, Volume 23, Issue 9, pp 1619–1628 | Cite as

Influence of transgenic rice expressing a fused Cry1Ab/1Ac protein on frogs in paddy fields

  • Jia-Mei Wang
  • Xiu-Ping Chen
  • Yu-Yong Liang
  • Hao-Jun Zhu
  • Jia-Tong Ding
  • Yu-Fa Peng


As genetic engineering in plants is increasingly used to control agricultural pests, it is important to determine whether such transgenic plants adversely affect non-target organisms within and around cultivated fields. The cry1Ab/1Ac fusion gene from Bacillus thuringiensis (Bt) has insecticidal activity and has been introduced into rice line Minghui 63 (MH63). We evaluated the effect of transgenic cry1Ab/1Ac rice (Huahui 1, HH1) on paddy frogs by comparing HH1 and MH63 rice paddies with and without pesticide treatment. The density of tadpoles in rice fields was surveyed at regular intervals, and Cry1Ab/1Ac protein levels were determined in tissues of tadpoles and froglets collected from the paddy fields. In addition, Rana nigromaculata froglets were raised in purse nets placed within these experimental plots. The survival, body weight, feeding habits, and histological characteristics of the digestive tract of these froglets were analyzed. We found that the tadpole density was significantly decreased immediately after pesticide application, and the weight of R. nigromaculata froglets of pesticide groups was significantly reduced compared with no pesticide treatment, but we found no differences between Bt and non-Bt rice groups. Moreover, no Cry1Ab/1Ac protein was detected in tissue samples collected from 192 tadpoles and froglets representing all four experimental groups. In addition, R. nigromaculata froglets raised in purse seines fed primarily on stem borer and non-target insects, and showed no obvious abnormality in the microstructure of their digestive tracts. Based on these results, we conclude that cultivation of transgenic cry1Ab/1Ac rice does not adversely affect paddy frogs.


Transgenic rice Bt protein Frog Safety assessment Non-target effect 



We thank Professor Yongjun Lin (Huazhong Agricultural University, Wuhan, China) for kindly providing transgenic rice seeds. We also thank Professor Fajun Chen (Nanjing Agricultural University, Nanjing, China) for his constructive comments on the early draft of this manuscript. This work was supported by the National GMO New Variety Breeding Program of the PRC (2012ZX08011-002 and 2014ZX08011-001).

