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Agrobacterium-mediated transformation of shoot apices of Kentucky bluegrass (Poa pratensis L.) and production of transgenic plants carrying a betA gene

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Abstract

Apical meristems of multiple shoots produced from axenic seedlings of Kentucky bluegrass (Poa pratensis L.) were used for Agrobacterium tumefaciens-mediated transformation. Transformation parameters were optimized for concentration of bacterial cells, duration of infection, and vacuum infiltration. The highest transformation frequency (1.42%) was obtained by infection with Agrobacterium suspension of OD600 = 0.6 for 5 min, under a negative pressure of 0.5 × 105 Pa. After co-cultivation, the herbicide-resistant plants were rooted and transplanted into flowerpots. Transgenic plants were confirmed by polymerase chain reaction (PCR) assay and Southern blot analysis. Using this transformation system, the betA gene encoding choline dehydrogenase and mutant als gene encoding the enzyme acetolactate synthase were introduced into three Kentucky bluegrass cultivars.

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Abbreviations

CTAB:

Cetyltrimethyl ammonium bromide

BA:

6-benzyladenine

2,4-D :

2,4-dichlorophenoxyacetic acid

AS:

Acetosyringone

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Acknowledgments

We would like to thank Dr. L. B. Han at the Turfgrass Institute of Beijing Forestry University for providing the seeds of Kentucky bluegrass. This research was supported by the National Project for Transgenic Plant Research and Industrialization of China (2008ZX08005-4), the Cotton Improved Variety Project of Shandong Province (2009GG10009010), and the High-Tech Research and Development Program of China (2007AA091701). We would also like to thank International Science Editing for their assistance in the language editing of the manuscript.

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Authors and Affiliations

  1. School of Life Science, Shandong University, 27 Shanda South Road, 250100, Jinan, Shandong, China

    Kewei Zhang, Juan Wang, Aifang Yang & Juren Zhang

  2. National Defense Technology Institute, Shandong University, 250100, Jinan, Shandong, China

    Xiaorui Hu

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  1. Kewei Zhang
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  2. Juan Wang
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  3. Xiaorui Hu
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Correspondence to Kewei Zhang.

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Zhang, K., Wang, J., Hu, X. et al. Agrobacterium-mediated transformation of shoot apices of Kentucky bluegrass (Poa pratensis L.) and production of transgenic plants carrying a betA gene. Plant Cell Tiss Organ Cult 102, 135–143 (2010). https://doi.org/10.1007/s11240-010-9713-9

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