Temporal and spatial changes in Bt toxin expression in Bt-transgenic poplar and insect resistance in field tests

Published Date
December 2016, Volume 27, Issue 6, pp 1249–1256

Original Paper

First Online: 

06 May 2016

DOI: 10.1007/s11676-016-0254-x

Cite this article as: 

Zhang, Y., Zhang, J., Lan, J. et al. J. For. Res. (2016) 27: 1249. doi:10.1007/s11676-016-0254-x

Author

• Yiwen Zhang
• Jun Zhang
• Jinping Lan
• Jinmao Wang
• Junxia Liu
• Minsheng YangEmail

Abstract

Extensive planting of Bacillus thuringiensis (Bt)-transgenic plants economically benefits society; however, the potential risk they pose is receiving increasing attention. This study used enzyme-linked immunosorbent assay and fluorescence quantitative PCR (RT-PCR) to monitor the temporal and spatial dynamics of the expression of Bt toxic protein in a forest of 6- to 8-year-old trees of transgenic insect-resistant poplar 741 for three consecutive years. The enrichment, distribution, and degradation of Bt toxic protein and the influence of transgenic poplars on the targeted insect population, Hyphantria cunea, were investigated. The content of Cry1Ac toxic protein dynamically changed in transgenic poplar. During the annual growth cycle, the content initially increased, then decreased in the long and the short branches of the crown and in the root system, peaking in August. During the study, the protein did not accumulate overtime. The mRNA transcription of gene Cry1Ac was almost consistent with the level of the protein, but transcription peaked in July. In the transgenic and control forestland, microscale levels of the Cry1Ac toxic protein were detected from the soil, but increased accumulation was not observed with the planting year of transgenic poplar. Meanwhile, Bt was isolated and detected molecularly from the soil in the experimental forestland. A systematic investigation of the density of H. cunea in the experimental transgenic poplar forest indicated that transgenic Pb29 poplar could resist insects to a certain degree. At peak occurrence of the targeted insects, the density of H. cuneain the experimental forest was significantly lower than in the nontransgenic poplar forest.

