Facella Paolo, Lopez Loredana, Carbone Fabrizio, Galbraith David W, Giuliano Giovanni, Perrotta Gaetano
ENEA, Trisaia Research Center, Rotondella (MT), Italy.
PLoS One. 2008 Jul 30;3(7):e2798. doi: 10.1371/journal.pone.0002798.
Circadian clocks are internal molecular time-keeping mechanisms that provide living organisms with the ability to adjust their growth and physiology and to anticipate diurnal environmental changes. Circadian clocks, without exception, respond to light and, in plants, light is the most potent and best characterized entraining stimulus. The capacity of plants to respond to light is achieved through a number of photo-perceptive proteins including cryptochromes and phytochromes. There is considerable experimental evidence demonstrating the roles of photoreceptors in providing light input to the clock.
In order to identify genes regulated by diurnal and circadian rhythms, and to establish possible functional relations between photoreceptors and the circadian clock in tomato, we monitored the temporal transcription pattern in plants entrained to long-day conditions, either by large scale comparative profiling, or using a focused approach over a number of photosensory and clock-related genes by QRT-PCR. In parallel, focused transcription analyses were performed in cry1a- and in CRY2-OX tomato genotypes.
We report a large series of transcript oscillations that shed light on the complex network of interactions among tomato photoreceptors and clock-related genes. Alteration of cryptochrome gene expression induced major changes in the rhythmic oscillations of several other gene transcripts. In particular, over-expression of CRY2 had an impact not only on day/night fluctuations but also on rhythmicity under constant light conditions. Evidence was found for widespread diurnal oscillations of transcripts encoding specific enzyme classes (e.g. carotenoid biosynthesis enzymes) as well as for post-transcriptional diurnal and circadian regulation of the CRY2 transcript.
生物钟是内部分子计时机制,使生物体能够调节其生长和生理机能,并预测昼夜环境变化。生物钟无一例外地对光做出反应,在植物中,光是最有效且特征最明确的同步刺激因素。植物对光的反应能力是通过包括隐花色素和光敏色素在内的多种光感受蛋白实现的。有大量实验证据证明光感受器在为生物钟提供光输入方面的作用。
为了鉴定受昼夜节律调节的基因,并确定番茄中光感受器与生物钟之间可能的功能关系,我们通过大规模比较分析,或使用定量逆转录聚合酶链反应(QRT-PCR)对多个光感受和生物钟相关基因采用聚焦方法,监测了适应长日照条件的植物中的时间转录模式。同时,在cry1a和CRY2过表达番茄基因型中进行了聚焦转录分析。
我们报告了一系列转录振荡,这为番茄光感受器和生物钟相关基因之间复杂的相互作用网络提供了线索。隐花色素基因表达的改变引起了其他几种基因转录本节律振荡的重大变化。特别是,CRY2的过表达不仅影响昼夜波动,还影响持续光照条件下的节律性。我们发现编码特定酶类(如类胡萝卜素生物合成酶)的转录本存在广泛的昼夜振荡,以及CRY2转录本的转录后昼夜和生物钟调节的证据。