Upregulation of pyrophosphate: fructose 6-phosphate 1-phosphotransferase (PFP) activity in strawberry.
Pyrophosphate: fructose 6-phosphate 1-phosphotransferase (PFP) is a cytosolic enzyme catalyzing the first committed step in glycolysis by reversibly phosphorylating fructose-6-phosphate to fructose-1,6-bisphosphate. The position of PFP in glycolytic and gluconeogenic metabolism, as well as activity patterns in ripening strawberry, suggest that the enzyme may influence carbohydrate allocation to sugars and organic acids. Fructose-2,6-bisphosphate activates and tightly regulates PFP activity in plants and has hampered attempts to increase PFP activity through overexpression. Heterologous expression of a homodimeric isoform from Giardia lamblia, not regulated by fructose-2,6-bisphosphate, was therefore employed to ensure in vivo increases in PFP activity. The coding sequence was placed into a constitutive expression cassette under control of the cauliflower mosaic virus 35S promoter and introduced into strawberry by Agrobacterium tumefaciens-mediated transformation. Heterologous expression of PFP resulted in an up to eightfold increase in total activity in ripe berries collected over two consecutive growing seasons. Total sugar and organic acid content of transgenic berries harvested during the first season were not affected when compared to the wild type, however, fructose content increased at the expense of sucrose. In the second season, total sugar content and composition remained unchanged while the citrate content increased slightly. Considering that PFP catalyses a reversible reaction, PFP activity appears to shift between gluconeogenic and glycolytic metabolism, depending on the metabolic status of the cell.
Basson
C E
CE
Institute for Plant Biotechnology, Genetics Department, Stellenbosch University, Private Bag X1 Matieland, Stellenbosch, 7602, South Africa.
Groenewald
J-H
JH
Kossmann
J
J
Cronjé
C
C
Bauer
R
R
D031985:Fragaria / Q000201:enzymology* / Q000235:genetics; D005635:Fructosediphosphates / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D005822:Genetic Vectors; D010732:Phosphofructokinase-1 / Q000235:genetics / Q000378:metabolism*; D012641:Selection, Genetic; D015854:Up-Regulation
D005635:Fructosediphosphates; C027652:fructose 2,6-diphosphate; D010732:Phosphofructokinase-1