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探讨了利用原生质体诱变育种技术选育富硒能力强的血芝优良菌株,确定其原生质体制备的最适条件为:以6d菌龄的菌丝体为基体,用3%裂解酶+0.5%蜗牛酶,0.6mol/L甘露醇为渗透压稳定剂,30℃下酶解3.5h。原生质体的产量最高可达到8.28×106cfu/mL。对其原生质体进行了紫外照射诱变以及再生培养,筛选获得了1株具有稳定遗传性的富硒血芝菌株UV37。该菌株液体发酵菌丝干重和菌丝体中含硒量分别为19.81mg/mL和42.87μg/g,与出发菌株相比分别提高了20.35%和58.31%。
In this study, we explored the use of protoplast mutagenesis breeding of elite strains of selenium with strong selenium-rich ability to determine the optimum conditions for the protoplast preparation: 6d mycelium age of mycelium as the substrate, with 3% lyase +0.5 % Snail enzyme, 0.6mol / L mannitol as osmotic pressure stabilizer, enzymatic hydrolysis at 30 ℃ 3.5h. Protoplast production up to 8.28 × 106cfu / mL. The protoplasts were subjected to UV irradiation mutagenesis and regeneration culture. One stable and hereditary selenium-enriched sesame strain UV37 was obtained. The liquid fermentation mycelium dry weight and mycelium selenium content of the strain were 19.81mg / mL and 42.87μg / g, respectively, compared with the starting strain increased by 20.35% and 58.31%.