• Molecules and Cells
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• 제39권3호
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• pp.250-257
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• 2016

Abiotic stresses such as drought and low temperature critically restrict plant growth, reproduction, and productivity. Higher plants have developed various defense strategies against these unfavorable conditions. CaPUB1 (Capsicum annuum Putative U-box protein 1) is a hot pepper U-box E3 Ub ligase. Transgenic Arabidopsis plants that constitutively expressed CaPUB1 exhibited drought-sensitive phenotypes, suggesting that it functions as a negative regulator of the drought stress response. In this study, CaPUB1 was over-expressed in rice (Oryza sativa L.), and the phenotypic properties of transgenic rice plants were examined in terms of their drought and cold stress tolerance. Ubi:CaPUB1 T3 transgenic rice plants displayed phenotypes hypersensitive to dehydration, suggesting that its role in the negative regulation of drought stress response is conserved in dicot Arabidopsis and monocot rice plants. In contrast, Ubi:CaPUB1 progeny exhibited phenotypes markedly tolerant to prolonged low temperature ($4^{\circ}C$) treatment, compared to those of wild-type plants, as determined by survival rates, electrolyte leakage, and total chlorophyll content. Cold stress-induced marker genes, including DREB1A, DREB1B, DREB1C, and Cytochrome P450, were more up-regulated by cold treatment in Ubi:CaPUB1 plants than in wild-type plants. These results suggest that CaPUB1 serves as both a negative regulator of the drought stress response and a positive regulator of the cold stress response in transgenic rice plants. This raises the possibility that CaPUB1 participates in the cross-talk between drought and low-temperature signaling pathways.

• Plant Biotechnology Reports
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• 제2권1호
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• pp.41-46
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• 2008

Salinity stress is a major limiting factor in cereal productivity. Many studies report improvements in salt tolerance using model plants, such as Arabidopsis thaliana or standard varieties of rice, e.g., the japonica rice cultivar Nipponbare. However, there are few reports on the enhancement of salt tolerance in local rice cultivars. In this work, we used the indica rice (Oryza sativa) cultivar BR5, which is a local cultivar in Bangladesh. To improve salt tolerance in BR5, we introduced the Escherichia coli catalase gene, katE. We integrated the katE gene into BR5 plants using an Agrobacterium tumefaciens-mediated method. The introduced katE gene was actively expressed in the transgenic BR5 rice plants, and catalase activity in $T_1$ and $T_2$ transgenic rice was approximately 150% higher than in nontransgenic plants. Under NaCl stress conditions, the transgenic rice plants exhibited high tolerance compared with nontransgenic rice plants. $T_2$ transgenic plants survived in a 200 mM NaCl solution for 2 weeks, whereas nontransgenic plants were scorched after 4 days soaking in the same NaCl solution. Our results indicate that the katE gene can confer salt tolerance to BR5 rice plants. Enhancement of salt tolerance in a local rice cultivar, such as BR5, will provide a powerful and useful tool for overcoming food shortage problems.

• 생명과학회지
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• 제14권2호
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• pp.368-373
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• 2004

광범위 제초제인 Bast $a^{(R)}$에 대해 저항성을 가지는 형질전환체 벼를 개발하였다. Bar유전자를 함유하고 있는 플라스미드 pCaMV35S::Bar를 embryogenic 현탁 배양체로부터 분리한 벼의 원형질체에 도입하였다. Phosphinotricin에 대해 저항성을 가지는 형질전환체 식물체들이 재분화되었고, 이들을 15 mg/l phosphinotricin이 함유한 배지에서 다시 선별하였다. 형질전환체 벼에서 bar유전자의 삽입과 발현을 Southern과 Northern blot분석으로 확인하였고, 또한 $R_1$ 형질전환 식물체들을 PAT 활성 assay로 재차 유전자 발현을 확인하였다. Bar 유전자는 다음 세대인 $R_1$ 식물체에서 3:1 멘델 유전 양상을 나타내었고, 형질전환체 $R_1$과 $R_2$ 식물체들은 fieild에서 살포되는 제초제 양만큼 Bast $a^{(R)}$ 를 살포했을 때 제초제 저항성을 나타내었다..

