• Korean Journal of Breeding Science
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• v.41 no.2
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• pp.168-172
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• 2009

'Punghan' (Avena sativa L.), a winter oat for forage use, was developed by the breeding team at the Department of Rice and Winter Cereal Crop, NICS, RDA in 2008. It was derived from a cross between 'Beltsville 61-150 (IT133501)' and $F_1$ between 'Sikyonggwiri' and 'PA202-210'. A oat cultivar from USA, 'Beltsville 61-150', has a high cold tolerance, while the $F_1$ between 'Sikyonggwiri' and 'PA202-210' has early heading and high yielding with large-size grain. Subsequent enerations followed by the cross were handled in bulk and pedigree selection programs at Suwon and Yeoncheon, respectively. A line, 'SO99013-B-YB-31', was selected for earliness and good agronomic characteristics and designated as a line name of 'Gwiri 60'. The line 'Gwiri 60' was subsequently evaluated for winter hardiness, earliness, and forage yield in five locations, Yesan, Cheongwon, Iksan, Kimjae, and Jeju, from 2006 to 2008 and finally named as 'Punghan'. Over 3 years, the average forage dry matter yield of 'Punghan' harvested at milk-ripe stage was $14.5\;ton\;ha^{-1}$, compared with $13.6\;ton\;ha^{-1}$ of check cultivar 'Samhan'. 'Punghan' is about 2 days earlier in heading date than check cultivar. 'Punghan' is adapted primarily for winter planting use in the mid-southern regions of Korean peninsula.

• Journal of Food Hygiene and Safety
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• v.38 no.6
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• pp.537-543
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• 2023

This study monitored the caffeine content of ready-to-drink coffee and verified the appropriateness of the labeling. The caffeine content was analyzed using HPLC. The average caffeine content of cold brew coffee was 0.31-1.04 mg/mL, with an average of 0.55 mg/mL. The average content of product was 147.27 mg/bottle, and taking into account the recommended daily intake, an adult can consume 2.7 bottles. Americano coffee was 0.15-0.38 mg/mL, with an average of 0.28 mg/mL. The average content of product was 110.42 mg/bottle, and considering the recommended daily intake, an adult can consume 3.6 bottles. The caffeine content of decaffeinated cold brew coffee was 5.14 mg/bottle and compared to Americano coffee, more than 95% of the caffeine was removed. In addition, we verified the tolerance level of the total caffeine content in ready-to-drink coffee, and none of them exceeded 120%, signifying that all commercial products were effectively managed.

• Journal of Microbiology and Biotechnology
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• v.9 no.2
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• pp.213-218
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• 1999

Fructans are polyfructose molecules that function as nonstructural storage carbohydrates in several plants. In addition, it has been suggested that, due to their solubility, they can play an important role in helping plants survive periods of osmotic stress. In order to study the effect of levan synthesis on plant growth, the coding region of the levansucrase gene, which was isolated from Zymomonas mobilis, was introduced into tobacco plants using Agrobacterium tumefaciens-mediated transformation. The presence of the levansucrase gene in transgenic plants was verified by genomic DNA gel blot analysis. RNA gel blot and immunoblot analyses showed an accumulation of the corresponding transcript and protein product of the bacterial levansucrase gene in transgenic plants. Furthermore, a thin layer chromatography analysis revealed that fructans were synthesized and deposited in transgenic tobacco plants. When $T_1$ seeds were germinated and grown under polyethylene glycol-mediated drought stress or cold stress, the transgenic seedlings displayed a substantially higher level of growth than that of untransformed plants. These results suggest that fructans may playa significant role in the tolerance of plants under osmotic stress.

