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Agrobacterium-medlated high-efficiency transformation of creeping bentgrass with herbicide resistance

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Abstract

We performedAgrobacterium-mediated genetic transformation of creeping bentgrass(Agrostis stolonifera L.) and produced herbicide-resistant transformants from commercial cultivars Crenshaw and Penncross. Seed-derived embryogenie calli were infected withA. tumefaciens EHA105 harboring pCAMBIA 3301, which includes an intron-containinggus reporter and abar selection marker. To establish a stable system, we examined various factors that could potentially influence transformation efficiency during the pre-culture, infection, and co-cultivation steps. The addition of kinetin to the callus pre-culture media increased efficiency about three-fold. Once the optimum infection and co-cultivation conditions were identified, this protocol was used successfully to bulk-produce herbicide-resistant transgenic plants whose herbicide resistance was confirmed using the BASTA® resistance test. Southern blot analysis demonstrated integration and low copy numbers of the integrated transgenes, and northern blot analysis verified their expression. Thus, we have established an efficient genetic transformation system for creeping bentgrass and confirmed a high frequency of single-copy transgene integration and functional gene expression.

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Authors and Affiliations

  1. Department of Molecular Biotechnology (BK21 Program) and Kumho Life Science Laboratory, Chonnam National University, 500-757, Gwangju, Korea

    Soo Jung Kim, Jee-Yeon Lee, Yong-Min Kim, Song-Sook Yang, Ok-Jin Hwang, Nam-Ju Hong & Jeong-II Kim

  2. Chcju National University, 690-756, Jeju, Korea

    Kyung-Moon Kim, Hyo-Yeon Lee & Pill-Soon Song

  3. Environmental Biotechnology National Core Research Center, Gyeongsang National University, 660-701, Jinju, Korea

    Pill-Soon Song & Jeong-II Kim

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  1. Soo Jung Kim
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  2. Jee-Yeon Lee
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  3. Yong-Min Kim
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  4. Song-Sook Yang
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  5. Ok-Jin Hwang
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  6. Nam-Ju Hong
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  7. Kyung-Moon Kim
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  8. Hyo-Yeon Lee
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  9. Pill-Soon Song
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  10. Jeong-II Kim
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Correspondence to Jeong-II Kim.

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Kim, S.J., Lee, JY., Kim, YM. et al. Agrobacterium-medlated high-efficiency transformation of creeping bentgrass with herbicide resistance. J. Plant Biol. 50, 577–585 (2007). https://doi.org/10.1007/BF03030712

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  • DOI: https://doi.org/10.1007/BF03030712

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