Impaired photoassimilate partitioning caused by phloem-specific removal of pyrophosphate can be complemented by a phloem-specific cytosolic yeast-derived invertase in transgenic plants.
Constitutive expression of the Escherichia coli ppa gene encoding inorganic pyrophosphatase resulted in sugar accumulation in source leaves and stunted growth of transgenic tobacco plants. The reason for this phenotype was hypothesized to be reduced sucrose utilization and loading into the phloem. To study the role of PPi in phloem cells, a chimeric gene was constructed using the phloem-specific rolC promoter of Agrobacterium rhizogenes to drive the expression of the ppa gene. Removal of cytosolic PPi in those cells resulted in photoassimilate accumulation in source leaves, chlorophyll loss, and reduced plant growth. From these data, it was postulated that sucrose hydrolysis via sucrose synthase is essential for assimilate partitioning. To bypass the PPi-dependent sucrose synthase step, transgenic plants were produced that express various levels of the yeast suc2 gene, which encodes cytosolic invertase, in their phloem cells. To combine the phloem-specific expression of the ppa gene and the suc2 gene, crosses between invertase- and pyrophosphatase-containing transgenic plants were performed. Analysis of their offspring revealed that invertase can complement the phenotypic effects caused by the removal of PPi in phloem cells.
Lerchl
J
J
Institut für Pflanzengenetik und Kulturpflanzenforschung, Gatersleben, Germany.
Geigenberger
P
P
Stitt
M
M
Sonnewald
U
U
D001483:Base Sequence; D003600:Cytosol / Q000201:enzymology; D017931:DNA Primers; D011756:Diphosphates / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015870:Gene Expression; D005816:Genetic Complementation Test; D006026:Glycoside Hydrolases / Q000378:metabolism*; D007150:Immunohistochemistry; D043564:Inorganic Pyrophosphatase; D008954:Models, Biological; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified / Q000502:physiology*; D010947:Plants, Toxic; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011755:Pyrophosphatases / Q000096:biosynthesis* / Q000235:genetics; D012231:Rhizobium / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D014026:Nicotiana / Q000502:physiology; D043324:beta-Fructofuranosidase
D017931:DNA Primers; D011756:Diphosphates; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase; D011755:Pyrophosphatases; D043564:Inorganic Pyrophosphatase