Bennett J, Jenkins G I, Hartley M R
J Cell Biochem. 1984;25(1):1-13. doi: 10.1002/jcb.240250102.
The photoregulation of chloroplast development in pea leaves has been studied by reference to three polypeptides and their mRNAs. The polypeptides were the large subunit (LSU) and the small subunit (SSU) of ribulose 1,5-bisphosphate carboxylase/oxygenase (RUBISCO), and the light-harvesting chlorophyll a/b protein (LHCP). The polypeptides were assayed by a sensitive radioimmune assay, and the mRNAs were assayed by hybridization to cloned DNA probes. LSU, LSU mRNA, and LHCP mRNA were detectable in etiolated seedlings but LHCP, SSU, and SSU mRNA were at or below the limit of detection. During the first 48 hr of de-etiolation under continuous white light, the mRNAs for LSU, SSU, and LHCP increased in concentration per apical bud by about 40-fold, at least 200-fold, and about 25-fold, respectively, while the total RNA content per apical bud increased only 3.5-fold. In the same period, the LSU, SSU, and LHCP contents per bud increased at least 60-, 100-, and 200-fold, respectively. The LHCP increased steadily in concentration during de-etiolation, whereas the accumulation LSU, SSU, and SSU mRNA showed a 24-hr lag. The accumulation of SSU, SSU mRNA, and LHCP mRNA showed classical red/far-red reversibility, indicating the involvement of phytochrome in the regulatory mechanism. LSU and LSU mRNA were induced equally well by red and far-red light. The LHCP failed to accumulate except under continuous illumination. These results indicate that the accumulation of SSU is controlled largely through the steady-state level of its mRNA, which is in turn almost totally dependent on light as an inducer and on phytochrome as one of the photoreceptors. The accumulation of LSU is largely but not totally determined by the level of its mRNA, which appears to be under strong photoregulation, which has yet to be shown to involve phytochrome. Phytochrome is involved in the regulation of LHCP mRNA levels but substantial levels of the mRNA also occur in the dark. LHCP accumulation is not primarily governed by the levels of LHCP mRNA but by posttranslational stabilization in which chlorophyll synthesis plays a necessary but not sufficient role.
通过研究三种多肽及其信使核糖核酸(mRNA),对豌豆叶片叶绿体发育的光调节作用进行了探讨。这三种多肽分别是1,5-二磷酸核酮糖羧化酶/加氧酶(RUBISCO)的大亚基(LSU)、小亚基(SSU)以及捕光叶绿素a/b蛋白(LHCP)。采用灵敏的放射免疫分析法检测多肽,通过与克隆的DNA探针杂交来检测mRNA。在黄化幼苗中可检测到LSU、LSU mRNA和LHCP mRNA,但LHCP、SSU和SSU mRNA的含量处于检测极限或低于检测极限。在连续白光下脱黄化处理的最初48小时内,每个顶芽中LSU、SSU和LHCP的mRNA浓度分别增加了约40倍、至少200倍和约25倍,而每个顶芽的总RNA含量仅增加了3.5倍。在同一时期,每个芽中LSU、SSU和LHCP的含量分别增加了至少60倍、100倍和200倍。在脱黄化过程中,LHCP的浓度稳步增加,而LSU、SSU和SSU mRNA的积累则有24小时的延迟。SSU、SSU mRNA和LHCP mRNA的积累表现出典型的红/远红光可逆性,表明光敏色素参与了调节机制。LSU和LSU mRNA受红光和远红光诱导的效果相同。除了在连续光照下,LHCP无法积累。这些结果表明,SSU的积累主要受其mRNA稳态水平的控制,而mRNA稳态水平反过来几乎完全依赖于光作为诱导剂以及光敏色素作为光受体之一。LSU的积累在很大程度上但并非完全由其mRNA水平决定,其mRNA水平似乎受到强烈的光调节,不过尚未证明这种调节涉及光敏色素。光敏色素参与了LHCP mRNA水平的调节,但在黑暗中也会出现相当水平的mRNA。LHCP的积累并非主要由LHCP mRNA水平决定,而是由翻译后稳定作用决定,其中叶绿素合成起到了必要但不充分的作用。