In this study, we established a novel somatic embryogenesis and plant regeneration system through cell suspension culture of white dandelion (Taraxacum coreanum NAKAI.). Embryogenic calli could be initiated from leaf and root explants of sterile seedlings on solid Murashige and Skoog (MS) medium sup...
In this study, we established a novel somatic embryogenesis and plant regeneration system through cell suspension culture of white dandelion (Taraxacum coreanum NAKAI.). Embryogenic calli could be initiated from leaf and root explants of sterile seedlings on solid Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) after 3-week cultures. To proliferate embryogenic calli rapidly, cell suspension culture was performed with transferred to liquid MS medium with various combinations of plant growth regulators (PGRs) including 2,4-D, ${\alpha}$-naphthalene acetic acid (NAA), indole-3-acetic acid (IAA), $N^6$-benzylamino purine (BAP), thidiazuron (TDZ), and kinetin. During suspension cultures, embryogenic calli not only greatly proliferated, but shoot organogenesis also simultaneously occurred from the surface of somatic embryos. Among them, TDZ at lower concentration, 0.1 mg/L produced the highest efficiency of somatic embryo formation and shoot organogenesis. Rooting of embryogenic calli with adventitious shoots was done on solid MS medium containing 0.1 mg/L NAA and 0.3% activated carbon. Nearly 80% of embryogenic calli with shoot organogenesis could be rooted normal. Well-rooted plantlets were transferred into pots under a greenhouse condition, and plants derived from this system appeared phenotypically normal.
In this study, we established a novel somatic embryogenesis and plant regeneration system through cell suspension culture of white dandelion (Taraxacum coreanum NAKAI.). Embryogenic calli could be initiated from leaf and root explants of sterile seedlings on solid Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) after 3-week cultures. To proliferate embryogenic calli rapidly, cell suspension culture was performed with transferred to liquid MS medium with various combinations of plant growth regulators (PGRs) including 2,4-D, ${\alpha}$-naphthalene acetic acid (NAA), indole-3-acetic acid (IAA), $N^6$-benzylamino purine (BAP), thidiazuron (TDZ), and kinetin. During suspension cultures, embryogenic calli not only greatly proliferated, but shoot organogenesis also simultaneously occurred from the surface of somatic embryos. Among them, TDZ at lower concentration, 0.1 mg/L produced the highest efficiency of somatic embryo formation and shoot organogenesis. Rooting of embryogenic calli with adventitious shoots was done on solid MS medium containing 0.1 mg/L NAA and 0.3% activated carbon. Nearly 80% of embryogenic calli with shoot organogenesis could be rooted normal. Well-rooted plantlets were transferred into pots under a greenhouse condition, and plants derived from this system appeared phenotypically normal.
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문제 정의
The objectives of this study were to establish reliable plant regeneration system from cell suspension culture with T. coreanum. In an attempt to demonstrate the application of suspension culture techniques to enhance reproductibility, an efficient plant regeneration system through cell suspension culture has been established.
coreanum. This study provides to develop novel tissue culture techniques for application to T. coreanum. This system would help further application of biotechnology to Taraxacum breeding programs through genetic modification.
To authors’ knowledge, this work is the first report about embryogenic callus and plant regeneration through cell suspension culture of white dandelion.
가설 설정
Threadlike adventitious leaves induced from embryogenic calli. C. Normal rooting of embryogenic calli with the removal of vitrified leaves. Bars = 2 mm.
Embryogenic calli with adventitious shoots during shoot organogenesis. H. Rooting of embryogenic calli with adventitious shoots on the halfstrength solid MS medium. I.
제안 방법
To authors’ knowledge, this work is the first report about embryogenic callus and plant regeneration through cell suspension culture of white dandelion. To determine the effects of various PRGs and their concentrations, three auxins, two cytokinins, and kinetin with both concentrations (0.1 mg/L and 1.0 mg/L) were investigated in this work.
대상 데이터
Seeds of T. coreanum used in this study were collected from Kangwon-Do, Korea from May to June in 2009. The seeds were sterilized by immersion in 4% sodium hypochlorite for 10 min followed in 70% ethanol for 30 sec.
5 mg/L NAA. Twenty explants per replication were used in this experiment. Means followed by the same letter are not significantly different at P < 0.
성능/효과
0 mg/L TDZ, respectively. Based on above results, 0.1 mg/L TDZ was considered to be the most efficient in the proliferation of embryogenic calli in cell suspension culture of T. coreanum.
2). From the results of weight changes of embryogenic calli after 4-week suspension culture (Fig. 2), it was suggested that cytokines such as BAP and TDZ, played more important roles than kinetin and some auxins such as NAA, and IAA. With the addition of 2,4-D, NAA, IAA or kinetin, the weights of embryogenic calli showed 5~14-fold higher than those of initial inoculated calli.
, 2009). In this study, both the proliferation rates and sizes of embryogenic calli varied depending on PGR type and concentration (Fig. 3). Liquid media with cytokinin produced greater amounts and larger sizes of embryogenic calli (Fig.
With the addition of 2,4-D, NAA, IAA or kinetin, the weights of embryogenic calli showed 5~14-fold higher than those of initial inoculated calli. With the addition of BAP and TDZ, the weights increased more largely compared to the addition of 2,4-D, NAA, IAA or kinetin, with 13.71 g and 17.07 g with 0.1 mg/L and 1.0 mg/L BAP, and 23.04 g and 19.79 g with 0.1 mg/L and 1.0 mg/L TDZ, respectively. Based on above results, 0.
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