• Journal of The Korean Society of Grassland and Forage Science
• /
• v.27 no.4
• /
• pp.235-240
• /
• 2007

Optimum tissue culture conditions for an efficient induction of embryogenic callus from mature seeds of perennial ryegrass (Lolium perenne L.) and regeneration of plants from callus tissues were investigated. MS medium containing 3 mg/L 2,4-D and 0.1 mg/L BA was optimal for embryogenic callus induction from mature seeds. The highest plant regeneration frequency (58.3%) was observed when the embryogenic callus tissues were cultured on N6 medium supplemented with 1 mg/L 2,4-D and 3 mg/L BA. Regenerated plants were grown normally when shoots transplanted to the soil. A short tissue culture period and high-frequency regeneration system would be helpful for molecular breeding of perennial ryegrass through Agrobacterium-mediated genetic transformation.

• Korean Journal of Plant Tissue Culture
• /
• v.21 no.4
• /
• pp.247-252
• /
• 1994

The effects of explant sources, plant growth regulators on callus induction and plantlet differentiation from leaf blade, petiole, and meristem tissue of Pelalgonium citrosa were investigated under illumination or in dark condition Leaf blade explants cultured on Murashige and Skoog's medium containing 2,4-D and kinetin did not form callus or organ. But those cultured on medium with NAA and BA produced callcus and shoots. Dark condition was more effective than light condition to callus induction and showed that some of shoot were differentiated directly from leaf blade explane. Callus proliferated vigorously on meristem tissue after 7 days of culture, and multiple shoots were obtained Sum callus on medium with 0.5 mg/L NAA and BA. Roots formed readily from about 80% of the shoots cultured on medium with 1.0 mg/L NAA. Regenerated plantlets regenerated had phenotypically normal leaves and roots.

• The Journal of the Convergence on Culture Technology
• /
• v.5 no.1
• /
• pp.409-412
• /
• 2019

We investigated the development of a micropropagation protocol for multiplication of calla 'Gagsi' by using shoots as explant. The callus was induced on Murashige and Skoog (MS) basal medium containing cefotaxime antibiotics (25, 50, 100 mg/L). Also, MS basal medium with NAA 0.5 mg/L and BA 1.0 mg/L was used. The callus induction and browning rates were compared by treatment supplemented cefotaxime 25, 50 and 100 mg/L in basal MS medium. The callus induction rate was 10.5 % and browning rate was also, 10.5 % on the MS containing 25 mg/L. In the MNB containing cefotaxime, the callus induction rate was 34.5 % and browning rate was 27.0 %. The cefotaxime experiment has been widely used in previous studies. It is thought that it will help establish the mass multiplication system by positively affecting the growth and browning reduction of calla plants.

• Plant Biotechnology Reports
• /
• v.4 no.4
• /
• pp.261-267
• /
• 2010

We describe culture conditions for a high-efficiency in vitro regeneration system of Papaver nudicaule through somatic embryogenesis and secondary somatic embryogenesis. The embryogenic callus induction rate was highest when petiole explants were cultured on Murashige and Skoog (MS) medium containing 1.0 mg $1^{-1}$ ${\alpha}$-naphthaleneacetic acid (NAA) and 0.1 mg $1^{-1}$ 6-benzyladenine (BA) (36.7%). When transferred to plant growth regulator (PGR)-free medium, 430 somatic embryos formed asynchronously from 90 mg of embryogenic callus in each 100-ml flask. Early-stage somatic embryos were transferred to MS medium containing 1.0 mg $1^{-1}$ BA and 1.0 mg $1^{-1}$ NAA to germinate at high frequency (97.6%). One-third-strength MS medium with 1.0% sucrose and 1.0 mg $1^{-1}$ $GA_3$ had the highest frequency of plantlet conversion from somatic embryos (91.2%). Over 90% of regenerated plantlets were successfully acclimated in the greenhouse. Secondary somatic embryos were frequently induced directly when the excised hypocotyls of the primary somatic embryos were cultured on MS medium without PGRs. Sucrose concentration significantly affected the induction of secondary embryos. The highest induction rate (89.5) and number of secondary somatic embryos per explant (9.3) were obtained by 1% sucrose. Most secondary embryos (87.2-94.3%) developed into the cotyledonary stage on induction medium. All cotyledonary secondary embryos were converted into plantlets both in liquid and on semisolid 1/3-strength MS medium with 1.0% sucrose.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.6 no.1
• /
• pp.51-55
• /
• 2001

