Cloning and characterization of a novel fructan 6-exohydrolase strongly inhibited by sucrose in Lolium perenne

MAIN CONCLUSION : The first 6-fructan exohydrolase (6-FEH) cDNA from Lolium perenne was cloned and characterized. Following defoliation, Lp6 - FEHa transcript level unexpectedly decreased together with an increase in total FEH activity. Lolium perenne is a major forage grass species that accumulates...

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Bibliographic Details
Published in:Planta 2014-09, Vol.240 (3), p.629-643
Main Authors: Lothier, Jérémy, Van Laere, André, Prud’homme, Marie-Pascale, Van den Ende, Wim, Morvan-Bertrand, Annette
Format: Article
Language:English
Subjects:
Agriculture
Amino Acid Sequence
Amino acids
Barley
Biomedical and Life Sciences
biosynthesis
cDNA libraries
Cell walls
Complementary DNA
Defoliation
Ecology
Enzymes
forage grasses
Forestry
Fructans
fructose
genes
Glycoside Hydrolases - genetics
Glycoside Hydrolases - metabolism
grasslands
Leaf sheaths
leaves
Life Sciences
Lolium - enzymology
Lolium - genetics
Lolium perenne
Molecular Sequence Data
Original Article
Pichia
Pichia pastoris
Plant Proteins - genetics
Plant Proteins - metabolism
Plant Sciences
Plants
Recombinant Proteins - metabolism
Regrowth
sucrose
Sucrose - metabolism
Vegetal Biology
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Summary:MAIN CONCLUSION : The first 6-fructan exohydrolase (6-FEH) cDNA from Lolium perenne was cloned and characterized. Following defoliation, Lp6 - FEHa transcript level unexpectedly decreased together with an increase in total FEH activity. Lolium perenne is a major forage grass species that accumulates fructans, mainly composed of β(2,6)-linked fructose units. Fructans are mobilized through strongly increased activities of fructan exohydrolases (FEHs), sustaining regrowth following defoliation. To understand the complex regulation of fructan breakdown in defoliated grassland species, the objective was to clone and characterize new FEH genes in L. perenne. To find FEH genes related to refoliation, a defoliated tiller base cDNA library was screened. Characterization of the recombinant protein was performed in Pichia pastoris. In this report, the cloning and enzymatic characterization of the first 6-FEH from L. perenne is described. Following defoliation, during fructan breakdown, Lp6-FEHa transcript level unexpectedly decreased in elongating leaf bases (ELB) and in mature leaf sheaths (tiller base) in parallel to increased total FEH activities. In comparison, transcript levels of genes coding for fructosyltransferases (FTs) involved in fructan biosynthesis also decreased after defoliation but much faster than FEH transcript levels. Since Lp6-FEHa was strongly inhibited by sucrose, mechanisms modulating FEH activities are discussed. It is proposed that differences in the regulation of FEH activity among forage grasses influence their tolerance to defoliation.
ISSN:0032-0935
1432-2048
DOI:10.1007/s00425-014-2110-6