• Korean Journal of Weed Science
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• v.7 no.2
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• pp.200-207
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• 1987

This study was conducted to select and develop herbicide resistant plant through tissue culture. Growth response of seedlings and callis of various rice varieties with Echinochloa species was assessed under the treatment of various rates of butachlor [N-(butoxy methyl)-2-chloro-2', 6'-diethyl acetamide] and simetryne [2,4-bis(ethyl amino)-6-methyl thio-1,3,5-triazine]. Further, succinate dehydrogenase activity was determined in herbicide treated callus to characterize different response of plants to herbicide. Rice variety like Sangpung showed relative resistance in both callus and seedling states against butachlor, indicating maintenance of resistance. However, in the simetryne treatment, the similar response was not observed in callus and seedling state, although there was a great different response among plant materials against simetryne. Rice variety which exhibited resistance in callus and seedling states showed low succinate dehydrogenase inhibition index. Succinate dehydrogenase inhibition index can be used as an important marker characters to differentiate varietal response of plant to herbicide. Rice plant was differentiated from butachlor and simetryne tolerant callus treated at $2.5{\times}10^{-5}$ M is growing under the growth chamber and can be used for resistant source.

• Korean Journal of Plant Tissue Culture
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• v.22 no.1
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• pp.35-39
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• 1995

In order to investigate the effects of gelling agent on rice anther culture, anthers of rice (Japonica cv Daecheongbyeo) were cultured on N$_{6}$ media supplemented with 0.8, 1.2 or 1.6% Junsei agar and 05, 0.4, 0.6, 0.8 or 1.0% Gelrite (Phytagel, Sigma). On Junsei agar media, the frequency of callus induction was decreased in proportion to agar concentration. The frequency of callus induction was more increased as 67.6% and 54.8% in media containing 0.4 and 0.6% Gelrite than in agar media. The frequency of plant regeneration and spontaneous doubled-diploid was directly proportional to Junsei agar and Gelrite concentration. The number of green and spontaneous doubled diploid plant was highest on 0.6% Gelrite medium. In order to optimize the concentration of growth regulators for the callus induction medium containing 0.6% Gelrite, anthers were cultured on N$_{6}$ media supplemented with 2mg/L NAA, 2 mg/L 2,4-D, 1mg/L NAA and 1mg/L 2, 4-D, or 1mg/L NAA, 1mg/L 2,4-D and 0.5mg/L kinetin. The maximum frequency of callus induction and plant regeneration was obtained from the medium supplemented with 2 mg/L NAA and 0.6% Gelrite. In conclusion the induction of embryogenic callus, the frequency of plant regeneration and in vivo chromosome doubling was more effective in Gelrite media than in Junsei agar media.dia.

• Journal of Plant Biology
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• v.36 no.3
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• pp.211-218
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• 1993

Plant regeneration was accomplished from protoplast culture of rice (Oryza sativa L. cv. Taebaeg). Embryogenic callus was induced from mature seed on MS medium containing 5 mM proline, 2.5 mg/L 2,4-D, 30 g/L sucrose in the dark at 28$^{\circ}C$ and used to establish embryogenic cell suspension culture. Suspension cells were subcultured every one week in N6 medium supplemented with 5 mM proline, 200 mg/L casein hydrolysate, 2.5 mg/L 2,4-D and amino acids of AA medium. Suspension cultures were composed of cells that were densely cytoplasmic, potentially embryogenic and were at least maintained for more than 6 months in liquid medium. Protoplasts were isolated from fast-growing suspension culture cells and cultured in a slightly modified KpR medium by mixed nurse culture. Isolated protoplasts began to divide within 5~7 days and thereafter, protoplast-derived calli were sequentially transferred to callus proliferating medium that soft agar MS medium contained 2 mg/L 2,4-D and produced distinct embryogenic cells. Microcolonies were then transferred to solid medium which consisted of MS medium containing 5 mg/L kinetin, 1 mg/L NAA, 1 mg/L ABA, 30 g/L sucrose and 10 g/L sorbitol under fluorescent light. Mulitple shoots of 4~5 per callus emerged and were transferred to hormone-free MS medium for root initiation. Thereafter, The plantlets were transferred to pots of soil to mature in the culture room.

• Proceedings of the Korean Society of Crop Science Conference
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• 1996.05a
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• pp.118-119
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• 1996

• Current Research on Agriculture and Life Sciences
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• v.1
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• pp.1-9
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• 1983

This study was conducted to obtain basic information on the rice anther culture. Materials used were (Inabawase X YR 2404-14-2-1) $F_1$ hybrid. Callus growth rate on various media, induction frequency of callus in spikelet and panicle, and the effect of treatment on anther and callus were evaluated. The results obtained were summarized as follows ; The growth rate of callus on N-6, M-S, P.E.agar media was 19.8, 13.1, 4.1 times respectively after 30 days inoculated, and on liquid media was 3.8, 5.1, 1.4 times, respectively. Organ differentiation on N-6, M-S, P.E.agar media was 37.5%, 12.5%, 17.5% respectively. The difference of induction frequency of callus per panicle was 0.14%-6.25% and per spikelet was 0-19.05%. Almost callus was induced 30-35 days after inoculation. Organ differentiation of induced callus was decreased by culture. Callus cultured for 13 days after induction did not make shoot. Anthers cold shocked at $8^{\circ}C$ for 5 days obtained 3.32% efficiencys of callus induction per number of anthers plated, and compared with 2.41% of no treated anthers. But anther treated at $8^{\circ}C$ for 7 days decreased 2.24%. Callus induction periods were shortened by cold treatment for about 5 days. Callus cultured on medium containing 2 mg/l of 2, 4-D showed 5% on root formation but medium containing 5 mg/l of 2, 4-D showed 30% of root formation after transfered on the medium without 2, 4-D. Callus cold shocked at $15-18^{\circ}C$ revealed poor efficiency for root formation, but 5 days treatment was good for shoot formation.

