• Title/Summary/Keyword: Plant Transformation

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Antioxidant and Anti-Inflammatory Activities of Ethanol Extract of Clematis trichotoma Nakai (할미밀망 에탄올 추출물의 항산화 및 항염증 활성 평가)

  • Jung, Jaemee;Shin, Mijoon;Jeong, Naeun;Hwang, Dahyun
    • Korean Journal of Clinical Laboratory Science
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    • v.53 no.2
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    • pp.165-173
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    • 2021
  • Clematis trichotoma Nakai (CTN) is a broad-leaved vine plant belonging to the family Ranunculus, native to Korea. Young leaves are used as food, and the stem and roots are used as medicinal materials. Antioxidant studies have been reported on the stems of CTN, but no studies have been conducted on the leaves. In this study, a 70% ethanol extract of CTN was prepared and its antioxidant and anti-inflammatory activities were investigated. For measuring the antioxidant activity, five assays (polyphenol and flavonoid content, reducing power, 2,2-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), and 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity) were performed and CTN showed a concentration-dependent effect in all assays. To investigate the anti-inflammatory activity, we used RAW 264.7 cells. The concentrations (from 31.25 to 250 ㎍/mL) of CTN did not show cytotoxicity. CTN (250 ㎍/mL) inhibited dendritic transformation (34.4%) and also inhibited inflammation as seen by reduced levels of NO (77.4%), IL-6 (85.5%) and TNF-α (41.2%) compared to lipopolysaccharide (LPS). CTN (250 ㎍/mL) also suppressed the expression of the following genes: COX-2 (79.8%), iNOS2 (93.9%), IL-6 (87.6%), and TNF-α (77.3%) compared to LPS. These results demonstrated that CTN has excellent antioxidant and anti-inflammatory activities and can therefore be used as a natural biological resource.

Construction of novel promoters based on the characteristics of drought stress specific cis-regulatory element (가뭄 스트레스 특이적인 cis-regulatory element의 특성을 기반으로 한 신규 프로모터 구축)

  • Kim, Kihwan;Kim, Byeonggyu;Shin, Juhyung;Kim, Won-Chan
    • Journal of Applied Biological Chemistry
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    • v.64 no.1
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    • pp.39-48
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    • 2021
  • Droughts are one of the abiotic stresses that hinders the growth and productivity of crop plants. Coping with abiotic stress is necessary to understand the molecular regulatory networks that makes plants respond to adverse environmental conditions. In our experiment to find a combination that can cope with abiotic stress (respond to drought), we screened 5 stress-inducible promoters that are expressed only under stress conditions. This founded 36 cis-elements in stress-inducible promoters. With the result we designed 2 synthetic promoters (BL1, BL2) for fine-controlled regulation by assembling cis-elements from the native promoters, which are expressed only under stress caused by droughts. Analysis of the transgenic plant (BL1-GUS, BL2-GUS) showed that the synthetic promoters increased the expression of β-glucuronidase (GUS) in transgenic plants under desiccation. Also in the transient activation assay demonstrated that synthetic promoters induced the co-transformation of effector DREB1A and DREB2C. These results expect that the synthetic promoter with a combination of drought-specific elements can be used to respond to various abiotic stress and is resistant to stress without causing growth retardation.

Technology Trends of Smart Abnormal Detection and Diagnosis System for Gas and Hydrogen Facilities (가스·수소 시설의 스마트 이상감지 및 진단 시스템 기술동향)

  • Park, Myeongnam;Kim, Byungkwon;Hong, Gi Hoon;Shin, Dongil
    • Journal of the Korean Institute of Gas
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    • v.26 no.4
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    • pp.41-57
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    • 2022
  • The global demand for carbon neutrality in response to climate change is in a situation where it is necessary to prepare countermeasures for carbon trade barriers for some countries, including Korea, which is classified as an export-led economic structure and greenhouse gas exporter. Therefore, digital transformation, which is one of the predictable ways for the carbon-neutral transition model to be applied, should be introduced early. By applying digital technology to industrial gas manufacturing facilities used in one of the major industries, high-tech manufacturing industry, and hydrogen gas facilities, which are emerging as eco-friendly energy, abnormal detection, and diagnosis services are provided with cloud-based predictive diagnosis monitoring technology including operating knowledge. Here are the trends. Small and medium-sized companies that are in the blind spot of carbon-neutral implementation by confirming the direction of abnormal diagnosis predictive monitoring through optimization, augmented reality technology, IoT and AI knowledge inference, etc., rather than simply monitoring real-time facility status It can be seen that it is possible to disseminate technologies such as consensus knowledge in the engineering domain and predictive diagnostic monitoring that match the economic feasibility and efficiency of the technology. It is hoped that it will be used as a way to seek countermeasures against carbon emission trade barriers based on the highest level of ICT technology.

