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《Journal of Marine Sciences》 2008-04
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The determination of sensitivities of Porphyra haitanensis female thalloid to antibiotics

LI Fu-hou,WANG Wei-xia,WANG Ping,GAO Fei(Key Lab of Marine Biotechnology of Jiangsu Province,College of Marine Science,Huaihai Institute of Technology,Lianyungang 222005,China)  
Sensitivities of Porphyra haitanensis female thalloid to four kinds of antibiotics were studied respectively.The thalloid was soaked in sterilized seawater for anabiosis at 20 ℃ for 1~2 d first.Then,the anabatic thalloid was spread on a piece of filter paper and the tissues differentiated by the sexual cells on the margin were removed.The thalloid was cut into 2 mm×2 mm lumps,which were cultured in 25 mL PES medium.Different antibiotics of different mass concentrations were added into the culture medium.The cell development was observed with microscope for 18 d,and the dead tissues were recorded and analyzed.The experiment with the same concentration was done for three times,and at the same time,the experiments without adding antibiotics was done as the contrast.The results show that Porphyra haitanensis had different sensitivities to four antibiotics.Porphyra haitanensis was sensitive to chloramphenicol.When chloramphenicol was 50 μg/mL,the thalloid cells grew slowly,did not differentiate but cell division could be observed.When chloramphenicol was 100 μg/mL, lethal effects were observed,and the higher concentration of the antibiotics,the stronger was the lethal effect.When chloramphenicol was 100~250 μg/mL,more than half of female thalloid lumps died in 14 d.The cleavage of female thalloid cells was restrained strongly by the antibiotics of increasing concentration.Porphyra haitanensis had very strong tolerance to Kanamycin.After cultured for 14 d,the control group was normal.The cells were dark green in color and rhizoid appeared on the margin of the thalloid.The cell development of thalloid of the treated groups was fine.The cells were brownish red,and chromatophores were distinct,at the same time,the cells were splitting and cell wall thickening,which shows that Kanamycin had no inhibiting effect to the development of thalloid cells.Porphyra haitanensis had certain tolerance to tetracycline.But tetracycline could restrain cleavage and differentiation of thalloid cell,and it could restrain the formation of rhizoid.After cultured for 14 d,the dark brown cell mass of the female thalloid cell appeared,and chromatophores were not distinct.Cell mass also appeared when tetracycline was 500 μg/mL,and it contained lots of cells,but division of cells was not distinct.G-418 had certain inhibiting effects on the development of female thalloid.When G-418 was 50 μg/mL,rhizoid appeared on the margin of the thalloid,chromatophores were distinct,and splitting cells could be observed.When G-418 was 100~300 μg/mL,the cell mass of the female thalloid appeared,and chromatophores were not distinct.At the same time,the thalloid cells grew slowly,division of cells was not distinct,but there were no dead cells.Results show that chloramphenicol should be appropriate selectable marker for Porphyra haitanensis genetic engineering,and CAT gene could be an ideal selectable marker gene.
【Fund】: 江苏省海洋生物技术重点建设实验室资助项目(2005 HS011)
【CateGory Index】: S968.431
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