雨生红球藻对4种抗生素的敏感性研究

Study of the sensitivity of Haematococcus pluvialis to four antibiotics

微藻对抗生素的敏感性研究是确定微藻遗传转化体系中筛选标记的基础.监测不同琼脂浓度下雨生红球藻的生长状况,确定该藻藻落生长的最优琼脂浓度;依据不同质量浓度庆大霉素、博莱霉素、氯霉素和壮观霉素对雨生红球藻生长的影响,确定该藻对4种抗生素的敏感性,每种抗生素均设置固体和液体培养实验.研究结果表明:(1)固体平板培养基上雨生红球藻生长的最佳琼脂浓度为0.8%.(2)实验浓度下,雨生红球藻对庆大霉素、博莱霉素敏感性高,可作为雨生红球藻遗传转化的选择抗生素,其对应的抗性基因可作为选择标记基因,所需质量浓度分别为5μg·m L-1和0.5μg·m L-1.雨生红球藻对氯霉素不敏感,可用于藻种无菌化处理.(3)在固体和液体培养基上,雨生红球藻对同一种抗生素的敏感性稍有不同,研究及应用需根据培养方式选择适宜的抗生素浓度.

Establishment of genetic transformation system is based on the sensitivity of algae to antibiotics.The current study investigated the optimal agar concentration for Haematococcus pluvialis growth.The sensitivities of four kinds of antibiotics were respectively determined in solid and liquid mediums.The results showed that the most optimal agar concentration for H.pluvialis growth was0.8%.At the experimental concentration,Gentamicin and Zeocin showed the strongest inhibitory effects on H.pluvialis,which could serve as the selective antibiotics in transformation system at5and0.5μg·mL-1,respectively.Chloramphenicol can be used to purify algae,due to its lowest inhibition to H.pluvialis.In the presence of the different effects of antibiotics in solid and liquid mediums,appropriate antibiotic concentration should be chosen for research or application of H.pluvialis.