椪柑原生质体分离与培养过程中活性氧动态变化及来源分析

The Dynamic Changes and Origin Analysis of Reactive Oxygen Species during Isolation and Culture of Citrus reticulata Blanco Protoplasts

分析原生质体分离与培养过程中活性氧(ROS)的动态变化与来源,对揭示原生质体再生能力差异的机制具有重要意义。本研究采用荧光标记法分析了ROS在椪柑原生质体分离与培养中的动态变化,同时通过ROS抑制剂处理探究了其ROS的来源。结果表明,ROS分布在原生质体的细胞质和细胞膜上;分离阶段愈伤组织原生质体的ROS水平显著低于叶肉原生质体,但在培养阶段显著高于叶肉原生质体;DPI与NaN3处理均显著降低了原生质体分离与培养过程中的ROS水平。因此,ROS在椪柑原生质体再生中具有重要调控作用。

In order to uncover the mechanisms underlying different regeneration capacity of protoplasts, we analyzed the dynamic changes and origin of reactive oxygen species (ROS) during isolation and culture of Citrus reticulata Blanco protoplasts. The dynamic changes of ROS were analyzed by flu-orescent markers, and ROS origin was investigated by ROS indicators. The results showed that ROS signals were visible in the cytoplasm and cell membrane of protoplasts;compared with mesophyll protoplasts, callus protoplasts contained significantly lower level of ROS during isolation, but significantly higher level of ROS during culture;a significant decrease in the level of ROS during isolation and culture of callus protoplasts was observed after DPI or NaN3 treatment. Our findings indicate ROS plays a crucial mediating role in protoplast regeneration of Citrus reticulata.