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Genome-wide survey of artificial mutations induced by ethyl methanesulfonate and gamma rays in tomato

机译:番茄中甲磺酸乙酯和γ射线诱导的人工突变的基因组调查

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Genome-wide mutations induced by ethyl methanesulfonate (EMS) and gamma irradiation in the tomato Micro-Tom genome were identified by a whole-genome shotgun sequencing analysis to estimate the spectrum and distribution of whole-genome DNA mutations and the frequency of deleterious mutations. A total of similar to 370 Gb of paired-end reads for four EMS-induced mutants and three gamma-ray-irradiated lines as well as a wild-type line were obtained by next-generation sequencing technology. Using bioinformatics analyses, we identified 5920 induced single nucleotide variations and insertion/deletion (indel) mutations. The predominant mutations in the EMS mutants were C/G to T/A transitions, while in the gamma-ray mutants, C/G to T/A transitions, A/T to T/A transversions, A/T to G/C transitions and deletion mutations were equally common. Biases in the base composition flanking mutations differed between the mutagenesis types. Regarding the effects of the mutations on gene function, >90% of the mutations were located in intergenic regions, and only 0.2% were deleterious. In addition, we detected 1 140 687 spontaneous single nucleotide polymorphisms and indel polymorphisms in wild-type Micro-Tom lines. We also found copy number variation, deletions and insertions of chromosomal segments in both the mutant and wild-type lines. The results provide helpful information not only for mutation research, but also for mutant screening methodology with reverse-genetic approaches.
机译:通过全基因组霰弹枪测序分析鉴定甲磺酸甲酯(EMS)和γ微基因组中γ辐射诱导的基因组突变,以估计全基因组DNA突变的光谱和分布和有害突变的频率。通过下一代测序技术获得了四种EMS诱导的突变体和三条γ射线照射线以及野生型线的370gB的配对读数。使用生物信息学分析,我们确定了5920个诱导的单一核苷酸变化和插入/缺失(Indel)突变。 EMS突变体中的主要突变是C / G至T / A的转变,而在γ射线突变体中,C / G至T / A转变,A / T至T / A横向,A / T至G / C.过渡和缺失突变同样常见。基础组成侧翼突变中的偏置在诱变类型之间不同。关于突变对基因官能的影响,突变的> 90%的突变位于非基因区域,并且仅0.2%是有害的。此外,我们在野生型微汤线中检测到140 687自发单核苷酸多态性和吲哚多态性。我们还发现突变体和野生型线路中的拷贝数变异,删除和染色体段的插入。结果不仅提供了有用的信息,不仅可以用于突变研究,而且还提供了具有反向遗传方法的突变体筛选方法。

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