• Asian Journal of Turfgrass Science
• /
• v.12 no.4
• /
• pp.195-202
• /
• 1998

The development of a protocol for high efficiency of embryogenic callus separation, maintenance and plant regeneration from the seeds of zoysiagrass cv. Zenith was studied. Embryogenic callus ratio is absolutely determined by genotype, but by adding high concentration of copper into medium, changing light condition and maintaining callus on initial induction medium for 8∼10 weeks, embryogenic callus can be easily distinguished and its growth can be promoted. There were significant differences among selected callus lines (each from one seed) according to their growth rates and regeneration percentages. Callus pre-treatment with activated charcoal inhibited callus growth, increased the level of precocious germination during culture and promoted shoot cluster formation after transfer to regeneration medium. For long-term callus maintenance, N6AA medium was better than MS medium, because the former inhibited non-embryogenic callus formation and kept vigor of embryogenic callus. The best callus lines Z-(5) has been successfully used for transformation and somaclonal variation selection in our laboratory.

• Journal of Plant Biotechnology
• /
• v.33 no.4
• /
• pp.257-262
• /
• 2006

The paper evaluated the behavior of in vitro culture responses from a diverse set of Indica rice (Oryza sativa L.) genotypes. Significant differences were found in embryogenic callus induction frequency, callus growth and plant regeneration frequency when mature embryos of 11 cultivars, breeding lines and land races were compared. Genotype as well as plant growth regulator influenced the plant regeneration frequency. Callus induction frequency was not correlated with callus growth as well as plant regeneration frequency. The regenerated plants could grow to normal, fertile plants after they were successfully established in soil.

• Asian Journal of Turfgrass Science
• /
• v.11 no.4
• /
• pp.311-320
• /
• 1997

This study was carried out to induce and maintain callus from 59 zoysiagrass lines, to know the effective disinfestation method for zoysiagrass stolon as explant and the difference in the response of callus induction among 59 lines, and to investigate the effect of medium, growth regulators, light, temperature, stolon part and internode position on callus induction and emhryogenic callus(E.C.) formation. The treatment of 0.lmg/L $HgCl_2$for 15 min resulted in no contamination and the highest callus induction(46.6%). Callus was induced from the 59 zoysiagrass lines. The callus growth of Z. japonica and Z. sinica was generally better than Z. matrella Ten cell lines whose callus and stolon grow fast in culture and in field, respectively were selected to he used for breeding. Callus induction was the most effective at 2.0mg /L of both 2, 4-D and picloram in MS medium. MS medium was the best for callus induction and growth while LS medium was the best for embryogenic callus and shoot formation. Callus induction and growth was better at 28, 31$^{\circ}C$. than 25$^{\circ}C$. and dark condition was better than light condition in MS me-dium containing 2mg/L 2,4-D. While callus induction was better with node part as explant than with internode part, callus growth and embryogenic callus formation was better with internode part. In 'Japonica 1', the first internode was the most effective in callus induction, but third internode was the best in '$M_2$ X $S_2$'.

• Journal of Plant Biotechnology
• /
• v.32 no.1
• /
• pp.31-35
• /
• 2005

To obtain regeneration system of onion, we analyzed the effects of 2,4-D and BA concentration on the embryogenic callus induction from mature embryos. The highest embryogenic callus induction ratio was shown on MS medium (Murashie and Skoog 1962) containing $2.5\;\cal{mg/L}\;or\;5\;\cal{mg/L}$ picloram after mature embryos were placed on medium. When induced callus were cultured on half strength of MS medium containing $1\;\cal{mg/L}$ Kinetin, the highest shoot formation ratio was observed on MS medium containing $1\;{mg/L}$ 2,4-D and $1\;{mg/L}$ BA. Embryogenic callus were cultured in MS liquid medium containing $1\;\ccal{mg/L}$ of 2,4-D and $1\;\cal{mg/L}$ BA. The suspension cultured cell clumps could be mass propagated. Embryogenic callus were friable, but non-embryogenic callus included a lot of moisture, hence the identification between embryogenic and non-embryogenic callus as easily achieved. When embryogenic callus as cultured on half strength of MS medium containing $1\;\cal{mg/L}$ Kinetin, shoots were induced. The whole plantlet was obtained on rooting medium containing $0.5\;\cal{mg/}$ of NAA.

