Molecular characterization of a putative sucrose:fructan 6-fructosyltransferase (6-SFT) of the cold-resistant Patagonian grass Bromus pictus associated with fructan accumulation under low temperatures.
Fructans are fructose polymers synthesized from sucrose in the plant vacuole. They represent short- and long-term carbohydrate reserves and have been associated with abiotic stress tolerance in graminean species. We report the isolation and characterization of a putative sucrose:fructan 6-fructosyltransferase (6-SFT) gene from a Patagonian grass species, Bromus pictus, tolerant to drought and cold temperatures. Structural and functional analyses of this gene were performed by Southern and Northern blot. Sugar content, quality and fructosyltransferase activity were studied using HPAEC-PAD (high-pH anion-exchange chromatography with pulsed amperometric detection), enzymatic and colorimetric assays. The putative 6-SFT gene had all the conserved motifs of fructosyl-transferase and showed 90% identity at the amino acid level with other 6-SFTs from winter cereals. Expression studies, and determination of sugar content and fructosyl-transferase activity were performed on five sections of the leaf. Bp6-SFT was expressed predominantly in leaf bases, where fructosyltransferase activity and fructan content are higher. Bp6-SFT expression and accumulation of fructans showed different patterns in the evaluated leaf sections during a 7 d time course experiment under chilling treatment. The transcriptional pattern suggests that the B. pictus 6-SFT gene is highly expressed in basal leaf sections even under control temperate conditions, in contrast to previous reports in other graminean species. Low temperatures caused an increase in Bp6-SFT expression and fructan accumulation in leaf bases. This is the first study of the isolation and molecular characterization of a fructosyltransferase in a native species from the Patagonian region. Expression in heterologous systems will confirm the functionality, allowing future developments in generation of functional markers for assisted breeding or biotechnological applications.
del Viso
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Instituto de Biotecnología, CICVyA, Instituto Nacional de Tecnología Agropecuaria (INTA, Castelar), (1686), Hurlingham, Buenos Aires, Argentina.
Puebla
A F
AF
Fusari
C M
CM
Casabuono
A C
AC
Couto
A S
AS
Pontis
H G
HG
Hopp
H E
HE
Heinz
R A
RA
D000595:Amino Acid Sequence; D031724:Bromus / Q000201:enzymology / Q000235:genetics*; D003001:Cloning, Molecular; D003080:Cold Temperature; D018076:DNA, Complementary / Q000235:genetics; D005630:Fructans / Q000096:biosynthesis*; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D006602:Hexosyltransferases / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D018749:RNA, Plant / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D005630:Fructans; D010940:Plant Proteins; D018749:RNA, Plant; D006602:Hexosyltransferases; C097181:sucrose - fructan 6-fructosyltransferase