• Journal of Plant Biology
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• v.31 no.4
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• pp.249-258
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• 1988

Changes in starch content and activities of ADPG- and UDPG-starch synthase and $\alpha$- and, $\beta$-amylase were studied in order to investigate effects of gibberellic aicd and abscisic acid on endogenous starch content during shoot differentiation and protocorm propagation in Cymbidium spp. (Jungfrau) protocorm. Shoot differentiation was promoted during the degradation of endogenous starch and protocorn propagation was promoted during starch accumulation in protocorm. The activities of ADPG- and, UDPG-starch synthase and $\alpha$- and $\beta$-amylase seemed to be related with starch content. Shoot differentiation and protocorm propagation were slightly inhibited in protocorm explants treated with 100$\mu$M gibberellic acid. The explants treated with 10$\mu$M abscisic acid lost the capacity for shoot differentiation and protocorm propagation, and that could not be overcome by 100$\mu$M gibberellic acid added to culture medium. Starch content fluctuated as the control even after 10$\mu$M abscisic acid. None the less, the treatment completely inhibited shoot differentiation and protocorm propagation.

• Journal of Plant Biology
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• v.33 no.1
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• pp.41-48
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• 1990

Changes in polamine titers during shoot differentiation in Cymbidium sp. (Jungfrau) protocorms were studied in order to investigate the mechanism of shoot differentiation by using auxin-inhibitors(PCIB, TIBA), hormones(GA3, ABA, BA), and phenolic compounds (2,4-dichlorophenol, catechol). The shoot differentiation and propagation of protocorms were promoted by PCIB or 2,4-dichlorophenol, and the growth of differentiated shoot were promoted by TIBA or catechol. In BA-treated protocorms, white or brown protocorms were observed. Putrescine was the most abundant polyamine during the propagation and differentiation processes. As compared with putrescine, spermidine did not show significant changes and spermine was not detected at all. Putrescine titers decreased after a temporary increase, and then again increased in the presence of GA3, ABA, 2,4-dichlorophenol, and then again increased in the presence of GA3, ABA, 2,4-dichlorophenol, catechol, or PCIB. But, in BA-treated protocorms, putrescine level was much lower than spermidine.

• Journal of Plant Biology
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• v.33 no.2
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• pp.105-110
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• 1990

Physiological gradient of IAA-oxidizing enzyme activities was investigated in order to elucidate the mechanism of shoot differentiation in Cymbidium sp. (‘Jungfrau’) protocorms by using phenolic compounds (2, 4-dichlorophenol, catechol), auxin-inhibitors (PCIB, TIBA), and hormones (GA3, ABA, BA). The activity of IAA oxidase was decreased in protocorms treated with catechol decreased the catalytic activity of IAA oxidase or TIBA but this enzyme activity was increased after a temporary decrease at initial stages in the presence of 2, 4-dichlorophenol or PCIB. The activity of IAA oxidase in BA-treated protocorms (white and crown gall-like) was the highest of all. However, the catalytic activity of peroxidase increased after a temporary decrease at initial period. These results suggest that shoot differentiation and growth may be influenced by effective IAA levels in the protocorms causing IAA-oxidizing enzyme and phenolic compounds.

• Journal of Plant Biotechnology
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• v.44 no.2
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• pp.164-170
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• 2017

A rapid and efficient in vitro regeneration system was established for Hibiscus syriacus L. The successful regeneration protocol employs induction of shoot organogenesis on leaf, petiole, and root explants. Among the various plant growth regulators evaluated, thidiazuron (TDZ) was the most effective for inducing rapid shoot formation. Most efficient shoot regeneration frequency was obtained from Murashige and Skoog (MS) media containing 0.01 mg/L TDZ. Regeneration efficiency was highest in the roots, and lowest in the leaves. A combination of 0.01 mg/L TDZ with benzyladenine (BAP) markedly improved the frequency of shoot differentiation from the root (up to 98%) and petiole (up to 88%) explants. Furthermore, leaf and petiole explants showed the highest frequency of shoot induction in half-strength MS media containing 0.01 mg/L TDZ and 1.0 mg/L BAP, while root explants formed the greatest number of shoots when 0.01 mg/L TDZ and 0.1 mg/L BAP were added to half-strength MS media. Although the frequency of shoot differentiation from leaf explants was only 50%, the leaf is considered the most efficient plant organ for use in tissue culture because leaves are easier to obtain than roots and petioles. Our findings show that various organs of H. syriacus can be used for plant regeneration, and the protocol developed in this study may be applicable in the horticulture industry.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.29 no.1
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• pp.102-107
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• 1984

This study was undertaken to know the possibility of in vitro propagation of Stevia through axillary bud culture and the results indicated that: (1) Addition of NAA (0.01-0.05 mg/l) alone on Murashige-Skoog basal medium promoted shoot differentiation and growth rate. And also additional of kinetin of 0.5-1.0 mg/1 alone showed the same trend as that of NAA: (2) Addition of both NAA (0.01-0.05 mg/l) and kinetin (0.5-1.0mg/l) to MS medium promoted better shoot formation. (3) Shoot differentiation and growth were better on the full salt strength of MS medium (1X MS) than that of half strength ( $\frac{1}{2}$MS), while their effects were reversed for root differentiation

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

• Korean Journal of Plant Tissue Culture
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• v.22 no.2
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• pp.83-87
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• 1995

