Korean Journal of Agricultural Science (농업과학연구)

Institute of Agricultural Science, CNU (충남대학교 농업과학연구소)
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Molecular biological analysis of Bt-transgenic (Bt-9) rice and its effect on Daphnia magna feeding

  • Oh, Sung-Dug (Department of Agricultural Biotechnology, National Institute of Agricultural Sciences, Rural Development Administration) ;
  • Yun, Doh-Won (Department of Agricultural Biotechnology, National Institute of Agricultural Sciences, Rural Development Administration) ;
  • Chang, Ancheol (Department of Agricultural Biotechnology, National Institute of Agricultural Sciences, Rural Development Administration) ;
  • Lee, Yu-jin (Korea Testing & Research Institute) ;
  • Lim, Myung-Ho (Department of Agricultural Biotechnology, National Institute of Agricultural Sciences, Rural Development Administration)
  • Received : 2018.09.18
  • Accepted : 2019.01.02
  • Published : 2019.03.01
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Abstract

Insect-resistant transgenic (Bt-9) rice was generated by inserting mCry1Ac1, a modified gene from the soil bacterium Bacillus thuringiensis, into the genome of a conventional variety of rice (Ilmi). With regard to potential problems such as safety, an evaluation of non-target organisms is necessary as an essential element of an environmental risk assessment of genetically modified (GM) crops. We studied the effects of the Bt-9 rice on the survival of cantor Daphnia magna, a commonly used model organism in ecotoxicological studies. D. magna fed on the Bt-transgenic rice (Bt-9) and its near non-GM counterparts (Ilmi) grown in the same environment (a 100% ground rice suspension). The Bt-9 rice was confirmed to have the inserted T-DNA and protein expression evident by the PCR and ELISA analyses. The feeding study showed a similar cumulative immobility and abnormal response of the Daphnia magna between the Bt-9 rice and Ilmi. Additionally, the 48 h-EC50 values of the Bt-9 and Ilmi rice were 4,400 mg/L (95% confidence limits: 3861.01 - 5015.01 mg/L) and 5,564 mg/L (95% confidence limits: 4780.03 - 6476.93 mg/L), respectively. The rice NOEC (No observed effect concentration) value for D. magna was suggested to be 1,620 mg/L. We conclude that the tested Bt-9 and Ilmi have a similar cumulative immobility for D. magna, a widely used model organism, and the growth of Bt-9 did not affect non-target insects.

Keywords

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Fig. 1. Confirmation of the T-DNA genes on the Bt-transgenic (Bt-9) rice and non-genetically modified rice (Ilmi). M, 100bp DNA ladder; N, non-GM rice (Ilmi); Bt-9, Bt-transgenic rice T6; Bar, Phosphinothricin acetyltransferase; SPS, Sucrose-phosphate synthase.

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Fig. 2. Confirmation of genes expression for Bt-transgenic (Bt-9) rice by using immunostrip. (a) immunostrip tests for the PAT (phosphinothricin acetyltransferase) detection, (b) immunostrip tests for the CryIAc1 detection. N, non-genetically modified rice (Ilmi); Bt-9, Bt-transgenic rice T6.

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Fig. 3. mCryIAc1 protein levels (μg/g fresh weight) in Ilmi and Bt-transgenic (Bt-9) rice. Values are the average ± SD of triplicate measures.

Table 1. Primers list used for PCR (Polymerase chain reaction) analysis.

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Table 2. Changes of water temperature (℃) during cumulative immobility tests of Daphnia magna in non-Genetically modified rice (Ilmi) and Bt-transgenic (Bt-9) rice.

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Table 3. Changes of pH during cumulative immobility tests of Daphnia magna in non-genetically modifed rice (Ilmi) and Bt-transgenic (Bt-9) rice.

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Table 4. Cumulative immobility of Daphnia magna in non-genetically modifed rice (Ilmi) and Bt-transgenic (Bt-9) rice.

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Table 5. EC50 (effective concentration 50) values of Daphnia magna after 24 and 48 hours in non-genetically modified rice (Ilmi) and Bt-transgenic (Bt-9) rice.

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