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Arabinogalactan proteins (AGPs), a superfamily of highly glycosylated hydroxyproline-rich glycoproteins, are widely implicated in plant growth and development. A gene (including its cDNA), designated GhAGP31, encoding a non-classical AGP protein was isolated from cotton (Gossypium hirsutum). The deduced GhAGP31 protein contains the conserved features of non-classical AGPs: a putative signal peptide, N-terminal histidine-rich stretch, middle repetitive proline-rich domain and a cysteine-containing 'PAC' domain. GFP fluorescence assay demonstrated that GhAGP31 protein was localised on cell walls. GhAGP31 transcripts were mainly detected in roots, hypocotyls and ovules, but little or almost none were detected in other tissues. In particular, expression of GhAGP31 was developmentally regulated in roots. Further study demonstrated that GhAGP31 expression in cotton roots was remarkably up-regulated by cold stress. Expression of the GUS gene driven by the GhAGP31 promoter was also dramatically enhanced in roots of transgenic Arabidopsis seedlings under cold treatment. Additionally, overexpression of GhAGP31 in yeast and Arabidopsis significantly improved the freezing tolerance of yeast cells and cold tolerance of Arabidopsis seedlings. These data imply that GhAGP31 protein may be involved in the response to cold stress during early root development of cotton., (© 2011 German Botanical Society and The Royal Botanical Society of the Netherlands.)