• Title/Summary/Keyword: plant transformation

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Fusaric Acid Production in Fusarium oxysporum Transformants Generated by Restriction Enzyme-Mediated Integration Procedure (Restriction Enzyme-Mediated Integration 방법으로 확보한 Fusarium oxysporum 형질전환체의 후자리산 생성능 분석)

  • Lee, Theresa;Shin, Jean Young;Son, Seung Wan;Lee, Soohyung;Ryu, Jae-Gee
    • Research in Plant Disease
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    • v.19 no.4
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    • pp.254-258
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    • 2013
  • Fusaric acid (FA) is a mycotoxin produced by Fusarium species. Its toxicity is relatively low but often associated with other mycotoxins, thus enhancing total toxicity. To date, biosynthetic genes or enzymes for FA have not been identified in F. oxysporum. In order to explore the genetic element(s) for FA biosynthesis, restriction enzyme mediated integration (REMI) procedure as an insertional mutagenesis was employed using FA producing-F. oxysporum strains. Genetic transformation of two F. oxysporum strains by REMI yielded more than 7,100 transformants with efficiency of average 3.2 transformants/${\mu}g$ DNA. To develop a screening system using phytotoxicity of FA, eleven various grains and vegetable seeds were tested for germination in cultures containing FA: Kimchi cabbage seed was selected as the most sensitive host. Screening for FA non-producer of F. oxysporum was done by growing each fungal REMI transformant in Czapek-Dox broth for 3 weeks at $25^{\circ}C$ then observing if the Kimchi cabbage seeds germinated in the culture filtrate. Of more than 5,000 REMI transformants screened, fifty-three made the seeds germinated, indicating that they produced little or fewer FA. Among them, twenty-six were analyzed for FA production by HPLC and two turned out to produce less than 1% of FA produced by a wild type strain. Sequencing of genomic DNA regions (252 bp) flanking the vector insertion site revealed an uncharacterized genomic region homologous (93%) to the F. fujikuroi genome. Further study is necessary to determine if the vector insertion sites in FA-deficient mutants are associated with FA production.

Effects of Plant Growth Regulators and Culture Medium Supplements on Embryogenic Callus Induction from Seeds of Zoysiagrass (들잔디 종자배양시 식물생장조절물질과 배지첨가물질이 배발생 캘러스 유도에 미치는 영향)

  • Jeon, Chan-Ho;Kim, Kyung-Hee;Park, Choong-Hoon;Kim, Yong-Gu;Lee, Hyo-Jin;Alam, Iftekar;Sharmin, Shamima;Lee, Ki-Won;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.29 no.1
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    • pp.1-6
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    • 2009
  • In order to optimize tissue culture conditions for genetic transformation of zoysiagrass (Zoysia japonica Stued.), the effect of plant growth regulators and culture medium supplements on embryogenic callus induction from mature seeds of a cultivar 'Zenith' were investigated. The optimal concentration and treatment period of NaOCl is 30% (v/v) for 60 minutes. Cultivation of mature seed on the callus Induction medium containing 3 mg/L 2,4-D and 3 mg/L dicamba showed 17.5% of embryogenic callus formation frequency. Supplementation of 1 g/L casein hydrolysate and 500 mg/L L-proline improved frequency of embryogenic callus induction. Audition of the medium with 5 mg/L $AgNO_3$ and 20 mg/L cysteine enhanced frequencies of embryogenic callus induction. Efficient callus induction system established in this study will be useful for molecular breeding of Boysiagrass through genetic transformation.

