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Plant Physiol, March 2000, Vol. 122, pp. 747-756

Genetic Engineering of Glycinebetaine Production toward Enhancing Stress Tolerance in Plants: Metabolic Limitations1

Jun Huang, Rozina Hirji, Luc Adam,2 Kevin L. Rozwadowski,3 Joe K. Hammerlindl, Wilf A. Keller, and Gopalan Selvaraj*

Plant Biotechnology Institute, National Research Council of Canada, Saskatoon, Saskatchewan, Canada S7N 0W9

Glycinebetaine (betaine) affords osmoprotection in bacteria, plants and animals, and protects cell components against harsh conditions in vitro. This and a compelling body of other evidence have encouraged the engineering of betaine production in plants lacking it. We have installed the metabolic step for oxidation of choline, a ubiquitous substance, to betaine in three diverse species, Arabidopsis, Brassica napus, and tobacco (Nicotiana tabacum), by constitutive expression of a bacterial choline oxidase gene. The highest levels of betaine in independent transgenics were 18.6, 12.8, and 13 µmol g-1 dry weight, respectively, values 10- to 20-fold lower than the levels found in natural betaine producers. However, choline-fed transgenic plants synthesized substantially more betaine. Increasing the choline supplementation further enhanced betaine synthesis, up to 613 µmol g-1 dry weight in Arabidopsis, 250 µmol g-1 dry weight in B. napus, and 80 µmol g-1 dry weight in tobacco. These studies demonstrate the need to enhance the endogenous choline supply to support accumulation of physiologically relevant amounts of betaine. A moderate stress tolerance was noted in some but not all betaine-producing transgenic lines based on relative shoot growth. Furthermore, the responses to stresses such as salinity, drought, and freezing were variable among the three species.

1 This paper is National Research Council of Canada Publication no. 43,775.

2 Present address: Mendel Biotechnology, 21375 Cabot Boulevard, Hayward, CA 94545.

3 Present address: Agriculture and Agri-Food Canada, 107 Science Place, Saskatoon, Saskatchewan, Canada S7N 0X2.

* Corresponding author; e-mail gopalan.selvaraj{at}nrc.ca; fax 306-975-4839.

© 2000 American Society of Plant Physiologists


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