• Journal of Plant Biology
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• v.15 no.1
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• pp.28-32
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• 1972

Young haploid leaf derived from the anthers of tobacco plant was cultuerd and plantlets of various ploidies were obtained. When the leaf was put on the medium supplemented with kinetin as growth regulator, plantlets developed directly from the leaf, and the plants coming out in early stage of culture were all haploid. Plants developing in later stage were mostly haploids with some exception of diploid and aneuploid. Leaves were also cultured on the callus-inducing media supplemented with 2,4-D and kinetiion, and the calluses were sub-cultured for six months. Plants developed from these calluses were mostly aneuploids of various chromosome numbers. In view of the fact that the plants directly developed from the leaf were all haploid, the tissue of the original leaf explant was assumed to be uniform as far as chromosome number was concerned. On the other hand, it seemed that the occurrence of various ploidies in the plants derived from the calluses of same origin was the result of the influence of in vitro culture. Apical meristem tissues and various multicellular bodies were formed in the epidermal and inner mesophyll tissues as well as in the sub-epidermal cells.

• Plant Resources
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• v.6 no.2
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• pp.89-93
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• 2003

This study was conducted to develop the clonal propagation technique through in vitro culture using basal stem explants in Phalaenopsis hybrid grown in vitro. The highest frequency of protocorm-like body (PLB) formation was obtained when basal stem explants were cultured on VW medium containing 30g/L sucrose, 500 mg/L activated charcoal, 150 ml/L coconut water, 1 mg/L NAA, 5 mg/L 2iP and 2.5g/L gel rite. PLBs transferred to Hyponex medium were regenerated to plantlets. Plantlets transferred to plastic pots containing spagnum moss were developed and successfully acclimatized under greenhouse. The flower was bloomingly opened in plants regenerated from basal stem explants. The flower was not different from both mother plant and plant induced through clonal propagation of Phalaenopsis hybrid.

• Journal of Plant Biology
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• v.14 no.2
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• pp.7-10
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• 1971

Haploid cell obta-ined from microspores of Solanum nigrum were cultured on two kinds of medium, "Callus-inducing medium" and "Differentiation medium", in order to conduct histological studies of callus and examine differentiation of plantlets. On the callus-inducing medium the calli grew rapidly. The bulk of callus mass was light brown colored "Wet callus" covered on the surface with thin layers of rough and gleaming "White callus". The wet callus was consisted of parenchyma and meristematic tissues, while the white callus had no meristematic tissues. Large parenchyma cells, by successive divisions, became multicellular or poly nucleate cells which developed later to be meristematic tissues. The calli embedded on the differentiation medium quickly turned to dark brown color. Plantlets, however, came out later from these blackened callus mass. In the callus sectioned about ten weeks after imbedding on the differentiation medium, radially elongated tissue, concentric tissue, epidermis, tracheid-like structure, and plant jprimordia were observed.ure, and plant jprimordia were observed.

• Korean Journal of Plant Resources
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• v.15 no.3
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• pp.221-226
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• 2002

This experiment was conducted to establish the mass propagation system of Calanthe discolor Lindley. When the Calanthe discolor seeds were sown in Murashige and Skoog medium, the percentage of germination was 65％. Seedlings grew more rapidly in the liquid medium than the solid medium. All regenerated plantlets were survived in acclimatized condition of 70% shade and more than 80％ humidity. Also, we found out that the 88% of survival ratio could be achieved in containing soil mixture of vermiculite and perlite as same as amount.

• Plant Biotechnology Reports
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• v.3 no.3
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• pp.199-203
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• 2009

Rugosa rose (Rosa rugosa) is cultivated as a garden flower and an important genetic resource for the breeding of roses (R. hybrida). This study describes culture conditions for high frequency plant regeneration from zygotic embryo explants via somatic embryogenesis in rugosa rose. Mature zygotic embryo, cotyledon, and radicle explants formed embryogenic calluses at frequencies of 38, 6.7, and 8.8% when cultured on half-strength Murashige and Skoog medium (${\frac{1}{2}}MS$) supplemented with 2.26, 9.05, and $9.05{\mu}M$ 2,4-dichlorophenoxyacetic acid, respectively. Embryogenic calluses produced numerous somatic embryos, which then developed into plantlets on ${\frac{1}{2}}MS$ without growth regulators. Regenerated plantlets were grown to whole plants in a growth chamber.

