[Cloning and functional analysis of trehalose -6-phosphate synthase gene from Selaginella pulvinata]

Trehalose-6-phosphate synthase, a key enzyme in trehalose synthesis pathway of plant, plays an important role in response to abiotic stress in xerophilous rock lily and other resurrection plants. In this study, homologous amplification and RACE technique were used to clone gene SpTPS1 for trehalose-6-phosphate synthase from Selaginella pulvinata, which is an endemic xerophilous plant in China. The full-length cDNA is 3223 bp long, containing an open reading frame (ORF) of 2790 bp. Protein sequence comparison showed that the pupative amino acid sequence of this ORF shares high similarity to trehalose-6-phosphate synthases of mode species, especially at the conserved sites of catalytic activity centers. Yeast functional complementation test showed that trehalose-6-phosphate synthase mutant (tps1 big up tri, open), transformed by the ORF of SpTPS1 gene, can restore growth on the medium supplemented with glucose as a sole carbon source. This result indicated that SpTPS1 of S. pulvinata encodes for an active protein and is hopeful to be applied in transgenic improvement of abiotic stress tolerance in plant.