• Journal of Bio-Environment Control
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• v.10 no.4
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• pp.244-250
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• 2001

This study was conducted to determine the change in freezing tolerance of Chinese cabbage (Brassica campestris L. ssp. penkinensis). Four-week old seedlings grown in a greenhouse at $25^{\circ}C$$\pm$0.5 were transferred to 5, 10 or 15$^{\circ}C$ condition for acclimation for one day, and then transferred to a lethal temperature of -3$^{\circ}C$ condition. Optimal acclimation temperatures were 5$^{\circ}C$ and 1$0^{\circ}C$. Freezing tolerance of leaf tissues was investigated during a week-long cold stress. Development of freezing tolerance was shown by survival ratio, and amounts of total RNA, soluble protein and soluble sugar. LT$_{50}$ value increased from -3 to -4$^{\circ}C$ after 5 days acclimation at 5 or 1$0^{\circ}C$, and this was considered to be the first indication of enhanced freezing tolerance. Plants, cold acclimated for five days, reached to a survival ratio of 60%. This increase in freezing tolerance was found to be associated with the increased levels of total RNA, soluble sugar and soluble protein. These metabolic changes imply the association of adjustment of growth and cell metabolism with low temperatures at the beginning of cold acclimation in chinese cabbage.e.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.50 no.2
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• pp.67-72
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• 2005

Overwintering capacity, closely related to winter hardiness, of Chinese milk vetch planted with different sowing times and sowing practices was investigated to improve the incorporation into cropping system in Korea. The tolerance to low temperature was evaluated with $LT_50$ using leaf disc leaching method. Dry weight of CMV was reduced remarkably with delayed planting from Sep. 5 to Oct. 20. The differences in tolerance to freezing temperature were not conspicuous among CMV genotypes, however, the differences between genotype (collections at different regions) were due to the plant architecture, mainly to the leaf angle. The crouching genotype collected at central region of Korean peninsula, which showed excellent freezing tolerant, has planophile leaves. The feature of internal constituents of CMV genotypes did not show any noticeable differences with respect to the freezing tolerance which evaluated by leaf disc leaching experiment. To overcome the poor overwintering capacity, tolerant genotype should be developed by selection with considering the plant architecture. The reduction of CMV growth during overwintering period was ameliorated with furrow-sowing under late-sown condition, therefore, when the CMV is inevitably sown late after recommended time, the seeds should be sown on furrow to overcome the cold stress.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.40 no.1
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• pp.69-76
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• 1995

Cold acclimation involves changes in gene expression. BN28 and BN115 are two genes which are regulated by cold temperature and assumed having roles in cold acclimation. The objectives of this experiment was to explore the expression of BN28 and BN115 under field conditions. Six winter cultivars were planted at three different dates during the fall. The expression of the genes was determined by northern blot analysis of total RNA taken from leaves 15 to 30 day-intervals after planting. The expression of the two genes was detected within 15 days after planting well before onset of freezing tolerance in plants. This suggestes either their expression was a prerequisite of the freezing tolerance or their expression was regulated by other environmental factors as well as temperature. Two genes showed a different expression pattern suggesting they had a different regulatory system. Although timecourse increase in expression of the cold-regulated genes was matched with increase in freezing tolerance, the difference of expression in cultivar level at specific times of measurement was not correlated with freezing tolerance at the moment.

• Korean Journal of Plant Tissue Culture
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• v.27 no.4
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• pp.339-343
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• 2000

When plants are exposed to subfreezing temperatures ice crystals are forming within extracelluar space in leaves. The growth of ice crystal is closely related to the degree of freezing injury. It was shown that an antifreeze protein binds to an ice nucleator through hydrogen bonds to prevent growth of ice crystal and also reduces freezing damage. The antifreeze proteins in plants are similar to PR proteins but only the PR proteins induced upon cold acclimation were shown to have dual functions in antifreezing as well as antifungal activities. Three of the genes encoded for CLP, GLP, and TLP were isolated from barley and Kentucky bluegrass based on amino acid sequence revealed after purification and low temperature-inducibility as shown in analysis of the protein. The deduced amino acid of the genes cloned showed a signal for secretion into extracellular space where the antifreezing activity sup-posed to work. The western analysis using the antisera raised against the antifreeze proteins showed a positive correlation between the amount of the protein and the level of freeze tolerance among different cultivars of barely. Besides it was revealed that TLP is responsible for a freeze tolerance induced by a treatment of trinexapac ethyl in Kentucky bluegrass. Analysis of an overwintering wild rice, Oryza rufipogon also showed that an acquisition of freeze tolerance relied on accumulation of the protein similar to CLP. The more direct evidence for the role of CLP in freeze tolerance was made with the analysis of the transgenic tobacco showing extracellular accumulation of CLP and enhanced freeze tolerance measured by amount of ion leakage and rate of photosynthetic electron transport upon freezing. These antifreeze proteins genes will be good candidates for transformation into crops such as lettuce and strawberry to develop into the new crops capable of freeze-storage and such as rose and grape to enhance a freeze tolerance for a safe survival during winter.

