• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.159-159
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• 2017

Drought, a common environmental constraint, induces a range of physiological, biochemical and molecular changes in plants, and can cause severe reductions in crop yield. Consequently, understanding the molecular mechanisms of drought tolerance is an important step towards crop biotechnology. Here, we report that the rice (Oryza sativa) homeodomain-leucine zipper class IV transcription factor gene, ${\underline{R}ice}$ ${\underline{o}utermost}$ ${\underline{c}ell-specific}$ gene 10 (Roc10), enhances drought tolerance and grain yield by increasing lignin accumulation in ground tissues. Overexpression of Roc10 in rice significantly increased drought tolerance at the vegetative stages of growth and promoted both more effective photosynthesis and a reduction in water loss rate, compared with non-transgenic controls or RNAi transgenic plants. Importantly, Roc10 overexpressing plants had a higher drought tolerance at the reproductive stage of growth and a higher grain yield compared with the controls under field-drought conditions. Roc10 is mainly expressed in outer cell layers including the epidermis and the vasculature of the shoots, which coincides with areas of cell wall lignification. Roc10 overexpression elevated the expression levels of lignin biosynthetic genes in shoots, with a concomitant increase in the accumulation of lignin, while the overexpression and RNAi lines showed opposite patterns of lignin accumulation. We identified downstream target genes of Roc10 by performing RNA-seq and chromatin immunoprecipitation (ChIP)-seq analyses of shoot tissues. Roc10 was found to directly bind to the promoter of PEROXIDASEN/PEROXIDASE38, a key gene in lignin biosynthesis. Together, our findings suggest that Roc10 confers drought stress tolerance by promoting lignin biosynthesis in ground tissues.

• Plant Biotechnology Reports
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• 제4권3호
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• pp.213-222
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• 2010

Within their natural habitat, crops are often subjected to drought and heat stress, which suppress crop growth and decrease crop production. Causing overaccumulation of glycinebetaine (GB) has been used to enhance the crop yield under stress. Here, we investigated the response of wheat (Triticum aestivum L.) photosynthesis to drought, heat stress and their combination with a transgenic wheat line (T6) overaccumulating GB and its wild-type (WT) Shi4185. Drought stress (DS) was imposed by controlling irrigation until the relative water content (RWC) of the flag leaves decreased to between 78 and 82%. Heat stress (HS) was applied by exposing wheat plants to $40^{\circ}C$ for 4 h. A combination of drought and heat stress was applied by subjecting the drought-stressed plants to a heat stress as above. The results indicated that all stresses decreased photosynthesis, but the combination of drought and heat stress exacerbated the negative effects on photosynthesis more than exposure to drought or heat stress alone. Drought stress decreased the transpiration rate (Tr), stomatal conductance (Gs) and intercellular $CO_2$ concentration (Ci), while heat stress increased all of these; the deprivation of water was greater under drought stress than heat stress, but heat stress decreased the antioxidant enzyme activity to a greater extent. Overaccumulated GB could alleviate the decrease of photosynthesis caused by all stresses tested. These suggest that GB induces an increase of osmotic adjustments for drought tolerance, while its improvement of the antioxidative defense system including antioxidative enzymes and antioxidants may be more important for heat tolerance.

• 한국약용작물학회지
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• 제12권2호
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• pp.123-128
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• 2004

Hyoscyamine and scopolamine are two most common tropane alkaloids found in the Solanaceae. The pEB expression vector carrying Nspmt gene was transformed to Agrobacterium rhizogenes. The growth of transgenic hairy roots were approximately to 80% of the wild type root. Transgenic hairy roots are less developed on only their branch roots than those of the wild type root. The extracts from Nspmt transgenic hairy root lines, 3 and 5 contained between 3.52 and 4.23 mg/g dry weight as hyoscyamine and did between 5.23 to 6.40 mg/g dry weight as scopolamine. These results showed that the overexpression of the pmt gene enhanced tropane alkaloids production of S. parviflora transformed roots and this improvement affected both hyoscyamine and scopolamine production.

• 한국작물학회지
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• 제46권5호
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• pp.375-379
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• 2001

Transgenic plants from hypocotyl segments of buckwheat were produced with the Agrobacterium strain LBA4404 harboring the binary vector pBI121 containing chimeric genes of neomycin phosphotransferase II (npt II) and $\beta$-glucuronidase (gus). Two weeks after co-cultivation with Agrobacterium, most of the hypocotyl segments gradually became brown and died on the selection medium containing 100mg/$\ell$ of kanamycin. Plants regenerated from the hypocotyl explants grown on selection medium were GUS-positive in the leaf, stem and vascular tissues by histochemical assay, and varied in gus activity (440-2568 pmol, 4-MU/mg protein) by fluorimetry. The plants showing GUS activity were confirmed of containing GUS and NPT-II genes by polymerase chain reaction (PCR). Within 3 months, transgenic buckwheat plants were able to obtained from the hypocotyl segments.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2022년도 추계학술대회
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• pp.189-189
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• 2022

