Discovery of Jasmin Virus H

Small RNA libraries were constructed from total RNA from Jasminum sambac plants exhibiting virus-like symptoms. After sequencing, small RNAs were assembled into contigs with MetaVelvet and assembled contigs were aligned against the NR database of NCBI using BLASTx. Top hits that reported a virus as subject were considered putative viral sequences. Based on such alignments, the whole genome of a virus, we tentatively name Jasmine Virus H was recovered and cloned. Two more small RNA libraries were made in a confirmatory experiment. One from Jasminum sambac and another one from Nicotiana benthamiana plants infected with the newly-cloned virus. The small RNA libraries were aligned against the full-length sequence of Jasmine Virus H to determine the spacial distribution of virus-derived small RNAs along the virus genome. Overall design: Symptomatic leaves were collected in the field and used for virus discovery and isolation as described in the summary. Collection was repeated once and the small RNAs libraries repeated. In addition an infectious clone was produced and used to infect Nicotiana benhtamina plants, from which total RNA was extracted and viral small RNAs characterized again. All libraries were sequenced in an Illumina HiSeq 2500 instrument.