【摘要】 将2016年7月在黑龙江省凉水国家级自然保护区采集的朱红栓菌Trametes cinnabarina子实体接种于PDA固体培养基平皿中,30℃下培养7 d,截取菌丝体接种于液体培养基中用于原生质体制备与再生实验。通过单因素试验和正交试验,研究酶解温度、酶解时间、菌丝质量分数、酶配比对原生质体制备的影响。结果表明,以0.6 mol·L-1甘露醇溶液作为高渗缓冲液,复合酶组成为1.5%溶壁酶﹢1.0%纤维素酶﹢1.0%蜗牛酶,菌丝质量分数为20%,30℃条件下酶解3.5 h为朱红栓菌原生质体制备的最适条件,产量达1.00×107个·m L-1;以0.6 mol·L-1蔗糖溶液作为再生高渗缓冲液,原生质体再生率最高达20.6%。更多还原

【Abstract】 Sporecarp of Trametes cinnabarina was collected in July 2016 in Liangshui National Nature Reserve of Heilongjiang province, and was cut out on PDA agarslantculture-medium under 30 ℃ for 7 days. Mycelium was inoculated on liquid nutrient medium for protoplast preparation and regeneration test. Single factor and orthogonal test was carried out on enzymolysis temperatures and enzymolysis time, mass fractions of hyphae and enzyme ratios. The result demonstrated that the best condition for protoplast preparation was 0.6 mol·L-1 mannitol solution as buffer, 1.5% of lywallzyme, 1.0% of cellulase and 1.0% of glusulase as enzume ratio, 20% of mass fraction of mycelia, 3.5 hours foren zymolysis under 30℃. The yield of protoplast topped to 1.0×107 mL-1. The highest regeneration rate was 20.6% with 0.6 mol·L-1 sucrose solution as the regeneration stabilizer.更多还原