Soil salinity negatively affects plant growth, productivity, and metabolism. Rice is known to have more sensitive phenotypes than other cereal crops, such as wheat, sorghum, and barley. We characterized the molecular function of rice C3HC4 as a really interesting new gene (RING). Oryza sativa RING finger protein H2-3 (OsRFPH2-3) was highly expressed in 100 mM NaCl. To identify the localization of OsRFPH2-3, we fused vectors that include C-terminal GFP protein (35S;;OsRFPH2-3-GFP). OsRFPH2-3 was expressed in the nucleus in rice protoplasts. An in vitro ubiquitin assay demonstrated that OsRFPH2-3 possessed E3-ubiquitin ligase activity. However, the mutated OsRFPH2-3 were not possessed any E3-ubiquitin ligase activity. Under normal conditions, there is no significant phenotypic difference between transgenic plants and WT plants. However, OsRFPH2-3-overexpressing plants exhibited higher fresh weight and length under saline conditions. Also, transgenic plants maintain higher chlorophyll, proline, and soluble sugar contents and lower H₂O₂ and MDA contents than the wild type; these results support transgenic plants with enhanced salinity tolerance phenotypes.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.191-191
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• 2017

Hydroxy fatty acid (HFA) is an important industrial resource that known to be extracted from seeds of Castor or Lesquerella. However, mass production of HFA from those crops are difficult because of their behavior or life cycle. In this study, we applied HFA synthesis related gene FAH12, RcPDAT1, RcLPCAT, RcDGAT2, and RcPDCT on bioenergy crop Camelina sativa. Furthermore, we determined NaCl or cold stress tolerance changes of transgenic Camelina. RcFAH12, RcPDAT1, RcLPCAT, RcDGAT2, and RcPDCT genes were cloned into multigene expression vector which is engineered with seed specific promoter of FAE1 or Napin. Combination of HFA genes multi-expression vector constructs were divided into Set3 (RcFAH12, RcPDAT1-2, RcLPCAT), Set4 (RcFAH12, RcDGAT2, RCPDAT1-2, RcLPCAT), and Set5 (RcFAH12, RcDGAT2, RCPDAT1-2, RcLPCAT, RcPDCT). Transgenic HFA synthesis Camelina plants were generated using agrobacterium-mediated vacuum infiltration system. Results of fatty acid composition of T1 transgenic Camelina seeds analyzed by GC-MS showed 9.5, 9.0, and 13.6 % of HFA contents in Set3#6, Set4#8, and Set5#10, respectively. Therefore, seeds of T2 generation were harvest from Set5#10 which is shown highest HFA contents, and, 17.7, 8.1 and 10.5 % of HFA contents were determined in Set5#10-5, Set5#10-8, and Set#10-10, respectively. However, 7.7% of C18:2 and 22.3 % of C18:3 among unsaturated fatty acids were decreased in Set5#10-5 than WT. Meanwhile, we confirmed abiotic stress responses in T2 transgenic Camelina Set5#10-5 and Set5#10-10 under 0, 100, 150, and 200 mM NaCl or 25, 15, and $10^{\circ}C$ temperature for 5 weeks. Both Set5#10-5 and Set5#10-10 showed lower growth in height than WT in control and NaCl condition. Growth of leaf length and width were similar in WT and Set5#10-10 but lower in Set5#10-5 under NaCl stress. Number of opened flowers showed that both transgenic Camelina were lower than WT under normal condition. But, WT and Set5#10-10 showed similar opened flower number in 100 and 200 mM NaCl. In cold stress, 15 and $10^{\circ}C$ treatment for 5 weeks did not showed significant changes in between WT and both transgenic lines even they showed different growth rate in control condition. Taken together, growth and development are delayed by expression of exogenous HFA related genes in transgenic lines but relative abiotic stress sensitivity is similar with WT. In conclusion, reduced C18:2 or C18:3 fatty acid composition of seed by HFA synthesis is resulted from lack of resource supplement for development at seedling stage but it is not affect NaCl and cold stress tolerance.

• Journal of Plant Biotechnology
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• 제44권3호
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• pp.296-302
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• 2017

