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

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Biologia Plantarum

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.

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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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Authors and Affiliations

  1. National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, Hubei, 430070, P.R. China

    X. Guo, C. Huang, S. Jin, S. Liang, Y. Nie & X. Zhang

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  1. X. Guo
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  2. C. Huang
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  3. S. Jin
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  4. S. Liang
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  5. Y. Nie
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  6. X. Zhang
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Correspondence to X. Zhang.

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Guo, X., Huang, C., Jin, S. et al. Agrobacterium-mediated transformation of Cry1C, Cry2A and Cry9C genes into Gossypium hirsutum and plant regeneration. Biol Plant 51, 242–248 (2007). https://doi.org/10.1007/s10535-007-0048-2

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  • DOI: https://doi.org/10.1007/s10535-007-0048-2

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