• Journal of Plant Biotechnology
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• 제7권4호
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• pp.233-240
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• 2005

Slices of embryonic axis of mature pea (Pisum sativum L. cv. Green Arrow) seeds were used as explant. Transformation of explants was done via Agrobacterium tumefaciens bearing vector pBI-P5CS construct. The best results for inoculation of explants were obtained when they were immersed for 90 s at a concentration of $6{\times}10^8$ cell $ml^(-1)$ of bacterial suspension. Transformed pea plants were selected on $50\;mg\;l^(-1)$ kanamycin and successful transformants were confirmed by PCR and blotting. Transgenic plants were further analyzed with RT-PCR to confirm the expression of P5CS. Transgenic plants and non-transgenic plants were treated with different concentrations of NaCl 0 (control), 100, 150 and 200 mM in culture medium. Measurement of proline content indicated that transgenic plants produced more amino acid proline in response to salt in comparison with non-transgenic plants. Photosynthetic efficiency in transgenic plants under salt-stress was more than that of non-transgenic plants.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2022년도 추계학술대회
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• pp.234-234
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• 2022

Gene-editing is currently one of the most popular technologies in recent years. Development of the new crop using the gene editing have advantage of improved accuracy and efficiency compared with conventional breeding. Soybean (Glycine max L.) is one of the most important crops worldwide used as food and forage. We tried to establish a system for breeding improvement of soybean through gene-editing technology. For the gene-editing system of soybean, i) selection of efficiency gRNA of targeted gene, ii) efficient genetic transformation of the selected gRNA, iii) selection of trans-clean mutant is essential. First of all, we investigated the selection conditions of gRNA with high editing efficiency of targeted gene using isolated protoplast of soybean. Furthermore, we performed the Agrobacterium-mediated genetic transformation of various soybean cultivars. We identified the tissue culture ability in 23 soybean cultivars for genetic transformation of soybean. The six cultivars with high tissue culture ability were selected and confirmed the transgenic plants in four cultivars. Finally, we established a speed-breeding system as a powerful tool for the fast selection of trans-clean mutants from transgenic plants. Our laboratory will provide the valuable system for improvement of soybean by the gene-editing technology.

• Genomics & Informatics
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• 제13권3호
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• pp.81-85
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• 2015

Molecular characterization technology in genetically modified organisms, in addition to how transgenic biotechnologies are developed now require full transparency to assess the risk to living modified and non-modified organisms. Next generation sequencing (NGS) methodology is suggested as an effective means in genome characterization and detection of transgenic insertion locations. In the present study, we applied NGS to insert transgenic loci, specifically the epidermal growth factor (EGF) in genetically modified rice cells. A total of 29.3 Gb (${\sim}72{\times}coverage$) was sequenced with a $2{\times}150bp$ paired end method by Illumina HiSeq2500, which was consecutively mapped to the rice genome and T-vector sequence. The compatible pairs of reads were successfully mapped to 10 loci on the rice chromosome and vector sequences were validated to the insertion location by polymerase chain reaction (PCR) amplification. The EGF transgenic site was confirmed only on chromosome 4 by PCR. Results of this study demonstrated the success of NGS data to characterize the rice genome. Bioinformatics analyses must be developed in association with NGS data to identify highly accurate transgenic sites.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.164-164
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• 2017

High throughput transcriptome investigations of immunity in plants highlight the complexity of gene networks leading to incompatible interaction. To identify genes crucial to resistance against Xanthomonas oryzae pv oryzae, functional genetic analysis of selected differentially expressed genes from our microarray data set was carried out. A total of 13 overexpression vector constructs were made using 35S CaMV promoter which drive constitutive expression in rice. Most of the genes are developmentally expressed especially during maximum tillering stage and are commonly highly expressed in the leaves. When screened against Xoo strain K2, the transgenic plants displayed shorter lesion length compared with wild type Dongjin which indicates partial resistance. The levels of ROS continuously magnified after inoculation which indicates robust cellular sensing necessary to initiate cell death. Elevated transcripts levels of several defense-related genes at the downstream of defense signal network also corroborate the phenotype reaction of the transgenic plants. Moreover, expression assays revealed regulation of these genes by cross-communicating signal-transductions pathways mediated by salicylic and jasmonic acid. These collective findings revealed the key immune signaling conduits critical to mount full defense against Xoo.

