• Plant Biotechnology Reports
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• v.4 no.1
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• pp.53-59
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• 2010

A specific deleted version of ARABIDOPSIS RESPONSE REGULATOR1 (ARR1) lacking the signal receiver domain (1.152 amino acids)-coding sequence, referred to as $ARR1{\Delta}DDK$, was amplified using Arabidopsis thaliana cDNA prepared from adult leaves and transferred into the genome of Nicotiana tabacum cv. Samsun under the transcriptional control of a ${\beta}$-estradiol-inducible expression system. The ectopic expression of $ARR1{\Delta}DDK$ affected the morphology of transgenic seedlings and their segments in vitro. In the presence of an inducer, ${\beta}$-estradiol, ectopic expression of $ARR1{\Delta}DDK$ induced only the formation of soft, pseudo-bulbous tissue in the root tip region of intact seedlings, which appeared similar to callus generated on a hypocotyl segment in the presence of 2,4-D and 6-benzyladenine (BA), both at $1\;{\mu}M$. Those callus tissues on the root tip region could not generate shoots unless $1\;{\mu}M$ BA was supplied. In segment culture, ectopic expression of $ARR1{\Delta}DDK$ induced calluslike tissue around the cut-end of cotyledon and hypocotyl segments with occasional shoot formation, suggesting that the expression of $ARR1{\Delta}DDK$ could substitute for the effects of cytokinin on these segments. Additionally, treatment with only ${\beta}$-estradiol induced NtWUS, a WUS ortholog in tobacco, which was detected during the process of callus tissue formation in the root tip region and also in cotyledon or hypocotyl segments. These findings suggest that the NtWUS might be associated in the transdifferentiation process caused by the functional regulation of $ARR1{\Delta}DDK$ in transgenic tobacco seedlings.

• Korean Journal of Plant Tissue Culture
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• v.21 no.4
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• pp.221-225
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• 1994

This experiment was conducted to identify the optimal in vitro propagation condition of Cnidii rhizoma (Cnidium officinale Makino). It was effective to reduce contamination and improve regeneration of shoot when shoot tips taken in July were cultured in 1/2 strength Murashige and Skoog medium supplemented with 500 mg/L carbenicillin disodium 1.0 mg/L BA and 1.0mg/L $GA_3$followed by surface sterilization of explant source in solution of 1% sodium hypochlorite for 20 minutes. When shoot tips were 쳐cultured in 1/2 strength MS medium with 0.5 mg/L BA and 60 g/L sucrose, shoot elogation and subsequent multiplication of the formed shoot were favorable than in other media. Regenerants were well rooted in 1/2 strength MS medium containing 3.0 mg/L NAA.

• Korean Journal of Weed Science
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• v.6 no.1
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• pp.25-32
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• 1986

This experiment was conducted to evaluate effect of various hormones on callus induction, and on plantlet formation on various media, and to detect of Londax [Methyl 2-[[[[[(4,6-dimethoxy pyrimidin-2-yl) amino] carbonyl] amono] sulfonyl) methyl] benzoate] and Basta[Ammonium-(3-amino-3-carboxy-propyl)-methyl phosphinate] on callus growth and reaction of succinate dehydrogenase in callus against TTC, using various species such as Eleocharis kuroguwai, Cyperus serotinus, Oryza sativa (samgangbyeo) and Echinochloa crusgalli P. Beauv. var. caudata Kitagawa. The optimal levels of 2,4-D in MS medium seems to be different among species tested, 2.0 ppm for rice and E. crusgalli, 1.0 ppm for Eleocharis kuroguwai, and 4.0 ppm for C. serotinus derived callus from shoot-tip. In case of combination of 2,4-D with BA, 1.0 plus 0.3 ppm appeared the most appropriate level to induce callus from rice and E. kuroguwai, and I.0 plus 0.1 ppm for C. serotinus and E. crusgalli. When 2,4-D treated with TIBA, 1.0 plus 0.5 ppm appeared the most appropriate rates to induce callus derived from seeds of rice, E. crusgalli, seeds of C. serotinus and E. kuroguwai, 1.0 plus 0.3 ppm for shoot-tip of C. serotinus. Positive reaction of succinate dehydrogenase against TTC was observed regardless of calli and herbicides tested, indicating that they all are alive, and these herbicides were not able to kill the calli tested within the short period of time 20 hrs treatment. Regardless of plant species used, the rate of plantlet formation from callus was very low. However, some plantlet formed from E. crusgalli at 0.8 ppm of 2,4-D plus 8.0 ppm of kinetin, and from E. kuroguwai at 1.6 ppm of 2,4-D plus 16.0 ppm of kinetin, showing effectiveness of 2,4-D with kinetin mixture treatment. No callus was induced from C. serotinus treated with Basta from $10^{-6}M$ to $10^{-3}M$. In general, rice was the least susceptible to Basta among plant species tested, followed by E. crusgalli, and E. kuroguwai. In Londax treatment, rice showed the least inhibition rate in callus growth. Callus was induced from rice even at $10^{-3}M$ of Londax. However, $10^{-3}M$ of Londax completely inhibited callus induction from the test species. Rice showed most tolerant to both herbicides, indicating the existence of different responses among plant species.

