Abstract
Salvia miltiorrhiza Bunge (Lamiaceae) hairy root cultures were inoculated (at 0.02 and 0.2% v/v) and co-cultured with Bacillus cereus bacteria. The root biomass growth was inhibited significantly by the bacteria inoculated to the root culture on the first day (day 0) but not by the bacteria inoculated on days 14 or 21 (in a 28-day overall period). On the other hand, the growth of the bacteria in the hairy root culture was also strongly inhibited by the hairy roots, partially because of the antibacterial activity of the secondary compounds produced by the roots. Most interestingly, the tanshinone production was promoted by the inoculation of bacteria at any of these days but more significantly by an earlier bacteria inoculation. With 0.2% bacteria inoculated on day 0, for example, the total tanshinone content of roots was increased by more than 12-fold (from 0.20 to 2.67 mg g−1 dry weight), and the volumetric tanshinone yield increased by more than sixfold (from 1.40 to 10.4 mg l−1). The tanshinone production was also stimulated by bacterial water extract and bacterial culture supernatant but less significantly than by the inoculation of live bacteria. The results suggest that the stimulation of tanshinone production by live bacteria in the root cultures may be attributed to the elicitor compounds originating from the bacteria, and the hairy root–bacteria coculture may be an effective strategy for improving secondary metabolite production in plant tissue cultures.
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References
Able AJ, Guest DI, Sutherland MW (2001) Relationship between transmembrane ion movements, production of reactive oxygen species and the hypersensitive response during the challenge of tobacco suspension cells by zoospores of Phytophthora nicotianae. Physiol Mol Plant Pathol 58:189–198
Able AJ, Sutherland MW, Guest DI (2003) Production of reactive oxygen species during non-specific elicitation, non-host resistance and field resistance expression in cultured tobacco cells. Funct Plant Biol 30:91–99
Atkinson MM, Huang JS, Knopp JA (1985) The hypersensitive reaction of tobacco to . Pseudomonas syringae pv. pisi. Plant Physiol 79:843–847
Atkinson MM, Keppler LD, Orlandi EW, Baker CJ, Mischke CF (1990) Involvement of plasma membrane calcium influx in bacterial induction of the K+/H+ and hypersensitive responses in tobacco. Plant Physiol 92:215–221
Barea JM, Pozo MJ, Azcón R, Azcón-Aguilar C (2005) Microbial co-operation in the rhizosphere. J Exp Bot 56:1761–1778
Chen H, Chen F, Zhang YL, Song JY (1999) Production of lithospermic acid B and rosmarinic acid in hairy root cultures of Salvia miltiorrhiza. J Ind Microbiol Biotechnol 22:133–138
Chen H, Chen F, Chiu FCK, Lo CMY (2001) The effect of yeast elicitor on the growth and secondary metabolism of hairy root cultures of Salvia miltiorrhiza. Enzyme Microbial Technol 28:100–105
De Wit PJGM (1995) Fungal avirulence genes and plant resistance genes: unravelling the molecular basis of gene for gene interactions. Adv Bot Res 21:148–177
Ebell LF (1969) Variation in total soluble sugars of conifer tissues with method of analysis. Phytochemistry 8:227–233
Ebel J, Cosio EG (1994) Elicitors of plant defense responses. Int Rev Cytol 148:1–36
Ge X, Wu J (2005) Tanshinone production and isoprenoid pathways in Salvia miltiorrhiza hairy roots induced by Ag+ and yeast elicitor. Plant Sci 168:487–491
Giri A, Narasu L (2000) Transformed hairy roots: recent trends and applications. Biotechnol Adv 18:1–22
Guillon S, Trémouillaux-Guiller J, Pati PK, Rideau M, Gantet P (2006) Hairy root research: recent scenario and exciting prospects. Curr Op Plant Biol 9:341–346
Hu ZB, Alfermann AW (1993) Diterpenoid production in hairy root cultures of Salvia miltiorrhiza. Phytochemistry 32:699–703
Keppler LD, Baker CJ, Atkinson MM (1989) Active oxygen production during a bacteria-induced hypersensitive reaction in tobacco suspension cells. Phytopathology 79:974–978
Kloepper JW, Ryu CM, Zhang SA (2004) Induced systemic resistance and promotion of plant growth by Bacillus spp. Phytopathology 94:1259–1266
Lee DS, Lee SH, Noh JG, Hong SD (1999) Antibacterial activities of cryptotanshinone and dihydrotanshinone I from a medicinal herb, Salvia miltiorrhiza Bunge. Biosci Biotechnol Biochem 63:2236–2239
Murashige T, Skoog FA (1962) A revised medium for rapid growth and biosynthesis with tobacco tissue cultures. Physiol Plant 15:473–497
Rao SR, Ravishankar GA (2002) Plant cell cultures: Chemical factories of secondary metabolites. Biotechnol Adv 20:101–153
Roos W, Evers S, Hieke M, Tschöpe M, Schumann B (1998) Shifts of intracellular pH distribution as a part of the signal mechanism leading to the elicitation of benzophenan thridine alkaloids. Plant Physiol 118:349–364
Sakano K (2001) Metabolic regulation of pH in plant cells: role of cytoplasmic pH in defense reaction and secondary metabolism. Int Rev Cytol 206:1–44
Tang W, Eisenbrand G (1992) Chinese drugs of plant origin: chemistry, pharmacology, and use in traditional and modern medicine. Springer, Berlin, pp 891–902
Wang JW, Wu JY (2005) Nitric oxide is involved in methyl jasmonate-induced defense responses and secondary metabolism activities of Taxus cells. Plant Cell Physiol 46:923–930
Wang X, Morris-Natschke SL, Lee KH (2007) New developments in the chemistry and biology of the bioactive constituents of Tanshen. Med Res Rev 27:133–148
Zhao J, Davis LC, Verpoorte R (2005) Elicitor signal transduction leading to production of plant secondary metabolites. Biotechnol Adv 23:283–333
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This work was supported by grants (G-U150 and A-PE94) from The Hong Kong Polytechnic University.
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Wu, JY., Ng, J., Shi, M. et al. Enhanced secondary metabolite (tanshinone) production of Salvia miltiorrhiza hairy roots in a novel root–bacteria coculture process. Appl Microbiol Biotechnol 77, 543–550 (2007). https://doi.org/10.1007/s00253-007-1192-5
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DOI: https://doi.org/10.1007/s00253-007-1192-5