Expression of Antimicrobial Peptide (AMP), Moricin Using SUMO Fusion Tag in Escherichia coli (대장균에서 SUMO fusion tag을 이용하여 항균펩타이드인 moricin의 발현)

  • Ahn, Dong-gyu;Park, Sun Ill;Kim, Soon Young
    • Journal of Life Science
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    • v.32 no.12
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    • pp.956-961
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    • 2022
  • Plant Chloroplast have several advantages as an expression platform of biopharmaceuticals over conventional expression platforms such as mammalian cells, yeast and bacteria. First, plants do not serve as a host for mammalian infectious virus and have endotoxin like bacteria which can cause anaphylactic shock. In addition, high copy number of chloroplast genome allows for chloroplast transformants to reach the high level of expression of heterologous genes. Moreover, the integration of transgenes into specific region of chloroplast genomes makes chloroplast transformants unaffected by positional effect which can be frequently observed from nuclear transformants, resulting in loss of transgene expressions. Antimicrobial peptides (AMPs) are a kind of innate immunity which is found from bacteria to humans. Unlike conventional antibiotics, very less dosage of AMPs can have catastrophic effect on bacterial survival. Further, the repeated use of AMPs does not trigger the development of bacterial resistance. Moricin, one of the AMPs, was isolated from Bombyx mori, a silkworm moth. The C-terminal of moricin consists largely of basic amino acids, and the N-terminal has an α-helix structure. Moricin was chosen and expressed in a SUMO/SUMOase without leaving any unwanted amino acids which could potentially affect the anti-bacterial activity of the moricin. The transformation vector used in this study has already been created in this lab for the expression in both prokaryotic systems such as E. coli and chloroplast. The expressed moricin was purified using Ni columns and SUMOase, and the antibacterial activity of the purified moricin was confirmed using an agar diffusion assay.

Evaluation of Planting Distance in Rice Paddies Using Deep Learning-Based Drone Imagery (딥 러닝 기반 드론 영상을 활용한 벼 포장의 재식거리 평가)

  • Hyeok-jin Bak;Dongwon Kwon;Woo-jin Im;Ji-hyeon Lee;Eun-ji Kim;Nam-jin Chung;Jung-Il Cho;Woon-Ha Hwang;Jae-Ki Chnag;Wan-Gyu Sang
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.69 no.3
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    • pp.154-162
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    • 2024
  • In response to the increasing impact of climate change on agriculture, various cultivation technologies have been recently developed to improve agricultural productivity and reduce carbon emissions for carbon neutrality. This study presents an algorithm for estimating rice planting density in agriculture using drone-captured images and deep learning-based image analysis technology. The algorithm utilizes images collected from various paddies; these images are processed through pre-processing steps and serve as training data for the YOLOv5x deep learning model. The trained model demonstrated high precision and recall, effectively estimating the position information of rice plants in each image. By accurately estimating the position of rice plants based on the central coordinates in diverse unpaved environments, the model allowed for estimation of rice plant density in each paddy, producing values closely aligned with actual measurements. Moreover, the algorithm proposed in this study provides a novel approach for precise determination of rice planting density based on the position information of rice plants in the images. Analysis of drone footage from different regions capturing portions of paddies revealed that the developed algorithm exhibited a significant correlation (R2 =0.877) with actual planting density. This finding suggests the potential effective application of the algorithm in real-world agricultural settings. In conclusion, we believe that this research contributes to the ongoing digital transformation in agriculture by offering a valuable technology that supports the goals of enhancing efficiency, mitigating methane emissions, and achieving carbon neutrality, in response to the challenges posed by climate change.

