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Alcohol-inducible gene expression in transgenic Populus

  • Genetic Transformation and Hybridization
  • Published:
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Abstract

We tested the efficiency and optimized the conditions for controlled alcohol-inducible transgene expression in Populus using gus as a reporter gene. Specificity of induction, efficiency in different organs, effect of three chemical inducers, and induction methods were tested using up to 10 independent transgenic events generated in two different Populus genotypes. The optimal inducer concentration and the duration of induction period were determined in dose–response and in time–course experiments. Under in vitro conditions, β-glucuronidase (GUS) induction was efficient both in the aerial parts and in the roots of regenerated plantlets. Among the chemical inducers tested, ethanol was the most effective activator with no apparent phytotoxicity when concentrations were at or below 2%. After 5 days of treatment, fluorometrically-determined the GUS activity could be detected when inducing with ethanol at concentrations as low as 0.5%. Prolonged induction by ethanol vapors significantly increased the GUS activity in leaves from both the tissue culture plants and greenhouse-grown plants.

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Abbreviations

Ac:

Acetaldehyde

2-B:

2-Butanone

BAP:

6-Benzylaminopurine

CaMV:

Cauliflower mosaic virus

CIM:

Callus-induction medium

dpi:

Days post-induction

IBA:

Indole-3-butyric acid

IM:

Agrobacterium-induction medium

LB:

Luria-Bertani medium

MS:

Murashige and Skoog medium

RCF:

Relative centrifugal force

SIM:

Shoot-induction medium

TDZ:

Thidiazuron

uidA :

β-Glucuronidase, gus reporter gene

X-Gluc:

5-Bromo-4-chloro-3-indolyl-β-d-glucuronic acid

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Acknowledgements

We thank Syngenta Biotechnology, Inc. for providing pJH0143, and Dr. Renee Arias for the critical discussions during manuscript preparation. This work was supported in part by grants from the Office of Science (BER), U.S. Department of Energy, grant no. DE-4000023558 “Genome-enabled discovery of carbon sequestration genes in poplar”), the National Science Foundation Industry/University Cooperative Research Centers (award no. 9980423), and by members of the Tree Biosafety and Genomics Research Cooperative based at Oregon State University (http://wwwdata.forestry.oregonstate.edu/tgbb/).

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Authors and Affiliations

  1. Department of Forest Science, Oregon State University, 97331, Corvallis, OR, US

    S. A. Filichkin, C. Ma, A. M. Brunner & S. H. Strauss

  2. Department of Forestry and Natural Resources, Purdue University, 47907, West Lafayette, IN, USA

    R. Meilan

  3. School of Forest Resources and Environmental Science, Michigan Technological University, 49931, Houghton, MI, USA

    V. B. Busov

Authors
  1. S. A. Filichkin
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  2. R. Meilan
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  3. V. B. Busov
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  4. C. Ma
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  5. A. M. Brunner
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  6. S. H. Strauss
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Corresponding author

Correspondence to S. H. Strauss.

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Communicated by: D.A. Somers

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Filichkin, S.A., Meilan, R., Busov, V.B. et al. Alcohol-inducible gene expression in transgenic Populus . Plant Cell Rep 25, 660–667 (2006). https://doi.org/10.1007/s00299-005-0112-3

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  • DOI: https://doi.org/10.1007/s00299-005-0112-3

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