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Accumulation of ginsenosides and caffeic acid derivatives as affected by co-cultures of Panax ginseng and Echinacea purpurea adventitious roots
Abstracts / Journal of Biotechnology 136S (2008) S147–S169
II6-P-051 Accumulation of ginsenosides and caffeic acid derivatives as affected by co-cultures of Panax ginseng and Echinacea purpurea adventitious roots Chun-Hua Wu, E.J. Hahn, K.Y. Paek Research Center for the Development of Advanced Horticultural Technology, Chungbuk National University, Cheongju 361-763, South Korea Panax ginseng C.A. Meyer and Echinacea purpurea (L.) are wellknown medicinal plants for their diverse efficacy on human health with anti-oxidative, anti-bacterial, anti-viral, immune and sedative properties, etc. The main active compounds are ginsenosides and caffeic acid derivatives, respectively. We have reported the optimal cultures for the maximal production of root biomass and secondary metabolites in the adventitious root cultures of these two species, which is successfully being applied to large-scale bioreactor cultures. Intra-species and inter-species co-culture systems constitute a new area in biotechnological research of higher plants, especially used for the production of secondary metabolites in medicinal plants. However, the applications have been limited to a small numbers of plants and therefore, not much information is available on co-cultures influencing secondary metabolism. To investigate the effect of co-cultures of P. ginseng C.A. Meyer and E. purpurea (L.) on biomass and secondary metabolite production, the adventitious roots of these two species were co-cultured in 5-L scale bioreactors with different proportions (P. ginseng:E. purpurea = 5:0, 4:1, 3:2, 2:3, 0:5), and with or without MJ elicitation. Cultures were maintained on 0.75 strength of MS medium supplemented with 5% sucrose and 5 mg L−1 IBA for 40 days in the dark. Biomass accumulation was higher in single cultures (P. ginseng:E. purpurea = 5:0 and 0:5) compared with co-cultures. Ginsenoside contents (diol, triol, total) were highest in the ginseng single culture with MJ elicitation, while caffeic acid derivatives contents were highest in the Echinacea single culture regardless of MJ elicitation. Higher contents of polyphenolics and flavonoids were detected in the Echinacea single culture or co-culture of ginseng and Echinacea at 2:3 ratios. MJ elicitation slightly increased flavonoid content but no changes were observed in polyphenolic contents. The inoculum ratio of ginseng and Echinacea also affected root morphology, resulting in differences in the development of lateral root and eventually root biomass. doi:10.1016/j.jbiotec.2008.07.360 II6-P-052 Adventitious root cultures and secondary metabolite production in Morinda citrifolia as affected by growth regulators and inoculum density Abdullahil Baque, E.J. Hahn, K.Y. Paek ∗ Research Center for the Development of Advanced Horticultural Technology, Chungbuk National University, Cheongju 361-763, South Korea E-mail address: [email protected] (K.Y. Paek). Morinda citrifolia L., commonly named as noni, contains medicinally active components that exhibit various therapeutic effects, such as anti-viral, anti-bacterial, and anti-cancer activities. To prevent the depletion of natural habitat and to produce bioactive compounds in large scale, plant cell and organ cultures have been applied to various medicinal plants. On the other hand, no reports are available in M. citrifolia. This study aims to improve
S167
root biomass and secondary metabolite contents in the adventitious roots of M. citrifolia by treatments of different types and concentrations of auxin and cytokinin. In the auxin treatment,the accumulation of root biomass was enhanced by high concentrations of IBA (5 mg l−1 ) and NAA (7.9 mg l−1 ). In contrast, high auxin concentrations decreased the contents of Anthraquinone, phenolics, and flavonoids. Secondary metabolites were highest at 1 mg l−1 IBA followed by 1 mg l−1 NAA. The combinations of auxin and cytokinin resulted in a significant decrease of root biomass, but increased the secondary metabolite contents more than two folds. DPPH activity also increased more than 10% in the combination of auxin and cytokinin compared with control (5 mg l−1 IBA). These results suggested that the adventitious root cultures of M. citrifolia require different types and concentrations of auxin and cytokinin for the accumulation of root biomass and secondary metabolites, respectively. In suspension cultures, higher root inoculum density (15 g l−1 ) was favorable to both root growth and the secondary metabolite accumulation, showing more than two-fold increases compared with lower inoculum density (5 g l−1 ). Keywords: Adventitious root; Biomass; Morinda citrifolia; Secondary metabolites doi:10.1016/j.jbiotec.2008.07.361 II6-P-053 Effects of media, BAP and IBA concentrations on proliferation and rooting of Gisela 6 rootstock Alireza Daneshvar Hosseini a,b , Ebrahim Ganji Sedighe Anahid a , Mohammdreza Bihamta b,∗
