Yoshida Takashi, Takashima Yukari, Tomaru Yuji, Shirai Yoko, Takao Yoshitake, Hiroishi Shingo, Nagasaki Keizo
Department of Marine Bioscience, Fukui Prefectural University, 1-1 Gakuencho, Obama, Fukui 917-0003, Japan.
Appl Environ Microbiol. 2006 Feb;72(2):1239-47. doi: 10.1128/AEM.72.2.1239-1247.2006.
We isolated a cyanophage (Ma-LMM01) that specifically infects a toxic strain of the bloom-forming cyanobacterium Microcystis aeruginosa. Transmission electron microscopy showed that the virion is composed of anisometric head and a tail complex consisting of a central tube and a contractile sheath with helical symmetry. The morphological features and the host specificity suggest that Ma-LMM01 is a member of the cyanomyovirus group. Using semi-one-step growth experiments, the latent period and burst size were estimated to be 6 to 12 h and 50 to 120 infectious units per cell, respectively. The size of the phage genome was estimated to be ca. 160 kbp using pulse-field gel electrophoresis; the nucleic acid was sensitive to DNase I, Bal31, and all 14 restriction enzymes tested, suggesting that it is a linear double-stranded DNA having a low level of methylation. Phylogenetic analyses based on the deduced amino acid sequences of two open reading frames coding for ribonucleotide reductase alpha- and beta-subunits showed that Ma-LMM01 forms a sister group with marine and freshwater cyanobacteria and is apparently distinct from T4-like phages. Phylogenetic analysis of the deduced amino acid sequence of the putative sheath protein showed that Ma-LMM01 does not form a monophyletic group with either the T4-like phages or prophages, suggesting that Ma-LMM01 is distinct from other T4-like phages that have been described despite morphological similarity. The host-phage system which we studied is expected to contribute to our understanding of the ecology of Microcystis blooms and the genetics of cyanophages, and our results suggest the phages could be used to control toxic cyanobacterial blooms.
我们分离出一种蓝藻噬菌体(Ma-LMM01),它能特异性感染形成水华的有毒蓝藻铜绿微囊藻菌株。透射电子显微镜显示,病毒粒子由不规则形头部和尾部复合体组成,尾部复合体包括一根中央管和一个具有螺旋对称的收缩鞘。形态特征和宿主特异性表明Ma-LMM01是蓝藻肌病毒群的成员。通过半一步生长实验,潜伏期和裂解量分别估计为6至12小时和每细胞50至120个感染单位。使用脉冲场凝胶电泳估计噬菌体基因组大小约为160 kbp;核酸对DNase I、Bal31和所有测试的14种限制酶敏感,表明它是一种甲基化水平较低的线性双链DNA。基于编码核糖核苷酸还原酶α亚基和β亚基的两个开放阅读框推导的氨基酸序列进行的系统发育分析表明,Ma-LMM01与海洋和淡水蓝藻形成一个姐妹群,明显不同于T4样噬菌体。对假定的鞘蛋白推导的氨基酸序列进行的系统发育分析表明,Ma-LMM01与T4样噬菌体或前噬菌体均未形成单系群,这表明尽管形态相似,但Ma-LMM01与已描述的其他T4样噬菌体不同。我们研究的宿主-噬菌体系统有望有助于我们理解微囊藻水华的生态学和蓝藻噬菌体的遗传学,我们的结果表明噬菌体可用于控制有毒蓝藻水华。