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Baque, Md. Abdullahil,Hahn, Eun-Joo,Paek, Kee-Yoeup The Korean Society of Plant Biotechnology 2010 Plant biotechnology reports Vol.4 No.2
Morinda citrifolia adventitious roots were cultured in shake flasks using Murashige and Skoog medium with different types and concentrations of auxin and cytokinin. Root (fresh weight and dry weight) accumulation was enhanced at 5 $mg\;l^{-1}$ indole butyric acid (IBA) and at 7 and 9 $mg\;l^{-1}$ naphthalene acetic acid (NAA). On the other hand, 9 $mg\;l^{-1}$ NAA decreased the anthraquinone, phenolic and flavonoid contents more severely than 9 $mg\;l^{-1}$ IBA. When adventitious roots were treated with kinetin (0.1, 0.3 and 0.5 $mg\;l^{-1}$) and thidiazuron (TDZ; 0.1, 0.3 and 0.5 $mg\;l^{-1}$) in combination with 5 $mg\;l^{-1}$ IBA, fresh weight and dry weight decreased but secondary metabolite content increased. The secondary metabolite content (including 1,1-diphenyl-2-picrylhydrazyl activity) increased more in TDZ-treated than in kinetin-treated roots. Antioxidative enzymes such as catalase (CAT) and guaiacol peroxidase (G-POD), which play important roles in plant defense, also increased. A strong decrease in ascorbate peroxidase activity resulted in a high accumulation of hydrogen peroxide. This indicates that adventitious roots can grow under stress conditions with induced CAT and G-POD activities and higher accumulations of secondary metabolites. These results suggest that 5 $mg\;l^{-1}$ IBA supplementation is useful for growth and secondary metabolite production in adventitious roots of M. citrifolia.
Cui, Hai-Yan,Baque, Md. Abdullahil,Lee, Eun-Jung,Paek, Kee-Yoeup 한국식물생명공학회 2013 Plant biotechnology reports Vol.7 No.3
In an attempt to scale-up of adventitious root cultures of Echinacea angustifolia for the production of biomass and caffeic acid derivatives, i.e. echinacoside, chlorogenic acid, cichoric acid, caftaric acid, and cynarin, the effects of Murashige and Skoog (MS) medium dilutions, and initial sucrose concentrations were investigated in a 5-L airlift bioreactor. In addition, the kinetics of adventitious root growth and accumulation of secondary metabolites were also studied. The greatest root dry weight ($6.50gL^{-1}$) and accumulation of total phenolics [$22.06mgg^{-1}$ DW (dry weight)], total flavonoids ($5.77mgg^{-1}$ DW) and total caffeic acid derivatives ($10.63mgg^{-1}$ DW) were obtained at quarter-strength MS medium. Of the various gradients of sucrose tested, 5 % sucrose supplementation was regarded as an optimal concentration for enhancing productivity of biomass and bioactive compounds. Neither higher salt strength (3/4-2 MS) nor sucrose concentrations (7 and 9 %) showed promotive effect on root growth and metabolite production. The kinetic studies revealed that 4 weeks of culture period is the optimal time to achieve highest productivity of metabolites. Based on these results, a largescale (20 L) and a pilot-scale (500 L) adventitious root culture system was established. In the pilot-scale bioreactor, adventitious roots were elicitor-treated with $100{\mu}M$ methyl jasmonate (MJ) on day 28. After 1 week of elicitation, 1.75 kg dry root biomass was harvested containing $60.41mgg^{-1}$ DW of total phenolics, $16.45mgg^{-1}$ DW of total flavonoids, and $33.44mgg^{-1}$ DW of total caffeic acid derivatives. Among the caffeic acid derivatives, the accumulation of echinacoside (the major bioactive compound) in MJ-treated adventitious roots grown in the 500-L bioreactor was the highest ($12.3mgg^{-1}$ DW), which is approximately threefold more than the non-MJ-treated roots cultured in 5- and 20-L bioreactors.