• Journal of The Korean Society of Grassland and Forage Science
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• v.42 no.3
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• pp.188-194
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• 2022

This study was conducted to investigate the forage characteristics of very early-maturing Italian ryegrass (IRG) three cultivars within September, 2021 to July, 2022 in Cheonan, Korea. We used three different cultivars of Italian ryegrass in this study "Greenfarm" "Greenfarm2ho" and "Greenfarm3ho" The results of heading date in this study, "Greenfarm" was recorded 3 days earlier than the other two cultivars, and its lodge tolerance score was the highest among the three. However, in case of plant length, there was no significantly difference in whole cultivars, in addition disease resistance, insect resistance and cold tolerance were not significantly different in three cultivars. The forage productivity of "Greenfarm2ho" was recorded the highest, especially the 1^st harvesting of "Greenfarm2ho" yielded significantly the highest and superior to other varieties. As a result of feed value analysis, three cultivars were generally superior to overseas varieties and in particular, "Greenfarm2ho" was recorded to have excellent value because of the lowest NDF and ADF content. On the other hand, there was no significant difference in crude protein content among three varieties. In case of the monosaccharides content "Greenfarm3ho" had significantly higher glucose content than other two varieties, therefore the "Greenfarm" has advantage for preparing high quality of silage. In contrast, there was no significant difference among three varieties in fructose content.

• Journal of Plant Biotechnology
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• v.35 no.1
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• pp.55-61
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• 2008

Previous studies on genetic transformation of chrysanthemum using cold regulated gene (BN115) have been conducted and the PCR and Real-Time PCR based method to determine the presence of the transferred cold regulated gene in the chrysanthemum was established. To check whether over-expression of BN115 gene in transgenic chrysanthemum will enhance their tolerance to cold stress, the transgenic chrysanthemum were grown under low temperature condition and several cold signalling including growth characteristics, stoma size and shape, SPAD value and ion leakage test were investigated. The transgenic chrysanthemum in the low temperature growth chamber grow much faster in term of the height, number and size of the leaves than those of wild-type plants and damage of transgenic plant caused by the low temperature was much less than that of wild-type plants. The stoma type and size of transgenic plant leaves grown at $5^{\circ}C$ were much similar to of wild-type plant cultured on $25^{\circ}C$ It has been found that SPAD value of transgenic plants was much higher than those of wild-type, but the EC density being lower under low temperature condition.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.20-20
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• 2017

Plant genomes include heterochromatic loci that consist of repetitive sequences and transposable elements. LTR retrotransposon is the major class of transposons in advanced plants in terms of proportion in plant genome. The elements contribute not only to genome size but also to genome stability and gene expression. A number of cases have been reported transposon insertions near genic regions affect crop traits such as fruit pigments, stress tolerance, and yields. Functional LTR retrotransposons produce extrachromosomal DNA from genomic RNA by reverse transcription that takes place within virus-like-particles (VLPs). DECREASED DNA METHYLATION 1 (DDM1) plays important roles in maintaining DNA methylation of heterochromatin affecting all sequence contexts, CG, CHG, and CHH. Previous studies showed that ddm1 mutant exhibits massive transcription of retrotransposons in Arabidopsis, but only few of them were able to create new insertions into the genome. RNA-dependent RNA POLYMERASE 6 (RDR6) is known to function in restricting accumulation of transposon RNA by processing the transcripts into 21-22 nt epigenetically activated small interfering RNA (easiRNA). We purified VLPs and sequence cDNA to identify functional LTR retrotransposons in Arabidopsis ddm1 and ddm1rdr6 plants. Over 20 LTR copia and gypsy families were detected in ddm1 and ddm1rdr6 sequencing libraries and most of them were not reported for mobility. In ddm1rdr6, short fragments of ATHILA gypsy elements were detected. It suggests easiRNAs might regulate reverse transcription steps. The highest enriched element among transposon loci was previously characterized EVADE element. It has been reported that active EVADE element is more efficiently silenced through female germline than male germline. By genetic analyses, we found ddm1 and rdr6 mutation affect maternal silencing of active EVADE elements. DDM1-GFP protein accumulated in megaspore mother cell but was not found in mature egg cell. The fusion protein was also found in early embryo and maternal DDM1-GFP allele was more dominantly expressed in the embryo. We observed localization of DDM1-GFP in Arabidopsis and DDM1-YFP in maize and found the proteins accumulated in dividing zone of root tips. Currently we are looking at cell cycle dependency of DDM1 expression using maize system. Among 10 AGO proteins in Arabidopsis, AGO9 is specifically expressed in egg cell and shoot meristematic cells. In addition, mutation of AGO9 and RDR6 caused failure in maternal silencing, implying 21-22 nt easiRNA pathway is important for retrotransposon silencing in female gametophyte or/and early embryo. On the other hand, canonical 24 nt sRNA-directed DNA methylation (RdDM) pathways did not contribute to maternal silencing as confirmed by this study. Heat-activated LTR retrotransposon, ONSEN, was not silenced by DDM1 but the silencing mechanisms require RdDM pathways in somatic cells. We will propose distinct mechanisms of LTR retrotransposons in germline and somatic stages.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.97-97
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• 2017

