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Title 

Amplification of uncultured single-stranded DNA viruses from rice paddy soil

Authors 

Kyung Ho KimHo Won ChangYoung Do NamSeong Woon RohMin-Su KimY SungC O JeonHee-Mock OhJin-Woo Bae

Publisher 

American Society for Microbiology

Issue Date 

2008

Citation 

Applied and Environmental Microbiology, vol. 74, no. 19, pp. 5975-5985

Keywords 

amplificationrice paddiesvirusesdouble stranded DNAsingle stranded DNAvirus DNAsoilsvirus proteinpaddy fieldsoil microorganism

Abstract 

Viruses are known to be the most numerous biological entities in soil; however, little is known about their diversity in this environment. In order to explore the genetic diversity of soil viruses, we isolated viruses by centrifugation and sequential filtration before performing a metagenomic investigation. We adopted multiple-displacement amplification (MDA), an isothermal whole-genome amplification method with φ29 polymerase and random hexamers, to amplify viral DNA and construct clone libraries for metagenome sequencing. By the MDA method, the diversity of both single-stranded DNA (ssDNA) viruses and double-stranded DNA viruses could be investigated at the same time. On the contrary, by eliminating the denaturing step in the MDA reaction, only ssDNA viral diversity could be explored selectively. Irrespective of the denaturing step, more than 60% of the soil metagenome sequences did not show significant hits (E-value criterion, 0.001) with previously reported viral sequences. Those hits that were considered to be significant were also distantly related to known ssDNA viruses (average amino acid similarity, approximately 34%). Phylogenetic analysis showed that replication-related proteins (which were the most frequently detected proteins) related to those of ssDNA viruses obtained from the metagenomic sequences were diverse and novel. Putative circular genome components of ssDNA viruses that are unrelated to known viruses were assembled from the metagenomic sequences. In conclusion, ssDNA viral diversity in soil is more complex than previously thought. Soil is therefore a rich pool of previously unknown ssDNA viruses.

ISSN 

0099-2240

Link 

http://dx.doi.org/10.1128/AEM.01275-08

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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