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Development of Transgenic CryIA(c) + GNA Cotton Plants via Pollen Tube Pathway Method Confers Resistance to Helicoverpa armigera and Aphis gossypii Glover

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Transgenic Cotton

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

Abstract

Elite cotton cultivar Sumian16 was transformed with p7RPSBK-mGNA-NPTII containing Bt (CryIA(c)), Galanthus nivalis agglutinin (GNA) resistance genes and selectable marker NptII gene via the pollen tube pathway method and two fertile transgenic Bt + GNA plants were obtained in the present study. The integration and expression of the Bt and GNA genes were confirmed by molecular biology techniques and insect bioassays. Insect bioassays showed that the transformed plants were highly toxic to bollworm larvae as well as obviously retarding development of aphid populations. PCR analyses and identification of resistance to Kanamycin and bollworm showed that the resistance to bollworm for the two transgenic plants was dominantly inherited in a Mendelian manner and the two resistance genes and selectable marker co-segregated from primary transformed parents to the first self-fertilized progeny plants.

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References

  1. Benedict JH, Sachs ES, Altman WR, Deaon RJ, Kohel RJ, Ring DR, Berberich SA (1996) Field performance of cottons expressing transgenic Cry1A insecticidal protein for resistance to Heliothis virescens and Helicoverpa zea (Lepidoptera: Nochxidae). J Econ Entomol 89:230–238

    Google Scholar 

  2. Fitt GP, Mares CL, Llewellyn DJ (1994) Field evaluation and potential ecological impact of transgenic cottons in Australia. Biocontrol Sci Technol 4:535–548

    Article  Google Scholar 

  3. Zhang TZ, Tang CM (2000) Commercial production of transgenic Bt insect resistant cotton varieties and the resistance management for bollworm (Helicoverpa armigera Hubner). China Sci Bull 45(14):1249–1257

    Article  CAS  Google Scholar 

  4. Powell KS, Gatehouse AMR, Hilder VA, Gatehouse JA (1993) Antimetabolic effects of plant and fungal enzymes on the nymphal stages of two important rice pest, Nilaparvata lugens and Nephotettix cinciteps. Entmol Exp Appl 66:119–126

    Article  CAS  Google Scholar 

  5. Hilder VA, Powell KS, Gatehouse AMR, Gatehouse JA, Gatehouse LN, Shi Y, Hamilton WDO, Merryweather A, Newell CA, Timans JC, Peumans WJ, Van Damme EJS, Boulter D (1995) Expression of snowdrop lectin in transgenic tobacco plant results in added protection against aphids. Transgenic Res 4:18–25

    Article  CAS  Google Scholar 

  6. Zhou Y, Tian YC, Wu B, Mang KQ (1998) Inhibition effect of transgenic tobacco plants expressing snowdrop lectin on the population of Myzus persicae. Chin J Biotechnol 14(1):13–19

    CAS  Google Scholar 

  7. Yuan ZQ, Zhao CY, Zhou Y, Tian YC (2001) Aphid-resistant transgenic tobacco plants expressing modified GNA gene. Acta Bot Sin 43(6):592–597

    CAS  Google Scholar 

  8. Liu Z, Yuan XL, Zhang TZ (2001) Strategies to develop transgenic crops expressing multiple genes. Hereditas 13(2):182–186

    Google Scholar 

  9. McGaughey WH, Gould F, Gelernter W (1998) Bt resistance management. Nat Biotechnol 16:144–146

    Article  PubMed  CAS  Google Scholar 

  10. Van der Salm T, Bosch D, Honee G, Feng LX, Munsterman E, Bakker P, Stiekema WJ, Visser B (1994) Insect resistance of transgenic plants that express modified Bacillus thuringiensis cryIA(b)and cryIC genes: a resistance management strategy. Plant Mol Bio 26:51–59

    Article  Google Scholar 

  11. Wang ZB, Guo SD (1999) Transgenic cryIA  +  GNA tobacco plants conferring resistance to bollworm and aphid. China Sci Bull 4(19):2068–2075

    Google Scholar 

  12. Boulter D, Edward GA, Gatehouse AMR, Hilder VA (1990) Additive protective effects of different plant-derived insect resistance genes in transgenic tobacco. Crop Prot 9:351–354

    Article  Google Scholar 

  13. Zhao JZ, Fan YL, Fan XL, Shi SP, Lu GM (1999) Evaluation of effect of transgenic tobacco plants expressing double resistance genes on delaying resistance of bollworm (Helicoverpa armigera Hubner). China Sci Bull 44(15):1635–1639

    Article  Google Scholar 

  14. Constable G (1998) Two gene technology in the Australian scene. The Australian Cotton Grower 5:27–29

    Google Scholar 

  15. Wang W, Zhu Z, Gao YF, Shi CL, Chen WX, Guo ZC, Li XH (1999) Obtaining a transgenic upland cotton harboring two insecticidal genes. Acta Bot Sin 41(4):384–388

    CAS  Google Scholar 

  16. Guo SD, Cui HZ, Xia LQ, Wu DL, Ni WC, Zhang ZL, Zhang BL, Xu YJ (1999) Development of bivalent insect-resistant transgenic cotton plants. Sci Agri Sin 32(3):1–7

    Google Scholar 

  17. Li FG, Cui JJ, Liu CL, Wu ZX, Li FL, Zhou Y, Li XL, Guo SD, Cui HZ (2000) The study of insect-resistant transgenic cotton harboring double-gene and its insect-resistance. Sci Agri Sin 33(1):46–52

