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Production of ribosome-inactivating protein from hairy root cultures of Luffa cylindrica (L.) Roem.

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Abstract

Transformed root lines of Luffa cylindrica (L.) Roem. (Cucurbitaceae) were established by inoculation of in vitro grown plantlets with wild type Agrobacterium rhizogenes strain 1855. Cloned lines of hairy roots were tested for the presence of ribosome-inactivating proteins; crude extracts inhibited protein synthesis in a reaction mixture based on rabbit reticulocyte lysate. Inhibitory activity increased during culture period, reaching a maximum value in the stationary phase. No activity could be detected in the culture medium, nor in extracts from callus and/or suspension cultures. A ribosome-inactivating protein having specific activity of 62,100 U mg protein−1 and a molecular mass of 26–28,000 Da was purified to homogeneity. The protein showed N-glycosidase activity on rat liver ribosomes. The results demonstrate that hairy root cultures can be successfully utilized for the in vitro production of ribosome-inactivating proteins.

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Abbreviations

BAP:

benzylaminopurine

2,4D:

2,4dichlorophenoxyacetic acid

HPLC:

high pressure liquid chromatography

MS:

Murashige and Skoog

NAA:

naphthaleneacetic acid

NCPPB:

National Collection of Phytopathogenic Bacteria

Ri:

root-inducing

RIP:

ribosome-inactivating protein

UV:

ultra-violet

YMB:

yeast mannitol broth

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Authors and Affiliations

  1. Dipartimento di Biologia di Base ed Applicata, Università degli Studi di L'Aquila, I-67100, Coppito L'Aquila, Italy

    Luigi Sanità di Toppi, Giuliana Properzi & Laura Spanò

  2. Dipartimento di Patologia Sperimentale, Università degli Studi di Bologna, via S. Giacomo 14, 1-40126, Bologna, Italy

    Paola Gorini & Luigi Barbieri

Authors
  1. Luigi Sanità di Toppi
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  2. Paola Gorini
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  3. Giuliana Properzi
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  4. Luigi Barbieri
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  5. Laura Spanò
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Communicated by M. R. Davey

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di Toppi, L.S., Gorini, P., Properzi, G. et al. Production of ribosome-inactivating protein from hairy root cultures of Luffa cylindrica (L.) Roem.. Plant Cell Reports 15, 910–913 (1996). https://doi.org/10.1007/BF00231586

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  • DOI: https://doi.org/10.1007/BF00231586

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