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Akhtar ZR, Tian JC, Chen Y, Fang Q, Hu C, Chen M, Peng YF, Ye GY (2010) Impacts of six bt rice lines on nontarget rice feeding thrips under laboratory and field conditions. Environ Entomol 39(2):715–726CrossRefGoogle Scholar
  2. Betz FS, Hammond BG, Fuchs RL (2000) Safety and advantages of Bacillus thuringiensis-protected plants to control insect pests. Regul Toxicol Pharmacol 32:156–173CrossRefGoogle Scholar
  3. Blaustein AR, Han BA, Relyea RA, Johnson PTJ, Buck JC, Gervasi SS, Kats LB (2011) The complexity of amphibian population declines: understanding the role of cofactors in driving amphibian losses. Ann N Y Acad Sci 1223:108–119CrossRefGoogle Scholar
  4. Bøhn T, Primicerio R, Hessen DO, Traavik T (2008) Reduced fitness of Daphnia magna fed a Bt-transgenic maize variety. Arch Environ Contam Toxicol 55:584–592CrossRefGoogle Scholar
  5. Bøhn T, Traavik T, Primicerio R (2010) Demographic responses of Daphnia magna fed transgenic Bt-maize. Ecotoxicology 19:419–430CrossRefGoogle Scholar
  6. Brookes G, Barfoot P (2013) Key environmental impacts of global genetically modified (GM) crop use 1996–2011. GM Crops Food 4(2):109–119CrossRefGoogle Scholar
  7. Carstens K, Anderson J, Bachman P, Schrijver AD, Dively G, Federici B, Hamer M, Gielkens M, Jensen P, Lamp W, Rauschen S, Ridley G, Romeis J, Waggoner A (2012) Genetically modified crops and aquatic ecosystems: considerations for environmental risk assessment and nontarget organism testing. Transgenic Res 21:813–842CrossRefGoogle Scholar
  8. Chen YM (2011) The preliminary analysis of sexual dimorphism and feeding habits of the main anurans in Zhongxiang and Tianmen of Jianghan plain. Dissertation, Central China Normal University (In Chinese with English abstract)Google Scholar
  9. Chen M, Shelton A, Ye GY (2011) Insect-resistant genetically modified rice in China: from research to commercialization. Annu Rev Entomol 56:81–101CrossRefGoogle Scholar
  10. García M, Ortego F, Castañera P, Farinós GP (2010) Effects of exposure to the toxin Cry1Ab through Bt maize fed-prey on the performance and digestive physiology of the predatory rove beetle Atheta coriaria. Biol Control 55:225–233CrossRefGoogle Scholar
  11. Han LZ, Wu KM, Peng YF, Wang F, Guo YY (2006) Evaluation of transgenic rice expressing Cry1Ac and CpTI against Chilo suppressalis and intrapopulation variation in susceptibility to Cry1Ac. Environ Entomol 35:1453–1459CrossRefGoogle Scholar
  12. Han LZ, Wu KM, Peng YF, Wang F, Guo YY (2007) Efficacy of transgenic rice expressing Cry1Ac and CpTI against Cnaphalocrocis medinalis. J Invertebr Pathol 96:71–79CrossRefGoogle Scholar
  13. Han LZ, Hou ML, Wu KM, Peng YF, Wang F (2011) Lethal and sub-lethal effects of transgenic rice containing cry1Ac and CpTI genes on the pink stem borer, Sesamia inferens (walker). J Integr Agric 10(3):384–393Google Scholar
  14. Hayes TB, Case P, Chui S, Chung D, Haeffele C, Haston K, Lee M, Mai VP, Marjuoa Y, Parker J, Tsui M (2006) Pesticide mixtures, endocrine disruption, and amphibian declines: are we underestimating the impact? Environ Health Perspect 114:40–50CrossRefGoogle Scholar
  15. High SM, Cohen MB, Shu QY, Altosaar I (2004) Achieving successful deployment of Bt rice. Trends Plant Sci 9(6):287–289Google Scholar
  16. Hirai T (2002) Ontogenetic change in the diet of the pond frog Rana nigromaculata. Ecol Res 17(6):639–644CrossRefGoogle Scholar
  17. Hofmann C, Vanderbruggen H, Hofte H, Van RJ, Jansens S, Van Mellaert H (1988) Specificity of Bacillus thuringiensis delta-endotoxins is correlated with the presence of high-affinity binding sites in the brush border membrane of target insect midgets. Proc Natl Acad Sci U S A 85:7844–7848CrossRefGoogle Scholar
  18. James C (2013) Global status of commercialized biotech/GM crops: 2013. ISAAA Brief 46. ISAAA, IthacaGoogle Scholar
  19. Jensen PD, Dively GP, Swan CM, Lamp WO (2010) Exposure and nontarget effects of transgenic Bt corn debris in streams. Environ Entomol 39:707–714CrossRefGoogle Scholar
  20. Jia HP (2010) Chinese green light for GM rice and maize prompts outcry. Nat Biotechnol 28(5):390–391CrossRefGoogle Scholar
  21. Jones DK, Hammond JI, Relyea RA (2009) Very highly toxic effects of endosulfan across nine speices of tadpoles: lag effects and family-level sensitivity. Environ Toxicol Chem 28(9):1939–1945CrossRefGoogle Scholar
  22. Li YH, Romeis J (2010) Bt maize expressing Cry3Bb1 does not harm the spider mite, Tetranychus urticae, or its ladybird beetle predator, Stethorus punctillum. Biol Control 53:337–344CrossRefGoogle Scholar
  23. Li DP, Hu GZ, Liu NJ, Zou Y, Tang XC, Li XY (2010) Census methods for counting tadpoles of anura species in static water. Chin J Zool 45(5):72–78 (In Chinese with English abstract.)Google Scholar
  24. Li YH, Peng YF, Hallerman EM, Wu KM (2014) Biosafety management and commercial use of genetically modified crops in China. Plant Cell Rep. doi: 10.1007/s00299-014-1567-x Google Scholar
  25. Liu W, Lu HH, Wu WX, Wei QK, Chen YX, Thies JC (2008) Transgenic Bt rice does not affect enzyme activities and microbial composition in the rhizosphere during crop development. Soil Biol Biochem 40:475–486CrossRefGoogle Scholar
