利用SCGE技术对植物盐胁迫下DNA损伤的研究

Study on DNA Damage of Plants under Salt Stress Using SCGE Technology

以北斗七星蚕豆为试验材料,分别采用0,6,12,18,24,30g/L的6个质量浓度的NaCl溶液处理蚕豆根尖,利用SCGE技术对蚕豆根尖细胞在不同质量浓度盐胁迫下DNA的损伤进行检测,探讨逆境胁迫对植物细胞DNA的损伤效应,并以此建立快速准确的植物DNA损伤检测流程。结果表明,盐胁迫会对蚕豆根尖细胞DNA造成损伤,并且随着盐胁迫质量浓度的升高,细胞拖尾加剧,30 g/L盐胁迫下的细胞拖尾最明显;此外,当NaCl溶液质量浓度大于18 g/L时,随着NaCl溶液质量浓度的增加,蚕豆根尖细胞拖尾率逐渐增大,根尖细胞损伤概率增加。通过利用多种比较分析细胞尾部DNA发现,随着盐胁迫质量浓度的增加,细胞的彗星尾长增加显著,同时利用软件分析可增加对DNA损伤的直观检测,具有较好的应用效果。最后,通过对实验过程中涂层琼脂糖溶液和包埋凝胶溶液的制备、载玻片涂胶处理方法等均进行了合理的改良,为后续大批量开展植物逆境胁迫条件的评估提供便利。

In this study,the Big Dipper broad bean was used as the experimental material.The root tips of broad bean were treated with six mass concentrations of NaCl solution of 0,6,12,18,24,and 30 g/L,respectively.The DNA damage of root tip cells under salt stress at different concentrations was detected by SCGE technology,and to explore the damage effect of stress on plant cell DNA and establish a fast and accurate process for detecting DNA damage in plants.The results showed that salt stress causes damage to the root tip cell DNA and the tail dragging increased with the salt stress concentration,and the cell tailing under 30 g/L salt stress was the most obvious.Moreover,when the NaCl solution concentration was greater than 18g/L,the tailing rate of the root tip cells gradually increased and the probability of root tip cell damage increased.By using multiple comparative analysis of the cell tail DNA,the comet tail length of the cells increased significantly with increasing salt stress concentration.At the same time,the software analysis can increase the intuitive monitoring of DNA damage,which has a good application effect.Finally,the preparation of coated agarose solution and embedded gel solution and the treatment method of slides were reasonably improved,so as to facilitate the subsequent evaluation of plant stress conditions in large quantities.