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, 35 (3), 283-93

Ginsenoside Production and Morphological Characterization of Wild Ginseng (Panax Ginseng Meyer) Mutant Lines Induced by γ-Irradiation ((60)Co) of Adventitious Roots

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Ginsenoside Production and Morphological Characterization of Wild Ginseng (Panax Ginseng Meyer) Mutant Lines Induced by γ-Irradiation ((60)Co) of Adventitious Roots

Jun-Ying Zhang et al. J Ginseng Res.

Abstract

With the purpose of improving ginsenoside content in adventitious root cultures of Korean wild ginseng (Panax ginseng Meyer), the roots were treated with different dosages of γ-ray (5, 10, 25, 50, 75, 100, and 200 Gy). The growth of adventitious roots was inhibited at over 100 Gy. The irradiated adventitious roots showed significant variation in the morphological parameters and crude saponin content at 50 to100 Gy. Therefore, four mutant cell lines out of the propagation of 35 cell lines treated with 50 Gy and 100 Gy were selected on the basis of phenotypic morphology and crude saponin contents relative to the wild type control. The contents of 7 major ginsenosides (Rg1, Re, Rb1, Rb2, Rc, Rf, and Rd) were determined for cell lines 1 and 3 from 100 Gy and lines 2 and 4 from 50 Gy treatments. Cell line 2 showed more secondary roots, longer length and superior growth rate than the root controls in flasks and bioreactors. Cell line 1 showed larger average diameter and the growth rate in the bioreactor was comparable with that of the control but greater in the flask cultured roots. Cell lines 1 and 2, especially the former, showed much more ginsenoside contents than the control in flasks and bioreactors. Therefore, we chose cell line 1 for further study of ginsenoside contents. The crude saponin content of line 1 in flask and bioreactor cultures increased by 1.4 and 1.8-fold, respectively, compared to the control. Total contents of 7 ginsenoside types (Rg1, Re, Rb1, Rb2, Rc, Rf, and Rd) increased by 1.8 and 2.3-fold, respectively compared to the control. Crude saponin and ginsenoside contents in the bioreactor culture increased by about 1.4-fold compared to that the flask culture.

Keywords: Adventitious root; Ginsenosides; Mutant; Panax ginseng; γ-irradiation.

Figures

Fig. 1.
Fig. 1.. Culture system of Panax ginseng adventitious roots. (A) Adventitious root formation on Murashige and Skoog (MS) solid medium on petridishes. (B) Proliferation of the adventitious root in MS liquid medium in flasks. (C) Production of the adventitious root in bioreactors. (D) Harvest adventitious roots. Bar=1 cm.
Fig. 2.
Fig. 2.. Efficiency of secondary root induction of Panax ginseng adventitious root at various concentration 1-naphthaleneacetic acid (NAA) and indole-3-acetic acid (IAA) combination in the flask test. Bar shown are means±standard errors of three replicates.
Fig. 3.
Fig. 3.. Survival rate of Panax ginseng adventitious roots with 7 different dosages, growth 5 weeks on petridish on Murashige and Skoog solid medium with 2 mg/L 1-naphthaleneacetic acid, 1 mg/L indole-3-acetic acid and 5% sucrose at 23℃ in light condition after irradiation. Bar shown are means±standard errors of three replicates.
Fig. 4.
Fig. 4.. Effect of γ-rays on adventitious root survival number on Murashige and Skoog solid medium with 2 mg/L 1-naphthaleneacetic acid, 1 mg/ L indole-3-acetic acid and 5% sucrose at 23℃ in light condition after irradiation. (A) Control, (B-H) irradiated with 5, 10, 25, 50, 75, 100, and 200 Gy γ-radiation. Bar=1 cm.
Fig. 5.
Fig. 5.. Phenotypic characteristics of secondary roots in the control and cell lines growth in the flask with 50 mL Murashige and Skoog (MS) medium (A) and bioreactor with 5 L MS medium (B). Bar=1 cm. CL, cell line.
Fig. 6.
Fig. 6.. Determinated growth ratio of Panax ginseng adventitious root in the control and cell lines after 10, 20, and 30 days of culture in the flask with 50 mL Murashige and Skoog (MS) medium (A) and bioreactor with 5 L MS (B). Bar shown are means±standard errors of three replicates. CL, cell line.
Fig. 7.
Fig. 7.. Effect of γ-ray on crude saponin content in the control and cell lines of Panax ginseng adventitious root after 30 days growth in the flask with 50 mL Murashige and Skoog (MS) medium (♦) and bioreactor with 5 L MS medium (▲). Bar shown are means±standard errors of three replicates. CL, cell line.
Fig. 8.
Fig. 8.. Typical chromatograms obtained from the standard solution, cell line 1 (A) and control (B) solution extracted from bioreactor by UV detection at 203 nm. IS, internal standard of digoxin.

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