  26. Lu C (2010) The first approved transgenic rice in China. GM crops 1(3):113–115CrossRefGoogle Scholar
  27. Lu YH, Wu KM, Jiang YY, Xia B, Li P, Feng HQ, Wyckhuys KAG, Guo YY (2010) Mirid bug outbreaks in multiple crops correlated with wide-scale adoption of Bt cotton in China. Science 328:1151–1154CrossRefGoogle Scholar
  28. Lu ZB, Tian JC, Wang W, Xu HX, Hu C, Guo YY, Peng YF, Ye GY (2014) Impacts of Bt rice expressing Cry1C or Cry2A protein on the performance of nontarget leafhopper, Nephotettix cincticeps (Hemiptera: Cicadellidae), under laboratory and field conditions. Environ Entomol 43(1):209–217CrossRefGoogle Scholar
  29. Mannakkara A, Niu L, Ma W, Lei C (2013) Zero effect of Bt rice on expression of genes coding for digestion, detoxification and immune responses and developmental performances of Brown Planthopper Nilaparvata lugens (Stål). J Insect Physiol 59(10):985–993CrossRefGoogle Scholar
  30. National Research Council (1996) NIH Guide for the Care and Use of Laboratory Animals. National Academy Press, Washington, DCGoogle Scholar
  31. Qaim M (2009) The economics of genetically modified crops. Annu Rev Resour Econ 1:665–693CrossRefGoogle Scholar
  32. Raybould A, Vlachos D (2011) Non-target organism effects tests on Vip3A and their application to the ecological risk assessment for cultivation of MIR162 maize. Transgenic Res 20:599–611CrossRefGoogle Scholar
  33. Ricroch A, Bergé JB, Kuntz M (2010) Is the German suspension of MON810 maize cultivation scientifically justified? Transgenic Res 19:1–12CrossRefGoogle Scholar
  34. Romeis J, Hellmich RL, Candolfi MP, Carstens K, Schrijver AD, Gatehouse AMR, Herman RA, Huesing JE, McLean MA, Raybould A, Shelton AM, Waggoner A (2011) Recommendations for the design of laboratory studies on non-target arthropods for risk assessment of genetically engineered plants. Transgenic Res 20:1–22CrossRefGoogle Scholar
  35. Romeis J, McLean MA, Shelton AM (2013) When bad science makes good headlines: Bt maize and regulatory bans. Nat Biotechnol 31:386–387CrossRefGoogle Scholar
  36. Rosi-Marshall EJ, Tank JL, Royer TV, Whiles MR, Evans-White M, Chambers C, Griffiths NA, Pokelsek J, Stephen ML (2007) Toxins in transgenic crop byproducts may affect headwater stream ecosystems. Proc Natl Acad Sci USA 104:16204–16208CrossRefGoogle Scholar
  37. Sparling DW, Fellers G (2007) Comparative toxicity of chlorpyrifos, diazinon, malathion and their oxon derivatives to larval Rana boylii. Environ Pollut 147:535–539CrossRefGoogle Scholar
  38. Tam Q, Avenant-Oldewage A (2009) The effect of starvation on the ultrastructure of the digestive cells of Dolops ranarum (Stuhlmann, 1891) (Crustacea: Branchiura). Arthropod Struct Dev 38(5):391–399CrossRefGoogle Scholar
  39. Tank JL, Rosi-Marshall EJ, Royer TV, Whiles MR, Griffiths NA, Frauendorf TC, Treering DJ (2010) Occurrence of maize detritus and a transgenic insecticidal protein (Cry1Ab) within the stream network of an agricultural landscape. Proc Natl Acad Sci USA 107:17645–17650CrossRefGoogle Scholar
  40. Tian JC, Liu C, Chen M, Chen Y, Chen XX, Peng YF, Hu C, Ye GY (2010) Laboratory and field assessments of prey-mediated effects of transgenic Bt rice on Ummeliata insecticeps (Araneida: Linyphiidae). Environ Entomol 39(4):1369–1377CrossRefGoogle Scholar
  41. Tu J, Zhang G, Datta K, Xu C, He Y, Zhang Q, Khush GS, Datta SK (2000) Field performance of transgenic elite commercial hybrid rice expressing Bacillus thuringiensis δ-endotoxin. Nat Biotechnol 18:1101–1104CrossRefGoogle Scholar
  42. Viktorov AG (2011) Transfer of Bt corn byproducts from terrestrial to stream ecosystems. Russ J Plant Physiol 58:543–548CrossRefGoogle Scholar
  43. Wang YM, Zhang GA, Du JP, Liu B, Wang MC (2010) Influence of transgenic hybrid rice expressing a fused gene derived from cry1Ab and cry1Ac on primary insect pests and rice yield. Crop Prot 29:128–133CrossRefGoogle Scholar
  44. Wang YY, Li YH, Romeis J, Chen XP, Zhang J, Chen HY, Peng YF (2012) Consumption of Bt rice pollen expressing Cry2Aa does not cause adverse effects on adult Chrysoperla sinica Tjeder (Neuroptera: Chrysopidae). Biol Control 61(3):246–251CrossRefGoogle Scholar
  45. Wang YM, Hu HW, Huang JC, Li JH, Liu B, Zhang GA (2013) Determination of the movement and persistence of Cry1Ab/1Ac protein released from Bt transgenic rice under field and hydroponic conditions. Soil Biol Biochem 58:107–114CrossRefGoogle Scholar
  46. Wu WX, Ye QF, Min H (2004) Effect of straws from Bt-transgenic rice on selected biological activities in water-flooded soil. Eur J Soil Biol 40:15–22CrossRefGoogle Scholar
  47. Yao HW, Jiang CY, Ye GY, Hu C, Peng YF (2008) Toxicological assessment of pollen from different Bt rice lines on Bombyx mori (Lepidoptera: Bombyxidae). Environ Entomol 37(3):825–837CrossRefGoogle Scholar
  48. Zeng Zeng, Jiang LQ, Li FJ, Li XM, Cao ZD, Zhang YG (2012) The effects of starvation on digestive tract function and structure in juvenile southern catfish (Silurus meridionalis Chen). Comp Biochem Physiol A 162(3):200–211CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijingChina
  2. 2.College of Animal Science and TechnologyYangzhou UniversityYangzhouChina
  3. 3.Institute of Plant ProtectionJiangxi Academy of Agricultural SciencesNanchangChina

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