Regulation of organ differentiation by growth regulators was investigated through the shoot-tip and bulb-scale cultures of Lilium longiflorum (cv Georgia). When shoot tips were placed on MS medium supplemented with 0.1 mg/L NAA alone or 0.1 mg/L NAA and 0.1 mg/L BA, axillary shoots were proliferated. Root diffentiation and growth were stimulated on the basal medium. Although growth regulation did not seem to be necessary when bulb scales were used as explants for shoot differentiation, its differentiation was promoted vigorously by 0.2 mg/L NAA, but suppressed by BA. Bulblets were formed from bulb-scale-derived plantlets cultured on MS medium supplemented with 0.1 mg/L IBA. And more bulblets were formed from the plantlet in MS medium supplying 0.2 mg/L NAA with 6% than3% sucrose

• Plant Biotechnology Reports
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• v.3 no.4
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• pp.333-340
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• 2009

Developmental anomalies in the plumule meristem of peanut (Arachis hypogaea L.) somatic embryos resulted in poor shoot differentiation and reduced plant recovery. Existing meristems with caulogenic potential have never been tested for embryogenesis in peanut. The present experiment was designed to test the mature zygotic embryo axis derived plumule with three meristems for somatic embryogenesis. Embryogenic masses and embryos developed from the caulogenic meristems in the axils. Exposure of 2 weeks in primary medium with $90.5{\mu}M$ 2,4-D suppressed the shoot tip differentiation temporarily which then regained the ability to form the shoot on withdrawal of 2,4-D. Exposure of 4 weeks in primary medium with $90.5{\mu}M$ 2,4-D suppressed the shoot tip differentiation irreversibly. No shoot formation was noted from the tips in any of the cultures which were in secondary medium with $13.6{\mu}M$ 2,4-D. Development of somatic embryos directly from axillary meristems was confirmed histologically. Conversion frequency of these embryos was 11%. Thus, in this report, we describe a method to obtain somatic embryos from the determined organogenic buds of the axillary meristem, by culturing the nodal explant vertically on embryo induction medium. It also displays the possibility of obtaining both embryogenic and organogenic potential in two parts of the same explant simultaneously. The possibility of extending this approach for genetic transformation in in vivo system through direct DNA delivery or Agrobacterium injection in meristems can also be explored. Using Agrobacterium rhizogenes, we have demonstrated the possibility of gene transfer in the axillary meristems of seed-derived plumule explant.

• Horticultural Science & Technology
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• v.19 no.4
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• pp.535-539
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• 2001

To establish the cultural practices of persimmon in heated plastic house, the growth and flower bud differentiation of 4-year old 'Nishimurawase' persimmon tree planted in 80 L pots were investigated. Temperature was kept at above $18^{\circ}C$. The earlier heating induced the earlier flower bud differentiation, while the number of days from bud burst to flower bud differentiation was 70-75 days, which was not affected by the date of heating. The date of flower bud differentiation in 'Nishimurawase' persimmon was May 1, May 21, and June 1 in the plastic house heated from Jan. 15, Feb. 15, and Mar. 15, respectively. The number of flower primordia per bud in the plastic house was 6.2-7.1, slightly fewer than that of 7.8 in the open-field, with the tendency of increasing the number by earlier heating. The days required for the differentiation of the last flower bud primordia from the beginning of differentiation of the first bud were 40-60, and that required for the flower bud differentiation of all buds on the same shoot was 30. The total carbohydrates in shoot increased and the contents of nitrogen decreased, resulting in increased C/N ratio just before flower bud differentiation, and it coincided with the time when the total carbohydates increased above 15%. During two weeks before flower bud differentiaton, The contents of zeatin per 1 g dry matter of shoot drastically increased from $27.2{\mu}g$ to $47.3{\mu}g$, while that of IAA slightly decreased from $188.6{\mu}g$ to $172.4{\mu}g$, and that of GA decreased from $2,225{\mu}g$ to $1,555{\mu}g$.

• Journal of Plant Biotechnology
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• v.35 no.2
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• pp.133-140
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• 2008

The modification of DNA and histone plays an important role for gene expression in plant development. The objective of this research is to observe the effects of methylation on the gene expression during dedifferentiation from rice mature seeds to callus and differentiation from callus to shoots. The embryogenic callus with ability to shoot regeneration was not induced on the N6A medium supplemented with 5-azacytidine and abnormal callus with brown color was formed. When the normal rice callus was placed on the regeneration MSRA medium supplemented with 5-azacytidine, the shoot regeneration was inhibited. The results showed that 5-azacytidine, DNA demethylating agent, had negative effects on normal embryogenic callus formation and shoot regeneration. This suggested that DNA methylation of some genes was required for normal cell dedifferentiation and differentiation in tissue culture. The microarray and $GeneFishig^{TM}$ DEG screening were used to observe the gene transcript profile in callus induction and regeneration on N6A (N6 medium + 5-azaC) and MSRA (MS regeneration medium + 5-azaC). Subsets of genes were up-regulated or down-regulated in response to 5-azaC treatments. The genes related with epigenetic regulation, electron transport, nucleic acid metabolism and response to stress were up and down regulated. The different expression of some genes (germin like protein etc.) during callus induction and shoot regeneration was confirmed using RT-PCR and northern blot analysis.