• 한국환경농학회지
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• 제21권2호
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• pp.156-161
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• 2002

냉해나 한발등의 환경스트레스에 대해 저항성을 유발하는 유전자를 환경스트레스에 강한 잡초성벼로부터 선발하고 이들 유전자를 재배벼에 도입하여 도입유전자 산물의 과량 발현을 통해 냉해나 한발 등에 대한 저항성이 향상된 벼를 선발하고자 하였다. 잡초성벼인 Bhutan 14Ad로부터 냉해 및 한발 저항성 유전자로 알려진 superoxide dismutase (SOD) cDNA를 분리하고자 mRNA를 분리하고 이 분리된 mRNA를 이용해 reverse transcriptase PCR방법으로 SOD cDNA를 cloning 하였다. 그 결과 2종의 SOD cDNA가 cloning되어 SOD-A, SOD-B로 명명하였다. 이들 cDNA의 염기서열을 결정한 결과 이들은 아미노산 서열 상동성이 88.4%를 나타내었으며, SOD-A는 Oryza sativa 계열의 Cu/Zn SOD유전자인 GenBank accession No. L36320와 99.3% 동일하였으며, SOD-B는 accession No. D01000과 100% 동일하였다. 이들 SOD-A와 SOD-B cDNA를 재배벼인 낙동벼에 형질전환하여 형질전환체 벼를 선발하였으며, 이들 형질전환체 벼의 냉해저항성및 한발저항성 검정을 통해 저항성이 향상된 형질전환체 벼를 선발하고 있다.

• Journal of Plant Biotechnology
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• 제34권2호
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• pp.145-152
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• 2007

Lactoferrin is an 80-kDa iron-binding glycoprotein known to exert many biological activities, such as facilitating iron absorption and having antimicrobial and anti-inflammatory effects. Rice can be a useful target for edible food plants to introduce human lactoferrin, because it has lower allergenicity and is likely to be safer than microorganisms or transgenic animals. A cDNA fragment encoding human lactoferrin (HLF) driven by the maize polyubiquitin promoter, along with herbicide resistance gene (bar) driven by CaMV 35S promoter, was introduced into rice (Oryza sativa L. cv. Dong Jin) using the Agrobacterium -mediated transformation system. Putative transformants were initially selected on the medium containing bialaphos. The stable integration of the bar and HLF genes into transgenic rice plants was further confirmed through polymerase chain reaction (PCR) and Southern blot analyses. The expression of the full length HLF protein from various tissues such as grains and young leaves of transgenic rice was verified by Western blot analysis. Analysis of progeny also demonstrated that introduced genes were stably inherited to the next generation at the Mendelian fashion.

• Plant Resources
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• 제7권1호
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• pp.39-46
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• 2004

To demonstrate the importance of transformation efficiency in independent event, molecular and cytogenetic analysis were conducted with genomic DNA and chromosome of transgenic plants produced by Agrobacterium tumefeciens LBA4404 (pSBM-PPGN: gusA and bar). Selection ratios of putative transgenic calli were similar in independent experiments, however, transformation efficiencies were critically influenced by the type of regeneration media. MSRK5SS-Pr regeneration mediun, which contains 5 mgL$^{-1}$ kinetin, 2% (w/v) sucrose in combination with 3% (w/v) sorbitol, and 500 mgL$^{-1}$ proline, was efficient to produce transgenic plant of rice from putative transgenic callus in the presence of L-phosphinotricin (PPT). With MSRK5SS-Pr medium, transformation efficincies of Nagdongbyeo were significantly enhanced from 3.7% to 6.3% in independent callus lines arid from 7.3% to 19.7% in plants produced, respectively. Stable integration and expression of bar gene were confirmed by basta herbicide assay, PCR amplification and Southern blotting of bar gene, and fluorescence in situ hybridization (FISH) analysis using pSBM-PPGN as a probe. In Southern blot analysis, diverse band patterns were observed in total 44 transgenic plants regenerated from 20 independent PPT resistant calli showing from one to five copies of T-DNA segments, however, the transformants obtained from one callus line showed the same copy numbers with the same fractionized band patterns.

