Polano Cesare, Firrao Giuseppe
Department of Agricultural, Food, Environmental and Animal Sciences (DI4A), University of Udine, Udine, Italy.
Methods Mol Biol. 2019;1875:203-211. doi: 10.1007/978-1-4939-8837-2_16.
Genome drafts for the phytoplasmas may be rapidly and efficiently assembled from NGS sequence data alone exploiting the proper bioinformatic tools and starting from properly collected samples. Here, we describe the use of the Phytoassembly pipeline ( https://github.com/cpolano/phytoassembly ), a fully automated tool that accepts as input row Illumina data from two samples (a phytoplasma infected sample and a healthy reference sample) to produce a phytoplasma genome draft, using the healthy plant host genome as a filter and profiting from the difference in reads coverage between the genome of the pathogen and that of the host. For phytoplasma infected samples containing >2% of pathogen DNA and an isogenic healthy reference sequence the resulting assemblies span the almost entire genomes.
利用适当的生物信息学工具,并从正确采集的样本开始,仅通过NGS序列数据就可以快速、高效地组装植原体的基因组草图。在这里,我们描述了Phytoassembly流程(https://github.com/cpolano/phytoassembly)的使用,这是一个全自动工具,它接受来自两个样本(一个感染植原体的样本和一个健康对照样本)的原始Illumina数据作为输入,以产生植原体基因组草图,该流程以健康植物宿主基因组作为筛选条件,并利用病原体基因组和宿主基因组之间的 reads 覆盖度差异。对于病原体DNA含量>2%且有同基因健康对照序列的感染植原体的样本,最终组装结果几乎覆盖了整个基因组。