Antisense repression of cytosolic phosphoglucomutase in potato (Solanum tuberosum) results in severe growth retardation, reduction in tuber number and altered carbon metabolism.
The aim of this work was to investigate the role of cytosolic phosphoglucomutase (PGM; EC 5.4.2.2) in the regulation of carbohydrate metabolism. Many in vitro studies have indicated that PGM plays a central role in carbohydrate metabolism; however, until now the importance of this enzyme in plants has not been subject to reverse-genetics investigations. With this intention we cloned the cytosolic isoform of potato PGM (StcPGM) and expressed this in the antisense orientation under the control of the CaMV 35 S promoter in potato plants. We confirmed that these plants contained reduced total PGM activity and that loss in activity was due specifically to a reduction in cytosolic PGM activity. These plants were characterised by a severe phenotype: stunted aerial growth combined with limited root growth and a reduced tuber yield. Analysis of the metabolism of these lines revealed that leaves of these plants were inhibited in sucrose synthesis whereas the tubers exhibited decreased levels of sucrose and starch as well as decreased levels of glycolytic intermediates but possessed unaltered levels of adenylates. Furthermore, a broader metabolite screen utilising GC-MS profiling revealed that these lines contained altered levels of several intermediates of the TCA cycle and of amino acids. In summary, we conclude that cytosolic PGM plays a crucial role in the sucrose synthetic pathway within the leaf and in starch accumulation within the tuber, and as such is important in the maintenance of sink-source relationships.
Fernie
Alisdair R
AR
Max-Planck-Institut für Molekulare Pflanzenphysiologie, Golm, Germany. fernie@mpimp-golm.mpg.de
Tauberger
Eva
E
Lytovchenko
Anna
A
Roessner
Ute
U
Willmitzer
Lothar
L
Trethewey
Richard N
RN
D000596:Amino Acids / Q000378:metabolism; D050260:Carbohydrate Metabolism; D002244:Carbon / Q000378:metabolism*; D003001:Cloning, Molecular; D003600:Cytosol; D016373:DNA, Antisense / Q000235:genetics*; D051599:Glucose-1-Phosphate Adenylyltransferase; D005964:Glucosyltransferases / Q000378:metabolism; D006019:Glycolysis; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D009713:Nucleotidyltransferases / Q000378:metabolism; D010641:Phenotype; D010733:Phosphoglucomutase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010788:Photosynthesis / Q000502:physiology; D018515:Plant Leaves / Q000201:enzymology / Q000378:metabolism; D018547:Plant Stems / Q000201:enzymology* / Q000235:genetics; D030821:Plants, Genetically Modified; D011755:Pyrophosphatases / Q000378:metabolism; D011198:Solanum tuberosum / Q000201:enzymology* / Q000235:genetics; D013213:Starch / Q000378:metabolism; D013395:Sucrose / Q000378:metabolism
D000596:Amino Acids; D016373:DNA, Antisense; D007527:Isoenzymes; D013395:Sucrose; D002244:Carbon; D013213:Starch; D005964:Glucosyltransferases; C022699:sucrose synthase; D009713:Nucleotidyltransferases; D051599:Glucose-1-Phosphate Adenylyltransferase; D011755:Pyrophosphatases; C023255:UDPglucose pyrophosphatase; D010733:Phosphoglucomutase