Illumina sequencing of RYMV variants that escape the resistance of Tog5307 accession, carrying the rymv3-R1 resistance gene

We sequenced RYMV genome from infected leaf samples of the Tog5307 African rice accession to investigate the molecular determinants of RYMV3 resistance-breaking. Different RYMV isolates have been inoculated to Tog5307 accession, carrying the resistance allele Rymv3-R1 of RYMV3 gene. Sporadic infection events were observed (Pidon et al, 2017, doi :10.1007/s00122-017-2853-0). Leaves from symptomatic Tog5307 plants were used for back-inoculation on Tog5307 to confirm that infection events resulted from evolution of the virus toward virulence. Eight samples used in the back-inoculation tests were selected for full RYMV genome sequencing using Illumina technology. Corresponding RYMV wild-type samples collected on the susceptible variety IR64 were sequenced as references. The detailed protocols of samples and libraries preparation, the sequencing conditions and the sequence analysis have been previously described in Pinel-Galzi et al. (2016, doi :10.3389/fpls.2016.01779).

本研究对非洲稻种质Tog5307的染病叶片样本中的水稻黄斑驳病毒（Rice Yellow Mottle Virus, RYMV）基因组进行测序，以解析RYMV3抗性突破的分子决定因子。将不同RYMV分离株接种至携带RYMV3基因抗性等位基因Rymv3-R1的Tog5307种质系，仅观察到零星的侵染事件（Pidon等，2017，DOI：10.1007/s00122-017-2853-0）。采集表现症状的Tog5307植株叶片进行回接种实验，以确认该侵染事件源于病毒向致病力方向的演化。选取回接种实验中的8份样本，采用Illumina测序技术完成RYMV全基因组测序。以感病品种IR64上采集的对应RYMV野生型样本作为参照进行测序。样本与文库制备的详细流程、测序条件及序列分析方法已在Pinel-Galzi等（2016，DOI：10.3389/fpls.2016.01779）中已有详述。