Real-time PCR detection of host-mediated cyanophage gene transcripts during infection of a natural Microcystis aeruginosa population.
Real-time PCR detection of host-mediated cyanophage gene transcripts during infection of a natural Microcystis aeruginosa population.
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DOI：
10.1264/jsme2.me10117
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发表时间：
2010
影响因子：
2.2
通讯作者：
Mitsuhiro Yoshida;Takashi Yoshida;Yukari Yoshida-Takashima;A. Kashima;S. Hiroishi
中科院分区：
文献类型：
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作者：
Mitsuhiro Yoshida;Takashi Yoshida;Yukari Yoshida-Takashima;A. Kashima;S. Hiroishi
本研究的目的是建立一种实时荧光定量RT-PCR（real-time RT-PCR）方法来检测和定量感染的铜绿微囊藻细胞中噬藻体的mRNA。实验数据表明，在培养物中噬藻体数量增加之前，宿主细胞内噬藻体g91 mRNA的相对丰度增加。这种转录模式表明了病毒感染的动力学，这表明实时RT-PCR方法是一种潜在的工具，用于环境监测噬藻体感染。在本次野外调查中，感染M.从噬藻体数量估计的铜绿假单胞菌细胞群体较低，为0.01-2.9个细胞mL（-1）。噬菌体g91 RNA的最高相对丰度（10（-2）/rnpB转录物）与Ma-LMM 01的基于实验室的生长数据的表达水平大致相同（感染宿主细胞的估计密度：10（5）个细胞mL（-1））;并且当噬藻体数量快速增加（以及宿主细胞数量减少）时观察到。噬藻体数量的定量对于理解噬菌体与其宿主之间的生态关系是重要的。我们的数据表明，在一个自然的宿主细胞群体内的噬菌体基因转录的定量是一个强大的工具，用于调查的噬菌体裂解在感染的宿主群体的定量影响。
The aim of this study was to develop a quantitative real-time reverse transcription-PCR (real-time RT-PCR) assay to detect and quantify mRNA of cyanophages within infected Microcystis aeruginosa cells in a freshwater pond. Laboratory-based data showed that the relative abundance of the cyanophage g91 mRNA within host cells increased before cyanophage numbers increased in culture. This transcriptional pattern indicated the kinetics of the viral infection suggesting the real-time RT-PCR method to be a potential tool for environmental monitoring of cyanophage infections. In this field survey, the numbers of infected M. aeruginosa cell populations estimated from cyanophage numbers were low at 0.01-2.9 cells mL(-1). The highest relative abundance of phage g91 RNA (10(-2) per rnpB transcript) was at about the same levels of expression as laboratory-based growth data for Ma-LMM01 (estimated density of infected host cells: 10(5) cells mL(-1)); and was observed when cyanophage numbers rapidly increased (as well as a decrease in host cell numbers). Quantification of cyanophage numbers is important to understand ecological relationships between the phage and its hosts. Our data suggest the quantification of phage gene transcripts within a natural host cell population to be a strong tool for investigating the quantitative effects of phage lysis during infection of the host population.