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一株海洋牟勒氏角毛藻(Chaetoceros muelleri)病毒的分离与初步鉴定
吴庆喜1,2, 程 凯2, 杨季芳1, 陈吉刚1, 赵以军2, 许 敏2
1.浙江万里学院 宁波市微生物与环境工程重点实验室;2.华中师范大学 城市水环境生态学湖北省重点实验室
摘要:
以海洋赤潮藻为材料, 从宁波市象山港附近海域采取水样, 经反冲超滤法对水样进行浓缩处理后, 用其对培养的赤潮藻细胞进行感染, 分离到了一株能裂解牟勒氏角毛藻的病毒, 经反复的液体感染, 建立了稳定的藻病毒-宿主藻系统, 命名为牟勒氏角毛藻病毒。该病毒具有很强的感染力, 稀释 108倍以后仍然具有明显的感染效果。病毒纯化后, 电镜观察显示此株病毒为球形多面体结构, 直径约为 58nm; 电镜切片观察显示, 感染后的藻细胞中充满了大量的游离的和组装好尚未释放的病毒粒子。经宿主范围鉴定,
关键词:  牟勒氏角毛藻, 病毒, 分离, 鉴定
DOI:10.11693/hyhz201103019019
分类号:
基金项目:国家海洋公益性行业科研专项, 200705014 号; 浙江省海洋与渔业局项目, 浙海渔计(2009)214 号
ISOLATION AND CHARACTERIZATION OF A MARINE CHAETOCEROS MUELLERI VIRUS
WU Qing-Xi1,2, CHENG Kai2, YANG Ji-Fang1, CHEN Ji-Gang1, ZHAO Yi-Jun2, XU Min2
1.Municipal Key Laboratory of Microorganism and Environmental Engineering, Zhejiang Wanli University;2.Hubei Key Laboratary of Urban Environmental Ecology, Central China Normal University
Abstract:
Water from Xiangshan harbor and the East China Sea were collected and concentrated by backflushing ultrafiltration, and then were used as infecting medium for growing marine red-tide algae as host. A novel virus named Chaetoceros muelleri virus was found that lysed C. muelleri. The virus was isolated by repeated liquid infection and a stable Phycoviruses-Host system of algae was established. Infection via Most-Probable-Number analysis revealed that the virus had a strong infectious power; it was found significantly infectious even after 108-fold dilution. The virus was purified through saccharose gradient ultracentrifugation. Phosphotungstic acid negative staining analysis revealed the viral capsid of C. muelleri virus had a distinct icosahedral shape with a diameter of 58nm. Electron microscopic section revealed that there were many free particles and some assembled viral particles ready to be released in the infected algae cells. Identification of the host range confirmed that the virus had a strict host specificity for C. muelleri cells but not others. This novel C. muelleri virus genome is about 23kb through molecular characterization. No known report for this virus, it is a novel marine phycovirus worthy to be studied.
Key words:  Chaetoceros muelleri, Virus, Isolation, Characterization
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