• Journal of Bio-Environment Control
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• v.29 no.1
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• pp.1-8
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• 2020

The influence of growth regulators (NAA and BA) and sucrose concentrations (0, 3, 5, 7, 9%) on in vitro rapid-propagation of virus-free sweet potato [Ipomoea batatas (L.) Lam.] was investigated with single-node or shoot-tip culture of two cultivars ('Matnami' and 'Shinhwangmi'). The survival rate and growth of shoot-tip explant was also investigated under the presence or absence of light (blue and red LED = 7:3, 150±5 μmol·m^-2·s^-1 PPFD) during minimal-growth in vitro conservation at 15℃. Vine length, vine diameter, fresh weight and dry weight were enhanced without callusing of explant in the MS medium supplemented with 0.2-0.5 mg·L^-1 BA. The growth of single-node and shoot-tip explants were significantly enhanced with the increase of vine length, number of leaf, number of root, fresh weight, and dry weight in the solid medium containing 5% sucrose and 0.2 mg·L^-1 BA. Vine elongation of shoot-tip explants were highest in the liquid medium containing 3% sucrose than the solid medium. The survival rate of minimal-growth in vitro conservation was 100% in 5 months under the presence of light (LED, 150±5 μmol·m^-2·s^-1 PPFD) at 15℃, but the explants in dark condition died in 3 months. The light was absolutely necessary for the in vitro conservation under minimal-growth conditions of virus-free sweet potato plantlets at 15℃, and the high density of explants (10 plantlets per Petri Dish) was increased the efficiency of mass conservation.

• Korean Journal of Plant Tissue Culture
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• v.25 no.3
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• pp.159-163
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• 1998

In vitro shoot tip culture technique was established in pear (Pyrus pyrifolia 'Niitaka') as related to tree vigor, sampling time, and plant growth regulators and sucrose supplemented to medium. Shoot tips excised in June from the tree having medium-vigor developed good shoots. BA (1.0 and 2.0 mg/L) without NAA produced shoots suitable for proliferation, and NAA supplemented to medium resulted in poor shoot growth and excessive callus formation. BA of 2.0 mg/L combined with 0.01 mg/L NAA provided shoots suitable for rooting and sucrose of 30 g/L was recommended for proliferation medium. A fourth strength MS medium supplemented with 0.1 mg/L NAA produced plantlets in good quality of root number and root length.

• Korean Journal of Plant Tissue Culture
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• v.27 no.1
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• pp.51-55
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• 2000

To investigate the effect of media and growth regulators in micropropagation of persimmon (Diospyros kaki Thunb.), dormant axillary buds taken from trees of persimmon cultivars such as Ichikikeijiro, Tonawase and Hiratenenashi were used. Shoot tips were successfully cultured in full or half of nitrogen strength of MS medium. The most effective cytokinins for shoot proliferation and elongation of persimmon cv. Ichikikeijiro were 5 mg/L and 2 mg/L zeatin, respectively. Shoots were successfully rooted in 1/2N-MS medium with 1 mg/L IBA.

• Korean Journal of Plant Resources
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• v.30 no.6
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• pp.612-617
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• 2017

The preservation of pear germplasm, like that of other clonal germplasms, is difficult because it requires conservation of whole plants or their tissues. Among the currently available methods for long-term conservation of clonal germplasm, cryopreservation of shoot tips is the most reliable and cost- and space-effective option. Alginate-coated axillary shoot tips from in vitro-grown pear were conserved successfully in liquid nitrogen (LN) following dehydration. Shoot recovery from cryopreserved shoot tips was improved greatly after 8 weeks of cold acclimation, but recovery decreased slightly after then. The highest regeneration rate was observed when in vitro shoot tips were preincubated in MS (Murashige and Skoog) medium with 0.3 M sucrose for 48 h, and when alginate-coated shoot tips were precultured in MS medium with increasing sucrose concentrations (0.5 M and 0.7 M) for 8 and 16 h, respectively. When the encapsulated beads were dehydrated for up to 7 h [25% water content (fresh weight basis)] under laminar flow, the highest regeneration rate was observed in "BaeYun No. 3" (55.7%) and "Whanggeum" (43.3%) after warming from LN. This technique is useful as a practical procedure to cryopreserve plant material that is sensitive to freezing of the surrounding cryoprotectant medium. Therefore, this technique appears to be promising for the cryopreservation of shoot tips from in vitro-grown plantlets of pear germplasm.