Moghadam a ,
a
Khorasan Agric. Research Center, Department of Horticulture, Mashhad, Iran b Tehran university, Department of Biotechnology, Tehran, Iran
E-mail address: [email protected] (M. Bihamta). Gisela 6 is one of the semi-dwarf rootstocks suitable for all types of sweet cherry. It is adapted to all kinds of soils especially heavy soils (Andersen et al., 1999). With respect to the importance of this rootstock, this study was conducted to determine the effects of media, BAP and IBA concentrations on proliferation and rooting of Gisela 6 rootstock. In the experiment related to proliferation for culture media (MS, 1/2MS, LS, Modify LS) (Murashige and Skoog, 1962; Linsmaier and Skoog, 1965) and three concentrations BAP (0, 1, 2 mg/l) were applied. Results showed that the best culture medium is MS (Murashige and Skoog, 1962) and the best BAP concentration is 1 mg/l. The average of shoot number and length are 5.11 and 1.70 cm, respectively. In the experiment related to rooting for culture media (MS, 1/2MS, LS, Modify LS) (Murashige and Skoog, 1962; Linsmaier and Skoog, 1965) and three concentrations IBA (0, 1, 2 mg/l) were applied. In this respect, results showed that the best rooting medium is MS (Murashige and Skoog, 1962) and the best IBA concentration is 1 mg/l. The average of root number and length are 3 and 3.25 cm, respectively. The percentage of rooted plantlets is 54 and the percentage of survival under in vivo condition is 76. Keywords: Media; BAP; IBA; Gisela 6 rootstock References Andersen, R.L., Robinson, T., Lang, G.A., 1999. Managing the Gisela cherry rootstocks. New York Fruit Quart. 7, 1–4. Linsmaier, E.M., Skoog, F., 1965. Organic growth factor requirements of tobacco tissue culture. Physiol. Plant. 18, 100–127.
II6-P-051 Accumulation of ginsenosides and caffeic acid derivatives as affected by co-cultures of Panax ginseng and Echinacea purpurea adventitious roots Chun-Hua Wu, E.J. Hahn, K.Y. Paek Research Center for the Development of Advanced Horticultural Technology, Chungbuk National University, Cheongju 361-763, South Korea Panax ginseng C.A. Meyer and Echinacea purpurea (L.) are wellknown medicinal plants for their diverse efficacy on human health with anti-oxidative, anti-bacterial, anti-viral, immune and sedative properties, etc. The main active compounds are ginsenosides and caffeic acid derivatives, respectively. We have reported the optimal cultures for the maximal production of root biomass and secondary metabolites in the adventitious root cultures of these two species, which is successfully being applied to large-scale bioreactor cultures. Intra-species and inter-species co-culture systems constitute a new area in biotechnological research of higher plants, especially used for the production of secondary metabolites in medicinal plants. However, the applications have been limited to a small numbers of plants and therefore, not much information is available on co-cultures influencing secondary metabolism. To investigate the effect of co-cultures of P. ginseng C.A. Meyer and E. purpurea (L.) on biomass and secondary metabolite production, the adventitious roots of these two species were co-cultured in 5-L scale bioreactors with different proportions (P. ginseng:E. purpurea = 5:0, 4:1, 3:2, 2:3, 0:5), and with or without MJ elicitation. Cultures were maintained on 0.75 strength of MS medium supplemented with 5% sucrose and 5 mg L−1 IBA for 40 days in the dark. Biomass accumulation was higher in single cultures (P. ginseng:E. purpurea = 5:0 and 0:5) compared with co-cultures. Ginsenoside contents (diol, triol, total) were highest in the ginseng single culture with MJ elicitation, while caffeic acid derivatives contents were highest in the Echinacea single culture regardless of MJ elicitation. Higher contents of polyphenolics and flavonoids were detected in the Echinacea single