Expression of E. coli inorganic pyrophosphatase in transgenic plants alters photoassimilate partitioning.
Transgenic plants were constructed expressing a novel cytosolic inorganic pyrophosphatase in order to reduce the cytosolic pyrophosphate content. To this end the Escherichia coli gene ppa encoding inorganic pyrophosphatase was cloned between the 35S CaMV promoter and the poly(A) site of the octopine synthase gene and transferred into tobacco and potato plants by Agrobacterium-mediated gene transfer. Regenerated plants were tested for the expression of the ppa gene by Northern blots and activity gels. Plants expressing active inorganic pyrophosphatase showed a dramatic change in photoassimilate partitioning. In both transgenic tobacco and potato plants the ratio between soluble sugars and starch was increased by about 3-4-fold in source leaves as compared with the wild-type. However, whereas source leaves of transgenic tobacco plants accumulated much higher levels of glucose (up to 68-fold), fructose (up to 24-fold), sucrose (up to 12-fold) and starch (up to 8-fold) this was not observed in potato plants where the change in assimilate partitioning in source leaves was due to an increase of about 2-fold in sucrose and a reduction in starch content. Expression of the cytosolic inorganic pyrophosphatase in tobacco results in stunted growth of vegetatively growing plants due to a reduced internode distance. Upon flowering the transgenic plants increase their growth rate, reaching almost the same height as control plants at the end of the growth period. Old source leaves accumulate up to 100-fold more soluble sugars than control leaves. This increase in soluble sugars is accompanied by a reduction in chlorophyll content (up to 85%). Transgenic potato plants showed a less dramatic change in their growth behaviour. Plants were slightly reduced in size, with stems more highly branched. Tuber number increased 2-3-fold, but tuber weight was lower resulting in no net increase in fresh weight.
Institut für Genbiologische Forschung Berlin GmbH, Germany.
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Sonnewald
D001483:Base Sequence; D050260:Carbohydrate Metabolism; D003001:Cloning, Molecular; D017931:DNA Primers; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D043564:Inorganic Pyrophosphatase; D008969:Molecular Sequence Data; D010788:Photosynthesis; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000378:metabolism*; D010947:Plants, Toxic; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011755:Pyrophosphatases / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D012231:Rhizobium; D011198:Solanum tuberosum / Q000201:enzymology / Q000235:genetics; D014026:Nicotiana / Q000201:enzymology / Q000235:genetics; D014162:Transfection
D017931:DNA Primers; D011755:Pyrophosphatases; D043564:Inorganic Pyrophosphatase