Abstract
Astragalus membranaceus is one of the most widely used traditional medicinal herbs in China, but the time required to generate a useful product in the field production is long. The growth of adventitious root cultures was compared between cultures grown in solid, liquid, or a 5-L balloon-type bubble bioreactor. The maximum growth ratio (final dry weight/initial dry weight) was determined for adventitious roots grown in the bioreactor. Studies carried out to optimize biomass production of adventitious roots compared adventitious root growth from various inoculum root lengths, inoculum densities, and aeration volume in the bioreactors. The maximum growth ratio occurred in treatments with a 1.5-cm inoculum root length, with 30 g (fresh weight) of inoculum per bioreactor or with an aeration volume of 0.1 vvm (air volume/culture medium volume per min). The polysaccharide, saponin, and flavonoid content of roots from bioreactor-grown cultures were compared to roots from field-grown plants grown for 1 and 3 yr. Total polysaccharide content of adventitious roots in the bioreactor (30.0 mg g−1 dry weight (DW)) was higher than the roots of 1-yr-old (13.8 mg g−1 DW) and 3-yr-old (21.1 mg g−1 DW) plants in the field. Total saponin (3.4 mg g−1 DW) and flavonoid (6.4 mg g−1 DW) contents were nearly identical to 3-yr-old roots and higher than that of 1-yr-old roots under field cultivation.



Similar content being viewed by others
References
Bhadra R.; Shanks J. V. Statistical design of the effect of inoculum conditions on growth of hairy root cultures of Catharanthus roseus. Biotechnol. Tech. 9: 681–686; 1995.
Choi S. M.; Son S. H.; Yun S. R.; Kwon O. W.; Seon J. H.; Paek K. Y. Pilot-scale culture of adventitious roots of ginseng in a bioreactor system. Plant Cell Tiss. Org. Cult. 62: 187–193; 2000.
Du M.; Wu X. J.; Ding J.; Hu Z. B.; White K. N.; Branford-White C. J. Astragaloside IV and polysaccharides production by hairy roots of Astragalus membranaceus in bioreactors. Biotechnol. Lett. 25: 1853–1856; 2003.
Dubois M.; Gilles K. A.; Hamilton J. K.; Rebers P. A.; Smith F. Colorimetric method of determination of sugars and related substances. Anal. Chem. 28: 350–356; 1956.
Gamborg O. L.; Miller R. A.; Ojima K. Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell Res. 50: 151–158; 1968.
Hirotani M.; Zhou Y.; Lui H.; Furuya T. Astragalosides from hairy root cultures of Astragalus membranaceus. Phytochemistry 36: 665–670; 1994.
Ionkova I.; Momekov G.; Proksch P. Effects of cycloartane saponins from hairy roots of Astragalus membranaceus Bge. on human tumor cell targets. Fitoterapia 81: 447–151; 2010.
Jeong C. S.; Chakrabarty D.; Hahn E. J.; Lee H. L.; Paek K. Y. Effects of oxygen, carbon dioxide and ethylene on growth and bioactive compound production in bioreactor culture of ginseng adventitious roots. Biochem. Eng. J. 27: 252–263; 2006.
Jeong C. S.; Murthy H. N.; Hahn E. J.; Lee H. L.; Paek K. Y. Inoculum size and auxin concentration influence the growth of adventitious roots and accumulation of ginsenosides in suspension cultures of ginseng (Panax ginseng C.A. Meyer). Acta. Physiol. Plant 31: 219–222; 2009a.
Jeong J. A.; Wu C. H.; Murthy H. N.; Hahn E. J.; Paek K. Y. Application of an airlift bioreactor system for the production of adventitious root biomass and caffeic acid derivatives of Echinacea purpurea. Biotechnol Bioproc. Eng. 14: 91–98; 2009b.
Kim Y. S.; Chakrabarty D.; Hahn E. J.; Paek K. Y. Methyl jasmonate increases saponin content in bioreactor culture of ginseng (Panax ginseng C. A. Meyer) adventitious roots. Acta. Hortic. 625: 289–292; 2004.
