ViromeofwildplantsontheriversideoftheancientcanalinZhenjiangcity,China."plant73,Sesamum indicum L."
From Jun. to Aug. 2018, to investigate the virome of plant species in the Ancient canal ecosystem in Zhenjiang City, 184 plant leaves samples were collected from 184 wild plants in the Ancient canal in Zhenjiang City, Jiangsu Province, China. All samples were collected by disposable materials. About 1g plant leaf were grounded using steel balls and re-suspended in ten volumes of phosphate-buffered saline (PBS) and vigorously vortexed for 5 min. The grounded samples were then frozen and thawed three times on dry ice. The supernatants were then collected after centrifugation (10 min, 15,000g). 500 l of each supernatant was filtered through a 0.45-m filter (Millipore) to remove eukaryotic and bacterial cell-sized particles. The filtrates enriched in viral particles were treated with DNase and RNase to digest unprotected nucleic acid at 37C for 60 min. Remaining total nucleic acid was then isolated using QIAamp MinElute Virus Spin Kit (Qiagen) according to manufacturer's protocol. 184 libraries were then constructed using Nextera XT DNA Sample Preparation Kit (Illumina) and sequenced using the Illumina Hiseq 2500 platform with 250 bases paired ends with dual barcoding for each pool. For bioinformatics analysis, paired-end reads of 250 bp generated by MiSeq were debarcoded using vendor software from Illumina.