지구온난화로 인해 온도 상승에 따른 고온 스트레스는 전세계 많은 지역에서 농업적으로 문제가 되어 세계 3대 곡물인 벼의 생산에 피해가 크게 나타나고 있다. 식물은 생장하면서 다양한 환경스트레스에 노출되며, 이러한 스트레스는 작물의 생장, 발달, 수확량 등에 영향을 미친다. 본 연구는 벼의 안정적인 생산성을 높이기 위해 벼 유래 OsOPT 유전자를 이용한 형질전환 후대에서 고온 조건하에서도 생육이 가능한 계통을 선발하여 그 특성을 살펴보았다. 먼저, OsOPT10 유전자 도입 형질전환 벼를 이용하여 고온 처리에 따른 저항성 계통을 선발하고, 선발된 계통의 생리적 특성을 분석하였으며, 분자적 특성을 qRT-PCR을 통해 유전자의 발현 양상을 분석하였다. 고온 스트레스에 의한 세포막 피해 정도를 알아보기 위해 전해질 누출(electrolyte leakage), 삼투조절제 역할을 하는 수용성 당 및 proline 함량 분석을 하여 대조구와 비교분석 하였다. 본 실험에서 고온 처리에 의한 가용성 당 함량의 변화는 OsOPT10-16 형질전환 벼를 제외하고 OsOPT10-1와 OsOPT10-7 계통이 WT 보다 당 함량이 높게 나타났다. 모품종 동진에 비해 형질전환 벼 계통의 EL 값이 낮게 나타난 것과 가용성 당 함량이 비슷하거나 높게 나타난 것으로 보아 OsOPT10 형질전환 벼가 고온 스트레스에서 저항성 반응을 나타낸 것으로 판단하였다.

• 생명과학회지
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• 제23권11호
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• pp.1317-1324
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• 2013

고분자 글루테닌 서버유닛(high molecular-weight glutenin subunit, HMW-GS)은 밀의 가공적성을 결정하는데 중요한 역할을 수행한다. 우리는 Agrobacterium 동시 형질전환법을 이용하여 한국 밀 품종인 '조경'으로부터 밀 HMW-GS을 암호화하는 Glu-1Bx7 유전자를 가지는 marker-free 형질전환 벼를 생산하였다. Glu-1Bx7 유전자의 종자 특이적 발현을 위하여 밀 Glu-1Bx7 유전자 자체 프로모터를 벡터 내에 삽입하였다. 동시 접종을 위해서 오직 Glu-1Bx7 유전자와 hygromycin phosphotransferase II (HPTII) 저항성 유전자만으로 구성된 두 종류의 발현 카셋트를 독립적으로 Agrobacterium EHA105에 도입하였고, Glu-1Bx7와 HPTII가 도입된 각각의 EHA105 Agrobacterium 을 3:1 비율로 혼합하여 벼 캘러스에 접종하였다. 216개의 HPTII 저항성 형질전환체 중에서 벼 게놈에 Glu-1Bx7과 HPTII가 모두 삽입된 24개의 형질전환 라인을 획득하였다. Glu-1Bx7와 HPTII가 벼 게놈에 도입된 것을 Southern blot을 통해서 다시 확인하였다. 형질전환 벼 $T_1$ 세대의 종자에서 밀 Glu-1Bx7 유전자가 전사와 번역되어 오직 Glu-1Bx7만을 가지는 marker-free 식물체를 $T_1$ 세대에서 성공적으로 선발할 수 있었다.

• 생명과학회지
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• 제22권2호
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• pp.177-183
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• 2012

Ralstonia solanacearum (Rs)에 의해 유발되는 풋마름병은 감자 재배 시 발병하는 주요 병 중의 하나이다. 감자에서 풋마름병 저항성관련 유전자를 찾기 위해 기존에 기능이 알려진 다른 가지과 작물의 기능 유사 유전체를 이용하여 StACRE (HM749652) 유전자를 분리하고 염기서열을 분석하였다. 분리한 StACRE의 발현양상을 분석하기 위해 병 저항성 유도 신호전달 물질인 SA와 풋마름병원균 Rs (KACC10722)를 처리한 감자에서 RNA를 추출하여 RT-PCR을 실시한 결과 이 유전자는 SA 처리에 의해 3시간 후부터, Rs에 의해서는 12시간 후부터 발현이 현저하게 증가하였다. 따라서, 감자에서 이 유전자의 생물학적인 기능을 분석하기 위해 Gateway System을 이용하여 과발현용 vector를 만든 후 과발현 형질전환 감자를 제작하고 풋마름병균인 Rs를 접종하여 병 저항성 기능을 검정 한 결과 대조구인 수미 감자에 비해 병 저항성이 증대하였다.

• 한국육종학회지
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• 제41권3호
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• pp.252-260
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• 2009

1. 본 연구에서는 공동배양 배지에 Agrobacterium 성장 억제물질인 silver nitrate를 첨가하고 변온과 여과지처리를 추가하여 공동배양 기간을 7일로 늘였으며, 또한 항산화 물질 3종을 공동배양 배지에 첨가하여 세포의 oxidative burst를 최소화함으로써 벼 형질전환효율을 높일 수 있었다. 또한 이 방법을 적용하여 형질전환이 어려운 품종을 대상으로도 형질전환 식물체를 작성할 수 있었다. 2. 벼 형질전환체의 70%에서 도입유전자 수가 1copy인 것으로 나타나, 적은 수의 유전자가 안정적으로 도입됨을 확인 하였다. 3. 이러한 결과를 바탕으로, 새로운 공동배양 방법을 사용하여 우수한 농업적 형질을 가진 벼 육종 소재 및 품종을 신속하게 개발할 수 있을 것으로 기대된다.