基于VBL增白剂细胞染色的水稻原生质体细胞壁变化检测

Detection of Rice Protoplast Cell Wall Stained with Fluorescent Whitening Agent VBL

为快速、准确地观察水稻原生质体制备及培养过程中水稻细胞壁的去除和再生情况,利用荧光增白剂VBL细胞壁染色液对细胞进行染色,制片后置于激发波长为345nm、发射波长为430nm的荧光显微镜下观察。结果发现,在原生质体制备过程中,细胞周围发出的蓝色荧光初期表现分布均匀且荧光强烈,之后逐渐变为分布不均匀且荧光暗淡,最后蓝色荧光消失,表明纤维素逐渐被酶解去除;在原生质体培养过程中荧光情况与制备过程的刚好相反。结论:利用荧光增白剂VBL细胞壁染色液染色通过荧光强弱变化对比,可以快速、准确地检测水稻原生质体制备及培养过程中细胞壁的变化。

The rice cells stained with fluorescent whitening agent VBL were observed by fluorescence microscope with 345 nm excitation wavelength and 430 nm emission wavelength to observe elimination and regeneration of rice cell walls during preparation and culture process of rice protoplasts rapidly and accurately.The blue fluorescence around cells represents distribution equality and intensity fluorescence at early stage,gradually mal-distribution and dim fluorescence,and finally the blue fluorescence disappears,which indicates that cellulose is gradually eliminated by enzymolysis,but the blue fluorescence performance in the protoplast culture process is just the opposite of the protoplast preparation process.In conclusion,the fluorescent whitening agent（VBL）can be used in rapid and accurate detection of rice protoplasts during the preparation and culture process.