Murai N, Armstrong D J, Taller B J, Skoog F
Institute of Plant Development, Birge Hall, University of Wisconsin, Madison, Wisconsin 53706.
Plant Physiol. 1978 Mar;61(3):318-22. doi: 10.1104/pp.61.3.318.
The distribution of incorporated synthetic cytokinins (N(6)-[8-(14)C]benzyladenine ([8-(14)C]bzl(6)Ade) and N(6)[8-(14)C]furfuryladenine ([8-(14)C]fr(6)Ade) in ribosomal RNA prepared from tobacco callus (Nicotiana tabacum L. var. Wis. No. 38) grown in the presence of one of these for 25 or 26 days has been studied. The rRNA of tissue supplied with [8-(14)C]bzl(6)Ade or [8-(14)C]fr(6)Ade was fractionated by methylated albumin-Kieselguhr column chromatography and preparative gel electrophoresis, respectively. In each case about 80% of the incorporated cytokinin was recovered as the ribonucleoside [8-(14)C]bzl(6)A or [8-(14)C]fr(6)A in the rRNA peak after the fractionations. [8-(14)C]fr(6)A was found associated with both the 18S and 25S rRNA components in quantities roughly proportional to their 260 nm absorbance. This pattern of apparently nonspecific association was not affected by prior denaturation of the RNA with formamide.The distribution of [8-(14)C]fr(6)A moieties in mono- and oligonucleotides derived from combined treatment of [8-(14)C]fr(6)Ade-labeled rRNA with ribonuclease T(1) and pancreatic ribonuclease A was measured by fractionating the digest on a DEAE-cellulose column in the presence of 7 m urea and determining the [8-(14)C]fr(6)A content in each fraction. The [8-(14)C]fr(6)A content in the oligonucleotides varied from 46 to 210 mumol/mol of adenosine (A). The mol ratio of [8-(14)C]fr(6)A to A was three to four times greater for oligonucleotides containing uridine or cytidine (AUp or ACp) than for those containing quanosine (AGp).
研究了在含有合成细胞分裂素之一的条件下培养25或26天的烟草愈伤组织(烟草Wis. No. 38变种)制备的核糖体RNA中掺入的合成细胞分裂素(N(6)-[8-(14)C]苄基腺嘌呤([8-(14)C]bzl(6)Ade)和N(6)[8-(14)C]糠基腺嘌呤([8-(14)C]fr(6)Ade)的分布情况。分别通过甲基化白蛋白-硅藻土柱色谱法和制备性凝胶电泳法对供应[8-(14)C]bzl(6)Ade或[8-(14)C]fr(6)Ade的组织的rRNA进行分级分离。在每种情况下,分级分离后,约80%掺入的细胞分裂素以核糖核苷[8-(14)C]bzl(6)A或[8-(14)C]fr(6)A的形式在rRNA峰中回收。发现[8-(14)C]fr(6)A与18S和25S rRNA组分均有关联,其数量大致与其在260nm处的吸光度成比例。这种明显非特异性关联的模式不受用甲酰胺对RNA进行预先变性的影响。通过在7m尿素存在下在DEAE-纤维素柱上对[8-(14)C]fr(6)Ade标记的rRNA与核糖核酸酶T(1)和胰核糖核酸酶A联合处理得到的单核苷酸和寡核苷酸中[8-(14)C]fr(6)A部分的分布进行测量,并测定每个级分中的[8-(14)C]fr(6)A含量。寡核苷酸中[8-(14)C]fr(6)A的含量在46至210μmol/mol腺苷(A)之间变化。含有尿苷或胞苷(AUp或ACp)的寡核苷酸中[8-(14)C]fr(6)A与A的摩尔比是含有鸟苷(AGp)的寡核苷酸的三至四倍。