Investigation of Original Landscaping in the Vicinity of Yongyun and Hwahongmun in Suwon Hwaseong (수원 화성 용연(龍淵)과 화홍문 일곽의 원형경관 탐색)

  • Rho, Jae-Hyun;Choi, Jong-Hee;Shin, Sang-Sup
    • Journal of the Korean Institute of Landscape Architecture
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    • v.38 no.2
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    • pp.94-108
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    • 2010
  • The purpose of this study is to provide data for the restoration of 'Yongyun(龍淵)' and 'Hwahongmun(華虹門)' through an investigation of the vicinity of their original landscapes at the time of construction of Hwaseong in Suwon and through tracing the transformative process of the environments of this vicinity. The results are as follows; As identified by 'Yongyunjung(龍淵亭)' and 'Yongdugak(龍頭閣)', other names of Yongyun, 'Banghwasuryujung(訪花隨柳亭)', which was built on 'Yongduam', is a facility whose place identity is highlighted with a sense of unity with Yongyun. The south lakefront of Yongyun, bordering Banghwasuryujung, has boundaries that make the best use of the natural geographical features of Yongduam while the current circumference of Yongyun is comparatively shorter than its original state. The size of 'Joongdo(中島)', however, seems to be an example of apparent over-design complement and reorganize 'Joongdo', which had been restored larger than its original state at the time of restoration in the 1970s. The depth of 'Yongyun' was created to be lower than the actual depth, without consideration for its initial depth, as soil was accumulated through continuous flooding after it was created. It is assumed that the original drains which were installed about 10m inside the lake were created facing the stream. As regards the planting environment, a circular planting of willows was made in the outskirts of 'Yongyun', except the 'Yongduam' which is a pure forest, and a mix of 'Pinus densiflora', shrubbery and deciduous broad leaf trees was planted in 'Joongdo'. Of the plants growing in the area of this study, plant species introduced to Korea after Hwaseong was constructed are found, most of which provide interest and attraction. The old pine trees growing in a group once grew in the castle areas of the vicinity even in the 1920s, the period of Japanese occupation, but they disappeared from the area in the aftermath of subsequent urban development and the Korean War. Although restored to the site, the number and space taken up by these trees are insignificant compared to those of the original environment. On the basis of these results, the following is considered necessary for the true restoration of the vicinity of 'Yongyun' and 'Hwahongmun': First, the grounds of 'Yongyun' should be dredged deeply enough to expose the bedrock and should be recreated in the rough outline of a half moon by extension to the southwest toward 'Yongduam' and 'Hwahongmun', and the size of 'Joongdo' should be significantly reduced. Secondly, considering that most plant species, except the pine trees and wild trees in 'Yongduam', are non-native plants introduced in order to provide such attractions such as the appreciation of scenic areas, they should be replaced with native species, mainly with the pine trees which were utilized during the construction of Hwaseong. The weeping willows planted in the 'middle-island' should be relocated to the outskirts of 'Yongyun', and replaced with pine trees as the major trees and maple trees or deciduous broadleaf trees to fill in the gaps. Thirdly, exotic species such as the 'Pinus rigida' planted in a group around 'Banghwasuryujung' and 'Bugammun' and 'Pinus strobus' planted in the vicinity of Hwahongmun' should be removed.

The Influence of Pretreatment Period, 2-Hydroxynicotinic Acid and Anther Co-pretreatment on Embryo Induction in Isolated Microspore Culture of Capsicum annuum L. (고추의 나출 소포자 배양시 전처리 기간, 2-Hydroxynicotinic Acid 및 약-공동전처리가 소포자배 발생에 미치는 영향)