Hai-Yan Cui,Md. Abdullahil Baque,Eun Jung Lee,백기엽 한국식물생명공학회 2013 Plant biotechnology reports Vol.7 No.3
In an attempt to scale-up of adventitious rootcultures of Echinacea angustifolia for the production ofbiomass and caffeic acid derivatives, i.e. echinacoside,chlorogenic acid, cichoric acid, caftaric acid, and cynarin,the effects of Murashige and Skoog (MS) medium dilutions,and initial sucrose concentrations were investigated in a 5-Lairlift bioreactor. In addition, the kinetics of adventitiousroot growth and accumulation of secondary metaboliteswere also studied. The greatest root dry weight (6.50 g L-l)and accumulation of total phenolics [22.06 mg g-1 DW(dry weight)], total flavonoids (5.77 mg g-1 DW) and totalcaffeic acid derivatives (10.63 mg g-1 DW) were obtainedat quarter-strength MS medium. Of the various gradients ofsucrose tested, 5 % sucrose supplementation was regardedas an optimal concentration for enhancing productivity ofbiomass and bioactive compounds. Neither higher saltstrength (3/4–2 MS) nor sucrose concentrations (7 and 9 %)showed promotive effect on root growth and metaboliteproduction. The kinetic studies revealed that 4 weeks ofculture period is the optimal time to achieve highestproductivity of metabolites. Based on these results, a largescale(20 L) and a pilot-scale (500 L) adventitious rootculture system was established. In the pilot-scale bioreactor,adventitious roots were elicitor-treated with 100 lMmethyljasmonate (MJ) on day 28. After 1 week of elicitation,1.75 kg dry root biomass was harvested containing60.41 mg g-1 DWof total phenolics, 16.45 mg g-1 DWoftotal flavonoids, and 33.44 mg g-1 DW of total caffeic acidderivatives. Among the caffeic acid derivatives, the accumulationof echinacoside (the major bioactive compound)in MJ-treated adventitious roots grown in the 500-L bioreactorwas the highest (12.3 mg g-1 DW), which isapproximately threefold more than the non-MJ-treated rootscultured in 5- and 20-L bioreactors.
Md. Humayun Kabir Shiragi,Md. Abdullahil Baque,Khandokar Md. Nasiruddin 한국원예학회 2010 Horticulture, Environment, and Biotechnology Vol.51 No.3
An efficient protocol was established to produce virus free in vitro banana plantlets through meristem culture of virus infected four months old banana suckers cv. Sabri. Murashige and Skoog (MS) medium supplemented with different levels of BAP (0, 3, 4 and 5 ㎎ · ℓ?¹) and IAA (0.0, 1.0, 1.5, 2.0 and 2.5 ㎎ · ℓ?¹) were used for shoot proliferation in Experiment-I. Half strength of MS medium supplemented with different levels of IBA (0, 1, 2, 3 ㎎ · ℓ?¹), NAA (0, 2, 3, 4 ㎎ · ℓ?¹) were treated for root formation in Experiment-II. ELISA test was performed to confirm virus eradication rate of regenerated plants and survival rate of regenerated plants in Experiment-III. The treatment combination of 4 ㎎ · ℓ?¹ BAP + 2.5 ㎎ · ℓ?¹ IAA produced the highest number of shoots (3.4 explant-1), while the longest shoot (3.49 cm), maximum number of leaves (3.37 plantlet-1) and longest leaves (2.22 cm) were obtained in the culture treated with 5 ㎎ · ℓ?¹ BAP. On the contrary, for plantlets cultured on half strength of MS medium supplemented with different levels of IBA and NAA, the highest number of vigorous roots (8.01 explant-1) and maximum root length (4.64 cm) were observed at treatment combinations of 2 ㎎ · ℓ?¹ IBA + 4 ㎎ · ℓ?¹ NAA and 2 ㎎ · ℓ?¹ IBA+ 3 ㎎ · ℓ?¹ NAA, respectively. The combinations of 3 ㎎ · ℓ?¹ IBA + 3 ㎎ · ℓ?¹ NAA produced more vigorous roots while less vigorous roots were observed at respective control. The rate of virus free plant regeneration from banana bunchy top virus (BBTV) infected sources was 57.14%. However, the highest plant survival rate (83.33%) was observed in a mixture of sand, soil and cow dung (1:1:1) and in a mixture of coir and soil (1:2). These results suggested that eradication of BBTV through meristem culture can be used as an effective tool for commercial exploitation of virus free banana production.