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

作　　者：柳琳[1,2,3,4] 陈越[2,3,4] 陈玲[2,3,4] 李维蛟[2,3,4] 程在全[2,3,4] 

机构地区：[1]云南大学,云南昆明650091 [2]云南省农业科学院生物技术与种质资源研究所,云南昆明650223 [3]云南省农业生物技术重点实验室,云南昆明650223 [4]云南昆明农业部西南作物资源与新种质创制重点实验室,云南昆明650223

出　　处：《贵州农业科学》2014年第11期70-72,共3页Guizhou Agricultural Sciences

基　　金：国家自然科学基金项目"云南疣粒野生稻免疫白叶枯病的抗性基因分析及定位研究"(NFSC31160067);云南省科技计划项目省应用基础研究计划资助项目"云南疣粒野生稻泛素结合酶相关基因的筛选和功能分析"(2012FA005);国家公益性行业科研专项"农业野生植物资源保护利用技术研究与示范"(201003021);云南省青年基金项目"疣粒野生稻与栽培稻杂种细胞的构建"(201301YC00011)

摘　　要：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.

关 键 词：水稻 原生质体 细胞壁 荧光增白剂 检测 

分 类 号：S511[农业科学—作物学]