桧状青霉原生质体制备和再生的研究  

Analysis of the Preparation and Regeneration of Protoplasts from Penicillium piceum

作　　者：张粲[1,2] 高乐 赫荣琳 王国坤 于俊杰[1,2] 武改红 张东远 

机构地区：[1]天津科技大学生物工程学院,天津300457 [2]天津工业生物技术研究所,天津市工业生物系统与过程工程重点实验室,天津300308

出　　处：《基因组学与应用生物学》2014年第1期57-62,共6页Genomics and Applied Biology

基　　金：国家863项目(2012AA023200)资助

摘　　要：本文对桧状青霉H16的原生质体制备和再生的相关影响因子进行了初步的研究。结果表明:于30℃,转速为200 r/min的SDYS培养基中培养了16 h的菌丝,经S1溶液洗涤后,在1 mol/L的NaCl为等渗溶液,采用浓度为10 mg/mL的酶液,酶解温度为35℃,转速为100 r/min的条件下处理1.5 h时原生质体量达到最佳,本实验范围内最高达到6×106个/mL;同时,选取含有25 mmol/L Ca2+的改良的察氏培养基培养经S2洗涤的原生质体时,原生质体可以达到较佳的再生率,本实验范围内最高达到24.3%。该研究为桧状青霉后期的菌株改造和遗传转化体系的建立奠定了基础。As there are some factors affecting the preparation and regeneration of protoplasts of Penicillium piceum H16, the optimum conditions for it were studied. The results showed that maximum amount of protoplasts were released from mycelia cultured in SDYS medium at 30℃, 200 r/mim for 16 hours and when digested by the lysing enzymes at 35℃ for 1.5 hours in a buffer containing 1 mol/L of NaCl used as an isotonic solution. In the range of this experiment, the peak value can reach 6× 10^6/mL. The maximum regeneration frequency of protoplasts was obtained when protoplasts were plated on modified Czapek＇s medium containing 25 mmol/L of calciumion, and the resoult showed that the highest regeneration rate of protoplasts was 24.3% on the optimal regeneration medium. The study can lay foundation for genetic modification of the strain and also will be the basis for genetic transformation.

关 键 词：桧状青霉 原生质体制备 原生质体再生 

分 类 号：Q933[生物学—微生物学]