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GBSSI 유전자 3' UTR 영역의 발현 억제 dsRNAi 벡터를 이용한 아밀로스함량 조절 벼 개발
박향미,최만수,천아름,이정희,김명기,김연규,신동범,이장용,김율호 한국육종학회 2010 한국육종학회지 Vol.42 No.5
본 연구에서는 RNAi 기작을 이용하여 식미에 중요한 영향을 미치는 아밀로스 함량을 다양화하기 위해 GBSSI 유전자의 3’-UTR 부위를 targeting하여 dsRNA를 생성시킬 수 있는 운반체를 제작하고, 벼에 형질전환 하였다. 작성된 형질전환체들을 대상으로 I2-KI 용액 반응과 아밀로스 함량을 분석한 결과, I2-KI 용액에 대한 반응은 waxy 타입으로 나타났으나 아밀로스 함량은 찰벼와 저아밀로스 벼 사이에 해당되는 범위를 보였다. 원품종과 The amylose content of starch is a major factor in the texture of cooked cereal grains. Therefore, down-regulation of amylose synthesis is one of the alternative method to improve eating quality of rice. We developed transgenic rice plants designed to suppress granule-bound starch synthase I(GBSSI) gene using RNA interference(RNAi) technology. Transgenic plants with RNAi vector containing the 3’-UTR region of GBSSI showed a lower amylose content in rice endosperm than that of wild-type. The range of amylose content was 5.9~9.0% in the transgenic plants, whereas that of wild-type was 17.7~18.0%. Transgenic rices showed the decrease of short chain and the increase of long chain by analyzing chain length distribution of amylopectin in the endosperm. In the SEM micrographs, we found that compound starch granules in whole grains of the wild-type rice were readily split during fracturing, while the starch granules in RNAi-transgenic lines showed small voluminous, non-angular rounded bodies.
엽록체에 과발현된 PsAPX1 유전자 형질전환 벼의 다양한 환경스트레스 내성 증진
박향미,김율호,최만수,이재은,최인배,최임수,신동범,이장용,곽상수,권석윤 한국육종학회 2009 한국육종학회지 Vol.41 No.3
1. 본 연구에서는 완두유래의 세포질성 PsAPX1 유전자를 대상으로 산화스트레스 유도성 프로모터를 연결하여 엽록체에 targeting 되는 과발현 운반체를 제작하고 벼에 도입한 결과 형질전환체에서 도입유전자 수가 1~3 copy인 것으로 나타나, 적은 수의 유전자가 안정적으로 도입되었음을 확인하였다. 2. 염, 오존, 자외선, 한발과 같은 다양한 환경스트레스 조건에서 내성이 증진된 우수 계통을 선발하기 위하여 작성된 형질전환 벼 계통들을 대상으로 생물 Oxidative stress is a major damaging factor for plants exposed to environmental stresses. In order to develop transgenic rice plants with enhanced tolerance to various environmental stresses, PsAPX1, the gene of ascorbate peroxidase isolated from Pisum sativum was expressed in chloroplast under the control of an oxidative stress inducible sweet potato peroxidase2 (SWPA2) promoter (referred to as PsAPX1 plants). PsAPX1 transgenic plants showed enhanced tolerance to various environmental stresses, such as 170 mM NaCl, UV-B, ozone, 20% PEG, and drought in compared with non-transgenic (NT) plants. These results suggest that chloroplast-targeted over-expression of PsAPX1 gene could be very useful strategy for developing transgenic rice plants with increased tolerance to environmental stresses.
제초제 저항성 형질전환 벼 계통의 농업적 특성과 분자생물학적 평가
박향미,김율호,서정필,최만수,김기종,신동범,박장환,이장용 한국육종학회 2007 한국육종학회지 Vol.39 No.2
Agronomic characteristics of transgenic herbicide-resistant rice lines were evaluated under field condition. Differences in agronomic traits and rice quality were observed between transgenic plants and the corresponding untransformed controls. Transgenic Ilpumbyeo lines flowered earlier than untransformed controls, whereas transgenic Nagdongbyeo flowered at the same time as untransformed control. With regard to the yield and rice quality, most of the transgenic lines showed lower yield than control except some selected transgenic lines. Selected Nagdongbyeo transgenic line also showed good eating quality comparable to the control although most selected Ilpumbyeo transgenic lines showed slightly lower eating quality and increased white center/belly in the rice grain. In order to find the main factor leading to somaclonal variation among transgenic lines, Tos17, a rice retrotransposon activated by prolonged tissue culture period was investigated. Although the transgenic lines carried only one or two of transformed bar gene, the copy number of Tos17 increased in most transgenic lines compared with control. The activation of Tos17 was not detected in selected promising transgenic lines such as ND115-15-1-B and IP23-3-3, suggesting that the increased copy number of Tos17 may have negatively affected agronomic characteristics of transgenic rice.