摘要
为探索筛选β-胡萝卜素高产菌株的诱变育种方法,采用低能氮离子对三孢布拉氏霉负菌孢子进行辐照.研究了不同剂量对三孢布拉氏霉存活率、菌丝长势、菌落形态、产孢特性和突变率的影响,并采用高效液相色谱法测定了β-胡萝卜素含量.结果表明:随着注入剂量的增加,其存活率呈现先减小后增大再减小的"马鞍型"剂量效应曲线;除14×1015N+/cm2外,其它剂量下平板培养菌丝均密度增大、长度减小、长速加快、颜色变黄且长出一定量褐色的隐性孢子;其总突变率和正突变率呈现先增加后减小的变化趋势,当剂量为12×1015N+/cm2时,总突变率和正突变率均达到最大,且筛选出1株β-胡萝卜素产量达到1.252g/L,比出发菌株1.044 g/L提高了25%的高产菌株BT12-12-05,通过传代实验发现其遗传稳定.
In order to explore the mutation breeding method of screening high-yield strains of β-carotene,the negative bacteria spores of Blakeslea trispora were irradiated by low-energy nitrogen ions. The effect of different implantation doses on the survival rate, mycelium growth, colony morphology, sporulation characteristics and mutation rate of Blakeslea trispora was investigated and the β-carotene content was detected by high-performance liquid chromatography. The results showed that with the increase of N+implantation dose, the survival rate showed the "saddle" dose-response curve. In addition to the dose of 14×1015N+/cm2,the flat mycelium density increased, the flat mycelium length decreased, the flat mycelium growth rate accelerated, the flat mycelium color became yellow and a certain amount of brown latent spores grew. The overall mutation rate and positive mutation rate showed the trends of increasing firstly and then decreasing.At the dose of 12× 1015N+/cm2, the total mutation rate and positive mutation rate reached maximum and a β-carotene high-yielding strain BT12-12-05 was selected and its β-carotene productivity reached 1.252g/L,which increased by 25% compared to that of initial strain of 1.044g/L. It was found that the mutant had a good genetic stability through passage experiment.
引文
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