不同甲基紫精诱导离体水稻叶片氧化胁迫方法的研究

Study on Different Methods of Inducing Oxidative Stress of Rice Leaves in vitro by Methyl Viologen

[目的]建立一种甲基紫精诱导完整水稻叶片产生氧化胁迫对叶片无损伤的处理方法。[方法]建立叶鞘引入法,分别用浸根引入法、叶鞘引入法和涂抹引入法处理水稻幼苗叶片后,测定叶片的叶绿素荧光和Rubisco含量,分析不同氧化胁迫诱导方法和不同处理液浓度对诱导效果的影响。[结果]通过叶鞘引入100μmol/LMV的氧化胁迫诱导方法对水稻叶片无损伤,其效果比涂抹引入法和浸根引入法要好。在此条件下,20～40μmol/L MV处理可使水稻第5叶的叶绿素荧光参数发生显著变化。当处理液浓度达到100μmol/L时,Rubisco含量下降约50%。[结论]叶鞘引入法可用于以叶绿素荧光参数或Rubisco含量为标准进行水稻抗氧化胁迫突变体的筛选。该方法的建立为水稻抗氧化胁迫的研究奠定了基础。

[Objective] The aim of the research was to establish a kind of treatment method of inducing full rice leaves to produce the oxidative stress by methyl viologen without harms to leaves. [Method] Leave -sheath introduction method was set up. The leaves of rice seedlings were treated by root -soaking introduction method, leave sheath introduction method and painting introduction method to determine the chlorophyll fluorescence and Rubisco content of leaves and analyze the influences of different induction method of oxidative stress and different concentrations of treatment solution on the inducted effect. [Result] The induction method of oxidative stress inducing 100μmol/L MV by leaf sheath introduction method had no harm to rice leaves and its effect was better than that of root -soaking introduction method and painting introduction method. Under this condition, treatment with 20-40 μmol/L MV could change chlorophyll fluorescence parameters in the 5th leaf of rice significantly. When the concentration of treatment solution reached 100 μmol/L, Rubisco content was reduced about by 50 %. [Conclusion] Leave sheath introduction method could be used to screen rice mutants with resistance to oxidative stress. The establishment of this method laid the foundation for research on rice resistance to oxidative stress.