• BMB Reports
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• 제42권1호
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• pp.16-21
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• 2009

Three novel Class A genes that encode heat shock transcription factor (HSF) were cloned from Oryza Sativa L using a yeast hybrid method. The OsHSF7 gene was found to be rapidly expressed in high levels in response to temperature, which indicates that it may be involved in heat stress reception and response. Over-expression of OsHSF7 in transgenic Arabidopsis could not induced over the expression of most target heat stress-inducible genes of HSFs; however, the transcription of some HSF target genes was more abundant in transgenic plants following two hours of heat stress treatment. In addition, those transgenic plants also had a higher basal thermotolerance, but not acquired thermotolerance. Collectively, the results of this study indicate that OsHSF7 might play an important role in the response to high temperature. Specifically, these findings indicate that OsHSF7 may be useful in the production of transgenic monocots that can over-express protective genes such as HSPs in response to heat stress, which will enable such plants to tolerate high temperatures.

• 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.

• 식물조직배양학회지
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• 제27권6호
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• pp.441-445
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• 2000

저인산 조건하에서 인산 흡수 효율이 높은 벼 품종을 개발하고자 담배의 인산수송자 유전자를 벼에 형질전환하였다. 동진벼로부터 유도된 callus를 담배의 인산수송자 유전자가 도입된 A. tumefaciens LBA 4404와 공동배 양한 후, 선발배지에서 증식된 callus를 250 mg/L carbenicillin, 2 mg/L kinetin, 0.1 mg/L NAA가 첨가된 MS배지에 옮겨 약 2주 후부터 소식물체를 얻었다. Carbenicillin 250 mg/L 첨가된 MS 기본배지에서 소식물체의 발근을 유도하여 재분화 식물체를 얻었다. 선발된 callus는 약 52%의 식물체 재력화율을 보였다. 선발된 재분화 식물체에 대한 PCR 분석을 수행하여 형질전환 벼 식물체에 담배의 인산수송자 유전자가 삽입되었음을 확인하였다. 형질전환체의 genonlic DNA로부터 PCR에 의해 증폭된 DNA에 대한 Southern blot분석 결과, 대조 식물체에서는 band가 검출되지 않았으나 형질전환 식물체에서는 인산수송자 유전자와 동일한 1.7kb의 band가 검출되어 외래유전자인 담배 인산수송자 유전자가 안전적으로 도입되었음이 확인되었다.

• 식물조직배양학회지
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• 제28권2호
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• pp.97-101
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• 2001

동화산물의 분배 효율 개선을 통한 생산성 향상 가능성을 조사하고자 감자의 sucrose 수송자 유전자를 벼에 형질전환 하였다. 동진벼로부터 유도된 callus를 감자의 sucrose 수송자 유전자가 도입된 A. tumefaciens LBA 4404와 공동배양한 후, 선발배지에서 증식된 callus를 250 mg/L carbenicillin, 2 mg/L kinetin, 0.1 mg/L NAA가 첨가된 MS 배지에 옮겨 약2주 후부터 소식물체를 얻었다. Carbenicillin 250 mg/L 첨가된 MS 기본 배지에서 소식물체의 발근을 유도하여 재분화 식물체를 얻었다. 선발된 callus는 약 150%의 높은 식물체 재분화율을 보였다. 재분화 식물체에 대한 PCR 분석을 수행하여 감자의 sucrose수송자 유전자가 삽입된 형질전환 벼 식물체를 선발하였다. 선발된 형질전환 식물체에 대한 Southern blot 분석을 통하여 외래유전자인 감질 sucrose 수송자 유전자가 벼 genome 내에 안정적으로 도입되었음을 확인하였다.