    CAS  Google Scholar 

  18. Guo SD, Ni WC, Zhao GZ (2001) Development of transgenic cotton plants harboring two insecticidal genes. In: Jia SR, Guo SD, An DQ (eds) Transgenic cotton. Series book of 863 biological high-tech. Science press, Beijing, pp 160–163

    Google Scholar 

  19. Xie YQ, Zhu Z, Wu Q, Xu HB, Meng M, Liu YL (2000) Isolation of a strong promoter and its functional study. Acta Bot Sin 42(1):50–54

    CAS  Google Scholar 

  20. 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. Chin Sci Bull 45(4):363–367

    Article  Google Scholar 

  21. Yuan XL, Sun J, Tang CM, Zhang TZ (2001) Temporary fluctuation of resistance of the transgenic cotton harboring Bt  +  CpTI double-gene to Helicoverpa armigera. High Tech Lett 10(2):9–12

    Google Scholar 

  22. Zhou GY, Weng J, Zheng YS, Huang JQ, Qian SY, Liu GL (1983) Introduction of exogenous DNA into cotton embryos. In: Wu R, Gnssman L, Moldave K (eds) Methods in enzymology, vol 101. Academic, New York, pp 433–481

    Google Scholar 

  23. Sambrook J, Friston EF, Maniatis T (eds) (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  24. 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 123:343–351

    Article  CAS  Google Scholar 

  25. Paterson AH, Brubaker CL, Wendel JF (1993) A rapid method for extraction of cotton (Gossypium spp.) genomic DNA suitable for RFLP and PCR analysis. Plant Mol Biol Rep 11:122–127

    Article  CAS  Google Scholar 

  26. Song P, Yamamoto E, Allen R (1995) Differential display of cotton transcripts. Plant Mol Biol Rep 13:174–181

    Article  CAS  Google Scholar 

  27. Chong K, Bao S, Xu T, Tan K, Liang T, Zeng J, Huang H, Xu J (1998) Functional analysis of the ver gene using transgenic wheat. Physiol Plant 102:87–92

    Article  CAS  Google Scholar 

  28. Luo ZX, Wu R (1988) Simple method for the transformation of rice via the pollen-tube pathway. Plant Mol Biol Rep 6(3):165–174

    Article  CAS  Google Scholar 

  29. Ni WC, Zhang ZL (1998) Development of transgenic insect-resistant cotton plants. Sci Agri Sin 31(2):8–13

    CAS  Google Scholar 

  30. Xie DX, Fan YL, Ni PZ (1991) Obtaining transgenic plants by introducing Bacillus thuringiensis insecticidal crystal protein gene into rice cultivar Zhonghua11 in China. Sci China B 8:830–834

    Google Scholar 

  31. Xie DX, Fan YL, Ni WC, Huang JQ (1991) Obtaining transgenic plants by introducing Bacillus thuringiensis insecticidal crystal ­protein gene into cotton. Sci China B 4:367–373

    Google Scholar 

  32. Zeng JZ, Wang DJ, Wu YQ, Zhang J, Zhou WJ, Zhu XP, Xu NZ (1993) Obtaining of transgenic wheat plants by pollen tube pathway. China Sci Bull 23(3):256–262

    Google Scholar 

  33. Zeng JZ, Wu YQ, Wang DJ, Zhang J, Ma ZR, Zhou ZY (1998) Discussion in transformation mechanism and inheritance of transgenic plants progeny by pollen tube pathway (or vector). China Sci Bull 43(6):561–566

    Google Scholar 

  34. Maqbool SB, Riazuddin S, Loc NT, Gatehouse AMR, Gatehouse JA, Christou P (2001) Expression of multiple insecticidal genes confers broad resistance against a range of different rice pests. Mol Breed 7:85–93

    Article  CAS  Google Scholar 

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Acknowledgments

This project was financially supported in part by the National Research and Development Project of Transgenic Crops of China (2010ZX08009-003, 2011ZX08005-001) and the Priority Academic Program Development of Jiangsu Higher Education Institutions.

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Authors and Affiliations

  1. National Key Laboratory of Crop Genetics and Germplasm Enhancement, Cotton Research Institute, Nanjing Agricultural University, Nanjing, P.R. China

    Zhi Liu

  2. Genetics and Developmental Biology Institute, Chinese Academy of Sciences, Beijing, China

    Zhen Zhu

  3. National Key Laboratory of Crop Genetics and Germplasm Enhancement, Cotton Research Institute, Nanjing Agricultural University, Nanjing, People’s Republic of China

    Tianzhen Zhang

Authors
  1. Zhi Liu
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  2. Zhen Zhu
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  3. Tianzhen Zhang
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Corresponding author

Correspondence to Tianzhen Zhang .

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Editors and Affiliations

  1. , Department of Biology, East Carolina University, Howell Science Complex S112, Greenville, 27858, North Carolina, USA

    Baohong Zhang

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Liu, Z., Zhu, Z., Zhang, T. (2013). Development of Transgenic CryIA(c) + GNA Cotton Plants via Pollen Tube Pathway Method Confers Resistance to Helicoverpa armigera and Aphis gossypii Glover. In: Zhang, B. (eds) Transgenic Cotton. Methods in Molecular Biology, vol 958. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-212-4_17

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  • DOI: https://doi.org/10.1007/978-1-62703-212-4_17

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-211-7

  • Online ISBN: 978-1-62703-212-4

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