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Pyrimidine Nucleoside Uptake by Petunia Pollen

Specificity and Inhibitor Studies on the Carrier-Mediated Transport

Department of Agricultural Biochemistry, Waite Agricultural Research Institute, University of Adelaide, Glen Osmond, South Australia 5064

Transport of pyrimidine nucleosides into germinating Petunia hybrida pollen is carrier-mediated, and, except for thymidine, is inhibited by the energy poisons N,N'-dicyclohexylcarbodiimide, 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole, 2,4-dinitrophenol, and carbonylcyanide-m-chlorophenylhydrazone. Kinetic studies with analogs deoxyuridine and 5-bromodeoxyuridine show that they too are taken up faster than thymidine and inhibited by the energy poisons. These and other analogs inhibit uridine and cytidine transport more than thymidine, as do the inhibitors parachloromercuribenzoic acid, N-ethylmaleimide, phenylarsine oxide, o-phenanthroline, ethylene diamenetetraacetate, and ethylene glycol-bis (-aminoethyl ether) N,N,N'N'-tetraacetic acid. Citrate, phosphate, succinate, and tartrate inhibited uptake of all pyrimidine nucleosides. The specific inhibitor of nucleoside transport in animal cells, nitrobenzylthioinosine, has little effect on pollen transport. Uridine and deoxyuridine accumulate against a concentration gradient, suggesting active transport. Except for thymidine, however, transported nucleosides were found to be extensively phosphorylated. Until mutant plants are found which do not phosphorylate uridine, it is not possible to decide unequivocally between active and nonactive transport for uridine. However, consistent with a low level of DNA synthesis in germinating Petunia pollen, it is clear that thymidine transport is nonactive and relatively slow. It is apparent from these experiments that a more sensitive way to study DNA repair in this pollen would be to use 5-bromodeoxyuridine or deoxyuridine instead of thymidine to label repaired DNA. The results show that pollen has the transport systems necessary to take up pyrimidine nucleosides from Petunia styles, where it is known that the concentration of free nucleosides increase after pollination.

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