Study on the axenic culture and application of Porphyra haitanensisthallus
Study on the axenic culture and application of Porphyra haitanensisthallus
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作者：
Wu Xiao-kai
优化坛紫菜菌体无菌工艺，研究坛紫菜附生微生物对菌体的人工侵染，先用0.7%KI和0.1%(w/v)氨苄青霉素对坛紫菜菌体进行预处理，去除外来微生物，然后加入氨苄青霉素、链霉素、新霉素、卡那霉素等5种抗生素。单独或联合应用庆大霉素去除紫菜菌体中的附生微生物,优化了适宜的组合、浓度和培养条件。氨苄青霉素(终浓度300μg/mL)、卡那霉素(终浓度100μg/mL)和庆大霉素(终浓度100μg/mL)联合处理对坛紫菜菌体细胞影响最小。培养18小时,对89.5%紫菜附生微生物的抑制率达到80%以上。用105/mL的真菌孢子和菌悬液感染灭菌后的紫菜菌体。结果显示,感染后的菌体与微生物生长良好,而灭菌对照菌体培养22d后颜色褪色。将健康菌体表面剪成1~2mm划痕，感染浓度为108/mL的真菌孢子悬液，分别在18℃、28℃和35℃培养。28℃、35℃和穿刺处理下培养的菌体状况较差，腐烂明显，而18℃培养的坛紫菜菌体生长良好。即使与微生物共培养后。
The sterile processing of Porphyra haitanensis thallus was optimized.The artificial infection of thallus by epiphytic microorganisms from Porphyra was studied.The thalli of P.haitanensis were pretreated by 0.7% KI and 0.1%(w/v)ampicillin firstly to remove the outside organisms.Five antibiotics,such as ampicillin,streptomycin,neomycin,kanamycin and gentamicin,were applied independently or combinationally to remove the epiphytic microorganisms from the thallus of Porphyra.The appropriate combination,concentration and culture conditions were optimized.The cells of P.haitanensis thallus were minimum affected under the treatment of the combination of ampicillin(final concentration 300μg/mL),kanamycin(final concentration 100μg/mL) and gentamicin(final concentration 100μg/mL) for 18 hours,and the inhibitory rate of 89.5% epiphytic microorganisms from Porphyra was more than 80%.The sterilized Porphyra thallus was infected by 105/mL fungal spores and bacteria suspensions.The results showed that the infected thallus grew well with the microorganisms,while the sterilized control thallus had colour fading after they were cultured for 22 days.The sterilized healthy thallus were cut into 1~2mm scratches on the surface and infected by the fungal spores suspensions under the concentration of 108/mL,then cultured at 18℃,28℃and 35℃,respectively.The cultured thalli were in bad condition and rotted obviously under both 28℃ and 35℃ and puncture treatments,while the P.haitanensis thallus cultured at 18℃ grew well even after being co-cultured with microorganisms.