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食用真菌核荧光染色技术的研究

STUDIES ON THE TECHNIQUE OF FLUORESCENCE STAINING OF NUCLEI IN EDIBLE FUNGI

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【作者】 罗信昌陈大年

【Author】 Lo XiNG-CHANG CHEN Da-NIAN(Huazhong Agricultural University,Wuhan)

【机构】 华中农业大学

【摘要】 本文报道荧光染料双苯并咪唑Hoechst 3325 8(bi sbenzimide),应用于香菇(Lentinusedodes)、平菇(pleurotus ostreatus)、金针菇(Flammulina velutipes)、木耳(Aurtculariaauricula),毛木耳(A.polytricha)等5种食用菌菌丝体。担孢子、分生孢子核的荧光染色研究。配制染液的McIlvaine缓冲液pH值、缓冲液含0.6M MgSO4·7H2O、菌丝生长期、孢子贮存期亦与染色效果密切相关。我们观察到,使用插片法,爬片2/5的菌丝体,pH7.0—7.4Hoechst 3325 8染液,染色3分钟为菌丝体核荧光染色的最佳效果。新鲜的担孢子,分生孢子、使用pH 6.8含0.6 M MgSO4·7 H:O Hoechst 3325 8染色液染色3分钟比使用pH 6.8Hoechst 3325 8的染液有明显增效作用。

【Abstract】 The application of Fluorescence Dye bisbenzimide (Hoechst 33258) in fluorescence staining of nuclei of mycelia,basidospores,and oidia,of Lentinus edodes,Pleurotus ostreatus,Flammulia velutipes,Auricularia auricula,and Auricularia polytricha was reported in his paper.pH of of Mcllvaine buffer (used for staining solution containing 0.6 M MgSO4 7H2O),mycelial growth period,storage times of spore were associated with the quality of fluorescence saining.We observed that two fifths of mycelial growth on the cover slide using the methol of cover slide inserting,unler the cinlition of pH 7.0—7.4 of Mcllvaine buffer, staining for three minutes,Hoechst 33258 would show the best effect for the fluorescence staining of mycelial nuclei. The staining of fresh basidospores and conidia with Hoechst 33258 which prepared with pH 6.8 Mcllvaine buffer containing 0.6M MgSO4 gH2O in 2 minutes was obviously effective than that of Hoechst Dye 33258 prepared with pH 6.8 Mcllvaine buffer.

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