Plant genomes include heterochromatic loci that consist of repetitive sequences and transposable elements. LTR retrotransposon is the major class of transposons in advanced plants in terms of proportion in plant genome. The elements contribute not only to genome size but also to genome stability and gene expression. A number of cases have been reported transposon insertions near genic regions affect crop traits such as fruit pigments, stress tolerance, and yields. Functional LTR retrotransposons produce extrachromosomal DNA from genomic RNA by reverse transcription that takes place within virus-like-particles (VLPs). DECREASED DNA METHYLATION 1 (DDM1) plays important roles in maintaining DNA methylation of heterochromatin affecting all sequence contexts, CG, CHG, and CHH. Previous studies showed that ddm1 mutant exhibits massive transcription of retrotransposons in Arabidopsis, but only few of them were able to create new insertions into the genome. RNA-dependent RNA POLYMERASE 6 (RDR6) is known to function in restricting accumulation of transposon RNA by processing the transcripts into 21-22 nt epigenetically activated small interfering RNA (easiRNA). We purified VLPs and sequence cDNA to identify functional LTR retrotransposons in Arabidopsis ddm1 and ddm1rdr6 plants. Over 20 LTR copia and gypsy families were detected in ddm1 and ddm1rdr6 sequencing libraries and most of them were not reported for mobility. In ddm1rdr6, short fragments of ATHILA gypsy elements were detected. It suggests easiRNAs might regulate reverse transcription steps. The highest enriched element among transposon loci was previously characterized EVADE element. It has been reported that active EVADE element is more efficiently silenced through female germline than male germline. By genetic analyses, we found ddm1 and rdr6 mutation affect maternal silencing of active EVADE elements. DDM1-GFP protein accumulated in megaspore mother cell but was not found in mature egg cell. The fusion protein was also found in early embryo and maternal DDM1-GFP allele was more dominantly expressed in the embryo. We observed localization of DDM1-GFP in Arabidopsis and DDM1-YFP in maize and found the proteins accumulated in dividing zone of root tips. Currently we are looking at cell cycle dependency of DDM1 expression using maize system. Among 10 AGO proteins in Arabidopsis, AGO9 is specifically expressed in egg cell and shoot meristematic cells. In addition, mutation of AGO9 and RDR6 caused failure in maternal silencing, implying 21-22 nt easiRNA pathway is important for retrotransposon silencing in female gametophyte or/and early embryo. On the other hand, canonical 24 nt sRNA-directed DNA methylation (RdDM) pathways did not contribute to maternal silencing as confirmed by this study. Heat-activated LTR retrotransposon, ONSEN, was not silenced by DDM1 but the silencing mechanisms require RdDM pathways in somatic cells. We will propose distinct mechanisms of LTR retrotransposons in germline and somatic stages.

• Journal of Microbiology and Biotechnology
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• v.24 no.2
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• pp.254-263
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• 2014

