• Journal of Plant Biotechnology
• /
• v.7 no.3
• /
• pp.183-186
• /
• 2005

Cryopreservation has been recognized as a practical and efficient tool for the long-term storage of vegetatively propagated plants. This study was conducted to investigate the effects of slow-freezing techniques on the cryopreservation of potato. In vitro plantlets of the potato genotypes of 'Atlantic', 'Superior’, 'Namseo', 'J138', and 'CTO5-5' were cold acclimated, and the excised axillary buds were precultured, osmoprotected, exposed to plant vitrification solution, frozen slowly to $-40^{\circ}C$ and then rapidly plunged into liquid nitrogen, thawed and finally plated on the regeneration medium. It was found that the higher the sucrose concentrations in the subculture medium of donor plantlets, the higher the survival rates of shoot tips after cryopreservation, and the highest survival (20%) was observed in the medium added with 0.25 M sucrose. As for the effect of cooling, $0.3^{\circ}C/min$ cooling speed showed the highest survival (25%). Different varieties showed different responses over different cryopreservation treatments. Survival rate was increased by slow-freezing technique method as compared with that of the basic cryopreservation method of vitrification alone in the diverse potato genotypes. Leaf and tuber morphologies of potatoes regenerated after cryopreservation using slow freezing technique were similar to those derived from the in vitro stock plantlets.

• Korean Journal of Plant Tissue Culture
• /
• v.24 no.3
• /
• pp.183-188
• /
• 1997

Callus from scale segments of Lilium longiflorum 'Gelia' was effectively induced and maintained from unorganized tissue on the semi-solid medium by 0.42% Bacto agar with MS basal salts and vitamins of SH medium supplemented with 0.5 mg/L 2, 4-D, 1.0 mg/L NAA, 0.3 mg/L BA, and 3% sucrose. More than 5% of high sucrose level had inhibiting effect on regeneration capacity of formed callus and decreased callus growth. Various combinations of nitrogen did not effective to proliferate the ELC (Embryogenic-like callus), but friability of callus was increased in the medium containing only nitrate as nitrogen source. 5 mL conditioned medium into 30 mL fresh medium was good for cell growth. However friable cell aggregates during suspension culture had to form hard callus which hindered to establish suspention culture system. Addition of 2 g/L casein hydrolysate increased callus growth and friability of the hard callus. As a result of anatomical observation of callus, organogenesis such as shoots, roots and bulblets was independently induced from callus tissue. Somatic embryogenesis from callus tissue could be observed with low frequency.

• Nutrition Research and Practice
• /
• v.10 no.1
• /
• pp.11-18
• /
• 2016

BACKGROUND/OBJECTIVES: Type 2 diabetes (T2D) is more frequently diagnosed and is characterized by hyperglycemia and insulin resistance. $\small{D}$-xylose, a sucrase inhibitor, may be useful as a functional sugar complement to inhibit increases in blood glucose levels. The objective of this study was to investigate the anti-diabetic effects of $\small{D}$-xylose both in vitro and stretpozotocin (STZ)-nicotinamide (NA)-induced models in vivo. MATERIALS/METHODS: Wistar rats were divided into the following groups: (i) normal control; (ii) diabetic control; (iii) diabetic rats supplemented with a diet where 5% of the total sucrose content in the diet was replaced with $\small{D}$-xylose; and (iv) diabetic rats supplemented with a diet where 10% of the total sucrose content in the diet was replaced with $\small{D}$-xylose. These groups were maintained for two weeks. The effects of $\small{D}$-xylose on blood glucose levels were examined using oral glucose tolerance test, insulin secretion assays, histology of liver and pancreas tissues, and analysis of phosphoenolpyruvate carboxylase (PEPCK) expression in liver tissues of a STZ-NA-induced experimental rat model. Levels of glucose uptake and insulin secretion by differentiated C2C12 muscle cells and INS-1 pancreatic ${\beta}$-cells were analyzed. RESULTS: In vivo, $\small{D}$-xylose supplementation significantly reduced fasting serum glucose levels (P < 0.05), it slightly reduced the area under the glucose curve, and increased insulin levels compared to the diabetic controls. $\small{D}$-xylose supplementation enhanced the regeneration of pancreas tissue and improved the arrangement of hepatocytes compared to the diabetic controls. Lower levels of PEPCK were detected in the liver tissues of $\small{D}$-xylose-supplemented rats (P < 0.05). In vitro, both 2-NBDG uptake by C2C12 cells and insulin secretion by INS-1 cells were increased with $\small{D}$-xylose supplementation in a dose-dependent manner compared to treatment with glucose alone. CONCLUSIONS: In this study, $\small{D}$-xylose exerted anti-diabetic effects in vivo by regulating blood glucose levels via regeneration of damaged pancreas and liver tissues and regulation of PEPCK, a key rate-limiting enzyme in the process of gluconeogenesis. In vitro, $\small{D}$-xylose induced the uptake of glucose by muscle cells and the secretion of insulin cells by ${\beta}$-cells. These mechanistic insights will facilitate the development of highly effective strategy for T2D.

