【作者】 李元文；

【导师】 夏志兰；

【作者基本信息】 湖南农业大学 ， 蔬菜学， 2006， 硕士

【摘要】 本论文对影响姬菇原生质体制备、再生的条件进行了研究。结果表明：以液体静置MYG培养基培养3天的菌丝体，以2％的溶壁酶作为酶解液，在30℃下，0.6m/moL硫酸镁作为渗透压稳定剂，酶解3h为制备姬菇原生质体的最佳条件，以菌龄为1d，酶解时间为1h，0.6mol／L蔗糖作为渗透压稳定剂的PDA再生培养基为姬菇原生质体再生的最佳条件，再生率达4.5％。对影响姬菇5号与署优1号原生质体PEG融合的条件进行了研究。结果表明：以PEG6000的浓度应为35％，Ca²⁺浓度应为0.010mol／L，融合时温度应在30℃，PEG处理时间为20min，PH为8.0为姬菇5号与署优1号原生质体PEG融合的最佳条件，融合率达3.3×10^-4。对影响姬菇5号与署优1号原生质体电融合的条件进行了研究。结果表明：交流电场频率为1500KHz，交流电场强为300V／cm，直流脉冲场强为2.5KV／cm的，脉冲宽幅为60μs时，脉冲次数为4～5次，电融合液甘露醇浓度10％，CaCl₂浓度0.4mmol／L浓度为姬菇5号与署优1号原生质体电融合的最佳条件，融合率达21％。更多还原

【Abstract】 The conditions of separation and regeneration of Pleurotuscornucopiae protoplasts were studied. The results showed the optimal forPleurotus cornucopiae protoplast separation were that the age of hyphe,lywallzyrne consistency and stabilizer was 3 days, 2% and 0.6mol/L MgSO₄respectively. The Pleurotus cornucopiae the age of hyphe was.1 day, thedigesting time was 1 hour, the osmotic stabilizer was 0.6mol/L scourer, thefrequency of regeneration was the best and reach up 4.5%.The conditions of PEG fusion between Pleurotus cornucopiaeprotoplasts and Pleurotus ostreatus protoplasts were studied. The resultsshowed the optimal PEG concentration, Ca²⁺ concentration, temperature,treatment time, and PH value was 35%, 0.010mol/L, 20mintius and 8respectively, the frequency was 3.3×10^-4.The conditions electro fusion between Pleurotus cornucopiae protoplastsand Pleurotus ostreatus protoplasts were studied. The results showed optimalAC frequency, AC strength, DC pulse strength of, DC pulse PW, number of DCpulse number of, mannitol density and CaCl₂ density was 1500KHz,300V/cm, 2.5KV/cm,60μs, 4～5times, 10% and 0.4mmol/L respectively, thefrequency was 21%.更多还原