Molecular cloning and functional analysis of DoUGE related to water-soluble polysaccharides from Dendrobium officinale with enhanced abiotic stress tolerance

Abstract

UDP glucose 4-epimerase (UGE), an enzyme with significant impacts on sugar metabolism, catalyzes the reversible inter-conversion between UDP-glucose and UDP-galactose. However, very little is known about whether UGE plays a critical role in the accumulation of water-soluble polysaccharide (WSP) and its relationship to abiotic stress tolerance. Here, DoUGE from D. officinale, encoding UGE localized in the cytoplasm, was initially cloned and analyzed. DoUGE exhibited highly tissue-specific expression patterns. The highest expression was in the stems of seedlings and adult plants. The content of WSPs ranged from 168.43 to 416.12 mg g−1 DW from developmental stages S1 to S4, the highest value being in S3. DoUGE was expressed throughout S1 to S4, with a maximum in S3. This trend was similar in three cultivated varieties (T10, T32-5 and T636). There was a positive correlation between DoUGE expression and the content of WSPs (R 2 = 0.94; p < 0.01). Furthermore, promoter analysis showed its possible role in responses to abiotic stresses. Transgenic Arabidopsis thaliana seedlings overexpressing DoUGE accumulated 34.84–44.78% more WSPs, showed 26.24–32.79% more UGE activity, and had a 1.19–1.31-fold higher chlorophyll content than the wild type. Transgenic plants also showed a 50.84 and 34.33% increase in the average content of glucose and galactose, respectively. Transgenic lines growing in half-strength Murashige and Skoog medium containing 150 mM NaCl or 200 mM mannitol displayed enhanced root length and fresh weight, as well as lower proline and malondialdehyde accumulation under salt and osmotic stresses, indicating that the DoUGE gene could be used to improve tolerance to abiotic stress in crops and medicinal or ornamental plants. Our results provide genetic evidence for the involvement of DoUGE in the regulation of WSP content during plant development in D. officinale, as well as in enhanced tolerance to salt and osmotic stresses.