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Pistil drip following pollination: a simple in planta Agrobacterium-mediated transformation in cotton

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Abstract

Transgenic cotton plants were developed by pistil drip inoculation in a solution containing Agrobacterium carrying a gene for resistance to the herbicide Basta (bar), 10% (w/v) sucrose, 0.05% (v/v) Silwet L-77 and 40 mg acetosyringone l−1. Pistil drip during 17:00–19:00 on the first day of flowering resulted in 0.07–0.17% Basta-resistant plants/number of viable seeds generated, and stigma excision prior to pistil drip during this time period gave rise to a transformation efficiency of 0.46–0.93%, in contrast with 0.04–0.06% generated from pistil drip during 9:00–11:00 on the second day of flowering. PCR and Southern blot analysis confirmed the integration of the bar gene into the cotton genome, and a T1 and T2 generation herbicide resistance test consistently revealed expression and stable heritability of the bar gene in the two generations.

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Acknowledgements

The authors specially thank Dr. Muhammad Saeed for a critical review of the manuscript. This work was supported by National High Technology Research and Development Program of China (Grant No. 2006AA100105), Science & Technology Pillar Program of Jiangsu Province (Grant No. BE2008310) and Programme of Introducing Talents of Discipline to Universities (Grant No. B08025).

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Authors and Affiliations

  1. National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing, 210095, China

    Chen TianZi, Wu ShenJie, Zhao Jun, Guo WangZhen & Zhang TianZhen

  2. Cotton Research Institute, Shanxi Academy of Agricultural Sciences, Yuncheng, 030006, China

    Wu ShenJie

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  1. Chen TianZi
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  2. Wu ShenJie
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  3. Zhao Jun
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  4. Guo WangZhen
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Correspondence to Zhang TianZhen.

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TianZi, C., ShenJie, W., Jun, Z. et al. Pistil drip following pollination: a simple in planta Agrobacterium-mediated transformation in cotton. Biotechnol Lett 32, 547–555 (2010). https://doi.org/10.1007/s10529-009-0179-y

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  • DOI: https://doi.org/10.1007/s10529-009-0179-y

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