Flow cytometric detection and enumeration of DNA and RNA viruses infecting marine eukaryotic microalgae |
| |
| Authors: | Yuji Tomaru Keizo Nagasaki |
| |
| Institution: | (1) National Research Institute of Fisheries and Environment of Inland Sea, 2-17-5 Maruishi, Hatsukaichi, Hiroshima 739-0452, Japan |
| |
| Abstract: | Sample preparation protocols for flow cytometry (FCM) analysis with five algal viruses were optimized: Heterocapsa circularisquama virus (HcV), Heterosigma akashiwo virus (HaV), Chaetoceros salsugineum nuclear inclusion virus (CsNIV), Rhizosolenia setigera RNA virus (RsRNAV) and H. circularisquama RNA virus (HcRNAV). The optimum staining protocols differed significantly among the viruses tested. FCM counts for the large
DNA algal viruses HaV and HcV (∼0.2 μm in diameter) were similar to numbers determined by epifluorescence microscopy (EFM).
In contrast, FCM counts of viruses smaller than 40 nm that harbor DNA (CsNIV) or RNA genomes (RsRNAV, HcRNAV) were comparable
to or lower than most probable number (MPN) values, which indicate only infectious virus number, suggesting that the FCM counts
were underestimates. This is presumably because their single particle fluorescence signals were below the detection limit
for the flow cytometer. These results indicate that a large portion of the smaller viruses in the aquatic plankton virus community
may be overlooked by FCM. |
| |
| Keywords: | Flow cytometry algal virus SYBR Green I SYBR Gold DNA RNA |
| 本文献已被 SpringerLink 等数据库收录！ |
|
