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Agrobacterium-mediated transformation of Perilla (Perilla frutescens)

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Abstract

Agrobacterium tumefaciens-mediated transformation system for perilla (Perilla frutescens Britt) was developed. Agrobacterium strain EHA105 harboring binary vector pBK I containing bar and γ-tmt cassettes or pIG121Hm containing nptII, hpt, and gusA cassettes were used for transformation. Three different types of explant, hypocotyl, cotyledon and leaf, were evaluated for transformation and hypocotyl explants resulted in the highest transformation efficiency with an average of 3.1 and 2.2%, with pBK I and pIG121Hm, respectively. The Perilla spp. displayed genotype-response for transformation. The effective concentrations of selective agents were 2 mg l−1 phosphinothricin (PPT) and 150 mg l−1 kanamycin, respectively, for shoot induction and 1 mg l−1 PPT and 125 mg l−1 kanamycin, respectively, for shoot elongation. The transformation events were confirmed by herbicide Basta spray or histochemical GUS staining of T0 and T1 plants. The T-DNA integration and transgene inheritance were confirmed by PCR and Southern blot analysis of random samples of T0 and T1 transgenic plants.

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Abbreviations

γ-tmt:

γ-tocopherol methyltransferase gene

bar :

bialaphos resistance gene

gusA:

β-glucu- ronidase gene

hpt :

hygromycin phosphotransferase gene

hpt II :

neomycin phosphotransferase gene

PPT:

phosphinothricin

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Authors and Affiliations

  1. Division of Crop Environment and Biotechnology, National Institute of Crop Science, Suwon, 441-857, South Korea

    Byoung-Kyu Lee, Seung-Hee Yu, Yul-Ho Kim, Byung-Ohg Ahn, Han-Sun Hur & Jang-Yong Lee

  2. Department of Agronomy, Kyungpook National University, Daegu, 702-701, South Korea

    Sang-Chul Lee

  3. Plant Transformation Core Facility and Department of Agronomy, University of Missouri-Columbia, Columbia, MO, 65211, USA

    Zhanyuan Zhang

  4. Department of Agronomy, University of Missouri-Columbia, Columbia, MO, 65211, USA

    Zhanyuan Zhang

Authors
  1. Byoung-Kyu Lee
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  2. Seung-Hee Yu
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  3. Yul-Ho Kim
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  4. Byung-Ohg Ahn
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  5. Han-Sun Hur
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  6. Sang-Chul Lee
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  7. Zhanyuan Zhang
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  8. Jang-Yong Lee
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Correspondence to Jang-Yong Lee.

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Lee, BK., Yu, SH., Kim, YH. et al. Agrobacterium-mediated transformation of Perilla (Perilla frutescens). Plant Cell Tiss Organ Cult 83, 51–58 (2005). https://doi.org/10.1007/s11240-005-3665-5

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  • DOI: https://doi.org/10.1007/s11240-005-3665-5

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