A Selaginella lepidophylla trehalose-6-phosphate synthase complements growth and stress-tolerance defects in a yeast tps1 mutant.
The accumulation of the disaccharide trehalose in anhydrobiotic organisms allows them to survive severe environmental stress. A plant cDNA, SlTPS1, encoding a 109-kD protein, was isolated from the resurrection plant Selaginella lepidophylla, which accumulates high levels of trehalose. Protein-sequence comparison showed that SlTPS1 shares high similarity to trehalose-6-phosphate synthase genes from prokaryotes and eukaryotes. SlTPS1 mRNA was constitutively expressed in S. lepidophylla. DNA gel-blot analysis indicated that SlTPS1 is present as a single-copy gene. Transformation of a Saccharomyces cerevisiae tps1Delta mutant disrupted in the ScTPS1 gene with S. lepidophylla SlTPS1 restored growth on fermentable sugars and the synthesis of trehalose at high levels. Moreover, the SlTPS1 gene introduced into the tps1Delta mutant was able to complement both deficiencies: sensitivity to sublethal heat treatment at 39 degrees C and induced thermotolerance at 50 degrees C. The osmosensitive phenotype of the yeast tps1Delta mutant grown in NaCl and sorbitol was also restored by the SlTPS1 gene. Thus, SlTPS1 protein is a functional plant homolog capable of sustaining trehalose biosynthesis and could play a major role in stress tolerance in S. lepidophylla.
Zentella
R
R
Departamento de Biología Molecular de Plantas, Instituto de Biotecnología-Universidad Nacional Autónoma de México, Avenida Universidad 2001, Colonia Chamilpa, 62210 Cuernavaca Morelos, Mexico.
Mascorro-Gallardo
J O
JO
Van Dijck
P
P
Folch-Mallol
J
J
Bonini
B
B
Van Vaeck
C
C
Gaxiola
R
R
Covarrubias
A A
AA
Nieto-Sotelo
J
J
Thevelein
J M
JM
Iturriaga
G
G
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D015870:Gene Expression; D005800:Genes, Fungal; D017343:Genes, Plant; D005816:Genetic Complementation Test; D005964:Glucosyltransferases / Q000235:genetics* / Q000378:metabolism*; D006358:Hot Temperature; D008969:Molecular Sequence Data; D009154:Mutation; D010944:Plants / Q000201:enzymology* / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D017386:Sequence Homology, Amino Acid; D014199:Trehalose / Q000096:biosynthesis
D017931:DNA Primers; D018076:DNA, Complementary; D018744:DNA, Plant; D014199:Trehalose; D005964:Glucosyltransferases; C023180:trehalose-6-phosphate synthase