Comprehensive identification of mutations induced by heavy-ion beam irradiation in Arabidopsis thaliana
Comprehensive identification of mutations induced by heavy-ion beam irradiation in Arabidopsis thaliana
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DOI：
10.1111/tpj.12793
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发表时间：
2015-04-01
期刊：
影响因子：
7.2
通讯作者：
Abe, Tomoko
中科院分区：
文献类型：
--
作者：
Hirano, Tomonari;Kazama, Yusuke;Abe, Tomoko
重离子束被广泛用于诱变育种和分子生物学。虽然重离子束辐照的诱变效应的特点是一些限制性染色体区域或位点的序列分析，有没有在全基因组水平或详细的基因组重排的突变体基因组的评价。在这项研究中，使用阵列比较基因组杂交（array-CGH）和重测序，我们全面的特点在拟南芥基因组中的突变与Ar或Fe离子照射。随后，我们利用这些信息来调查重离子束的诱变效应。Array-CGH结果表明，Ar离子和Fe离子辐照后，每个基因组平均缺失面积分别为1.9和3.7个，缺失大小在149 ~ 602180 bp之间，81%的缺失伴有基因组重排。为了提供进一步的详细分析，对Ar离子束照射诱导的突变体的基因组进行重新测序，并使用三种算法检测总突变，包括碱基置换、重复、插入/缺失、倒位和易位。所有三个重新测序的突变体都有基因组重排。在22个参与重排的DNA片段中，19个片段参与染色体内重排，在染色体的局部区域形成多个重排。在多重重排区域检测到染色体间重排。这些结果表明，重离子束导致染色体中的成簇DNA损伤，并且它们具有诱导复杂的染色体内重排的巨大潜力。重离子束将被证明是植物育种和建立突变系的独特诱变剂。
Heavy-ion beams are widely used for mutation breeding and molecular biology. Although the mutagenic effects of heavy-ion beam irradiation have been characterized by sequence analysis of some restricted chromosomal regions or loci, there have been no evaluations at the whole-genome level or of the detailed genomic rearrangements in the mutant genomes. In this study, using array comparative genomic hybridization (array-CGH) and resequencing, we comprehensively characterized the mutations in Arabidopsis thaliana genomes irradiated with Ar or Fe ions. We subsequently used this information to investigate the mutagenic effects of the heavy-ion beams. Array-CGH demonstrated that the average number of deleted areas per genome were 1.9 and 3.7 following Ar-ion and Fe-ion irradiation, respectively, with deletion sizes ranging from 149 to 602180bp; 81% of the deletions were accompanied by genomic rearrangements. To provide a further detailed analysis, the genomes of the mutants induced by Ar-ion beam irradiation were resequenced, and total mutations, including base substitutions, duplications, in/dels, inversions, and translocations, were detected using three algorithms. All three resequenced mutants had genomic rearrangements. Of the 22 DNA fragments that contributed to the rearrangements, 19 fragments were responsible for the intrachromosomal rearrangements, and multiple rearrangements were formed in the localized regions of the chromosomes. The interchromosomal rearrangements were detected in the multiply rearranged regions. These results indicate that the heavy-ion beams led to clustered DNA damage in the chromosome, and that they have great potential to induce complicated intrachromosomal rearrangements. Heavy-ion beams will prove useful as unique mutagens for plant breeding and the establishment of mutant lines.