黑木耳菌丝原生质体的制备﹑再生及单核鉴定研究

Study on protoplast isoulation and regeneration and monokaryotization of Auricularia auricula

探讨了黑木耳原生质体的制备、再生及单核化的问题。结果表明,培养了8 d的黑木耳菌丝体用硫酸镁做稳渗剂,2.0%的溶壁酶酶解,31℃下水浴4 h所产生的原生质体数量最多,可以达到11.5×107个.mL-1,0.5 mol.mL-1硫酸镁渗透压稳定剂,RM5再生培养基条件下再生率最高,最高可达1.63%。原生质体再生的菌丝接种到PDA综合培养基的培养皿上,25℃静止培养。待菌丝长满整个平皿后,用剪刀剪边长1 cm正方形大小的透明胶布,用其粘平皿内的菌丝并在显微镜下观察。

The highest yield of protoplast could be obtained by using 2% lywallzyme made by 0.5 mol·L-1 MgSO4·7H2O,digesting for 4 h at the condition of 31 ℃,the production of protoplast reached 11.5×106 number·mL-1,0.5 mol·L-1 magnesium sulfate osmotic stabilizer,RM5 regeneration medium under the conditions of the highest recycling rate up to 1.63%.Regeneration of mycelial protoplasts inoculated Petri dishes on PDA medium comprehensive,25 ℃ static culture.After the plate to be covered with mycelium,cut with scissors the size of the square side length 1 cm transparent tape,with its sticky plate and the mycelium was observed under the microscope.