秀珍菇单孢子菌株原生质体制备条件的优化.PDFVIP

生物技术通报 ·技术与方法· BIOTECHNOLOGY BULLETIN ( ) 2018, 34 4 :70-76 秀珍菇单孢子菌株原生质体制备条件的优化 1 2 2 1，3 孙晓瑞  陈博文  张小林  孟俊龙 （1. 山西农业大学食品科学与工程学院，太谷 030801 ；2. 山西农业大学园艺学院，太谷 030801 ；3. 黄土高原食用菌提质增效协同创新中心， 太谷 030801） 摘 要 ： 以秀珍菇单核菌丝为材料，用溶壁酶进行原生质体的制备，考察各因素对原生质体产量的影响，用单因素试验和 正交试验确定最佳条件。结果表明，菌龄为8 d 菌丝体，0.6 mol/L 甘露醇+10 mmol/L Tris-HCl 为稳渗剂，酶解温度29℃、酶浓度1.6% 的条件下酶解160 min。原生质体产量达到3.5×107 CFU/mL，原生质体再生率达到1.4%。 关键词 ： 秀珍菇；单核菌丝 ；原生质体 ；溶壁酶 DOI ：10.13560/ki.biotech.bull.1985.2017-0797 Optimization of Preparation Conditions of Pleurotus geesterani Monocaryon Mycelia Protoplast 1 2 2 1，3 SUN Xiao-rui CHEN Bo-wen ZHANG Xiao-lin MENG Jun-long （1. College of Food science and Engineering ，Shanxi Agricultural University ，Taigu 030801 ；2. College of horticulture ，Shanxi Agricultural University，Taigu 030801 ；3. Collaborative Innovation Center of Quality and Efficiency of Loess Plateau Edible Fungi for Universities in Shanxi ，Taigu 030801 ） Abstract: The optimal conditions for preparing protoplast from lywallzyme by cultured monocaryon mycelia of Pleurotus geesterani were investigated via single factor and orthogonal array design. The results showed ：the yield of protoplast and regeneration rate of protoplast reached 3.5×107 CFU/mL and 1.4% respectively，under the conditions of 8 days mycelium in the enzyme solution of 1.6% lywallzyme digested for 160 min at temperature of 29℃ with 0.6 mol/L mannitol +10 mmol/L Tris-HCl as osmotic pressure stabilizer. Key words: Pleurotus gees