Development of System-Wide Functional Analysis Platform for Pathogenicity Genes in Magnaporthe oryzae

  • Park, Sook-Young;Choi, Jaehyuk;Choi, Jaeyoung;Kim, Seongbeom;Jeon, Jongbum;Kwon, Seomun;Lee, Dayoung;Huh, Aram;Shin, Miho;Jung, Kyungyoung;Jeon, Junhyun;Kang, Chang Hyun;Kang, Seogchan;Lee, Yong-Hwan
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.9-9
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    • 2014
  • Null mutants generated by targeted gene replacement are frequently used to reveal function of the genes in fungi. However, targeted gene deletions may be difficult to obtain or it may not be applicable, such as in the case of redundant or lethal genes. Constitutive expression system could be an alternative to avoid these difficulties and to provide new platform in fungal functional genomics research. Here we developed a novel platform for functional analysis genes in Magnaporthe oryzae by constitutive expression under a strong promoter. Employing a binary vector (pGOF1), carrying $EF1{\beta}$ promoter, we generated a total of 4,432 transformants by Agrobacterium tumefaciens-mediated transformation. We have analyzed a subset of 54 transformants that have the vector inserted in the promoter region of individual genes, at distances ranging from 44 to 1,479 bp. These transformants showed increased transcript levels of the genes that are found immediately adjacent to the vector, compared to those of wild type. Ten transformants showed higher levels of expression relative to the wild type not only in mycelial stage but also during infection-related development. Two transformants that T-DNA was inserted in the promotor regions of putative lethal genes, MoRPT4 and MoDBP5, showed decreased conidiation and pathogenicity, respectively. We also characterized two transformants that T-DNA was inserted in functionally redundant genes encoding alpha-glucosidase and alpha-mannosidase. These transformants also showed decreased mycelial growth and pathogenicity, implying successful application of this platform in functional analysis of the genes. Our data also demonstrated that comparative phenotypic analysis under over-expression and suppression of gene expression could prove a highly efficient system for functional analysis of the genes. Our over-expressed transformants library would be a valuable resource for functional characterization of the redundant or lethal genes in M. oryzae and this system may be applicable in other fungi.

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Current Status and Perspective and Future Task in Korea of Crop Genetic Transformation (작물형질전환의 현황과 한국내의 발전전망 및 과제)

  • Harn, Chee-Hark
    • Journal of Plant Biotechnology
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    • v.33 no.3
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    • pp.171-184
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    • 2006
  • According to ISAAA report, the global area of genetically modified (GM) crops increased more than 50 fold during the ten-year period from 1996 to 2005 with a sustained double-digit growth rate of 10%. This biotechnology adoption is one of the highest rates of technology adoption in agriculture history and this phenomenon indicates that the industrial value of the GM crops is highly perspective. In addition, the year 2010, 60% of cereal seeds in the global market would be GM or biotechnology related seeds so that the GM crop regards as the second green revolution that could provide a huge impact to food and agriculture. Nevertheless, there has not been any GM variety ever successfully commercialized in Korea and even none of the GM crops has ever been approved for safety testing by risk assessment. This seems that Korean agriculture industry might be indeed lost in the war of future seed market. However, lots of evidence show that Korean scientists have established advanced technologies and protocols to develop GM crops for last 20 years. Actually there have been many cases of successful transformation of crops that were previously known very difficult in transforming. Therefore, Korean agbiotechnology arena firmly holds an infrastructure for developing GM crops with a superior technology. Then what were the problems? Why has even a single GM crop not been commercialized in Korea? The tardiness shown by business in adopting the GM crop is caused by many factors: academical weakness, poor research funding, short knowledge of risk assessment, public concern, no successful experience, lack of professional leaders on GM variety development, lack of systems toward industrialization and inappropriate target transgenes from the beginning. In order to catch up in the race for the new green industry, each one of us in private sectors alongside academia and national research institutes needs to focus altogether on what can be done best in terms of choosing crops, investing fund and establishing a road map for commercialization of GM crops.

Generation of Transgenic Rice without Antibiotic Selection Marker through Agrobacterium-mediated Co-transformation System (아그로박테리움 동시 형질전환 시스템을 통한 항생제 선발 마커가 없는 형질전환벼의 생산)

