韩增华; 张丕奇; 戴肖东; 孔祥辉; 马庆芳; 张介驰
黑龙江省科学院微生物研究所
【中文摘要】 利用正交试验研究了黑木耳原生质体制备、再生的最佳条件,结果表明,原生质体制备最佳条件是酶解温度31℃、酶浓度1.0%、菌龄5 d、酶解时间4 h,原生质体再生的最佳条件是酶解温度27℃、酶解时间4 h、酶浓度1.5%、菌龄11 d。稳渗剂种类对黑木耳原生质体再生率的试验结果表明,采用0.5 mol/L蔗糖为稳渗剂,各黑木耳菌株的原生质体再生率最高。HW2号菌株原生质体经核染色其单核化比率最高;在荧光显微镜下观察菌丝核相,单、双核清晰可辨。
【英文摘要】 Conditions for the preparation and regeneration of Auricularia auricula protoplasts were optimized using orthogonal testing.Highest protoplast yields were obtained from 5-day-old mycelium after treatment with 1.0%(w/v) lywallzyme for 4 h at 31 ℃,while the highest protoplast regeneration rate was obtained from 11-day-old mycelium after treatment with 1.5%(w/v) lywallzyme for 4 h at 27 ℃.Of four osmotic stabilizers tested(sucrose,mannitol,MgSO4 and KCl),the highest rate of protoplast regeneration was obtained using 0.5 mol/L sucrose.Monokaryotic and dikaryotic mycelia were readily distinguishable using fluorescence microscopy.The highest rate of monokaryotic protoplast formation was recorded with A.auricula,strain HW2.
【中文关键词】 黑木耳; 原生质体; 再生; 单核体; 荧光核染色
【英文关键词】 Auricularia auricula; protoplast; regeneration; monokaryotic protoplast; fluorescence staining of nuclei
【基金】黑龙江省科学院基金"应用分子生物学技术进行黑木耳单核原生质体杂交育种"(编号:03BCCAHZH08)的部分研究内容
【文献出处】 食用菌学报,Acta Edulis Fungi,编辑部邮箱,2008年03期 【DOI】CNKI:SUN:SYJB.0.2008-03-003