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The Effects of He-Ne Laser and Enhanced Ultraviolet-B Radiation on Proliferating-Cell Nuclear Antigen in Wheat Seedlings

机译:He-Ne激光和增强的紫外线-B辐射对小麦幼苗增殖细胞核抗原的影响

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The level of ultraviolet-B (UV-B) radiation on the Earth’s surface has increased due to depletion of the ozone layer. Here, we explored the effects of continuous wave He-Ne laser irradiation (632 nm, 5 mW·mm-2, 2 min·d-1) on proliferating-cell nuclear antigen (PCNA) damage repair function of wheat seedlings exposed to enhanced UV-B radiation (10.08 kJ·m-2·d-1) at the early growth stages. Wheat seedlings were irradiated with enhanced UV-B, He-Ne laser treatment or a combination of the two. We explored the transcripts of PCNA in each treatment group using RT-PCR. In addition, total proteins were extracted from the 7-day-old wheat leaves, analyzed by SDS-PAGE and identified by western blot. The results showed that the transcription of PCNA was weakened following UV-B radiation compared to the control. However, when seedlings were subjected to elevated UV-B-damaging radiation followed by He-Ne laser irradiation, the expression of PCNA was signifi-cantly higher than UV-B radiation alone. These results suggest that He-Ne laser has an active role in repairing the UV-B damaging effects. In order to further investigate the function of PCNA, dynamic arrangements of PCNA in wheat root-tip cells were observed with confocal laser scanning microscopy (CLSM). The PCNA was marked fluorescent dimming and strength weakened in en-hanced UV-B radiation (UV-B) compared with the control group (CK) during processing. It shows that PCNA may be involved in the separation of chromosomes.
机译:由于臭氧层的消耗,地球表面的紫外线B(UV-B)辐射水平有所提高。在这里,我们探讨了连续波氦氖激光辐照(632 nm,5 mW·mm-2,2 min·d-1)对暴露于增强胁迫下的小麦幼苗增殖细胞核抗原(PCNA)损伤修复功能的影响。生长初期的UV-B辐射(10.08 kJ·m-2·d-1)。小麦幼苗用增强的UV-B,He-Ne激光处理或两者结合的方法进行辐照。我们使用RT-PCR探索了每个治疗组中PCNA的转录本。另外,从7天大的小麦叶片中提取总蛋白,通过SDS-PAGE分析并通过蛋白质印迹法鉴定。结果显示,与对照相比,UV-B辐射后PCNA的转录减弱。但是,当幼苗受到高强度的破坏UV-B的辐射后再进行He-Ne激光辐射时,PCNA的表达明显高于单独的UV-B辐射。这些结果表明,氦氖激光器在修复UV-B破坏作用方面具有积极作用。为了进一步研究PCNA的功能,共聚焦激光扫描显微镜(CLSM)观察了PCNA在小麦根尖细胞中的动态排列。在加工过程中,与对照组(CK)相比,增强型UV-B辐射(UV-B)中PCNA的荧光显着减弱,强度减弱。它表明PCNA可能参与了染色体分离。

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