Biologia Plantarum

, Volume 51, Issue 2, pp 242–248 | Cite as

Agrobacterium-mediated transformation of Cry1C, Cry2A and Cry9C genes into Gossypium hirsutum and plant regeneration

  • X. Guo
  • C. Huang
  • S. Jin
  • S. Liang
  • Y. Nie
  • X. Zhang
Original Papers

Abstract

Three constructs harbouring novel Bacillus thuringiensis genes (Cry1C, Cry2A, Cry9C) and bar gene were transformed into four upland cotton cultivars, Ekangmian10, Emian22, Coker201 and YZ1 via Agrobacterium-mediated transformation. With the bar gene as a selectable marker, about 84.8 % of resistant calli have been confirmed positive by polymerase chain reaction (PCR) tests, and totally 50 transgenic plants were regenerated. The insertions were verified by means of Southern blotting. Bioassay showed 80 % of the transgenic plantlets generated resistance to both herbicide and insect. We optimized conditions for improving the transformation efficiency. A modified in vitro shoot-tip grafting technique was introduced to help entire transplantation. This result showed that bar gene can replace antibiotic marker genes (ex. npt II gene) used in cotton transformation.

Additional key words

Agrobacterium tumefaciens bar gene cotton novel Bt genes 

Abbreviations

AS

acetosyringone

EC

embryogenic callus

IBA

indole-3-butyric acid

npt II

neomycin phosphotransferase

L-L-PPT

L-phosphinothricin

AS

acetosyringone

PCR

polymerase chain reaction

MSB

Murashige and Skoog (1962) medium with B5 (Gamborg et al. 1968) vitamins

PAT

phosphinothricin acetyltransferase

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Copyright information

© Institute of Experimental Botany, ASCR 2007

Authors and Affiliations

  • X. Guo
    • 1
  • C. Huang
    • 1
  • S. Jin
    • 1
  • S. Liang
    • 1
  • Y. Nie
    • 1
  • X. Zhang
    • 1
  1. 1.National Key Laboratory of Crop Genetic ImprovementHuazhong Agricultural UniversityWuhan, HubeiP.R. China

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