• Journal of Plant Biology
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• v.15 no.4
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• pp.13-17
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• 1972

Rice callus was treated with 0.56M sucrose solution mixed with 5% pectinase and 10% cellulase, and the protoplasts isolated were transferred to 0.25 M sodium nitrate to induce protoplasmic fusion. Callus tissues were macerated well and degradation of cell walls also proceeded satisfactorily. When the protoplasts were transferred to sodium nitrate solution, many giant roundish protoplasts and some multilobed complex protoplasmic bodies were observed. Most of the fusions took place immediately after the protoplasts were transferred to sodium nitrate. Some multilobed protoplasts which failed to fuse in the initial stage took longer time, about two hours, to get completely fused and rounded-off. Multilobed protoplasmic bodies were invariably multinucleate, while giant round protoplasts had either several nuclei or had one nucleus of large size. Nuclear fusion, also, seemed to occur immediately after the protoplasts were transferred to sodium nitrate.

• Korean Journal of Plant Tissue Culture
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• v.28 no.4
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• pp.205-208
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• 2001

This study was conducted to improve the regeneration efficiency from seed-derived calli of rice by optimizing the copper concentrations in the media. Mature seeds were cultured on MS medium supplemented with copper sulphate (0 to 5.0 mg/L) and 2 mg/L 2,4-D. Callus growth was influenced by the levels of copper sulphate containing with medium, The addition of copper sulphate (2.5 mg/L) in regeneration medium enhanced dramatically the ability of plant regeneration from seed-derived calli. The mean frequency of plant regeneration of 6 indica rices was 27.4% on medium containing copper sulphate, whereas that of the cultivars on copper-free medium was 2.4%. These results suggest that copper sulphate may have an important role in improving regeneration ability of indica rices.

• Korean Journal of Plant Tissue Culture
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• v.25 no.5
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• pp.295-299
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• 1998

To detect the effects of gelling agent brands and concentration on rice anther culture, anthers of rice(O. sativa L. japonica, cv, Nagdongbyeo) were inoculated on N6-Y1 basic media supplemented with 0.4~1.6% Bacto agar(Difco, 04140-01), Agarose(Sigma, Type 1) and 0.2~0.8% Gelrite(Kelco, 143364) as gelling agents. On 0.4% Bacto agar and Agarose media, the frequency of callus formation which was significantly decreased in proportion to gelling agent's concentration was 39% and 55%, respectively. On 0.6% Gelrite media, the frequency of callus formation which was not statistically significant among the 0.2~0.8% concentration was 44%. Calli derived from the higher concentration of gelling agents showed embryogenic with slow growth, small, whitish and hard shape compare to that of the lower concentration. The frequency of green plant regeneration was high not only in calli derived from the higher concentration but also in plant regeneration medium with the higher concentration after callus transfer. Calli derived from the higher concentration was effective to maintain the frequency of green plant regeneration up to 60 days after anther inoculation. Introduction of 0.6~0.8% Geltite for callus formation, then transferred 1.6% Bacto agar and Agarose or 0.8% Gelrite for green plant regeneration was effective to increase anther culture efficiency.

• Journal of Plant Biotechnology
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• v.32 no.1
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• pp.15-21
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• 2005

The expression patterns of cyclin genes, which play a crucial role on cell cycle control, were analyzed with rice calli and leaf blades from seedlings. When callus was transferred from media containing the combinations of 2,4-D and kinetin under the dark conditions to medium supplemented with cytokinin-only on 7 days after the cultures, the expression levels of A-, B- and C-type cyclins from callus were increased significantly. Despite the fact that cyclin genes were well expressed on leaf blades rather than other organs in rice seedlings, rice leaf blades grown on the medium containing various combinations and concentrations of cytokinin for 24 hours had no major effect on the expression patterns of cyclins except zeatin. The relation between cytokinin regulation and the expression of cyclins of rice is discussed.

• Journal of Life Science
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• v.31 no.4
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• pp.385-388
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• 2021

Anther cultivation for crop breeding is a method of rapid production of homozygosities by greatly reducing the time required for at least six generations to develop new varieties using conventional breeding methods. This technique of producing anther culture provides an opportunity to obtain more green plants from a methodological point of view, and the techniques that save time and effort in anther culture are also important because they increase the efficiency of culture. This study compared the callus induction rate and green plant regeneration rate of a one-step and a two-step culture that differ in their culture media and culture methods. One-step culture allows callus induction and plant regeneration in one medium, whereas two-step culture requires induction and plant regeneration in two different media. In this study, we compared the callus induction and plant regeneration rates of rice anthers as one-step and two-step cultures. The callus formation rate was 13.0% for one-step cultures and 8.6% for two-step cultures, so the rate was 4.4% higher for one-step cultures than for two-step cultures. The plant regeneration rate was 1.0% in one-step cultures and 3.0% in two-step cultures, so the regeneration rate was three times higher for the two-step cultures than for one-step cultures. This suggests that the two-step cultures are more efficient than the one-step cultures for haploid production.