Kuo Y. H.; Tsai W. J.; Loke S. H.; Wu T. S.; Chiou W. F. Astragalus membranaceus flavonoids (AMF) ameliorate chronic fatigue syndrome induced by food intake restriction plus forced swimming. J. Ethnopharmacol. 122: 28–34; 2009.
Kwok K. H.; Doran P. M. Kinetic and stoichiometric analysis of hairy roots in a segmented bubble column reactor. Biotechnol. Prog. 11: 429–435; 1995.
Ma X.; Shi Q.; Duan J.; Dong T.; Tsim K. Chemical analysis of radix astragali (Huangqi) in China: a comparison with Its adulterants and seasonal variations. J. Agric. Food Chem. 50: 4861–4866; 2002.
Ma X. Q.; Duan J. A.; Zhu D. Y.; Dong T. X.; Tsim K. W. K. Chemical comparison of Astragali Radix (Huangqi) from different regions of China. Nat. Med. 54: 213–218; 2000.
McClelland M. T.; Smith M. A. L. Vessel type, closure, and explant orientation influence in vitro performance of five woody species. Hortscience 25: 797–800; 1990.
Min J. Y.; Jung H. Y.; Kang S. M.; Kim Y. D.; Kang Y. M.; Park D. J.; Prasad D. T.; Choi M. S. Production of tropane alkaloids by small-scale bubble column bioreactor cultures of Scopolia parviflora adventitious roots. Biores. Technol. 98: 1748–1753; 2007.
Murashige T.; Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497; 1962.
Paek K. Y.; Hahn E. J. Increase of saponin contents via elicitor treatments in bioreactor culture of ginseng (Panax ginseng C.A. Meyer) adventitious roots. Acta. Hortic. 679: 145–148; 2005.
Paek K. Y.; Murthy H. N.; Hahn E. J. Establishment of adventitious root cultures of Echinacea purpurea for the production of caffeic acid derivatives. Methods Mol. Biol. 547: 3–16; 2009.
Sinclair S. Chinese herbs: a clinical review of Astragalus, Ligusticum, and Schizandrae. Altern. Med. Rev. 3: 338–344; 1998.
Sivakumar G.; Kim S. J.; Hahn E. J.; Paek K. Y. Optimizing environmental factors for large-scale multiplication of chrysanthemum (Chrysanthemum grandiflorum) in balloon-type bioreactor culture. In Vitro Cell Dev. Biol-Plant 41: 822–825; 2005.
Sivakumar G.; Yu K. W.; Paek K. Y. Enhanced production of bioactive ginsenosides from adventitious roots of Panax ginseng in bioreactor culture. J. Hortic. Sci. Biotechnol. 81: 549–552; 2006.
Song J. Z.; Yiu H. H. W.; Qiao C. F.; Han Q. B.; Xu H. X. Chemical comparison and classification of Radix Astragali by determination of isoflavonoids and astragalosides. J. Pharm. Biomed. Anal. 47: 399–406; 2008.
Wagner H.; Bauer R.; Xiao P. G.; Chen J. M.; Michler G. Radix astragali (Huang Qi). Chin. Drug Minogue. Anal. 1: 1–17; 1997.
Wu C. H.; Murthy H.; Hahn E. J.; Paek K. Y. Large-scale cultivation of adventitious roots of Echinacea purpurea in airlift bioreactors for the production of chichoric acid, chlorogenic acid and caftaric acid. Biotechnol. Lett. 29: 1179–1182; 2007.
Wu F.; Chen X. A review of pharmacological study on Astragalus membranaceus (Fisch.) Bge. J. Chin. Med. Mat. 27: 232–234; 2004.
Acknowledgments
This research was supported by the National Science Foundation of China (30860176 and 30860036). We thank Professors Jules Janick and Paul Michael Hasegawa of Purdue University for the assistance with the manuscript.
Author information
Authors and Affiliations
Corresponding author
Additional information
Editor: John Finer
Rights and permissions
About this article
Cite this article
Wu, S.Q., Lian, M.L., Gao, R. et al. Bioreactor application on adventitious root culture of Astragalus membranaceus . In Vitro Cell.Dev.Biol.-Plant 47, 719–724 (2011). https://doi.org/10.1007/s11627-011-9376-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11627-011-9376-1


