Journal
MOLECULAR ECOLOGY
Volume 19, Issue -, Pages 81-88Publisher
WILEY
DOI: 10.1111/j.1365-294X.2009.04470.x
Keywords
plant virus; virus biodiversity; metagenomics; wild viruses
Funding
- National Science Foundation [EF-0627108, FPS-0447262]
- The United States Department of Agriculture [OKLR-2007-01012]
Ask authors/readers for more resources
Environmental samples have been analysed for viruses in metagenomic studies, but these studies have not linked individual viruses to their hosts. We designed a strategy to isolate double-stranded RNA, a hallmark of RNA virus infection, from individual plants and convert this to cDNA with a unique four nucleotide Tag at each end. Using 96 different Tags allowed us to pool samples and still retain the link to the original sample. We then analysed the sequence of pooled samples using massively parallel sequencing with Roche 454 pyrosequencing such that 384 samples could be assessed per picotiter plate. Using this method we have been able to analyse thousands of plants, and we have discovered several thousand new plant viruses, all linked to their specific plant hosts. Here we describe the method in detail, including the results and analysis for eight pools of samples. This technology will be extremely useful in understanding the full scope of plant virus biodiversity.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available