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Enhanced cold tolerance in transgenic tobacco expressing a chloroplast ω-3 fatty acid desaturase gene under the control of a cold-inducible promoter

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Abstract

A new cold-inducible genetic construct was cloned using a chloroplast-specific omega-3-fatty acid desaturase gene (FAD7) under the control of a cold-inducible promoter (cor15a) from Arabidopsis thaliana. RT-PCR confirmed a marked increase in FAD7 expression, in young Nicotiana tabacum (cv. Havana) plants harboring cor15a–FAD7, after a short-term exposure to cold. When young, cold-induced tobacco seedlings were exposed to low-temperature (0.5, 2 or 3.5°C) for up to 44 days, survival within independent cor15a–FAD7 transgenic lines (40.2–96%) was far superior to the wild type (6.7–10.2%). In addition, the major trienoic fatty acid species remained stable in cold-induced cor15a–FAD7 N. tabacum plants under prolonged cold storage while the levels of hexadecatrienoic acid (16:3) and octadecatrienoic acid (18:3) declined in wild type plants under the same conditions (79 and 20.7% respectively). Electron microscopy showed that chloroplast membrane ultrastructure in cor15a–FAD7 transgenic plants was unaffected by prolonged exposure to cold temperatures. In contrast, wild type plants experienced a loss of granal stacking and disorganization of the thylakoid membrane under the same conditions. Changes in membrane integrity coincided with a precipitous decline in leaf chlorophyll concentration and low survival rates in wild type plants. Cold-induced double transgenic N. alata (cv. Domino Mix) plants, harboring both the cor15a–FAD7 cold-tolerance gene and a cor15a–IPT dark-tolerance gene, exhibited dramatically higher survival rates (89–90%) than wild type plants (2%) under prolonged cold storage under dark conditions (2°C for 50 days).

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Abbreviations

16:0:

Hexadecanoic acid

18:0:

Octadecanoic acid

16:1:

Hexadecenoic acid

18:1:

Octadecenoic acid

16:2:

Hexadecadienoic acid

16:3:

Hexadecatrienoic acid

18:2:

Octadecadienoic (linoleic) acid

18:3:

Octadecatrienoic (linolenic) acid

MS:

Murashige-Skoog medium

ω-3:

Position of the double bond from methyl end of a fatty acid

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Acknowledgements

The authors thank Dr. Richard Mercier (Plant Science Department, UCONN) for help in non-radioactive detection of FAD7 gene in transgenic tobacco and Dr. Marie Cantino and the staff at the University of Connecticut Electron Microscope Laboratory for their expertise in microscopic analysis. This research was supported by Connecticut Innovation Inc. (CII).

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Authors and Affiliations

  1. Plant Science Department, University of Connecticut, Storrs, CT, 06269, USA

    Mariya Khodakovskaya, Richard McAvoy, Hao Wu & Yi Li

  2. Institute of Biology and Soil Science, Far-Eastern Branch, Russian Academy of Sciences, 690022, Vladivostok, Russian Federation

    Mariya Khodakovskaya

  3. Plant Biology and Pathology Department, Rutgers, The State University of New Jersey, New Brunswick, NJ, 08901, USA

    Jeanne Peters

Authors
  1. Mariya Khodakovskaya
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  2. Richard McAvoy
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  3. Jeanne Peters
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  4. Hao Wu
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  5. Yi Li
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Correspondence to Richard McAvoy.

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Khodakovskaya, M., McAvoy, R., Peters, J. et al. Enhanced cold tolerance in transgenic tobacco expressing a chloroplast ω-3 fatty acid desaturase gene under the control of a cold-inducible promoter. Planta 223, 1090–1100 (2006). https://doi.org/10.1007/s00425-005-0161-4

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