• Current Research on Agriculture and Life Sciences
• /
• v.10
• /
• pp.137-145
• /
• 1992

An efficient transformation system was established for Ailanthus altissima utilizing the binary system of A. tumefaciens strain LBA4404. Callus was initiated from small portions of cambium tissue of A. altissima in vitro. Optimum regeneration was achieved with Murashige and Skoog(MS) medium containing 0.01mg/${\ell}$ 2, 4-D, 0.5mg/${\ell}$ BAP, 3%(w/v) sucrose and 0.75% agar. The multiplication of explants remarkably showed up on medium containing 1.0mg/${\ell}$ BAP. Leaf discs or internodal stem segments were inoculated with A. tumefaciens strain LBA 4404 containing the binary vector pPMB 101, which has both ${\beta}$-glucuronidase (GUS) marker gene and neomycin phosphotransferase II (NPT II) gene. Shoots had been regenerated from 24 lines out of inoculative 50 lines. Transformants were selected by their ability to grow on medium containing kanamycin sulphate (100mg/${\ell}$). Putative transformation was confirmed by GUS assays. Five GUS-positive plantlets were obtained which confirmed that this marker gene has been transferred into A. altissima.

• Korean Journal of Plant Tissue Culture
• /
• v.27 no.2
• /
• pp.125-131
• /
• 2000

Protoplasts of Arabidopsis thaliana were easily isolated from the shoot-forming (SF) suspension-cultured cell clusters with 4 hours-shaking condition (40 rpm) on CPD enzyme solution containing 1% cellulase R-10, 0.25% pectolyase Y-23 and 0.5% driselase. Protoplasts were cultured on liquid KAO medium supplemented with 1 mg/L 2,4-D, 0.5 mg/L kinetin, 200 mg/L spermidine and 68 g/L glucose. Also, protoplasts were cultured on 0.2 $\mu$M membrane filter placed onto CP solid medium containing the suspension cells as feeder cells in the dark at $25^{\circ}C$ for 4 weeks. Protoplast-derived-SF calli were cultured on MS medium containing 0.05 mg/L IAA, 7 mg/L 2 ip and 30 g/L sucrose under the continuous illumination for four weeks. The frequency of shoot formation was about 60%. The regenerants were transferred into potting soil to grow mature plants. The regenerants formed the silques with seeds after 8 weeks of cultures.

• Korean Journal of Plant Tissue Culture
• /
• v.24 no.5
• /
• pp.291-294
• /
• 1997

The experiments were carried out to determine the optimum conditions for the direct embrogenesis from the cotyledon derived zygotic embryo culture of Paeonia lactiflora Pall. Different peony tissues derived from zygotic embryos were cultured on MS medium with and without 2,4-D. Somatic embryos were formed from the cotyledons cultured on the medium without 2,4-D. The somatic embryogenesis from cotyledons was promoted in the growth regulator-free MS medium containing 1.65~3.3 g/L $NH_4NO_3$ and 30~40 g/L sucrose. The maximum frequency (80.0%) of somatic embryo formation was obtained from the cotyledons excised from zygotic embryos that cultured on MS medium containing 3.3 g/L $NH_4NO_3$. Epicotyl and roots were elongated from a somatic embryo by adding 0.3 mg/L GA$_3$ in the medium or the cold treatment at 4$^{\circ}C$ more than three weeks at 4$^{\circ}C$.

• Journal of Korean Society of Forest Science
• /
• v.78 no.4
• /
• pp.372-380
• /
• 1989

The widely cultivated hybrid poplar Populus alba ${\times}$ P. glandulosa in Korea was transformed with Agrobacterium rhizogenes agropine type strain A4. Genetic transformation was confirmed by the presence of agropine. Alteration in growth rate of hairy roots was seen following changes in the dilution rate of medium and concentration of sucrose, suggesting that improved growth might be achieved by more precise manipulation of the nutrient medium. Plant regeneration occurred from transformed hairy roots on MS medium supplemented with 0.5 mg/l BAP. Transformed plantlets grown in vitro exhibited a more developed root system characterized by fast growth behavior in comparison to normal plantlets. This work demonstrates that the root transformation would be useful in improving plantlet establishment and growth through the effective root system.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2023.04a
• /
• pp.33-33
• /
• 2023

