捕食性真菌Duddingtonia flagrans原生质体获得与再生的荧光标记评价
发布时间：2018-10-22 06:35
【摘要】：为了更准确快捷地评价利用酶解法酶解捕食性真菌Duddingtonia flagrans菌丝产生的原生质体数量及菌丝细胞壁降解情况,采用活体荧光染料羧基荧光素乙酰乙酸(carboxyfluorescein diacetate succinimidyl ester,CFSE),对该捕食性真菌菌丝酶解后产生的原生质体进行荧光标记,分别考察了标记浓度、标记时间、孵育温度对原生质体标记效果的影响,并观察CFSE标记后的原生质体再生情况。结果表明CFSE终浓度为10μmol/L,标记时间为15min,孵育温度为36℃,是CFSE标记捕食性真菌原生质体的理想条件,该试验同时表明CFSE不影响原生质体的再生率。CFSE作为一种活细胞示踪荧光探针,可以快速高效地标记捕食性真菌原生质体,该方法为捕食性真菌原生质体制备质量的快速评价提供了新的思路。
[Abstract]:In order to evaluate the number of protoplasts produced by enzymatic hydrolysis of predatory fungi Duddingtonia flagrans hypha and the degradation of hyphal cell wall more accurately and quickly. The protoplasts produced by enzymatic hydrolysis of the predatory fungus mycelium were labeled with fluorescent dye carboxyl fluorescein acetoacetate (carboxyfluorescein diacetate succinimidyl ester,CFSE) in vivo. The labeling concentration and labeling time of the protoplasts were investigated. The effect of incubation temperature on the effect of protoplast marker and the regeneration of protoplast after CFSE labeling were observed. The results showed that the final concentration of CFSE was 15 min and incubation temperature was 36 鈩，
本文编号：2286373
[Abstract]:In order to evaluate the number of protoplasts produced by enzymatic hydrolysis of predatory fungi Duddingtonia flagrans hypha and the degradation of hyphal cell wall more accurately and quickly. The protoplasts produced by enzymatic hydrolysis of the predatory fungus mycelium were labeled with fluorescent dye carboxyl fluorescein acetoacetate (carboxyfluorescein diacetate succinimidyl ester,CFSE) in vivo. The labeling concentration and labeling time of the protoplasts were investigated. The effect of incubation temperature on the effect of protoplast marker and the regeneration of protoplast after CFSE labeling were observed. The results showed that the final concentration of CFSE was 15 min and incubation temperature was 36 鈩，
本文编号：2286373
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