Enteromorpha polysaccharides (EP) extracted from green algae have displayed a wide variety of biological activities. However, their high molecular weight leads to a high viscosity and low solubility, and therefore, greatly restrains their application. To solve this problem, bacteria from the surface of Enteromorpha were screened, and an Alteromonas macleodii strain B7 was found to be able to decrease the molecular weight of EP in culture media. Proteins harvested from the supernatant of the A. macleodii B7 culture were subjected to native gel electrophoresis, and a band corresponding to the Enteromorpha polysaccharide lyase (EPL) was detected by activity staining. The enzyme identity was subsequently confirmed by MALDI-TOF/TOF mass spectrometry as the putative ${\alpha}$-amylase reported in A. macleodii ATCC 27126. The amylase gene (amySTU) from A. macleodii B7 was cloned into Escherichia coli, resulting in high-level expression of the recombinant enzyme with EP-degrading activity. AmySTU was found to be cold-adapted; however, its optimal enzyme activity was detected at $40^{\circ}C$. The ${\alpha}$-amylase was highly stable over a broad pH range (5.5-10) with the optimal pH at 7.5-8.0. The highest enzyme activity was detected when NaCl concentration was 2%, which dropped by 50% when the NaCl concentration was increased to 16%, showing an excellent nature of halotolerance. Furthermore, the amylase activity was not significantly affected by tested surfactants or the presence of some organic solvents. Therefore, the A. macleodii strain B7 and its ${\alpha}$-amylase can be useful in lowering EP molecular weight and in starch processing.

• Molecules and Cells
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• v.26 no.6
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• pp.595-605
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• 2008

Genome wide transcription analysis in response to stresses is essential to provide the basis of effective engineering strategies to improve stress tolerance in crop plants. In order to perform transcriptome analysis in Brassica rapa, we constructed a B. rapa oligo microarray, KBGP-24K, using sequence information from approximately 24,000 unigenes and analyzed cold ($4^{\circ}C$), salt (250 mM NaCl), and drought (air-dry) treated B. rapa plants. Among the B. rapa unigenes represented on the microarray, 417 (1.7%), 202 (0.8%), and 738 (3.1%) were identified as responsive genes that were differently expressed 5-fold or more at least once during a 48-h treatment with cold, salt, and drought, respectively. These results were confirmed by RT-PCR analysis. In the abiotic stress responsive genes identified, we found 56 transcription factor genes and 60 commonly responsive genes. It suggests that various transcriptional regulatory mechanisms and common signaling pathway are working together under the abiotic stresses in B. rapa. In conclusion, our new developed 24K oligo microarray will be a useful tool for transcriptome profiling and this work will provide valuable insight in the response to abiotic stress in B. rapa.

• Korean Journal of Agricultural and Forest Meteorology
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• v.3 no.4
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• pp.220-237
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• 2001

Over the 20th century global temperature increase has been 0.6$^{\circ}C$. The globally averaged surface temperature is projected to increase by 1.4 to 5.8$^{\circ}C$ over the period 1990 to 2100. Nearly all land areas will have higher maximum temperature and minimum temperature, and fewer cold days and frost days. More intense precipitation events will take plate over many areas. Over most mid-latitude continental interiors will have increased summer continental drying and associated risk of drought. By 2100, if the annual surface temperature increase is 3.5$^{\circ}C$, we will have 15.9$^{\circ}C$ from 12.4$^{\circ}C$ at present. Also the annual precipitation will range 1,118-2,447 mm from 972-1,841 mm at present in Korea. Consequently the average crop periods for summer crops will be 250 days that prolonged 32 days than at present. In the case of gradual increase of global warming, an annual crop can be adapted to the changing climate through the selection of filial generations in breeding process. The perennial crops such as an apple should be shifted the chief producing place to northern or high latitude areas where below 13.5$^{\circ}C$ of the annual surface temperature. If global warming happens suddenly over the threshold atmospheric greenhouse gases, then all ecosystems will have tremendous disturbance. Agricultural land-use plan, which state that farmers decide what to plant, based on their climate-based advantages. Therefore, farmers will mitigate possible negative imparts associated with the climate change. The farmers will have application to use agricultural meteorological information system, and agricultural long-range weather forecast system for their agroecosystems management. The ideal types of crops under $CO_2$ increase and climate change conditions are considered that ecological characteristics need indispensable to accomplish the sustainable agriculture as the diversification of genetic resources from yield-oriented to biomass-oriented characteristics with higher potential of $CO_2$ absorption and primary production. In addition, a heat-and-cold tolerance, a pest resistance, an environmental adaptability, and production stability should be also incorporated collectively into integrated agroecosystem.