• Journal of Plant Biotechnology
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• v.41 no.1
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• pp.33-37
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• 2014

Lychnis wilfordii (Regel) Maxim is a rare and valued ornamental plant. Germination rate reached 46.6% when seeds were treated with $100mg{\cdot}l^{-1}$ GA (Gibberellic acid). The highest callus induction was observed in the leaf explants of the seedlings on MS medium containing specific concentrations of $0.5mg{\cdot}l^{-1}$ BA ($N^6$-benzyladenine) and $3.0mg{\cdot}l^{-1}$ NAA (a-naphthalene acetic acid). The adventitious shoot was formed in 97.3% of calli on 1/2 WPM (Woody Plant Medium) medium. Shoot elongation of in vitro propagated plantlets was no difference among various medium. The plantlets grew well after transferring to the pot. This in vitro propagation protocol should be useful for conservation of this endangered plant.

• Plant Resources
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• v.2 no.1
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• pp.16-21
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• 1999

An efficient regeneration system was established by using in vitro plantlets of germinated seedlings from different cultivars of lettuce (Lactuca sativa cv. Chongchima, Chongchuckmyun, Jeokchima, Jeokchuckmyun). Shoot formation were observed from all cultivars on MS medium supplemented with 0.1 mg/L NAA and 0.5 mg/L BA. In all cultivars, when cotyledon was cultured, the number of shoot per explant was more greater than that hypocotyl and leaf disc were cultured. Shoot formation rate (91.7%) was high in a cotyledon culture of cultivar, Chongchukmyun. The growth of multiple shoots derived from the cultivar, Chongchukmyun, was most effective on medium containing 0.5 mg/L BA and 1.0 mg/L GA$_3$. When shoots were transferred on MS medium without plant growth regulators, roots were effectively differentiated. Rooted plantlets were acclimated on pots for further propagation.

• Journal of Plant Biotechnology
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• v.6 no.1
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• pp.55-61
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• 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 Biotechnology
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• v.7 no.4
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• pp.259-265
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• 2005

Indian citrus ringspot virus (ICRSV)-free plants of Kinnow mandarin (Citrus nobilis Lour x C. deliciosa Tenora) were raised from virus-infected plants using unfertilised ovules as explants. Plants were tested by indirect ELISA and RT-PCR before using their explant. An amplified product of 539 bp was obtained by RT- PCR in ICRSV infected plants. Unfertilized ovules were excised from unopened flower buds of plants tested postive for virus and were cultured on Murashige and Skoog's (MS) basal medium supplemented with various concentrations of kinetin (KN) or malt extract (ME). Maximum induction (31.94%) of embryogenic callus was observed on MS medium supplemented with KN ($9.29\;{\mu}M$). Transfer of embryogenic calli to similar media composition resulted in somatic embryogenesis in all cultures, with an average number of 60.36 globular, 17.39 heart and 7.71 cotyledonary-shaped somatic embryos per culture. All cotyledonary shaped embryos developed into complete plantlets within 60 days on transfer to similar medium. Embryogenic callus induction, somatic embryo formation, maturation, germination and plantlet formation were achieved on MS medium supplemented with KN ($9.29\;{\mu}M$) alone. The plantlets derived from somatic embryos were transferred to sterilized soil, sand and vermiculite (3:1:1) mixture. After acclimatization, the plantlets were transferred to screen house and were indexed for ICRSV employing indirect ELISA and RT-PCR and found free of virus. A distinct feature of this study is the induction of somatic embryogenesis from unfertilised ovules to produce virus-free plants.

• Korean Journal of Agricultural Science
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• v.5 no.1
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• pp.1-5
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• 1978

Aseptic meristem of Fragaria ananassa 'Hokowase' were inoculated on Murashige and Skoog medium containing various levels of Benzylamiro purine(BA), IAA, and 2.4-D. Formations of shoots plantlets, roots and callus depend on hormone levels used. On medium containing high level of BA 1.0mg/l, multiple plantlets were formed, however, elongation of shoots was inhibited than on BA 0.5 mg/l. 1.0mg/l IAA induced root formation and 1.0mg/l+1.0mg/l BA inhibited root formation. Callus formation was occurred on the medium added 2.4-D. When plantlets were subcultured, formation of callus or shoot depend on BA/2.4-D ratio. 2.0mg/l 2.4-D+0.2mg/l BA and 0.5 mg/l 2.4-D+0.2mg/l BA induced callus formation and 2.0mg/l BA induced plantlet and shoot vigorously.