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Aluminum borate whisker-mediated production of transgenic tobacco plants

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Abstract

An aluminum borate whiskers-mediated transformation system for calluses of tobacco (Nicotiana tabacum, cv. SR-1) has been developed. A total of 50 small pieces of calluses were vigorously agitated in a liquid medium containing aluminum borate whiskers, pBI221 plasmid carrying the β-glucuronidase (GUS) gene, and pBI222 plasmid carrying the hygromycin phosphotransferase (HPT) gene. After treatment, calluses were cultured to select for hygromycin resistance, and three resistant calluses were obtained. Adventitious shoots were produced from each hygromycin-resistant callus and were transferred to rooting medium. A total of three plantlets obtained from each hygromycin-resistant callus were acclimatized and established in soil. Polymerase chain reaction analysis revealed that all the plantlets were cotransformed with both the GUS and HPT genes. Detached leaves of transgenic individuals showed clear hygromycin resistance when cultured in liquid medium. Histochemical assay for GUS revealed that one of these transgenic plants expressed the GUS gene, indicating coexpression of foreign genes.

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Abbreviations

ABW:

aluminum borate whiskers

BA:

6-benzyladenine

CaMV:

cauliflower mosaic virus

GUS:

β-glucuronidase

HPT:

hygromycin phosphotransferase

NAA:

1-naphthaleneacetic acid

PCR:

polymerase chain reaction

SCW:

silicon carbide whiskers

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Author notes

  1. Wataru Takahashi

    Present address: Forage Crop Research Institute, Japan Grassland Farming and Forage Seed Association, 388-5 Higashiakada, Nishinasuno, Tochigi, 329-2742, Japan

Authors and Affiliations

  1. Department of Biology, Faculty of Science and Engineering, Konan University, Okamoto, Higashinada-ku, Kobe, 658-8501, Japan

    Kiyotaka Mizuno, Wataru Takahashi & Osamu Tanaka

  2. Department of Bioscience, Faculty of Science and Engineering, Teikyou University of Science and Technology, 2525 Uenohara, Yatsuzawa, Yamanashi, 409-0193, Japan

    Toshio Beppu

  3. Research Institute of Agricultural Resource, Ishikawa Agricultural College, 1-308 Suematsu, Nonoichi, Ishikawa, 921-8836, Japan

    Takiko Shimada

Authors
  1. Kiyotaka Mizuno
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  2. Wataru Takahashi
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  3. Toshio Beppu
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  4. Takiko Shimada
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  5. Osamu Tanaka
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Corresponding author

Correspondence to Osamu Tanaka.

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Mizuno, K., Takahashi, W., Beppu, T. et al. Aluminum borate whisker-mediated production of transgenic tobacco plants. Plant Cell Tiss Organ Cult 80, 163–169 (2005). https://doi.org/10.1007/s11240-004-9542-9

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  • DOI: https://doi.org/10.1007/s11240-004-9542-9

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