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。
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） lywalhyme 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） lywalhyme for 4 h at 27℃. Of four osmotic stabilizers tested （sucrose, Q 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.
出处
《食用菌学报》
2008年第3期13-17,共5页
Acta Edulis Fungi
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
关键词
黑木耳
原生质体
再生
单核体
荧光核染色
Auricularia auricula
protoplast
regeneration
monokaryotic protoplast
fluorescence staining of nuclei