culture or co-culture of ginseng and Echinacea at 2:3 ratios. MJ elicitation slightly increased flavonoid content but no changes were observed in polyphenolic contents. The inoculum ratio of ginseng and Echinacea also affected root morphology, resulting in differences in the development of lateral root and eventually root biomass. doi:10.1016/j.jbiotec.2008.07.360 II6-P-052 Adventitious root cultures and secondary metabolite production in Morinda citrifolia as affected by growth regulators and inoculum density Abdullahil Baque, E.J. Hahn, K.Y. Paek ∗ Research Center for the Development of Advanced Horticultural Technology, Chungbuk National University, Cheongju 361-763, South Korea E-mail address: [email protected] (K.Y. Paek). Morinda citrifolia L., commonly named as noni, contains medicinally active components that exhibit various therapeutic effects, such as anti-viral, anti-bacterial, and anti-cancer activities. To prevent the depletion of natural habitat and to produce bioactive compounds in large scale, plant cell and organ cultures have been applied to various medicinal plants. On the other hand, no reports are available in M. citrifolia. This study aims to improve
S167
root biomass and secondary metabolite contents in the adventitious roots of M. citrifolia by treatments of different types and concentrations of auxin and cytokinin. In the auxin treatment,the accumulation of root biomass was enhanced by high concentrations of IBA (5 mg l−1 ) and NAA (7.9 mg l−1 ). In contrast, high auxin concentrations decreased the contents of Anthraquinone, phenolics, and flavonoids. Secondary metabolites were highest at 1 mg l−1 IBA followed by 1 mg l−1 NAA. The combinations of auxin and cytokinin resulted in a significant decrease of root biomass, but increased the secondary metabolite contents more than two folds. DPPH activity also increased more than 10% in the combination of auxin and cytokinin compared with control (5 mg l−1 IBA). These results suggested that the adventitious root cultures of M. citrifolia require different types and concentrations of auxin and cytokinin for the accumulation of root biomass and secondary metabolites, respectively. In suspension cultures, higher root inoculum density (15 g l−1 ) was favorable to both root growth and the secondary metabolite accumulation, showing more than two-fold increases compared with lower inoculum density (5 g l−1 ). Keywords: Adventitious root; Biomass; Morinda citrifolia; Secondary metabolites doi:10.1016/j.jbiotec.2008.07.361 II6-P-053 Effects of media, BAP and IBA concentrations on proliferation and rooting of Gisela 6 rootstock Alireza Daneshvar Hosseini a,b , Ebrahim Ganji Sedighe Anahid a , Mohammdreza Bihamta b,∗
Moghadam a ,
a
Khorasan Agric. Research Center, Department of Horticulture, Mashhad, Iran b Tehran university, Department of Biotechnology, Tehran, Iran
E-mail address: [email protected] (M. Bihamta). Gisela 6 is one of the semi-dwarf rootstocks suitable for all types of sweet cherry. It is adapted to all kinds of soils especially heavy soils (Andersen et al., 1999). With respect to the importance of this rootstock, this study was conducted to determine the effects of media, BAP and IBA concentrations on proliferation and rooting of Gisela 6 rootstock. In the experiment related to proliferation for culture media (MS, 1/2MS, LS, Modify LS) (Murashige and Skoog, 1962; Linsmaier and Skoog, 1965) and three concentrations BAP (0, 1, 2 mg/l) were applied. Results showed that the best culture medium is MS (Murashige and Skoog, 1962) and the best BAP concentration is 1 mg/l. The average of shoot number and length are 5.11 and 1.70 cm, respectively. In the experiment related to rooting for culture media (MS, 1/2MS, LS, Modify LS) (Murashige and Skoog, 1962; Linsmaier and Skoog, 1965) and three concentrations IBA (0, 1, 2 mg/l) were applied. In this respect, results showed that the best rooting medium is MS (Murashige and Skoog, 1962) and the best IBA concentration is 1 mg/l. The average of root number and length are 3 and 3.25 cm, respectively. The percentage of rooted plantlets is 54 and the percentage of survival under in vivo condition is 76. Keywords: Media; BAP; IBA; Gisela 6 rootstock References Andersen, R.L., Robinson, T., Lang, G.A., 1999. Managing the Gisela cherry rootstocks. New York Fruit Quart. 7, 1–4. Linsmaier, E.M., Skoog, F., 1965. Organic growth factor requirements of tobacco tissue culture. Physiol. Plant. 18, 100–127.