Effects of light and UV-C radiation on the transcriptional activity of COP1 and HY5 gene homologues in barley.

Irina Boycheva, R. Georgieva, L. Stoilov, V. Manova
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Abstract

Abstract Photomorphogenic regulators COP1 (Constitutive Photomorphogenic 1) and HY5 (Elongated Hypocotyl 5) play a key role in plant development by guiding the transition from dark to light growth. In Arabidopsis they are also implicated in the transcriptional control of photolyase genes. Here we characterize the transcript abundance of COP1 and HY5 gene homologues in barley in relation to light-grown conditions and UV-damage response. Etiolated and green 6-day-old seedlings were UV-C irradiated and exposed to light or kept in darkness. The abundance of barley COP1 and HY5 transcripts was assessed by real-time RT-PCR. In etiolated leaves we found several-fold lower levels of COP1 transcripts which reached the levels of the green ones after 1 h of light exposure. Barley HY5 transcripts were very low in the dark-grown seedlings and after 1 h of illumination they increased drastically to levels significantly exceeding those measured in the green leaves. Both genes were upregulated by light in the irradiated plants as well, but to a lesser extent compared with their controls, probably due to the presence of non-repaired DNA damage in the etiolated leaves soon after irradiation. The enhanced transcription of barley COP1 under light is unexpected in view of the well-known function of COP1 as a negative regulator of plant photomorphogenesis but conforms to the positive role reported for AtCOP1 in UV-B signalling. HY5 is recognized as a stimulator of light-inducible genes and our data support such a role for the barley HY5 homologue as well. Our study shows that, in barley seedlings, the regulation of COP1 and HY5 gene expression is achieved through light-positive transcriptional modulation, suggesting that both genes contribute to the de-etiolation phase in barley. According to our knowledge, this is the first quantitation of the COP1 and HY5 mRNAs in barley that also regards the UV-damage response of this crop.
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光照和UV-C辐射对大麦COP1和HY5基因同源物转录活性的影响
光形态发生调控因子COP1 (Constitutive phophophogenic 1)和HY5 (Elongated Hypocotyl 5)在植物发育过程中起关键作用,引导植物从暗生长向光生长过渡。在拟南芥中,它们也参与光解酶基因的转录控制。在这里,我们描述了COP1和HY5基因同源物在大麦中的转录丰度与光照条件和紫外线损伤反应的关系。黄化和绿色的6日龄幼苗进行UV-C照射,暴露在光照下或保存在黑暗中。利用实时RT-PCR技术检测大麦COP1和HY5转录本的丰度。光照1 h后,黄化叶片的COP1转录本水平降低了几倍,达到了绿色叶片的水平。大麦HY5转录本在暗生苗中含量极低,光照1 h后急剧增加,显著高于绿叶。在受光照的植物中,这两个基因也被光上调,但与对照相比,上调幅度较小,这可能是由于照射后不久黄化叶片中存在未修复的DNA损伤。大麦COP1在光照下的转录增强是出乎意料的,因为众所周知,COP1是植物光形态发生的负调节因子,但与AtCOP1在UV-B信号传导中的积极作用一致。HY5被认为是光诱导基因的刺激物,我们的数据也支持大麦HY5同源物的这种作用。我们的研究表明,在大麦幼苗中,COP1和HY5基因表达的调控是通过光正转录调控实现的,这表明这两个基因都参与了大麦的去黄化阶段。据我们所知,这是大麦中COP1和HY5 mrna的第一次定量,也涉及了这种作物的紫外线损伤响应。
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