  • Park, Soo-Kwon;Kwon, Tack-Min;Lee, Jong-Hee;Shin, Dong-Jin;Hwang, Woon-Ha;Song, You-Chun;Cho, Jun-Hyun;Nam, Min-Hee;Jeon, Seung-Ho;Lee, Sang-Yeol;Park, Dong-Soo
    • Journal of Life Science
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    • v.22 no.9
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    • pp.1152-1158
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    • 2012
  • Development of transgenic plant increasing crop yield or disease resistance is good way to solve the world food shortage. However, the persistence of marker genes in crops leads to serious public concerns about the safety of transgenic crops. In the present paper, we developed marker-free transgenic rice inserted high molecular-weight glutenin subunit (HMW-GS) gene ($D{\times}5$) from the Korean wheat cultivar 'Jokyeong' using Agrobacterium-mediated co-transformation method. Two expression cassettes comprised of separate DNA fragments containing only the $D{\times}5$ and hygromycin resistance (HPTII) genes were introduced separately into Agrobacterium tumefaciens EHA105 strain for co-infection. Each EHA105 strain harboring $D{\times}5$ or HPTII was infected into rice calli at a 3: 1 ratio of EHA105 with $D{\times}5$ gene and EHA105 with HPTII gene expressing cassette. Then, among 66 hygromycin-resistant transformants, we obtained two transgenic lines inserted with both the $D{\times}5$ and HPTII genes into the rice genome. We reconfirmed integration of the $D{\times}5$ and HPTII genes into the rice genome by Southern blot analysis. Wheat $D{\times}5$ transcripts in $T_1$ rice seeds were examined with semi-quantitative RT-PCR. Finally, the marker-free plants containing only the $D{\times}5$ gene were successfully screened at the $T_1$ generation. These results show that a co-infection system with two expression cassettes could be an efficient strategy to generate marker-free transgenic rice plants.

Uptake and Transformation of RDX by Perennial Plants in Poaceae Family (Amur Silver Grass and Reed Canary Grass) under Hydroponic Culture Conditions (수경재배조건에서 다년생 벼과식물(물억새 및 갈풀)에 의한 RDX 흡수 및 분해)

  • Park, Jieun;Bae, Bumhan
    • Journal of Korean Society of Environmental Engineers
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    • v.36 no.4
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    • pp.237-245
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    • 2014
  • Amur silver grass (Miscanthus sacchariflorus) and reed canary grass (Phalaris arundinacea) were selected for RDX removal experiments in hydroponic culture conditions based on vegetation survey at three shooting ranges in northern Kyunggi province. Seedling of two plants were grown in 1/4 strength Hoagland solution in quadruplicates containing 10, 20, 30, 40 mg/L RDX for 15 days along with control and blank treatments. During the 15 days of incubation, pH and RDX concentration in medium were routinely analyzed and RDX contents in the shoot and the root were determined after solvent extraction at the end of the experiments. Both plant species showed no symptoms of RDX phyto-toxicity. The pseudo first order RDX-removal constants for amur silver grass and reed canary grass were in the range of $0.0143{\sim}0.0484day^{-1}$ and $0.0971{\sim}0.1853^{-1}$, respectively. Plant biomass normalized RDX removal rates, which decreased with the increase of initial RDX concentration, were in the range of $0.27{\sim}1.01mL{\cdot}g^{-1}day^{-1}$ and $0.87{\sim}1.66mL{\cdot}g^{-1}day^{-1}$ for amur silver grass and reed canary grass, respectively. After 15 days of treatment, RDX removal from the medium decreased from 49.0% to 23.7% with increase in the initial RDX concentration in amur silver grass and 7.3% of the initial RDX remained in the plant. In reed canary grass planted medium, less than 16.8% and 5% of the initial RDX remained in the medium and in the plant, respectively. Large quantities of unidentified polar compound, which was not detected in amur silver grass, accumulated in the root and shoot of reed silver grass.

