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

Protocol
Part of the Methods in Molecular Biology book series (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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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 UniversityNanjingP.R. China
  2. 2.Genetics and Developmental Biology Institute, Chinese Academy of SciencesBeijingChina
  3. 3.National Key Laboratory of Crop Genetics and Germplasm Enhancement, Cotton Research InstituteNanjing Agricultural UniversityNanjingPeople’s Republic of China

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