• Asian Journal of Turfgrass Science
• /
• v.12 no.4
• /
• pp.189-194
• /
• 1998

The effect of proline, glutamine, aspartic acid and their combinations on callus induction and embriogenic callus formation from 3 creeping bentgrass (Agrostis palustris) cv. Regent, Mariner, Cato and 1 colonial bentgrass (Agrostis tenuis) cv. Tiger was estimated in both light and dark condition. The addition of amino acids to the growth medium did not have a significant stimulatory effect on the induction of embryogenic callus, instead, they were inhibitory, particularly at higher concentration (40 mM). But supplement of amino acids at lower concentrations (5 or 10mM) to basal medium was beneficial in inhibiting the formation of hairy outgrowth on the surface of embryogenic callus.

• Journal of Plant Biotechnology
• /
• v.6 no.1
• /
• pp.55-61
• /
• 2004

Callus induction and somatic embryogenesis are fundamental to cotton tissue culture biotechnology. An efficient protocol for callus induction, somatic embryogenesis and their maturation have been developed to regenerate plantlets from cotton (Gossypium hirsutum L.) variety coker 312. Embryogenic callus was initiated from hypo-cotyl region that was used as an explant at seedling stage when it was about 7-8 days old. Callus induction was achieved through culturing hypocotyls (5-7mm) on $MS_{1a} medium supplemented with 2,4-D (0.1 mg/L) and KT (0.5 mg/L) for six weeks. A friable, colorless, bulky and well proliferating callus becomes greenish with the addition of NAA (2.0 mg/L), ZT (0.1 mg/L) and removal of 2,4-D (M $S_{1b}$) cultured for two weeks then again transferred to $MS_{1a}. 2,4-dichlorophenoxyacetic acid (2,4-D) promoted the proliferation of embryogenic callus, but had a negative effect on the differentiation and germination of somatic embryos. ZT (0.1mg/L) and activated charcoal (2g/L), both hormones play an important role in differentiation and germination of somatic embryos in hypocotyls derived embryogenic callus but in case of cotton, such a capability have been observed on MS medium with 1.92 g/L $KNO_3$, but it is considered to attain somewhat more improvement. High embryogenesis frequency was achieved through nutrient deficient stress treatment. The frequency of globular embryogenesis (two-three folds) was achieved when well proliferating callus was (from $MS_{1a}$ media) cultured on MS (1/5 strength) medium for four weeks. Here the development of anthocyanins is the best indicator for somatic embryogenesis. However, when embryoid callus was cultured on MS (full strength) medium, the globular embryos were developed into normal plantlets immediately. In this procedure 27.49% cotyledenary embryos were developed. Of that 70% cotyledenary embryos were developed not only into normal plantlets but rooted simultaneously, when cultured on MS (with 0.05 mgg/L giberrelic acid) medium. So complete plants could be regenerated through somatic embryogenesis from hypocotyl explants within 6 months.s.

• Journal of Plant Biology
• /
• v.31 no.1
• /
• pp.41-49
• /
• 1988

Several cultivars of rice were examined for induction of embryogenic callus on a medium containing MS salts, vitamins and 2, 4-D under darkness. Embryogenic callus was obtained from cultivar Cheonma with high ratio and embryo-like structures were formed from the callus on a medium with or without reduced 2, 4-D. Somatic embryoids with a plumule and radicle axis surrounded by a scutellum were observed. These embryoids germinated and produced plantlets in 30 days on the same medium. Protoplasts isolated from an embryogenic cell suspension culture derived from embryogenic callus were cultured either in liquid or in agar medium and protoplast derived cell colonies were obtained in 3-4 weeks.

