【摘要】：為了更準(zhǔn)確快捷地評(píng)價(jià)利用酶解法酶解捕食性真菌Duddingtonia flagrans菌絲產(chǎn)生的原生質(zhì)體數(shù)量及菌絲細(xì)胞壁降解情況,采用活體熒光染料羧基熒光素乙酰乙酸(carboxyfluorescein diacetate succinimidyl ester,CFSE),對(duì)該捕食性真菌菌絲酶解后產(chǎn)生的原生質(zhì)體進(jìn)行熒光標(biāo)記,分別考察了標(biāo)記濃度、標(biāo)記時(shí)間、孵育溫度對(duì)原生質(zhì)體標(biāo)記效果的影響,并觀察CFSE標(biāo)記后的原生質(zhì)體再生情況。結(jié)果表明CFSE終濃度為10μmol/L,標(biāo)記時(shí)間為15min,孵育溫度為36℃,是CFSE標(biāo)記捕食性真菌原生質(zhì)體的理想條件,該試驗(yàn)同時(shí)表明CFSE不影響原生質(zhì)體的再生率。CFSE作為一種活細(xì)胞示蹤熒光探針,可以快速高效地標(biāo)記捕食性真菌原生質(zhì)體,該方法為捕食性真菌原生質(zhì)體制備質(zhì)量的快速評(píng)價(jià)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，

本文編號(hào)：2286373