Characterization of highly efficient heavy-ion mutagenesis in Arabidopsis thaliana

doi:10.1186/1471-2229-11-161

	Characterization of highly efficient heavy-ion mutagenesis in Arabidopsis thaliana
	Kazama,Yusuke 1; Hirano,Tomonari 1,2; Saito,Hiroyuki 1; Liu,Yang1 ; Ohbu,Sumie 1; Hayashi,Yoriko 1; Abe,Tomoko 1,2
刊名	BMC Plant Biology
	2011-11-15
卷号	11
ISSN号	1471-2229
DOI	10.1186/1471-2229-11-161
英文摘要	AbstractBackgroundHeavy-ion mutagenesis is recognised as a powerful technology to generate new mutants, especially in higher plants. Heavy-ion beams show high linear energy transfer (LET) and thus more effectively induce DNA double-strand breaks than other mutagenic techniques. Previously, we determined the most effective heavy-ion LET (LETmax: 30.0 keV μm-1) for Arabidopsis mutagenesis by analysing the effect of LET on mutation induction. However, the molecular structure of mutated DNA induced by heavy ions with LETmax remains unclear. Knowledge of the structure of mutated DNA will contribute to the effective exploitation of heavy-ion beam mutagenesis.ResultsDry Arabidopsis thaliana seeds were irradiated with carbon (C) ions with LETmax at a dose of 400 Gy and with LET of 22.5 keV μm-1 at doses of 250 Gy or 450 Gy. The effects on mutation frequency and alteration of DNA structure were compared. To characterise the structure of mutated DNA, we screened the well-characterised mutants elongated hypocotyls (hy) and glabrous (gl) and identified mutated DNA among the resulting mutants by high-resolution melting curve, PCR and sequencing analyses. The mutation frequency induced by C ions with LETmax was two-fold higher than that with 22.5 keV μm-1 and similar to the mutation frequency previously induced by ethyl methane sulfonate. We identified the structure of 22 mutated DNAs. Over 80% of the mutations caused by C ions with both LETs were base substitutions or deletions/insertions of less than 100 bp. The other mutations involved large rearrangements.ConclusionsThe C ions with LETmax showed high mutation efficiency and predominantly induced base substitutions or small deletions/insertions, most of which were null mutations. These small alterations can be determined by single-nucleotide polymorphism (SNP) detection systems. Therefore, C ions with LETmax might be useful as a highly efficient reverse genetic system in conjunction with SNP detection systems, and will be beneficial for forward genetics and plant breeding.
语种	英语
出版者	BioMed Central
WOS记录号	BMC:10.1186/1471-2229-11-161
内容类型	期刊论文
源URL	[http://119.78.100.186/handle/113462/24453]
专题	中国科学院近代物理研究所
通讯作者	Abe,Tomoko
作者单位	1.RIKEN Nishina Center 2.RIKEN Innovation Center
推荐引用方式 GB/T 7714	Kazama,Yusuke,Hirano,Tomonari,Saito,Hiroyuki,et al. Characterization of highly efficient heavy-ion mutagenesis in Arabidopsis thaliana[J]. BMC Plant Biology,2011,11.
APA	Kazama,Yusuke.,Hirano,Tomonari.,Saito,Hiroyuki.,Liu,Yang.,Ohbu,Sumie.,...&Abe,Tomoko.(2011).Characterization of highly efficient heavy-ion mutagenesis in Arabidopsis thaliana.BMC Plant Biology,11.
MLA	Kazama,Yusuke,et al."Characterization of highly efficient heavy-ion mutagenesis in Arabidopsis thaliana".BMC Plant Biology 11(2011).