  • Park Eun-Joon;Kim Jin-Ae;Lee Jong-Suk;Jang In-Chang;Yoon Michung;Chung Sang-Ho;Kim Moonza
    • Journal of Plant Biotechnology
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    • v.32 no.1
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    • pp.37-44
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    • 2005
  • Microspores were isolated from pepper (Capsicum annuum L.) anthers by using a micro-blender and cultured in modified NLN medium at $25^{\circ}C$. The influence of pretreatment period at $32^{\circ}C$, adding the 2-hydroxynicotinic acid to a pretreatment medium, and co-pretreatment anthers with microscopes on the induction of embryo were examined. Globular and torpedo embryos were observed from 3 weeks after culture. Embryo development was not synchronized within culture. After 4 weeks in culture, in addition to globular and torpedo embryos, cotyledonary embryos were observed. Normal cotylodonary embryos developed into plantlets when transferred to a solid hormone free B5 medium containing $2\%$ sucrose. Embryo yields were significantly higher after 1- and 2-day pretreatment at $32^{\circ}C$. However the development of embryo ceased at the globular or heart stage. In contrast, embryo yields were lower after 3- to 6-day pretreatment at $32^{\circ}C$ and embryo developed at the cotyledonary stage. After adding the 2-hydroxynicotinic acid to anther pretreatment solution, embryo yields were slightly increased. However most embryos occurred were at the globular or heart stage. Co-pretreatment of microspores with anthers was deleterious for embryo induction and development. AS far as we know, this is the first report of success in obtaining high frequency of embryogenesis and plantlets formation from isolated microspores of pepper. Although the culture conditions have to be optimized further, this promising microspore culture system can be used for genetic transformation, selection for dominant and recessive traits as well as for the production of homozygous doubled haploid plants.

Effect of Trehalose on the Viability of Fluorescent Pseudomonas, Strain SSL3 (형광성 Pseudomonas, SSL3 균주의 생존율에 미치는 Trehalose의 효과)

  • Seong, Ki-Young;Ryu, Ok-Ran;Choi, Won-Yeol
    • Korean Journal of Soil Science and Fertilizer
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    • v.33 no.4
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    • pp.292-301
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    • 2000
  • To make a better use of the beneficial bacterial inoculants in the agricultural practice, dry forms of bacterial fertilizer or pesticides are prepared with carrier materials. During the drying process of bacterial inoculant, most of the cells face a severe osmotic pressure and dehydration, and die off. Our study describes the effect of osmoprotectants such as trigonelline and trehalose on the survival of bacterial cells in high salt concentration and drying conditions. A fluorescent Pseudomonas, strain SSL3, used in this study, could grow in high salt concentration of upto 5% but the cells could not overcome the growth retardation at over 7% of salt concentration. The addition of trigonelline, even on small amount, in liquid medium containing 4% NaCl was detrimental to the cell. However, the addition of trehalose of upto 10 mM to the liquid medium containing 4% NaCl, enhanced cell growth. The cell growth was retarded when 150mM trehalose was added to the medium. Upon dry formulation of cells, trehalose was added. And the dry cells were inoculated into the soil to determine the effect of osmoprotectants on the survival of the cells. The survival of the cells, both in wet or dry soil, was improved by the addition of trehalose during the dry cell formulation. The positive effect of trehalose on the cell survival at $-20^{\circ}C$ and $-70^{\circ}C$ was oven more pronounced. The FTIR (Fourier transformation infra-red) spectroscopic analysis showed that the change of the 2nd amide group was reduced by adding trehalose to the medium containing 4% NaCl. These results suggest that trehalose can protect the cell membrane from dryness or high concentration of salt, thereby diminishing the sudden change of the protein structure of the cell membrane and, as a consequence, improving the cell survival.

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Studies on the Physiological Chemistry of Flower Organ and Seed in Ginseng Plant. IV. Variation of Free Amino Acids in the Flower and Seeds of the $F_1$ Plants of the Combinations Panax ginseng ${\times}$ Panax quinquefolium and Panax ginseng ${\times}$ Panax japonicus. (인삼종자형성에 대한 생리화학적 연구 IV. 고려인삼과 미국인삼 및 고려인삼과 죽절인삼 $F_1$의 화기 및 종자 형성과정에 있어서의 유리아미노산의 소장)