• Korean Journal of Plant Resources
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• v.31 no.6
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• pp.684-694
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• 2018

This study describes the successful establishment of a cryopreservation protocol for Citrus limon cultivars: 'Frost Eureka limon' and 'Cook Eureka limon', using a droplet-vitrification method. The shoot tips that were excised from in vitro grown seedlings of the two cultivars were preserved in liquid nitrogen (LN) and successfully regenerated into whole plants. Excised shoot tips were pre-cultured for 1 or 2 days in 0.3 M and 0.5 M sucrose solutions at $25^{\circ}C$ and incubated in a loading solution (LS) composed of 17.5% glycerol + 17.5% sucrose in Murashige and Skoog (MS) medium for 40 min at $25^{\circ}C$. Prior to direct immersion in LN for 1 h, the shoot tips were dehydrated with plant vitrification solution 2 (PVS2) at $0^{\circ}C$ or PVS3 at $25^{\circ}C$. The frozen shoot tips were re-warmed and unloaded with 1.2 M sucrose in $\text\tiny{^1/_2}$ MS for 30 min at $25^{\circ}C$. Shoot tips were post-cultured overnight on survival medium and then micrografted onto 'trifoliate orange' (Poncirus trifoliate (L.) Raf. seedling rootstocks for recovery and to produce whole plants. The highest regrowth rates were 53.5% and 50.3% for cryopreserved shoot tips of 'Frost Eureka limon' and 'Cook Eureka limon', respectively, when pre-cultured in 0.3 M and 0.5 M sucrose concentrations in a sequencing manner, with LS and treated with PVS2 for 60 min at $0^{\circ}C$. We also investigated whether the ammonium ion concentration on post-culture medium affected the viability of the cryopreserved Citrus shoot tips. The viability of cooled samples, following culturing on woody plant media (WPM) containing $\text\tiny{^1/_4}$ ammonium nitrate overnight before micrografting, was the highest (70.3%) in 'Frost Eureka limon'. The study described here is a cost-effective and safe method to conserve Citrus fruit cultivars, for the improvement and large-scale multiplication of fruit plants and for breeding disease resistance.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.4
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• pp.206-211
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• 2002

Calli obtained from a shoot-tip of garlic, Allium sntivum L., were encapsulated using a calcium alginate gel. Some of the encapsulated calli were cultured on a 1/2 MS medium supplemented with 3% sucrose, 10$\^$-5/ kinetin, and 5 ${\times}$ 10$\^$-6/ M NAA whereas the remainder was stored for 40 days at 4$^{\circ}C$. All the naked calli regenerated on the solid medium, while 95% of the encapsulated calli regenerated, and 88% of the encapsulated calli regenerated after 40 days of storage at 4$^{\circ}C$. The capsule matrix delayed the germination time of the encapsulated calli, yet activated the shoot formation of the artificial garlic seeds. The shoot length of the encapsulated garlic calli was much longer than that of the naked garlic calli. The encapsulated garlic calli were dried in a laminar airflow cabinet and the conversion frequency of the dried artificial garlic seeds on a 1/2 MS medium remained at 93% with a water Loss of Less than 50%.

• Korean Journal Plant Pathology
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• v.14 no.5
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• pp.507-512
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• 1998

Molecular analysis has been done for characterization of the interactions between three beet curly top virus (BCTV) strains and two Arabidopsis ecotypes in terms of virus inducible disease symptoms and infectivities. The total DNA was isolated from three tissues (shoot tips, infection origins and roots) of virus infected plants and this DNA was analyzed by quantitatively and qualitatively to elucidate virus movement and symptom development. CTV-Worland infected Col-O and Sei-O showed only symptom shown in hypersusceptible ecotype Sei-O by BCTV-worland was shoot tip stunting. Kinetics of virus DNA accumulation of three different viruses indicated that roots contained more virus DNA than shoot tips or infection origins, and that disease symptom severity was strongly correlated with virus DNA accumulation. These results suggest that the mild and Worland-specific symptoms shown in Sei-O by BCTV-worland are caused by the interactions of host factors provided by hypersusceptible ecotype and viral factors of mild strain.