This study was conducted to establish a plant cell culture system for the production of medically important secondary metabolites from Xanthium strumarium. The effects of plant growth regulators including NAA, 2,4-D, kinetin, and ABA were examined in terms of callus induction, maintenance of callus and suspension cultures. It was shown that callus was induced upon treatment with NAA while embryo was induced after treatment with 2,4-D. Callus formation was further improved by treatment with ABA and NAA. The level of callusing increased by 17-29% for the seed case, cotyledon, leaf, and hypocotyl and by 96% in the case of the root. Suspension cell lines were established using calli produced from cotyledon, hypocotyl and root and cultured at 25$\^{C}$ under light conditions. The cells grew up to 15g/L with NAA 2ppm, BA 2ppm, and ABA 1ppm treatment. Supernatants of suspension cultures of cell lines derived from coyledon and hypocotyl produced some distinctive secondary metabolites, one of which was identified as 8-epi-tomentosin, which belongs to the xanthanolides. The amounts of 8-epi-tomentosin produced by the cotyledon- and hypocotylderived cell lines were 13.4mg/L and 11.0mg/L, respectively.

• Journal of Plant Biotechnology
• /
• v.5 no.2
• /
• pp.95-99
• /
• 2003

The leaf segments of garlic (Allium sativum L.) were cultured in vitro and determined optimal concentration of plant growth regulators and sugars for callus induction, multiple shoots regeneration and in vitro bulblet formation. Highest yield of callus was observed in the leaf segment culture on Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-D, 30 g/L sucrose and 8 g/L agar. Regeneration rate of multiple shoots from callus was high in the MS medium supplemented with kinetin 3.0 + NAA 3.0 mg/L or SA 1.0 + NAA 3.0 mg/L, containing 30 g/L sucrose. High rate of bulblet formation was observed as the concentration of jasmonic acid increased from 0.5 to 2.0 mg/L in medium, whereas addition of gibberellic acid significantly suppressed bulblet formation. The rate of in vitro bulbing was as high as $96\%$ in MS medium supplemented with 2.0 mg/L jasmonic acid and 120 g/l sucrose after two month culture at $25{\pm}1^{\circ}C$ under 16 hours day light.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.20 no.1
• /
• pp.25-30
• /
• 2000

The conditions for callus formation and plant regeneration were confirmed in Italian ryegrass (Lolium mulfiorum Lam.). Among SH (Schenk and Hildebrandt), MS (Murashige and Skoog) and N6 medium (Chu) MS medium was highest degree of efficiencies respectively in callus formation and plant regeneration. In this study, we determined volume of hormones and other compounds appended in media. For callus formation, only $5\;mg/\;{\ell}$ of 2,4-D (2,4-dichlorophenoxy acetic acid) was appended in their media. For plant regeneration, we used MS medium containing $1.0\;mg/\;{\ell}$ of BA and $0.1\;mg/\;{\ell}$ of NAA.

• Korean Journal of Plant Tissue Culture
• /
• v.22 no.1
• /
• pp.35-39
• /
• 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 Life Science
• /
• v.31 no.4
• /
• pp.385-388
• /
• 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.

• Journal of Plant Biotechnology
• /
• v.50
• /
• pp.19-26
• /
• 2023

Korean ginseng (Panax ginseng Meyer) is an economically important plant because of it is rich in saponins. It is mainly cultivated in Asia, including Korea and China. Since ginseng requires a long breeding period due to juvenility, homozygote production techniques, such as anther culture, must be urgently established. In the present study, callus induction and embryogenesis through anther culture were observed in P. ginseng. Murashige and Skoog medium was used as the basal medium suitable for callus induction. When the medium was supplemented with 3% sucrose, the callus induction rate was high and the callus size was large. Cold pretreatment did not significantly affect callus induction and embryogenesis. Embryogenesis was the most efficient when the embryo-formation medium was supplemented with 1.0 or 3.0 mg/L 2,4-dichlorophenoxyacetic acid. Cultivar significantly affected anther culture efficiency. Specifically, 'Cheongseon' showed the highest embryo-formation efficiency, whereas no embryogenesis occurred in 'Sunun'. Ploidy assessment revealed the haploid status of the induced calli. Embryos derived from anther culture formed shoots upon transfer to germination medium, although no difference in ploidy was noted between the induced callus and control. Overall, the anther culture conditions established in the present study may contribute to the production of homozygous P. ginseng plants in the future.