A method of batch-purifying microalgae with multiple antibiotics at extremely high concentrations  

作　　者：韩吉昌 王松 张琳 杨官品 赵璐 潘克厚 

机构地区：[1]Laboratory of Applied Microalgae Biology,Key Laboratory of Aquaculture of Ministry of Education,Ocean University of China [2]College of Marine Life Sciences,Ocean University of China

出　　处：《Chinese Journal of Oceanology and Limnology》2016年第1期79-85,共7页中国海洋湖沼学报（英文版）

基　　金：Supported by the National Basic Research Program of China(973 Program)(No.2011CB200901);the National Technical Supporting Project Foundation(No.2011BAD14B01);the National High Technology Research and Development Program of China(863 Program)(No.2013AA065801)

摘　　要：Axenic microalgal strains are highly valued in diverse microalgal studies and applications. Antibiotics, alone or in combination, are often used to avoid bacterial contamination during microalgal isolation and culture. In our preliminary trials, we found that many microalgae ceased growing in antibiotics at extremely high concentrations but could resume growth quickly when returned to an antibiotics-free liquid medium and formed colonies when spread on a solid medium. We developed a simple and highly efficient method of obtaining axenic microalgal cultures based on this observation. First, microalgal strains of different species or strains were treated with a mixture of ampicillin, gentamycin sulfate, kanamycin, neomycin and streptomycin （each at a concentration of 600 mg/L） for 3 days; they were then transferred to antibiotics-free medium for 5 days; and finally they were spread on solid f/2 media to allow algal colonies to form. With this method, five strains ofNannochloropsis sp. （Eustigmatophyceae）, two strains of Cylindrotheca sp. （Bacillariophyceae）, two strains of Tetraselmis sp. （Chlorodendrophyceae） and one strain ofAmphikrikos sp. （Trebouxiophyceae） were purified successfully. The method shows promise for batch- purifying microalgal cultures.A bs tract Axenic microalgal strains are highly valued in diverse microalgal studies and applications. Antibiotics,alone or in combination,are often used to avoid bacterial contamination during microalgal isolation and culture. In our preliminary trials,we found that many microalgae ceased growing in antibiotics at extremely high concentrations but could resume growth quickly when returned to an antibiotics-free liquid medium and formed colonies when spread on a solid medium. We developed a simple and highly efficient method of obtaining axenic microalgal cultures based on this observation. First,microalgal strains of different species or strains were treated with a mixture of ampicillin,gentamycin sulfate,kanamycin,neomycin and streptomycin(each at a concentration of 600 mg/L) for 3 days; they were then transferred to antibiotics-free medium for 5 days; and finally they were spread on solid f/2 media to allow algal colonies to form. With this method,five strains of Nannochloropsis sp.(Eustigmatophyceae),two strains of Cylindrotheca sp.(Bacillariophyceae),two strains of T etraselmis sp.(Chlorodendrophyceae) and one strain of Amphikrikos sp.(Trebouxiophyceae) were purified successfully. The method shows promise for batchpurifying microalgal cultures.

关 键 词：antibiotic AXENIC batch operation bacterium removal MICROALGA 

分 类 号：Q949.2[生物学—植物学]