• 한국육종학회지
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• 제42권5호
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• pp.540-546
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• 2010

콩의 PR10 유전자(GmPR10)를 벼에 형질전환하여 GmPR10 전이 유전자의 발현 정도와 내염성 관련 형질의 반응 사이의 인과관계를 조사하여 염 스트레스에 대한 GmPR10 생리적 기능을 분석하고 내염성 유전자원을 개발하였다. 1. 전이 유전자는 형질전환 계통에 따라 게놈 내에 1 ~ 6개의 사본이 도입되었고, 선발된 8개의 형질전환 계통 모두에서 전이 유전자가 발현되었으며, 발현 정도는 계통에 따라 변이를 보였다. 2. $T_1$세대 2계통의 형질전환 식물체와 비 형질전환 식물체에 125mM NaCl을 시간별로 처리한 결과, 전이 유전자 GmPR10의 전사체 검출양은 2계통의 형질전환체에서 모두 염처리 6시간까지 증가하였고, 12시간 이후에는 감소하였다. 3. 세포의 전해질 누출율은 형질전환체가 비 형질전환체에 비해 낮았고, 뿌리가 잎보다 낮았다. 또한, 전이 유전자 전사체의 검출량이 높을수록 전해질 누출율은 낮았다. 4. NaCl 용액에서의 생육 정도는 형질전환체가 비 형질전환체보다 현저히 양호하였으며 GmPR10 전이 유전자의 발현이 높을수록 생육 정도가 더 좋았다. 결론적으로 GmPR10 은 내염성을 증진시키는 기능이 있으며, GmPR10 전이유전자의 발현이 높은 계통은 내염성 벼 육성용 소재로 이용할 수 있을 것으로 평가된다.

• 생명과학회지
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• 제26권10호
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• pp.1121-1129
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• 2016

밀의 고분자 글루테닌 서브유닛[high molecular-weight glutenin subunit (HMW-GS)]은 밀가루의 성질을 결정하는데 가장 중요한 요소이며 가공적성을 나타내는데 중요한 역할을 수행한다. 우리는 Agrobacterium 동시 형질전환법을 이용하여 한국 밀 품종인 ‘조경’으로부터 밀 HMW-GS을 암호화하는 TaGlu-Ax1 유전자를 가지는 marker-free 형질전환 벼를 생산하였다. TaGlu-Ax1 유전자의 종자 특이적 발현을 위하여 밀에서 존재하는 TaGlu-Bx7 유전자의 자체 프로모터를 벡터 내에 삽입하였다. 동시 접종을 위해서 오직 TaGlu-Ax1 유전자와 hygromycin phosphotransferase II (HPTII) 저항성 유전자만으로 구성된 두 종류의 발현 카세트를 독립적으로 Agrobacterium EHA105에 도입하였고, TaGlu-Ax1와 HPTII가 도입된 각각의 EHA105 Agrobacterium을 3:1 비율로 혼합하여 벼 캘러스에 접종하였다. 210개의 HPTII 저항성 형질전환체 중에서 벼 게놈에 TaGlu-Ax1과 HPTII가 모두 삽입된 20개의 형질전환 라인을 획득하였다. TaGlu-Ax1와 HPTII가 벼 게놈에 도입된 것을 Southern blot을 통해서 다시 확인하였다. 형질전환 벼 T₁ 세대의 종자에서 밀 TaGlu-Ax1 유전자가 전사와 번역되어 오직 TaGlu-Ax1만을 가지는 marker-free 식물체를 T₁세대에서 성공적으로 선발할 수 있었다. TaGlu-Ax1 유전자가 발현되는 marker-free 형질전환 식물체는 야생형(wild type)과의 표현형 차이는 없었다. 형질전환 벼의 쌀가루의 제빵적성을 비교하였을 때 TaGlu-Ax1 유전자만이 발현되어서는 제빵적성이 더 나아지지 않았다. 그러므로 더 많은 밀 고분자 및 저분자 글루테닌, 글리아딘의 유전자의 집적과 조합이 쌀가루 가공적성을 증진시키는데 필요하다. 결론적으로 TaGlu-Ax1 marker-free 형질전환 벼는 쌀가루 가공적성을 증진시키는데 좋은 재료로 사용될 것이다.

