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Cyanobacteria, commonly known as blue-green algae, are important primary producers in aquatic ecosystems and common dominant algae causing algal blooms in freshwater. Cyanophages, especially procyanophages, are important planktonic ecological factors that affect the evolution of blue-green algae and aquatic microbial communities. Yet studies on procyanophages in cyanobacteria remain scarce. So far, few studies have reported the procyanophages in Microcystis. [Objective] To investigate the prevalence of lysogeny in Microcystis and characterize the genomic features of procyanophages in Microcystis. [Methods] All the 354 genome sequences of Microcystis spp. in GenBank were downloaded. PHASTER was used to predict procyanophage regions in the Microcystis genomes. Resistance and virulence factors in intact procyanophages and questionable procyanophages were annotated via the virulence factors of bacterial pathogens (VFDB) and comprehensive antibiotic resistance database (CARD). Bioinformatics tools were used for gene annotation and phylogenetic analysis of the procyanophages in Microcystis. Microcystis flos-aquae FACHB-1344 and M. aeruginosa FACHB-1326 each was predicted to harbor an intact procyanophage by PHASTER. To evaluate the infection activities of these two procyanophages, mitomycin C induction assays and dot-ELISA were conducted. [Results] Among all the 354 Microcystis genomes, 98.3% were predicted to harbor intact procyanophages, questionable or imcomplete procyanophages. A total of 13 intact procyanophages, 5 questionable procyanophages and 725 incomplete procyanophages were predicted by PHASTER. The 13 intact procyanophages and 5 questionable procyanophages were named as WZ1-WZ13 and YS1-YS5, respectively. No antibiotic resistance or virulence gene was detected in them. The phylogenomic tree displayed distant evolutionary relationships between the 18 procyanophages and other known viruses. Bioinformatics analysis suggested that YS5 revealed a previously unknown novel genus. WZ2, WZ3, WZ4, WZ5, WZ6, WZ7, WZ9, WZ10, WZ11, WZ12 and YS3 together revealed a novel family. WZ1 and YS1 together revealed a novel family. YS2, YS4, WZ8 and WZ13 each revealed a novel family. The procyanophages in M. flos-aquae FACHB-1344 and FACHB-1326 were verified to be activated by mitomycin C. [Conclusion] Lysogeny widely exists in Microcystis spp. The novel procyanophages in Microcystis spp. unlock novel viral evolutionary lineages previously unknown. This study enriches the understanding about cyanobacterium-virus interactions and the diversity of aquatic viruses.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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