Rapid accumulation of polyphosphate in extraradical hyphae of an arbuscular mycorrhizal fungus as revealed by histochemistry and a polyphosphate kinase/luciferase system

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The definitive version is available at www.blackwellsynergy.comThe rate of polyphosphate accumulation in extraradical hyphae of an arbuscularmycorrhizal fungus was investigated by conventional histochemistry and a newenzymatic method using a bacterial enzyme, polyphosphate kinase. Marigold (Tagetespatulacv. Bonanza Orange) was inoculated withArchaeospora leptotichaandgrown under P-deficient conditions. Extraradical hyphae were harvested at 0, 1, 3and 24 h after 1 mMP-application. PolyP levels were assessed by both metachromasyof Toluidine blue O and polyphosphate kinase which converted polyP to ATPfollowed by the ATP-luciferase assay. Percentage of hyphae with metachromaticgranules was increased from 25 to 44% from 0 to 1 h, and a maximum of 50% wasreach by 3 h. Polyphosphate content was doubled from 1 to 3 h after P-application(4.8–10.0 μmol as Pi mg−1protein) at a rate of 46.4±15.1 nmol min−1mg−1. Therate of polyphosphate accumulation in the hyphae was surprisingly rapid as thoseof polyphosphate-hyper accumulating microorganisms. The enzymatic methodemployed in the present study allows highly specific and sensitive assessment ofpolyphosphate in the mycorrhizal system.
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