Potatoes are the world's 4th major food crop after maize, rice, and wheat and also are a staple food for 1.3 billion people. Due to their wide adaptability to various environmental conditions, their yeild capacity, and high commercial value, potatoes have contributed to global food security. Many potato germplasms are commonly preserved as whole plants in fields or in storage to maintain their particular genetic combinations. However, field maintenance is expensive and has the risk of potential losses from diseases, pests, plant ageing and climate change. Over the past four decades, meaningful efforts have been made toward the safe long-term conservation of potatoes through cryopreservation methods such as droplet-vitrification. In this study, we tested 4 Korean potato varieties('Golden Egg', 'Golden Ball', 'Ja-Young' and 'Ha-Ryeong') with the modified potato droplet -vitrification protocol. Potato shoot tips are precultured in a sucrose-enriched medium(0.3 and 0.7M for 7 and 17hrs, respectively) and submitted to a loading step with C4 solution for osmoprotection. The treated explants were dehydrated with Plant Vitrification Solution(PVS)2 which is 80% A3 solution in ice for 30 minutes. Thawing and unloading steps were performed with 0.8M sucrose solution for 30 sec(40℃) followed by 30min(25℃, room temperature). In a potato post-culture medium(MS+0.1 mg·L^-1 GA₃+0.1 mg·L^-1 kinetin), we obtained a survival rates of post-thawed explants ranging 16.1-82.2%. The results suggest that modified and optimized protocols are required dependinig on every cultivar, genetic and ecological types. To achieve higher survival and regeneration rates, each step within the cryoprocedure must be carefully optimized.

• Horticultural Science & Technology
• /
• v.31 no.3
• /
• pp.359-365
• /
• 2013

Chrysanthemum is one of the most popular ornamental plants worldwide. Recently, lots of new and novel chrysanthemum varieties have been developed using mutagenesis. However, there was no study for comparison of tissue culture condition among the mutant varieties derived from one original variety, until now. This study was conducted to compare the efficient regeneration condition of the two chrysanthemum mutant varieties, 'ARTI-purple' and 'ARTI-queen'. Two different flower parts (disk and ray florets) at the unopened and early blooming stages were used for comparison of regeneration condition on MS medium supplemented with combinations of three growth regulators (BA, NAA, and IAA). The highest regeneration rate was identified on the NAA and BA combination when the disk florets at unopened blooming stage are used. The best optimum combinations of growth regulators were identified as NAA $1.0mg{\cdot}L^{-1}$ and BA $0.5mg{\cdot}L^{-1}$ at 'ARTI-purple', which displayed 47.9% regeneration. However, regeneration of 'ARTI-queen' was the highest as 25.6% at NAA $2.0mg{\cdot}L^{-1}$ and BA $1.0mg{\cdot}L^{-1}$. There results indicate that there is a difference for the optimum regeneration condition between the mutant varieties derived from one original variety. These results will be useful for construction of efficient regeneration system of diverse chrysanthemum mutants developed by mutation breeding.

• Journal of Plant Biotechnology
• /
• v.7 no.4
• /
• pp.247-257
• /
• 2005

An efficient, rapid and large-scale in vitro clonal propagation of agronomically important Indian cereal crop genotypes (NSH27 & K5) of Sorghum bicolor (L.) Moench. by enhanced shoot proliferation in shoot tip segments was designed. MS medium fortified with plant growth regulators and coconut water markedly influenced in vitro propagation of Sorghum bicolor. In vitro plantlet production system has been investigated on Murashige and Skoog (MS) medium with the synergistic combination of 6-benzyladenine ($22.2\;{\mu}M$), kinetin ($4.6\;{\mu}M$), adenine sulphate ($2.8\;{\mu}M$), 5% coconut water and 3% sucrose which promoted the maximum number of shoots as well as beneficial shoot length. Subculturing of shoot tip segments on a similar medium enabled continuous production of more than 100 healthy shoots with similar frequency. When the healthy shoot clumps were cultured on MS medium fortified with 6-benzyladenine ($22.2\;{\mu}M$), kinetin ($4.6\;{\mu}M$), adenine sulphate ($2.8\;{\mu}M$), ${\alpha}$-naphthaleneacetic acid ($2.7\;{\mu}M$), ascorbic acid ($30.0\;{\mu}M$) and 5% coconut water, a rapid production of axillary and adventitious buds was developed after 8 wk culture. More than 300 shoots were produced 10 wk after culture. Rooting was highest (100%) on half strength MS medium containing 22.8 mM IAA. Micropropagated plants established in garden soil, farmyard soil and sand (2:1:1) were uniform and identical to the donor plant with respect to growth characteristics. These plants grew normally without showing any traits.