  • Jong-Kyu Hwang;Hee-Chun Yang
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.14
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    • pp.165-172
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    • 1973
  • The sterile phenomenon is frequently found in the inter-species hybrids of ginseng as in other plants. It is known that among the hybrids between Panax Ginseng (PG) and Panax Quinquefolium (PQ), and between Panax Ginseng and Paxax Japonicus (PI), PG${\times}$PI is fertile only very rarely, while PG ${\times}$ PQ is always sterile. Therefore, in order to clarify the relationship between this sterility phenomenon and the metabolism of free amino acids, the changes of free amino acids through the formation of the flower organs and seeds of two hybrids, PG ${\times}$ PQ and PG ${\times}$ PI were investigated by thin layer chromatography. The results are summarized as follows: 1. Distinct differences in the quantity and number of free amino acids were recognized between PG ${\times}$ PQ, PG ${\times}$ PI and their parent plants. From the hybrid PG ${\times}$ PQ, 19 kinds of ninhyrin sensitive substances were detected in all. They were (1) 17 amino acids: alanine, valine, leucine, phenylalanine, proline, hydroxy-proline, serine, threonine, tyrosine, aspartic acid, glutamic acid, lysine, arginine, ${\gamma}$-amino butyric acid, ${\beta}$-alanine, cysteic acid and tryptophan, and (2) two amides: asparagine and glutamine. From the hybrid PG ${\times}$ PI, in addition to the above 19 substances, methionine and one unknown substance were detected. 2. Generally, alanine, as partie acid, glutamic acid, cysteic acid and asparagine were detected in large amounts in the two hybrids as in PG, PG and PJ but it was a noticeable fact concerning these two hybrids that the largest quantity of asparagine was found at microspore satge and pollen mature stage. 3. The decrease of cysteic acid in the two hybrids at the red ripened stage was the same as in PQ and PJ but opposite to the change in PG. The detection of methionine in PG ${\times}$ PJ was worthy of notice. 4. The change of proline was conspicuously different from that in their parent plants. It was detected as a trace of color at the micros pore stage while asparagine was detected in the greatest amount at that time. It is well known that the quantity of proline is closely related to the sterility of plant. This fact was also found true in the formation of ginseng seeds. It was reported as well that asparagine accumulated when proline decreased. 5. The deficiency of proline seemed to be closely related with the sterility of hybrids and with the degradation of pollen in anther. 6. The difference in the changes of free amino acids between the selfed lines of PG, PQ and PJ, and their hybrids seemed to be caused by the transformation of gene-action system by hybridization. On these phenomena along with proline metabolim and its physiological role in seed formation further studies are required.

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Analysis of Mutant Chinese Cabbage Plants Using Gene Tagging System (Gene Tagging System을 이용한 돌연변이 배추의 분석)

  • Yu, Jae-Gyeong;Lee, Gi-Ho;Lim, Ki-Byung;Hwang, Yoon-Jung;Woo, Eun-Taek;Kim, Jung-Sun;Park, Beom-Seok;Lee, Youn-Hyung;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.28 no.3
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    • pp.442-448
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    • 2010
  • The objectives of this study were to analyze mutant lines of Chinese cabbage ($Brassica$ $rapa$ ssp. $pekinensis$) using gene tagging system (plasmid rescue and inverse polymerase chain reaction) and to observe the phenotypic characteristics. Insertional mutants were derived by transferring DNA (T-DNA) of $Agrobacterium$ for functional genomics study in Chinese cabbage. The hypocotyls of Chinese cabbage 'Seoul' were used to obtain transgenic plants with $Agrobacterium$ $tumefaciens$ harboring pRCV2 vector. To tag T-DNA from the Chinese cabbage genomic DNA, plasmid rescue and inverse PCR were applied for multiple copies and single copy insertional mutants. These techniques were successfully conducted to Chinese cabbage plant with high efficiency, and as a result, T-DNA of pRCV2 vector showed distinct various integration patterns in the transgenic plant genome. The polyploidy level analysis showed the change in phenotypic characteristics of 13 mutant lines was not due to variation in somatic chromosome number. Compared with wild type, the $T_1$ progenies showed varied phenotypes, such as decreased stamen numbers, larger or smaller flowers, upright growth habit, hairless leaves, chlorosis symptoms, narrow leaves, and deeply serrated leaves. The polyploidy level analysis showed the change in phenotypic characteristics of 13 mutant lines was not due to variation in somatic chromosome number. To tag T-DNA from the Chinese cabbage genomic DNA, plasmid rescue and inverse PCR were applied for multiple copies and single copy insertional mutants. Mutants that showed distinct phenotypic difference compared to wild type with 1 copy of T-DNA by Southern blot analysis, and with 2n = 20 of chromosome number were selected. These selected mutant lines were sequenced flanking DNA, mapped genomic loci, and the genome information of the lines is being recorded in specially developed database.