가뭄저항성벼(CaMsrB2-8)의 환경위해성 평가 : 3세대의 후대안정성 연구

국문 초록 (Abstract)

가뭄저항성벼의 복수세대에 대한 후대안정성을 서던 블롯과 PCR로 분석한 결과, 가뭄저항성벼의 CaMsrB2-8 T4 ~ T6 세대에서는 도입된 모든 유전자들이 안정적으로 도입되어 있으며, T-DNA 구성요소 이외의 backbone DNA는 가뭄저항성벼에 삽입되지 않았음을 확인하였다. 목적 유전자인 CaMsrB2와 제초제 저항성 선발 마커인 bar가 가뭄저항성벼의 CaMsrB2-8 T4 ~ T6 세대에서 안정적으로 발현됨을 검증하였다. 제초제 저항성 선발 마커로 도입된 PAT 단백질의 발현 분석 결과에서도 CaMsrB2-8 T4 ~ T6의 3세대에서 생육시기별, 부위별로 안정적으로 발현됨을 입증하였다. 도입유전자의 삽입 위치를 확인한 결과, 가뭄저항성벼 CaMsrB2-8의 도입유전자가 벼 1번 염색체 내에서 intergenic한 상태로 안정적으로 유지되고있음을 확인하였다. 이상의 분석 기법을 통해 복수세대에서 가뭄저항성벼의 도입 유전자들이 안정적으로 유지되고 목적 단백질들이 안정적으로 발현되고 있음을 확인하였다.

다국어 초록 (Multilingual Abstract)

The purpose of this research is a molecular characterization of introduced T-DNA in transgenic rice T4~T6 generation lines (#8) harboring a pepper MsrB2 gene under the control of stress inducible Rab21 promoter, as a part of biosafety evaluation for drought-tolerant transgenic rice (CaMsrB2-8). We identified the structure and sequence of transformation vector of T-DNA and analyzed insertion sites, flanking sequences, and generational stability of inserted T-DNA in transgenic rice. The transformation vector was consisted of right border, a drought-tolerant CaMsrB2-8 gene unit (Rab21 promoter::CaMsrB2::PinII terminator), a selectable marker herbicide resistance unit (CaMV 35S promoter::bar::Nos terminator), and left border in sequential order. We confirmed that T-DNA was introduced at the position of 41,737,284 bp of chromosome No. 1. based on the adaptor-ligation PCR and whole genome sequence database. T-DNAs were stably inherited through the T4 to T6 generations, and also stable expression of bar gene from T-DNA was confirmed. It was also confirmed that the backbone DNA of transformation vector containing antibacterial gene (aadA) was not present in CaMsrB2-8 rice genome. These results will be filed to biosafety assessment document of CaMsrB2-8 rice.

참고문헌 (Reference)

1 이기종, "비타민A 강화벼의 환경위해성 평가 : 4세대의 후대안정성 연구" 한국육종학회 44 (44): 579-587, 2012

2 이현숙, "대규모 GMO 포장에서 비타민 A 강화 벼의 농업특성 검정" 한국육종학회 42 (42): 56-60, 2010

3 GLAAS, "UN-water Global Analysis and Assessment of Sanitation and Drinking-Water" WHO 2012

4 James C, "The global status of commercialized biotech/ GM crops: 2012" 44 : 2012

5 Boyer JS, "Plant productivity and environment" 218 : 443-448, 1982

6 Xia H, "Normal expression of insect-resistant transgene in progeny of common wild rice crossed with genetically modiWed rice: its implication in ecological biosafety assessment" 119 : 635-644, 2009

7 UNESCO, "Managing Water under Uncertainty and Risk" 1 : 22-69, 2012

8 Ju Sung Im, "Growth, quality, and yield characteristics of transgenic potato (Solanum tuberosum L.) overexpressing StMyb1R-1 under water deficit" 한국식물생명공학회 39 (39): 154-162, 2012

9 Jang IC, "Expression of a bifunctional fusion of the Escherichia coli genes for trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase in transgenic rice plants increases trehalose accumulation and abiotic stress tolerance without stunting growth" AMER SOC PLANT BIOLOGISTS 131 : 516-524, 2003

10 Kim HJ, "Expression of PAT and NPT II Proteins during the Developmental Stages of a Genetically Modified Pepper Developed in Korea" AMER CHEMICAL SOC 58 (58): 10906-10910, 2010

11 FAO, "Economic valuation of water resources in agriculture"

12 Cruz de Carvalho MH, "Drought stress and reactive oxygen species: Production, scavenging and signaling" 3 : 156-165, 2008

13 백광현, "CaMsrB2, Pepper Methionine Sulfoxide Reductase B2, Is a Novel Defense Regulator against Oxidative Stress and Pathogen Attack" AMER SOC PLANT BIOLOGISTS 154(1) : 2010

14 Cho SK, "Arabidopsis PUB22 and PUB23 are homologous U-box E3 ubiquitin ligases that play combinatory roles in response to drought stress" 20 (20): 1899-1914, 2008

15 Ha SH, "Application of two bicistronic systems involving 2A and IRES sequences to the biosynthesis of carotenoids in rice endosperm" WILEY-BLACKWELL PUBLISHING 8 (8): 928-938, 2010