References 

1. An XL, Dong JG, Han JM (2001) RAPD analysis of Exserohilum turcicumI. CTAB method of isolating DNA from Exserohilum turcicum for RAPD. J Agric Univ Hebei 24(1):38–41 (in chinese)Google Scholar
2. Axelsson EP, Hjältén J, LeRoy CJ, Julkunen-Tiitto R, WennstrÖm A, Pilate G (2010) Can leaf litter from genetically modified trees affect aquatic ecosystems? Ecosystems 13(7):1049–1059CrossRefGoogle Scholar
3. Axelsson EP, Hjältén J, LeRoy CJ (2012) Performance of insect-resistant Bacillus thuringiensis (Bt)-expressing aspens under semi-natural field conditions including natural herbivory in Sweden. For Ecol Manag 264:167–171CrossRefGoogle Scholar
4. Danielsen L, Thurmer A, Meinicke P, Buee M, Morin E, Martin F, Pilate G, Daniel R, Polle A, Reich M (2012) Fungal soil communities in a young transgenic poplar plantation form a rich reservoir for fungal root communities. Ecol Evol 2(8):1935–1948CrossRefPubMedPubMedCentralGoogle Scholar
5. Dively GP (2005) Impact of transgenic VIP3A × Cry1Ab lepidopteran-resistant field corn on the nontarget arthropod community. Environ Entomol 34:1267–1291CrossRefGoogle Scholar
6. Fladung M, Ewald D (2006) Tree transgenesis: recent developments. Springer, Berlin-HeidelbergCrossRefGoogle Scholar
7. Gao BJ, Wang YF, Wang JM, Liang HY, Yang MS, Zhang JH (2004) Researches on the insect-resistant effect of transgenic hybrid poplar 741 with two insect-resistant genes. Acta Ecol Sin 02:297–301 (in chinese)Google Scholar
8. Génissel A, Leplé JC, Millet N, Augustin S, Jouanin L, Pilate G (2003) High tolerance against Chrysomela tremulae of transgenic poplar plants expressing a synthetic cry3Aa gene from Bacillus thuringiensis ssp tenebrionis. Mol Breed 11(2):103–110CrossRefGoogle Scholar
9. Head G, Surber JB, Watson JA, Martin JW, Duan JJ (2002) No detection of Cry1Ac protein in soil after multiple years of transgenic Bt cotton (Bollgard) use. Environ Entomol 31(1):30–36CrossRefGoogle Scholar
10. Head G, Moar W, Eubanks M, Freeman B, Ruberson J, Hagerty A, Turnipseed S (2005) A multiyear, large-scale comparison of arthropod populations on commercially managed Bt and non-Bt cotton fields. Environ Entomol 34(5):1257–1266Google Scholar
11. Hiwasa-Tanase K, Nyarubona M, Hirai T, Kato K, Ichikawa T, Ezura H (2011) High-level accumulation of recombinant miraculin protein in transgenic tomatoes expressing a synthetic miraculin gene with optimized codon usage terminated by the native miraculin terminator. Plant Cell Rep 30(1):113–124CrossRefPubMedGoogle Scholar
12. Hjältén J, Axelsson EP, Whitham TG, LeRoy CJ, Julkunen-Tiitto R, Wennstoem A, Pilate G (2012) Increased resistance of Bt aspens to Phratora vitellinae (Coleoptera) leads to increased plant growth under experimental conditions. PLoS One 7(1):e30640CrossRefPubMedPubMedCentralGoogle Scholar
13. Hou YJ, Su XH, Jiao RZ, Huang QJ, Chu YF (2009) Effects of transgenic Populus alba × P. glandulosa on soil microorganism. Sci Silvae Sin 45(5):148–152 (in chinese)Google Scholar
14. Hu JJ, Tian YC, Han YF, Li L, Zhang BE (2001) Field evaluation of insect-resistant transgenic Populus nigra trees. Euphytica 121(2):123–127CrossRefGoogle Scholar
15. Hu JJ, Yang MS, Lu MZ (2010) Advances in biosafety studies on transgenic insect-resistant poplars in China. Biodivers Sci 18(4):336–345CrossRefGoogle Scholar
16. Li GW, Xie XS (2011) Central dogma at the single-molecule level in living cells. Nature 475(7356):308–315CrossRefPubMedPubMedCentralGoogle Scholar
17. Liu Z, Ye G, Hu C (2004) Effects of Bacillus thuringiensis transgenic rice and chemical insecticides on arthropod communities in paddy-fields. Chin J Appl Ecol 15(12):2309–2314Google Scholar
18. Lu MZ (2008) Advance in transgenic research and its biosafety of forest trees in China. In: Proceeding of the 8th symposium in agricultural biochemistry and molecular biology of the Chinese Society of Biochemistry and Molecular Biology, September 2008, Guiyang, China, pp 1–2
19. Perlak FJ, Fuchs RL, Dean DA, McPherson SL, Fischhoff DA (1991) Modification of the coding sequence enhances plant expression of insect control protein genes. Proc Natl Acad Sci USA 88(8):3324–3328CrossRefPubMedPubMedCentralGoogle Scholar
20. Tian YC, Zheng JB, LI CQ (2000) Studies of transgenic hybrid poplar 741 carring tow insect-resistant genes. Acta Bot Sin 42(3):263–268Google Scholar
21. Wang LR, Yang MS, Akinnagbe A, Liang HY, Wang JM, Ewald D (2012) Bacillus thuringiensis protein transfer between rootstock and scion of grafted poplar. Plant Biol 14:745–750CrossRefPubMedGoogle Scholar
22. Yang MS, Lang HY, Gao BJ, Wang JM, Zheng JB (2003) Insecticidal activity and transgene expression stability of transgenic hybrid poplar clone 741 carrying two insect-resistant genes. Silvae Genet 52(5–6):197–201Google Scholar
23. Yang MS, Gao BJ, Wang YF, Wang JM, Du GJ (2005) Analysis of main characteristic of white poplar hybrid 741 transformed two insect-resistant genes. Sci Silvae Sin 01(1):91–97 (in chinese)Google Scholar
24. Zhang HC, Liu ZS (2006) Agricultural statistics of China 2006. China Agricultural Press, Beijing, pp 230–257Google Scholar
25. Zhang BY, Chen M, Zhang XF, Luan HH, Diao S, Tian YH, Su XH (2011a) Laboratory and field evaluation of the transgenic Populus alba × Populus glandulosa expressing double coleopteran-resistance genes. Tree Physiol 31(5):567–573CrossRefPubMedGoogle Scholar
26. Zhang BY, Chen M, Zhang XF, Luan HH, Tian YH, Su XH (2011b) Expression of Bt-Cry3A in transgenic Populus alba × P. glandulosa and its effects on target and non-target pests and the arthropod community. Transgenic Res 20(3):523–532CrossRefPubMedGoogle Scholar
27. Zhen ZX, Wang JM, Yang MS (2011) Effects of transgenic insect-resistance hybrid poplar 741 groves on soil microorganisms. J Agric Univ HeBei 34(1):78–81 (in chinese)Google Scholar
28. Zheng JB, Liang HY, Gao BJ, Wang YF, Tian YC (2000) Selection and insect resistance of transgenic hybrid poplar 741 carrying two insect-resistant genes. Sci Silvae Sin 36(2):13–19Google Scholar
29. Zwahlen C, Hilbeck A, Gugerli P, Nentwig W (2003) Degradation of the Cry1Ab protein within transgenic Bacillus thuringiensis corn tissue in the field. Mol Ecol 12(3):765–775CrossRefPubMedGoogle Scholar

For further details log on website :
http://link.springer.com/article/10.1007/s11676-016-0254-x