• 한국약용작물학회지
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• 제15권2호
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• pp.100-104
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• 2007

A protocol for the production of transgenic Panax ginseng C.A. Meyer was established via Agrobacterium tumefaciens-mediated genetic transformation of direct somatic embryos. A number of conditions related to the co-cultivation were tested with respect to maximizing transformation efficiency. The results showed that pH of the co-cultivation medium (5.7), the bacterial growth phase (optical density; $OD_{600}$ = 0.8), co-cultivation period (3 days), and acetosyringone concentration $(100\;{\mu}M)$ had positive effects on transformation. Selected plantlets were cultured on the medium at an elevated hygromycin level(30 mg/l). Integration of the transgenes into the P. ginseng nuclear genome was confirmed by PCR analysis using hpt primers and by Southern hybridization using hpt-specific probe. The transgenic plantlets were obtained after 3-month cultivation and did not show any detectable variation in morphology or growth characteristics compared to wild-type plants.

• 한국약용작물학회:학술대회논문집
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• 한국약용작물학회 2009년도 심포지엄 및 춘계학술발표회
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• pp.257-258
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• 2009

• 한국환경농학회지
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• 제23권3호
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• pp.123-127
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• 2004

In many organisms, trehalose has been Down as an energy source and a protectant against various environmental stresses such as desiccation, freezing, heat and osmotic pressure. Previously, we have isolated and characterized the genes encoding trehalose-6-phosphate synthase (ZrTPS1) and trehalose-6-phosphate phosphatase (ZrTPS2) from one of the most osmotolerant yeasts, Zygosaccharomyces rouxii. We have also generated transgenic plants by co-introduction of ZrTPS2 and ZrTPS2 into potato plant (ZrTPS2-2A-ZrTPS1 plant) in an attempt to metabolically engineer trehalose in the transgenic plant using the foot-and-mouth disease virus(FMDV) 2A system and to generate drought resistant crop plants. In this research, we assayed previously generated the ZrTPS2-2A-ZrTPS1 plant biofunctionally by drought treatment, and measured the amount of trehalose in the ZrTPS2-2A-ZrTPS1 transgenic plants. The ZrTPS2-2A-ZrTPS1 transgenic plant showed strong drought resistance in spite of little or no accumulation of transgenic in he transgenic plant compared with control plant.

• Journal of Plant Biotechnology
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• 제38권3호
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• pp.215-220
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• 2011

This study was conducted to investigate how to enhance resistance to oxidative stress in petunia progeny obtained by a crossing between transgenic plants, MnSOD (SOD2) ($T_4$) and NDPK2 ($T_2$), to develop transgenic petunia much more resistant to environmental stress. At the treatment of MV 200 ${\mu}M$, the progeny was significantly less damaged than its parental plants (SOD2- or NDPK2-transgenic lines) as well as wild type plants, implying its resistance to oxidative stress was enhanced compare to that of SOD2- or NDPK2- transgenic plants. In an expression of 11 quantitative traits, the progeny remained similar to control plants, although it infrequently displayed slightly longer or wider than either parental or wild type plants. In the expression of 6 qualitative traits, there was no significant difference between parental or non-transgenic control plants.