Downregulation of pyrophosphate: D-fructose-6-phosphate 1-phosphotransferase activity in sugarcane culms enhances sucrose accumulation due to elevated hexose-phosphate levels.
Analyses of transgenic sugarcane clones with 45-95% reduced cytosolic pyrophosphate: D-fructose-6-phosphate 1-phosphotransferase (PFP, EC 2.7.1.90) activity displayed no visual phenotypical change, but significant changes were evident in in vivo metabolite levels and fluxes during internode development. In three independent transgenic lines, sucrose concentrations increased between three- and sixfold in immature internodes, compared to the levels in the wildtype control. There was an eightfold increase in the hexose-phosphate:triose-phosphate ratio in immature internodes, a significant restriction in the triose phosphate to hexose phosphate cycle and significant increase in sucrose cycling as monitored by (13)C nuclear magnetic resonance. This suggests that an increase in the hexose-phosphate concentrations resulting from a restriction in the conversion of hexose phosphates to triose phosphates drive sucrose synthesis in the young internodes. These effects became less pronounced as the tissue matured. Decreased expression of PFP also resulted in an increase of the ATP/ADP and UTP/UDP ratios, and an increase of the total uridine nucleotide and, at a later stage, the total adenine nucleotide pool, revealing strong interactions between PPi metabolism and general energy metabolism. Finally, decreased PFP leads to a reduction of PPi levels in older internodes indicating that in these developmental stages PFP acts in the gluconeogenic direction. The lowered PPi levels might also contribute to the absence of increases in sucrose contents in the more mature tissues of transgenic sugarcane with reduced PFP activity.
van der Merwe
Margaretha J
MJ
Institute of Plant Biotechnology, University of Stellenbosch, Merriman Avenue, Stellenbosch 7602, South Africa. mvdm@sun.ac.za
Groenewald
Jan-Hendrik
JH
Stitt
Mark
M
Kossmann
Jens
J
Botha
Frederik C
FC
D002851:Chromatography, High Pressure Liquid; D015536:Down-Regulation; D008401:Gas Chromatography-Mass Spectrometry; D006600:Hexosephosphates / Q000378:metabolism*; D019906:Nuclear Magnetic Resonance, Biomolecular; D010770:Phosphotransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000378:metabolism; D031786:Saccharum / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D013395:Sucrose / Q000378:metabolism*
D006600:Hexosephosphates; D010940:Plant Proteins; D013395:Sucrose; D010770:Phosphotransferases; C022502:pyrophosphate-fructose 6-phosphate 1-phosphotransferase