Research status of the development of genetically modified papaya (Carica papaya L.) and its biosafety assessment (GM 파파야 개발 및 생물안전성 평가 연구 동향)

  • Kim, Ho Bang;Lee, Yi;Kim, Chang-Gi
    • Journal of Plant Biotechnology
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    • v.45 no.3
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    • pp.171-182
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    • 2018
  • Papaya (Carica papaya L.) is one of the crops widely planted in tropical and subtropical areas. The papaya fruit has low calories and are plentiful in vitamins A and C and in minerals. A major problem in papaya production is a plant disease caused by the papaya ringspot virus (PRSV). The first PRSV-resistant GM papaya expressing a PRSV coat protein gene was developed by USA scientists in 1992. The first commercial GM papaya cultivars derived from the event was approved by the US government in 1997. Development of transgenic papayas has been focused on vaccine production and limited agricultural traits, including insect and pathogen resistance, long shelf life, and aluminum and herbicide tolerance. Approximately 17 countries, including the USA and China, produced transgenic papayas and/or commercialized them, which provoked studies on biosafety assessment and development of GM-detection technologies. For the biosafety assessment of potential effects on human health, effects of long-term feeding to model animals have been studied in terms of toxicity and allergenicity. Studies on environmental safety assessment include influence on soil-microbial biodiversity and transfer to soil bacteria of GM selection markers. Many countries, such as Korea, the European Union, and Japan, that have strict regulations for GM crops have serious concerns about unintended introduction of GM cultivars and food commodities using unauthorized GM crops. Transgene- and/or GM event-specific molecular markers and technologies for genomics-based detection of unauthorized GM papaya have been developed and have resulted in the robust detection of GM papayas.

Converting Lands that are damaged by Graveyards into Tree Burial Sites in order to Restore Green Areas (산지묘지의 훼손지 복원을 위한 수목장지로의 전환)

  • Woo, Jae-Wook;Byun, Woo-Hyuk;Kim, Hak-Beom;Park, Won-Kyoung;Kim, Min-Su;Norsyuhada, Norsyuhada
    • Journal of the Korean Institute of Landscape Architecture
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    • v.40 no.3
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    • pp.69-80
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    • 2012
  • The purpose of this paper was to study the issues related to converting the graveyards within forests into spaces intended for tree burials by means of planting, given the situation that the graveyards have encroached on land and damaged the environment. For the reason, a field survey was performed to determine the width, length, and distance to the nearest tree of 205 graveyards in the capital area. Through this, it was determined that the domestic lands damaged by graveyards amounted to $862km^2$, including the areas that were deforested to manage the graves. This only confirms that land encroachment by graveyards is a serious issue. The methods for making tree burial sites were examined from the perspective of how to meet public demands given the graveyard's spatial distinctiveness. As a result, this study suggested different methods to establish tree burial sites according to the degree of transformation and the term of its formation. This study also classified the graveyards into three types, and identified the planting methods that harmonized the safe growth of trees and the scenic beauty of memorial places based on the standard. This is in order to plant trees that are shade-tolerant and suitable to the forest line, along with which other tree line was and also, to plant aesthetic trees around the empty space. Through applying the developed methods, this study established and monitored two exemplary sites in Yongin and Boryeng. Aesthetic trees were planted in Yongin site which was located in an open area, aod the shade-tolerant trees were planted in Boryeong, which was located in a forest area. As a result, the image of a garden appeared at Yongin site and the image of a tree colony harmonized with the near forest emerged at Boryeong site. Therefore, it is confirmed that the method of planting according to the distribution status of neighboring trees was effective. As a result of monitoring, mulching wood chips were suitable for sites that were small or easy to approach. This is because the weeds were controlled in Yongin site by mulching. Furthermore, by monitoring the growth of 11 species of vegetation, this study confirmed that low and cover-type vegetations were suitable for tree burial sites. In Boryeong site, the wild cherry trees, which were planted as adult trees, all died, and the tilling of snake's beard, which were planted as cover vegetation, was slow. Therefore, this study found that seedlings were more suitable to plant in forest graveyards than adult trees, which were large and difficult to approach, and it was effective to use the remaining lawn and form a low vegetation after the crown of trees had expanded to such places.

