紫外辐射诱导植物细胞DNA损伤的彗星检测

COMET ASSAY ON THE DNA DAMAGE INDUCED BY UV RADIATION IN PLANT CELLS

单细胞凝胶电泳(彗星检测)已广泛应用于动物细胞DNA损伤检测.该文报道了单细胞凝胶电泳应用于UV-B诱导植物细胞DNA损伤的检测.通过对动物细胞的单细胞凝胶电泳实验方法的改进,获得了在植物细胞中应用的最佳条件.以植物细胞原生质体为材料,并结合T4 Endonu lease V的运用,显著提高了UV-B诱导植物细胞DNA损伤提高彗星检测的敏感性和特异性.植物细胞DNA损伤的彗星图像经CASP软件处理并量化,结果表明:UV-A和UV-B均能诱导发生DNA单链断裂;UV-A在一定的剂量下诱导少量嘧啶二聚体的形成,而UV-B则强烈诱导嘧啶二聚体的产生.

The single cell gel electrophoresis or comet assay is now widely used to detect DNA damage in animal cells induced by radiation or chemicals. In the present study, the application of comet assay on UV-B induced DNA damage in the plant cell is reported. The standard protocol of the comet assay for the animal cell was modified for optimizing conditions for the plant ceils. The specificity and sensitivity of comet assay on plant cell was improved by using protoplasts and T4 Endonuclease V. CASP software was used to analyze the comet image. Our results indicated that both UV-A and UV-B radiation can induce DNA single - strand breaking in the plant cell, however UV-B was more effective than UV-A for inducing pyrimidine dimers.