• Journal of Veterinary Clinics
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• v.17 no.2
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• pp.420-430
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• 2000

These studies were preformed to investigate the freezing conditions to achieve good post-thaw viability of spend and the practical methods of artificial insemination frozen canine semen. Semen were collected from nine male dogs which had been proved to be fertile in the past and the semen were treated for freezing procedure. Post-thaw motility and viability of canine sperm were evaluated to investigate individual tolerance of freezing, difference among freezing extenders, dif-ference among freezing equipments and freezing conditions, difference between fast and slow cooling rate, difference according to different glycerol concentration, effect of seeding on post-thaw viability, difference according to cutting part of straw, difference according to thawing temperatures, and dif-ference according to media added to thawed semen. Thawed semen for insemination were added with equal volnme of canine capacitation medium (CCM) and the volume of semen and the number per insemination were adjusted as 2-3 ml and $20-30 {\times}10^7,$ respectively. The semen were inseminated in vagina using balloon catheter and en17ryos were cellected from 9 to 11 days after the second Al to d determine fertilization.

• Reproductive and Developmental Biology
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• v.34 no.3
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• pp.253-256
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• 2010

This study was performed to investigate the characteristics within ages and freezing tolerance of spermatozoa in Jindo Dog. Experimental animals were selected 12 herds within 1~8 year's old and collected semen for 2 times in a week. Collected semen was evaluated whole volume and sperm number with CASA system (SIAS, Medical Supply, Korea). Then seminal plasma were separated and diluted with modified Tris-egg yolk extender and added 4, 6 and 8% glycerol for 4 times to final concentration and equilibrated for 1.5 hrs. Before and after freezing, equilibrated semen were evaluated the survival rates. Total volume of sperm at 1~2 year old group is as $5.2{\times}10^8\;cells/ml$ largest and there were no significance among groups. The motility of 1~2 year old group is highest as 90.9% and there were significance among groups. Abnormal sperm showed similar among groups. The survival rate in terms of pre-freezing and post-freezing were decreased all levels of glycerol and reveled 87.0% to 64.5% in 4%, 87.5% to 51.9% in 6% and 73.4% to 29.7% in 8%, there were significant difference among the groups (p<0.05). These results suggest that the optimal sperm-freezing methods in Jindo Dog are utilized with modified Tris egg-yolk extender with 4% glycerol and were improve the reproductive activity by these methods.

• Journal of Plant Biotechnology
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• v.30 no.4
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• pp.315-321
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• 2003

Freezing and drought tolerances in plants are very important for survival in the desert. In an effort to reduce desertifcation in Gobi, a molecular breeding of Artemisia adamsii using the CLP (chitinase like protein, antifreeze protein) and Dhn5 (dehydrin5) genes from barley is performed by introducing them into Artemisia adamsii via Agrobacteria. We had found an optimal combinatorial concentration of hormones at 0.05mg/L of NAA and 0.5mg/L of BA for growth of callus in Artemisia adamsii. In addition, the higher rate of callus induction using hypocotyl as explant was observed comparing to explants of stem and leaf. There were some variations in the level of the proteins expressed among the transgenic lines such that the lines of CLP(CS1-5, 1-7,4-4) and Dhn5(DS2-2, 2-3) lines produce the protein to higher levels. The transgenic lines showing a higher level of Dhn5 exhibited better growth than nontransgenic callus in presence of 10 and 20％ PEG. In case of the CLP tansgenic lines, both CS1-5 and CS1-7 showed a higher level of freezing tolerance determined by ion leakage test.

• Journal of Bio-Environment Control
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• v.21 no.4
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• pp.354-361
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• 2012

This study was conducted to evaluate on the freezing tolerance of introduced blueberry cultivars in Korea and to investigate availability of portable timber moisture meter for simple and rapid diagnosis of blueberry-shoot damage by freezing during wintering. Frost tolerance of blueberry cultivars showed big difference that rates of blueberry-shoot death were widely distributed from about 0% to 100% after wintering. Optical density in TTC reduction of blueberry twig treated low temperature was low in order of $-40^{\circ}C$ < $-21^{\circ}C$ < $4^{\circ}C$. Hardiness evaluation of visible injury in the cross-sectional surface color did not agree with that of rates of blueberry-shoot death during wintering. Lowest water content of blueberry stem measured by timber moisture tester during wintering was about 15%. During wintering, water contents of blueberry stems were higher at lower part of tree, but were low at end part of stems, and then when the blueberry grew again for spring, the water content gradually increased to 20~40%. Water content of blueberry stem with freezing injury during wintering decreased to under 5% by desiccation. Therefore it is assummed that the moisture content of blueberry stem injured by freezing during wintering was about under 14%, and it is expected that portable timber moisture meter could be available for rapid diagnosis of blueberry freezing injury in field.

• Proceedings of the Korean Society of Crop Science Conference
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• 1994.10a
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• pp.3-4
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• 1994

• Proceedings of the Korean Society of Crop Science Conference
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• 1994.10a
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• pp.24-25
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• 1994