Watanabe A, Imaseki H
Institute for Biochemical Regulation, Faculty of Agriculture, Nagoya University, Nagoya, Japan.
Plant Physiol. 1973 Apr;51(4):772-6. doi: 10.1104/pp.51.4.772.
Incorporation of (32)P-orthophosphate was found in the DNA fraction of aerobically incubated potato discs when examined by methylated albumin kieselguhr column chromatography. The estimation of DNA content of the discs was by a method developed for starchy tissues and showed that the incorporation of (32)P was due to net synthesis of DNA. The DNA content of a disc rapidly increased after a lag period of about 12 hours. The increase continued during the entire test period although at a lower rate during the later period of aging. DNA synthesis was further examined by measuring the rate of incorporation of (3)H-thymidine. The striking similarity which was found between changes in the rate of DNA accumulation and in the rate of (3)H-thymidine incorporation indicates that the incorporation of (3)H-thymidine actually represents the net synthesis of DNA. Although the experiments with microautoradiography revealed that DNA synthesis occurred exclusively in nuclei, no signs of cell division were detected by microscopic observation. DNA synthesis in potato discs was further examined by using inhibitors of protein and RNA synthesis and was sensitive to those inhibitors. The significance of the present results is discussed in relation to the role of wounding in the induction of DNA synthesis.
通过甲基化白蛋白硅藻土柱色谱法检测发现,在需氧培养的马铃薯圆片中,DNA部分含有掺入的(32)P-正磷酸盐。圆片DNA含量的测定采用了一种针对淀粉质组织开发的方法,结果表明(32)P的掺入是由于DNA的净合成。在约12小时的延迟期后,圆片的DNA含量迅速增加。在整个测试期间,这种增加持续存在,尽管在老化后期速率较低。通过测量(3)H-胸腺嘧啶核苷的掺入速率进一步研究了DNA合成。DNA积累速率和(3)H-胸腺嘧啶核苷掺入速率变化之间的显著相似性表明(3)H-胸腺嘧啶核苷的掺入实际上代表了DNA的净合成。尽管显微放射自显影实验显示DNA合成仅发生在细胞核中,但通过显微镜观察未检测到细胞分裂的迹象。通过使用蛋白质和RNA合成抑制剂进一步研究了马铃薯圆片中的DNA合成,结果表明其对这些抑制剂敏感。本文就创伤在DNA合成诱导中的作用对当前结果的意义进行了讨论。