目的 考察杜氏盐藻对氯酸盐的敏感性及其在分离盐藻氯酸盐抗性株中的作用。方法 测定了不同浓度的氯酸钾和氯酸钠对杜氏盐藻的抑制作用,并利用确定浓度的氯酸盐从盐藻化学诱变库中分离氯酸盐抗性株。结果 杜氏盐藻对氯酸钾的敏感浓度(细胞致死浓度,下同)为100 mmol/L,对氯酸钠的敏感浓度为200 mmol/L。利用200 mmol/L氯酸钠作为筛选剂,从化学诱变库中筛选了3株氯酸盐抗性突变株。结论 盐藻对氯酸盐中度敏感,可以利用氯酸盐抗性分离杜氏盐藻硝酸盐还原酶突变株,从而建立盐藻筛选标记体系,为利用转基因盐藻生产抗癌药物血管抑素奠定基础。
Objective To investigate the sensitivity of Dunaliella salina to chlorate and isolate chlorate-resistant mutants of D. salina. Methods Wild type D. salina cells were collected and used to study their sensitivity to the chlorate. After KClO3 and NaClO3 at different concentrations were respectively added to the cells suspensions, chlorate sensitivity of D. salina was evaluated by measuring OD685 values to represent the eel/densities and observing morphological change through microscope in different days. The wild type cells without chlorate served as controls. The chlorate-resistant mutants of D. salina were screened with 200 mmol/L NaClO3 from the mutant pool treated with mutagen ENU. Results D. salina is sensitive to KClO3 and NaClO3 with sensitive concentration being 100 mmol/L and 200 mmol/L respectively. Three chlorate-resistant mutants of D. salina were screened and showed the resistance to chlorate. Conclusions Chlorate sensitivity of D. sa- lina can be used to isolate chlorate-resistant mutants and prepare nitrate reduetase (NR)-deficient mutants of D. salina,whieh founded solid foundation for producing angiostatin using transgenie D. salina as bioactor.