Coutts R H, Barnett A, Wood K R
Nucleic Acids Res. 1975 Jul;2(7):1111-21. doi: 10.1093/nar/2.7.1111.
Aspects of the metabolism of RNA have been studied in enzymatically isolated protoplasts from cotyledon and first leaf mesophyll tissue of two cultivars of cucumber. The first leaf mesophyll protoplasts incorporated (3H)-uridine into ribosomal RNA at a constant rate for up to 25 hr in a simple salts medium and for up to 45 hr in a growth medium. Pulse-chase labelling experiments on such preparations showed a rapid dilution of the intracellular (3H)-uridine pool(s) and a high metabolic rate in the cells in one cultivar but not in another. Gel electrophoretic analysis of the RNA from both cotyledon and first leaf protoplasts showed that both protoplast types incorporated either (14C)- or (3H)-uridine into ribosomal RNA species. Incorporation of (3H)-uridine into chloroplasts RNA was minimal in cotyledon protoplasts, but significant in leaf protoplasts. Greater incorporation into the chloroplast RNA species could be achieved by longer pulses. Synthesis of all of the ribosomal RNA species was sensitive to actinomycin D at 10 and 25 mug/ml concentrations in all protoplasts tested.
在从两个黄瓜品种的子叶和第一片叶子叶肉组织中酶解分离得到的原生质体中,对RNA代谢的各个方面进行了研究。在简单盐溶液培养基中,第一片叶子叶肉原生质体以恒定速率将(3H)-尿苷掺入核糖体RNA中长达25小时,在生长培养基中长达45小时。对这些制剂进行的脉冲追踪标记实验表明,在一个品种的细胞中,细胞内(3H)-尿苷池迅速稀释且代谢率高,而在另一个品种中则不然。对子叶和第一片叶子原生质体的RNA进行凝胶电泳分析表明,两种原生质体类型均将(14C)-或(3H)-尿苷掺入核糖体RNA种类中。在子叶原生质体中,(3H)-尿苷掺入叶绿体RNA的量极少,但在叶子原生质体中则很显著。通过更长时间的脉冲可以实现更多地掺入叶绿体RNA种类中。在所有测试的原生质体中,所有核糖体RNA种类的合成在浓度为10和25微克/毫升的放线菌素D作用下均敏感。