Rapid accumulation of polyphosphate in extraradical hyphae of an arbuscular mycorrhizal fungus as revealed by histochemistry and a polyphosphate kinase/luciferase system
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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DOI：
10.1046/j.1469-8137.2003.00966.x
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发表时间：
2004-02-01
期刊：
影响因子：
9.4
通讯作者：
Ohtomo, R
中科院分区：
文献类型：
--
作者：
Ezawa, T;Cavagnaro, TR;Ohtomo, R
用常规组织化学和一种新的利用细菌酶--聚磷酸激酶的酶促方法研究了丛枝菌根真菌根外菌丝中聚磷酸盐的积累速率。万寿菊(万寿菊Patula cv.)Bonanza Orange)接种细小孢子菌(Archaeospora Leupticha)，在缺磷条件下生长。分别于施磷后0、1、3、24 h采集根外菌丝。息肉水平通过甲苯胺蓝O异染色法和将息肉转化为三磷酸腺苷的多聚磷酸酶结合三磷酸腺苷荧光素酶测定来评估。含异色颗粒的菌丝百分率从0~1h增加到25%~44%，3h达到最大值50%，聚磷酸盐含量在施磷后1~3h以46.4+/-15.1nmolmin(-1)mg(-1)的速率增加一倍。菌丝中聚磷酸盐的积累速度比聚磷酸盐超积累微生物的速度快得多。本研究中采用的酶促方法可以对菌根系统中的聚磷酸盐进行高度特异和灵敏的评估。
The rate of polyphosphate accumulation in extraradical hyphae of an arbuscular mycorrhizal fungus was investigated by conventional histochemistry and a new enzymatic method using a bacterial enzyme, polyphosphate kinase. Marigold (Tagetes patula cv. Bonanza Orange) was inoculated with Archaeospora leptoticha and grown under P-deficient conditions. Extraradical hyphae were harvested at 0, 1, 3 and 24 h after 1 mM P-application. PolyP levels were assessed by both metachromasy of Toluidine blue O and polyphosphate kinase which converted polyP to ATP followed by the ATP-luciferase assay. Percentage of hyphae with metachromatic granules was increased from 25 to 44% from 0 to 1 h, and a maximum of 50% was reach by 3 h. Polyphosphate content was doubled from 1 to 3 h after P-application (4.8-10.0 mol as Pi mg(-1) protein) at a rate of 46.4 +/- 15.1 nmol min(-1) mg(-1). The rate of polyphosphate accumulation in the hyphae was surprisingly rapid as those of polyphosphate-hyper accumulating microorganisms. The enzymatic method employed in the present study allows highly specific and sensitive assessment of polyphosphate in the mycorrhizal system.