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Polyester synthesis in transplastomic tobacco (Nicotiana tabacum L.): significant contents of polyhydroxybutyrate are associated with growth reduction

  • Genetic Transformation and Hybridization
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Abstract

The pathway for synthesis of polyhydroxybutyrate (PHB), a polyester produced by three bacterial enzymes, was transferred to the tobacco plastid genome by the biolistic transformation method. The polycistronic phb operon encoding this biosynthetic pathway was cloned into plastome transformation vectors. Following selection and regeneration, the content and structure of plant-produced hydroxybutyrate was analysed by gas chromatography. Significant PHB synthesis was limited to the early stages of in vitro culture. Within the transformants, PHB synthesis levels were highly variable. In the early regeneration stage, single regenerates reached up to 1.7% PHB in dry weight. At least 70% of plant-produced hydroxybutyric acid was proven to be polymer with a molecular mass of up to 2,500 kDa. PHB synthesis levels of the transplastomic lines were decreasing when grown autotrophically but their phb transcription levels remained stable. Transcription of the three genes is divided into two transcripts with phbB being transcribed separately from phbC and phbA. In mature plants even low amounts of PHB were associated with male sterility. Fertility was only observed in a mutant carrying a defective phb operon. These results prove successful expression of the entire PHB pathway in plastids, concomitant, however, with growth deficiency and male sterility.

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Abbreviations

3HB:

3-Hydroxybutyric acid

d.w.:

Dry weight

CoA:

Coenzyme A

GC:

Gas chromatography

GPC:

Gel permeation chromatography

HPLC:

High performance liquid chromatography

Mn:

Molecular number average

Mw:

Molecular weight average

nt:

Nucleotide

PHA:

Polyhydroxyalkanoate

PHB:

Polyhydroxybutyrate

UTR:

Untranslated region

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Acknowledgements

This work was supported through the Fachagentur für nachwachsende Rohstoffe (grant no. 99NR074). The excellent technical assistance by Katharina Yeiser, Stefan Kirchner and Petra Winterholler is gratefully acknowledged.

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Author notes

  1. C. Eibl

    Present address: Icon Genetics, 85354 , Freising, Germany

Authors and Affiliations

  1. Department of Botany, Ludwig-Maximilians-Universität, Menzinger Strasse 67, 80638 , Munich, Germany

    A. Lössl, C. Eibl & H.-U. Koop

  2. Institut für Pflanzenbau und Pflanzenzüchtung, Christian-Albrechts-Universität zu Kiel, Olshausenstrasse 40, 24098, Kiel, Germany

    H.-J. Harloff & C. Jung

Authors
  1. A. Lössl
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  2. C. Eibl
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  3. H.-J. Harloff
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  4. C. Jung
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  5. H.-U. Koop
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Correspondence to A. Lössl.

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Communicated by H. Lörz

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Lössl, A., Eibl, C., Harloff, HJ. et al. Polyester synthesis in transplastomic tobacco (Nicotiana tabacum L.): significant contents of polyhydroxybutyrate are associated with growth reduction. Plant Cell Rep 21, 891–899 (2003). https://doi.org/10.1007/s00299-003-0610-0

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