secondary metabolism.…”

“…(1.375-fold) [ 65 ] Panax ginseng Mayer MSL + colchicine soln. ➔ shaking at 100rpm ➔ dark Adventitious root Colchicine; 100mg/l; 1 d Tetraploid ➔ mutant (tetraploid) Increase in ginsenoside production (4.85-fold) [ 66 ] Stevia rebaudiana Soaked in colchicine soln. Seed Colchicine; 0.2%; 24 h Diploid➔tetraploid (2n=4x=44) Increase in rebaudioside-A content (2.84%) [ 67 ] Artemisia cina Soaked in colchicine soln.…”