Inhülsen D, Niemeyer R
Institut für Botanik der Technischen Universität Hannover, Herrenhäuser Str. 2, D-3000, Hannover, Federal Republic of Germany.
Planta. 1975 Jan;124(2):159-67. doi: 10.1007/BF00384758.
After previous incubation in a P-free culture medium whole plants of Lemna minor L. were given [(32)P]-orthophosphate in the logarithmic and in the stationary growth phase. The condensed phosphates could be separated from the nucleic acids by fractionation on methylated serum albumin and Kieselgur (MAK). The various inorganic condensed phosphates were eluted before the nucleic acids. The former were dialyzed and separated by two-dimensional thin layer chromatography on a mixture of cellulose and microcrystalline cellulose using an acid and an alkaline solvent in succession. (32)P-radioactivity was present in all linear oligophosphates containing 2 to 7 residues, in cyclic metaphosphates (tri-, tetra-, penta- and hexametaphosphates) and in high-molecular condensed phosphates which remained at the starting point.Among the low-molecular inorganic oligophosphates the trimetaphosphate contained significantly more radioactivity than the other compounds. The labeling rate of (32)P-incorporation depended on the previous depletion of phosphate. In the logarithmic phase maximal labeling occurred after 4 h following a 10 h period of phosphate depletion; in the stationary phase the maximum rate was reached 5 h after the end of a 4.5 h period of phosphate depletion. The results indicate that the phosphate pool was restored first and that the nucleic acids were labeled subsequently. That is to say: in contrast to previous findings by other authors, our results show that net synthesis of inorganic condensed phosphates does takes place during the logarithmic phase.After cultivation of Lemna for 24 h or longer in a P-free medium the DNA incorporated more (32)P than the other nucleic acids during the logarithmic phase. During the stationary phase [(32)P]-orthophosphate incorporation into nucleic acids was markedly lower than during the logarithmic phase.
在无磷培养基中预培养后,给处于对数生长期和稳定生长期的小浮萍(Lemna minor L.)全株施加[³²P] - 正磷酸盐。通过在甲基化血清白蛋白和硅藻土(MAK)上进行分级分离,可将缩合磷酸盐与核酸分离。各种无机缩合磷酸盐在核酸之前被洗脱。将前者进行透析,并在纤维素和微晶纤维素的混合物上先后使用酸性和碱性溶剂进行二维薄层色谱分离。在所有含有2至7个残基的线性寡磷酸盐、环状偏磷酸盐(三聚、四聚、五聚和六聚偏磷酸盐)以及留在起始点的高分子缩合磷酸盐中都存在³²P放射性。在低分子无机寡磷酸盐中,三聚偏磷酸盐含有的放射性明显高于其他化合物。³²P掺入的标记率取决于先前的磷耗尽情况。在对数生长期,经过10小时的磷耗尽后,4小时后出现最大标记;在稳定生长期,在4.5小时的磷耗尽期结束后5小时达到最大速率。结果表明,首先恢复的是磷库,随后核酸被标记。也就是说:与其他作者先前的发现相反,我们的结果表明,无机缩合磷酸盐的净合成确实发生在对数生长期。在无磷培养基中培养小浮萍24小时或更长时间后,在对数生长期,DNA比其他核酸掺入更多的³²P。在稳定生长期,[³²P] - 正磷酸盐掺入核酸的量明显低于对数生长期。