Journal of the Korean Society of International Agriculture (한국국제농업개발학회지)

The Korean Society of International Agriculture (한국국제농업개발학회)
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Molecular Biological Analysis of Carotenoid-Biofortified Rice and Its Effect on Daphnia magna Feeding

비타민A 강화 벼 분자생물학적 분석 및 물벼룩 급이 효과

  • Oh, Sung-Dug (National Academy of Agricultural Science, Rural Development Administration) ;
  • Lee, Kijong (National Academy of Agricultural Science, Rural Development Administration) ;
  • Park, Soo-Yun (National Academy of Agricultural Science, Rural Development Administration) ;
  • Sohn, Soo-In (National Academy of Agricultural Science, Rural Development Administration) ;
  • Ryu, Tae-Hun (National Academy of Agricultural Science, Rural Development Administration) ;
  • Kim, Jae-Kwang (National Academy of Agricultural Science, Rural Development Administration) ;
  • Kim, Jinseo (Division of Plant Biosciences, School of Plant Biosciences, Kyungpook National University) ;
  • Ahn, Hong-Il (National Academy of Agricultural Science, Rural Development Administration) ;
  • Ha, Sun-Hwa (National Academy of Agricultural Science, Rural Development Administration) ;
  • Park, Jong-Sug (National Academy of Agricultural Science, Rural Development Administration) ;
  • Ahn, Byung-Ohg (National Academy of Agricultural Science, Rural Development Administration) ;
  • Cho, Hyun Suk (National Academy of Agricultural Science, Rural Development Administration) ;
  • Suh, Sang Jae (National Academy of Agricultural Science, Rural Development Administration)
  • 오성덕 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 이기종 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 박수윤 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 손수인 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 류태훈 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 김재광 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 김진서 (경북대학교 농업생명과학대학 응용생명과학부) ;
  • 안홍일 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 하선화 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 박종석 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 안병옥 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 조현석 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 서상재 (농촌진흥청 국립농업과학원 농업생명자원부)
  • Published : 2012.12.31
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Abstract

A carotenoid-biofortified (PAC) rice was generated by inserting phytoene synthase (Psy) and carotene desaturase (CrtI) genes isolated from Capsicum annuum cv. Nockwang and Pantoea ananatis into the genome of a conventional variety of rice (Nakdongbyeo). For biosafety assessment, we evaluated the effects on survival of Daphnia magna which is a commonly used as a model organism in ecotoxicological studies. D. magna fed on PAC rice and non-GM rice (Nakdong) grown in the same environment (100% ground rice suspension). The PAC rice was confirmed to have the insertion of T-DNA and protein expression by the Southern blot and HPLC analysis. Feeding study showed similar cumulative immobility and abnormal response of Daphnia magna between PAC rice and non-GM counterparts. $48hr-EC_{50}$ values of PAC rice and non-GM rice showed 3,311 mg/L (95% confidence limits: 2,901.39 - 3,779.23 mg/L) and 3,655 mg/L (95% confidence limits: 3,156.71 - 4,232.86 mg/L) respectively, but there were not statistically significant.

물벼룩 급성 독성 평가를 위한 비타민A 강화벼의 분자생물학적 특성을 분석한 결과, Southern blot에서 베타-카로틴 생합성을 위한 Psy와 CrtI 유전자들이 one-copy로 도입됨을 확인하였으며, 선발마커인 Bar 유전자의 단백질 검출 immunostrip 분석에서도 비타민A 강화벼에서만 검출되었다. 비타민A 강화 벼의 목적하는 최종 산물인 베타-카로틴 함량도 낙동벼에 비해 8.9배 증가됨을 확인하였다. 비타민A 강화벼와 낙동벼의 농업환경 생물지표종인 물벼룩(Daphniamagna)에 대한 급성독성시험을 실시한 결과, 비타민A 강화벼의 48시간-EC50은 3,311.40 mg/L(95% 신뢰한계 : 2,901.39 ~ 3,779.23 mg/L), 무영 향농도(NOEC)는 1,800 mg/L였고, 낙동벼는 48시간-$EC_{50}$은 3,655.23 mg/L(95% 신뢰한계 : 3,156.71 ~ 4,232.86 mg/L), 무영 향농도는 1,800 mg/L였다. 따라서 Psy와 CrtI 유전자가 형질 전환된 비타민A 강화벼 및 낙동벼가 환경 지표생물종인 물벼룩에 미치는 영향 평가 결과 상대적 동등성을 보였으며, 이는 Psy와 CrtI 유전자의 단백질 노출이 물벼룩에 부정적인 영향을 미치지 않은 것으로 판단된다.

Keywords

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