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Genetic transformation of Begonia tuberhybrida by Ri rol genes

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Abstract

We have developed an Agrobacterium -mediated transformation system for commercial Begonia species. The leaf explants of Begonia semperflorens, Begonia x hiemalis and B. tuberhybrida were inoculated with Agrobacterium tumefaciens LBA4404 harboring a binary vector pBI121 which contains rolA, B and C genes of an agropine type Ri plasmid (pRiA4b). Kanamycin resistant shoots of B. tuberhybrida were obtained on MS agar medium supplemented with 0.1 mg/l NAA, 0.5 mg/l BA, 500 mg/l claforan and 100 mg/l kanamycin. These shoots exhibited GUS activity and Southern analysis showed a single copy insertion into the genome. When the transgenic plants were transferred to soil, they displayed the phenotype specific to the transgenic plants by A. rhizogenes such as dwarfness, delay of flowering, and wrinkled leaves and petals.

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Abbreviations

NAA:

1-naphthaleneacetic acid

BA:

benzyladenine

MS:

Murashige and Skoog (1962)

GUS:

β-glucuronidase (EC 3.2.1.31)

NPT II:

neomycin phosphotransferase (EC 2.7.1.95)

NOS:

nopaline synthase

CaMV:

Cauliflower Mosaic Virus

CTAB:

cetyl triethylammonium bromide

X-GLUC:

5-bromo-4-chloro-3-indolyl-β-D-glucuronide

PCR:

polymerase chain reaction

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Authors and Affiliations

  1. Institute of Bioscience and Biotechnology, Aomori University, 030, Aomori, Japan

    Shigeto Kiyokawa

  2. Tsukuba Research Laboratory, Kyowa Hakko Kogyo Co., Miyukigaoka, Tsukuba, 305, Ibaraki, Japan

    Yasuhiro Kikuchi

  3. Institute of Biological Sciences, University of Tsukuba, Tsukuba, 305, Ibaraki, Japan

    Hiroshi Kamada & Hiroshi Harada

Authors
  1. Shigeto Kiyokawa
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  2. Yasuhiro Kikuchi
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  3. Hiroshi Kamada
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  4. Hiroshi Harada
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Communicated by A. Komamine

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Kiyokawa, S., Kikuchi, Y., Kamada, H. et al. Genetic transformation of Begonia tuberhybrida by Ri rol genes. Plant Cell Reports 15, 606–609 (1996). https://doi.org/10.1007/BF00232462

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  • DOI: https://doi.org/10.1007/BF00232462

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