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Agrobacterium-mediated genetic transformation of Perilla frutescens

  • Genetic Transformation and Hybridization
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Abstract

A reproducible plant regeneration and an Agrobacterium tumefaciens-mediated genetic transformation protocol were developed for Perilla frutescens (perilla). The largest number of adventitious shoots were induced directly without an intervening callus phase from hypocotyl explants on MS medium supplemented with 3.0 mg/l 6-benzylaminopurine (BA). The effects of preculture and extent of cocultivation were examined by assaying β-glucuronidase (GUS) activity in explants infected with A. tumefaciens strain EHA105 harboring the plasmid pIG121-Hm. The highest number of GUS-positive explants were obtained from hypocotyl explants cocultured for 3 days with Agrobacterium without precultivation. Transgenic perilla plants were regenerated and selected on MS basal medium supplemented with 3.0 mg/l BA, 125 mg/l kanamycin, and 500 mg/l carbenicillin. The transformants were confirmed by PCR of the neomycin phosphotransferase II gene and genomic Southern hybridization analysis of the hygromycin phosphotransferase gene. The frequency of transformation from hypocotyls was about 1.4%, and the transformants showed normal growth and sexual compatibility by producing progenies.

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Abbreviations

BA :

6-Benzylaminopurine

GUS:

β-Glucuronidase

hpt :

Hygromycin phosphotransferase

NAA:

α-Naphthaleneacetic acid

nptII:

Neomycin phosphotransferase II

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Authors and Affiliations

  1. National Institute of Agricultural Biotechnology, Rural Development Administration, Suwon, 441-707, South Korea

    Kyung-Hwan Kim, Yeon-Hee Lee, Donghern Kim, Yong-Hwan Park, Young-Soo Hwang & Yong-Hwan Kim

  2. Department of Microbiology, Kyungpook National University, Taegu, 702-701, South Korea

    Jai-Youl Lee

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  1. Kyung-Hwan Kim
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  2. Yeon-Hee Lee
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  3. Donghern Kim
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  4. Yong-Hwan Park
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  5. Jai-Youl Lee
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  6. Young-Soo Hwang
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Correspondence to Yong-Hwan Kim.

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Communicated by I.S. Chung

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Kim, KH., Lee, YH., Kim, D. et al. Agrobacterium-mediated genetic transformation of Perilla frutescens . Plant Cell Rep 23, 386–390 (2004). https://doi.org/10.1007/s00299-004-0825-8

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  • DOI: https://doi.org/10.1007/s00299-004-0825-8

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