Inheritance of Transgenes in Transgenic Bt Lines Resistance to Helicoerpa armigera in Upland Cotton

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 958)

Abstract

Six transgenic Bt cotton cultivars (lines) including GKsu12, GK19, MR1, GK5, 109B, and SGK1 are highly resistant to bollworm from the seedling to boll-setting stages in bioassays with detached cotton leaves, though there are differences in resistant level and Bt toxin content in these transgenic cottons. Genetics analysis reveals that the resistance to Helicoverpa armigera in these six transgenic Bt cotton cultivars (lines) are controlled by one pair of dominant genes. Allelic tests further demonstrate some populations are in Mendel segregation for two nonallelic genes, i.e., the inserted Bt gene in GKsu12 is nonallelic to that of SGK1, GK5, 109B, and GK19 and Bt genes in GK19 and SGK1 are likely inserted in the same or in close proximity (genetically closely linked), while some F2 produce abnormal segregation patterns, with a segregation of resistance to Helicoerpa armigera vary between 15:1 and 3:1, though their Bt segregation fit into 15:1 by PCR analysis, suggesting Bt gene silence in these populations. Two genes silence may occur in these populations due to the homologous sequence by crossing since the silenced individuals accounted for 1/16 of the F2 populations for allelic test. To those silenced populations, one of their parents all showed high resistance to bollworm.

References

  1. 1.
    Hofte H, Whiteley HR (1989) Insecticidal crystal proteins of Bacillus thuringiensis. Microbiol Rev 53(242):255Google Scholar
  2. 2.
    Benedict JH, Sachs ES, Altman DW, Ring DR, Stone TB, Sims SR (1993) Impact of endoxin-producing transgenic cotton on insect-plant interaction with Heliothis virescens and Helicoverpa zea (Lepidoptera: Noctuidae). Environ Entomol 22(1):9Google Scholar
  3. 3.
    James C (2010) Global status of commercialized biotech/GM crops: 2010. ISAAA brief no. 42. ISAAA, Ithaca, NYGoogle Scholar
  4. 4.
    Sun J, Tang CM, Zhu XF, Guo WZ, Zhang TZ, Zhou WJ, Meng FX, Sheng JL (2002) Characterization of resistance to Helicoverpa armigera in three lines of transgenic Bt upland cotton. Euphytica 123(343):351Google Scholar
  5. 5.
    Wu K, Lu Y, Feng H, Jiang Y, Zhao J (2008) Suppression of cotton bollworm in multiple crops in China in areas with Bt toxin-containing cotton. Science 321:1676–1678PubMedCrossRefGoogle Scholar
  6. 6.
    Dong H, Li W (2007) Variability of endotoxin expression in Bt transgenic cotton. J Agron Crop Sci 193:21–29CrossRefGoogle Scholar
  7. 7.
    Tabashnik BE, Sisterson MS, Ellsworth PC, Dennehy TJ, Antilla L, Liesner L, Whitlow M, Staten RT, Fabrick JA, Unnithan GC (2010) Suppressing resistance to Bt cotton with sterile insect releases. Nat Biotechnol 28:1304–1307PubMedCrossRefGoogle Scholar
  8. 8.
    Register JC, Peterson DJ, Bell PJ, Bullock WP, Evans IJ, Frame B, Greenland AJ, Higgs NS, Jepson I, Jiao S, Lewnau CJ, Sillick JM, Wilson HM (1994) Structure and function of selectable and non-selectable transgenes in maize after introduction by particle bombardment. Plant Mol Biol 25(951):961Google Scholar
  9. 9.
    Cheng M, Fry JE, Pang S, Zhou H, Hironaka CM, Duncan DR, Conner TW, Wan Y (1997) Genetic transformation of wheat mediated by Agrobacterium tumfaciens. Plant Physiol 115(971):980Google Scholar
  10. 10.
    Spencer TM, O’Brien JV, Start WG, Adams TR, Gordon-Kamm WJ, Lemaux PG (1992) Segregation of transgenes in maize. Plant Mol Biol 18(201):210Google Scholar
  11. 11.
    Pawlowski WP, Somers DA (1996) Transgene inheritance in plants genetically engineered by microprojectile bombardment. Mol Biotechnol 6(17):30Google Scholar
  12. 12.
    Pawlowski WP, Torbert KA, Rines HW, Somers DA (1998) Irregular patterns of transgene silencing in allohexaploid oat. Plant Mol Biol 38:597–607PubMedCrossRefGoogle Scholar
  13. 13.
    Morino K, Olsen OA, Shimamoto K (1999) Silencing of an aleurone-specific gene in transgenic rice is caused by a rearranged transgene. Plant J 17(275):85Google Scholar
  14. 14.
    Tang CM, Sun J, Zhu XF, Guo WZ, Zhang TZ, Shen JL, Gao CF, Zhou WJ, Chen ZX, Guo SD (2000) Inheritance of resistance to Helicoverpa armigera of 3 kinds of transgenic Bt strains available in upland cotton in China. Chinese Sci Bull 45(363):367Google Scholar
  15. 15.
    Bao-hong Z, Teng-long G, Qing-lian W (2000) Inheritance and segregation of exogenous genes in transgenic cotton. J Genet 79:71–75CrossRefGoogle Scholar
  16. 16.
    Sachs ES, Bendict JH, Stelly DM, Taylor JF, Altman DW, Berbrich SA, Devis SK (1998) Expression and segregation of gene encoding Bt insecticidal proteins in cotton. Crop Sci 38:1–11CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  1. 1.National Key Laboratory of Crop Genetics and Germplasm Enhancement, Cotton Research InstituteNanjing Agricultural UniversityNanjingPeople’s Republic of China
  2. 2.National Key Laboratory of Crop Genetics and Germplasm EnhancementNanjing Agricultural UniversityNanjingP.R. China

Personalised recommendations