Antisense inhibition of cytosolic phosphorylase in potato plants (Solanum tuberosum L.) affects tuber sprouting and flower formation with only little impact on carbohydrate metabolism.
To determine the function of cytosolic phosphorylase (Pho2; EC 2.4.1.1), transgenic potato plants were created in which the expression of the enzyme was inhibited by introducing a chimeric gene containing part of the coding region for cytosolic phosphorylase linked in antisense orientation to the 35S CaMV promotor. As revealed by Northern blot analysis and native polyacrylamide gel electrophoresis, the expression of cytosolic phosphorylase was strongly inhibited in both leaves and tubers of the transgenic plants. The transgenic plants propagated from stem cuttings were morphologically indiscernible from the wild-type. However, sprouting of the transgenic potato tubers was significantly altered: compared with the wild-type, transgenic tubers produced 2.4 to 8.1 times more sprouts. When cultivated in the greenhouse, transgenic seed tubers produced two to three times more shoots than the wild-type. Inflorescences appeared earlier in the resulting plants. Many of the transgenic plants flowered two or three times successively. Transgenic plants derived from seed tubers formed 1.6 to 2.4 times as many tubers per plant as untransformed controls. The size and dry matter content of the individual tubers was not noticeably altered. Tuber yield was significantly higher in the transgenic plants. As revealed by carbohydrate determination of freshly harvested and stored tubers, starch and sucrose pools were not noticeably affected by the antisense inhibition of cytosolic phosphorylase; however, glucose and fructose levels were markedly reduced after prolonged storage. These results favour the view that cytosolic phosphorylase does not participate in starch degradation. The possible links between the reduced levels of cytosolic phosphorylase and the observed changes with respect to sprouting and flowering are discussed.
Duwenig
E
E
Institut für Genbiologische Forschung Berlin GmbH, Berlin, Germany.
Steup
M
M
Willmitzer
L
L
Kossmann
J
J
D016960:Agrobacterium tumefaciens; D001483:Base Sequence; D050260:Carbohydrate Metabolism; D003600:Cytosol / Q000201:enzymology; D004926:Escherichia coli; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D006005:Phosphorylases / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000378:metabolism; D018517:Plant Roots; D018520:Plant Shoots; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011198:Solanum tuberosum / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*
D016376:Oligonucleotides, Antisense; D011993:Recombinant Fusion Proteins; D006005:Phosphorylases