The Chemical Composition and Working Techniques of the Glass Beads Excavated in the Jisan-dong No. 73-74 Ancient Tombs, Goryeong (고령 지산동 73~74호분 출토 유리구슬의 제작 기법과 화학 조성)

  • Kim Nayoung;Kim Euna;Kim Gyuho
    • Conservation Science in Museum
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    • v.31
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    • pp.21-37
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    • 2024
  • This paper sought to garner an understanding of Daegaya glass culture by observing the micro-structure and analyzing the chemical composition of 43 glass beads excavated from the No.73 and 74 ancient tombs in Jisan-dong, Goryeong, which are estimated to have a central age of 5th century CE. The visible characteristics and micro-structure of these artifacts were observed with a optical microscope and an scanning electron microscope, while their chemical composition was analyzed with an energy-dispersing spectrometer attached to the scanning electron microscope. As a result, the glass beads of Jisan-dong, Goryeong were identified to have been formed using various methods such as drawing, casting, and folding techniques, with the majority molded by the drawing technique. In terms of chemical composition, 32.6% were in the potash glass group and 67.4% in the soda glass group, with the latter divided into various fluxes such as high alumina glass, netron glass, and plant ash glass. Compared to Baekje's cultural region in the same age, the composition of these ancient glass artifacts demonstrates a high share of the potash glass group. This shows that, despite the shift from the potash glass group to the soda glass group in ancient Korean glass culture, glass composition differs from region to region or depending on the cultural sphere of influence. In the soda glass group, high-alumina glass comprised 23.3%, natron glass 43.0%, and plant ash glass was 1.2%. Among them, the main type of Korean soda glass is high-alumina glass, as natron glass and plant ash glass are known to have appeared later, but the results of scientific analysis of the glass beads excavated in Jisan-dong can be expected to provide important clues about the inflow and transformation of ancient glass on the Korean Peninsula. In the No. 73, 74, and 74-1 ancient tombs, which were found to have been built in chronological order by the excavation survey, the glass beads showed only slight variations depending on their production period. Nonetheless, the chemical composition of glass is deemed to have a close correlation to color.

A Study on the Acculturation of Guǐmok(槐木) Plantings through the Remaining Species of Guǐjeong(槐亭) (괴정(槐亭)의 잔존 수종을 통해 본 괴목(槐木) 식재의 문화변용)

  • Rho, Jae-Hyun;Han, Sang-Yub;Choe, Seung-Heuy
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.37 no.4
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    • pp.81-97
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    • 2019
  • The purpose of this study is to examine what people in Korea recognize the cultural symbolism and the planting patterns of 'Guǐmok', pagoda tree(Sophora japonica). The species planted in the 'Guǐjeong' was empirically investigated and analyzed to determine which species of pagoda tree or Zelkova tree(Zelkova serrata) was taken through literature surveys, field surveys, and interviews with persons. This 'Guǐjeong' was combined to track how the culture of the 'Guǐmok' planting introduced in China was ultimately accepted and transformed in Korea. In this study, we tried to analyze the meaning implicit in the mystery while checking the distribution of the mystery and the form of the mystery, the name of the pavilion and its relevance to the contrast medium. Essentially, the trees that govern the characterization of the nectar plant, regardless of the region, are considered to be a pagoda tree, which is considered an internal factor in which the pagoda tree culture was not completely transformed into a zelkova tree. It was recognized throughout the Joseon Dynasty that the species representing 'Prime ministers(三公)' was judged from all the Joseon Dynasty periods, based on the builder of Guǐjeong's Aho(雅號) and Dangho(堂號). It was confirmed that the tree was very likely to be planted in place of the painting tree. But now is selectively zelkova tree is in accordance with the preparation of planting site conditions and areas on behalf of the Change is very high probability that is planted. Cultural variables that led to the cultural transformation of the 'Guǐmok' seem to have been deeply involved in the geographical space of China and Korea, Confucian practices of the Choson society, comings and goings and letter bridge, and network strength with China through the book spread. In addition, the culture of 'Guǐmok' is presumed to have led to cultural custom of the upper class, not the whole class, in the Yeongnam region, it can be said that the independent adaptation to act to recognize 'Guǐmok' as a pagoda tree, that is a Sophora japonica, has occurred very strongly. The difference between the cultural areas of Yeongnam and non-Yeongnam is also considered to be an internal factor that has played a major role in the cultural transformation of planting of 'Guǐmok'.