• The Journal of the Convergence on Culture Technology
• /
• v.7 no.3
• /
• pp.605-608
• /
• 2021

Lily is one of the most important 5 cut flowers in international flower market and lilies are distributed in Asia, Eurasia and North America. To develop a new lily cultivar, in addition to hybridization, mutation and selection methods, biotechnological techniques including tissue culture are also required. Establishment of tissue culture system is one of the requirement for the breeding program in Lily. Among many fields of plant tissue culture, establishment of regeneration system via embryogenic calluses are studied in many crops. In this study, research was carried out to decide the proper concentration of picloram which is used for the induction of embryogenic calluses. As a result, 3 different types of callused were observed after 3-4 weeks. They were CEC (compact embryogenic callus), FEC (friable embryogenic callus) and white callus type. 1.0 mg /l of picloram showed the best result for the production of embryogenic callus, however, due to its higher rate of browning in this concentration, 0.75 mg/l of picloram was selected as a proper concentration of picloram for the induction of CEC and FEC in Lily. These results can be contributed to the establishment of both regeneration system and mass propagation in lily in the future.

• Korean Journal of Plant Tissue Culture
• /
• v.28 no.5
• /
• pp.277-281
• /
• 2001

Embryogenic callus induction and plant regeneration methods were developed for native Kentucky bluegrass (Poa pratenes L.) ecotypes. Mature caryopses and immature inflorescences (20 mm in length) of 4 native ecotypes and 5 foreign cultivars were plated on MS medium (30 g/L sucrose, 3 g/L Phytagel) supplemented with 1 mg/L 2,4-D, and cultured in the dark at 24$^{\circ}C$. Most explants formed calli, but more embryogenic calli were induced from the explants of immature inflorescences than caryopses which produced mostly non-embryogenic rooty calli. In P77 ecotypes, immature inflorescence explants formed embryogenic calli with the rate of 62~95%, and those of field-grown plants were more efficient than greenhouse-grown ones in embryogenic callus induction. Plantlets were regenerated from the embryogenic calli when they were transferred to hormone-free MS medium, and grew to maturity without morphological variations in greenhouse.

• Plant Biotechnology Reports
• /
• v.3 no.3
• /
• pp.243-250
• /
• 2009

In the present study, a simple one medium formulation protocol for callus culture, somatic embryogenesis and in vitro production of ${\beta}-carboline$ alkaloids and diosgenin in Tribulus terrestris L. was developed. Extensive callus induction and proliferation was obtained in leaf explant on Murashige and Skoog (MS) medium supplemented with $5.0{\mu}M$ 6 benzyl adenine (BA) and $2.5{\mu}M$ ${\alpha}-naphthaleneacetic$ acid (NAA). The embryogenic callus was maintained on subculture to fresh parental medium at 4-week intervals over a period of 28 months. The frequency of embryo formation was at a maximum ($18.1{\pm}0.9$ per g of callus) on MS medium containing $5.0{\mu}M$ BA and $2.5{\mu}M$ NAA together with $75mg\;1^{-1}$ casein hydrolysate. Globular embryo developed into torpedo stage embryo under the influence of starvation. The accumulation of ${\beta}-carboline$ alkaloids (harmaline and harmine) and steroidal saponin (diosgenin) in non-embryogenic and embryogenic callus culture derived from leaf explant was compared with root, leaf, stem, and fruit of the mother plant. The embryogenic callus accumulated equivalent amounts of harmaline ($66.4{\pm}0.5{\mu}g/g$ dry weight), harmine ($82.7{\pm}0.6{\mu}g/g$ dry weight), and diosgenin ($170.7{\pm}1.0{\mu}g/g$ dry weight) to that of the fruit of T. terrestris. The embryogenic callus culture of this species might offer a potential source for production of important pharmaceuticals.