Molecular nature of mutations induced by high-LET irradiation with argon and carbon ions in Arabidopsis thaliana

Hirano, Tomonari; Kazama, Yusuke; Ohbu, Sumie; Yuki, Satoshi; Yang, Lu; Kambara, T.; Fukunishi, Nobuhisa; Abe, Tomoko

doi:10.1016/j.mrfmmm.2012.04.010

Cited by 55 publications

(69 citation statements)

References 46 publications

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“…Following this, they were washed 5 times with sterilised water, and then incubated on 0.7% agar-containing Murashige and Skoog (MS) medium that was supplemented with MS vitamins and 3% sucrose at 4°C in the dark for 4 d to induce vernalisation. The M 2 seeds were collected as described previously (Kazama et al, 2011;Hirano et al, 2012). In short, eleven seedlings were grown in a plastic tray, from which M 2 seeds of all plants were corrected and were treated as one batch.…”

Section: Methodsmentioning

confidence: 99%

“…From the M 2 generation, hy and gl mutants were screened, whose phenotypes were long-hypocotyl (Koornneef et al, 1980) and non trichome (Walker et al, 1999), respectively. The putative responsible genes of hy and gl mutants (HY1, HY2, HY3, HY4, and HY5 for hy mutants; GL1, GL2, and TTG1 for gl mutants) were amplified by performing PCR, and then the amplified fragments were sequenced as described previously (Kazama et al, 2011;Hirano et al, 2012). When the whole region or a part of the coding region of the putative responsible genes could not be amplified, flanking sequence analysis by using TAIL-PCR was performed (Liu et al, 1995).…”

Section: Methodsmentioning

confidence: 99%

“…The mutation frequencies of the hy and gl mutants for Fe-ion irradiation were 0.13‰ and 0.03‰, respectively (Table 1). These mutation frequencies were lower than that for any other ions that were previously tested (Kazama et al, 2011;Hirano et al, 2012). Because of the low mutation frequencies for Fe-ion irradiation, we also isolated one bp mutant (Venglat et al, 2002) and one nl mutant (Kim et al, 2002;Horiguchi et al, 2005) from the M 2 generation, and included them for the characterisation of the DNA mutations.…”

Section: Mutant Screeningmentioning

confidence: 99%

“…The methods of screening and calculation of mutation frequencies were the same as those of previous studies (Kazama et al, 2011;Hirano et al, 2012). The mutation frequencies of the hy and gl mutants for Fe-ion irradiation were 0.13‰ and 0.03‰, respectively (Table 1).…”

Section: Mutant Screeningmentioning

confidence: 99%

“…This highly efficient LET was designated as LETmax. The value of the LET also affects the size of the induced deletion: it becomes large with increasing LET Kazama et al, 2011;Hirano et al, 2012). Taken together, it is possible to select the LET value depending on the experimental purpose or target genes.…”

Section: Introductionmentioning

confidence: 99%

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Effect of high-LET Fe-ion beam irradiation on mutation induction in <i>Arabidopsis thaliana</i>

Kazama¹,

Hirano²,

Nishihara

et al. 2013

Genes Genet. Syst.

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Heavy-ion beams are powerful mutagens. They cause a broad spectrum of mutation phenotypes with high efficiency even at low irradiation doses and short irradiation times. These mutagenic effects are due to dense ionisation in a localised region along the ion particle path. Linear energy transfer (LET; keV·μm -1 ), which represents the degree of locally deposited energy, is an important parameter in heavy-ion mutagenesis. For high LET radiation above 290 keV•μm -1 , however, neither the mutation frequency nor the molecular nature of the mutations has been fully characterised. In this study, we investigated the effect of Fe-ion beams with an LET of 640 keV•μm -1 on both the mutation frequency and the molecular nature of the mutations. Screening of well-characterised mutants (hy and gl) revealed that the mutation frequency was lower than any other ion species with low LET. We investigated the resulting mutations in the 4 identified mutants. Three mutants were examined by employing PCR-based methods, one of which had 2-bp deletion, another had 178 bp of tandemly duplication, and other one had complicated chromosomal rearrangements with variable deletions in size at breakpoints. We also detected large deletions in the other mutant by using array comparative genomic hybridisation. From the results of the analysis of the breakpoints and junctions of the detected deletions, it was revealed that the mutants harboured chromosomal rearrangements in their genomes. These results indicate that Fe-ion irradiation tends to cause complex mutations with low efficiency. We conclude that Fe-ion irradiation could be useful for inducing chromosomal rearrangements or large deletions.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Mutant Screeningmentioning

confidence: 99%

Section: Mutant Screeningmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Effect of high-LET Fe-ion beam irradiation on mutation induction in <i>Arabidopsis thaliana</i>

Kazama¹,

Hirano²,

Nishihara

et al. 2013

Genes Genet. Syst.

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Mutants as a Genetic Resource for Future Crop Improvement

Gulfishan

Bhat²,

Oves

2015

Advances in Plant Breeding Strategies: Breeding, Biotechnology and Molecular Tools

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Characterization of a heavy-ion induced white flower mutant of allotetraploid Nicotiana tabacum

Kazama¹,

Fujiwara

Takehisa

et al. 2012

Plant Cell Rep

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KEY MESSAGE : We characterized a white flower mutant of allotetraploid N. tabacum as a DFR-deficient mutant; one copy of DFR has a cultivar-specific frameshift, while the other was deleted by heavy-ion irradiation. In most plants, white-flowered mutants have some kind of deficiency or defect in their anthocyanin biosynthetic pathway. Nicotiana tabacum normally has pink petals, in which cyanidin is the main colored anthocyanidin. When a relevant gene in the cyanidin biosynthetic pathway is mutated, the petals show a white color. Previously, we generated white-flowered mutants of N. tabacum by heavy-ion irradiation, which is accepted as an effective mutagen. In this study, we determined which gene was responsible for the white-flowered phenotype of one of these mutants, cv. Xanthi white flower 1 (xwf1). Southern blot analysis using a DNA fragment of the dihydroflavonol 4-reductase (DFR) gene as a probe showed that the xwf1 mutant lacked signals that were present in wild-type genomic DNAs. Sequence analysis demonstrated that one copy of the DFR gene (NtDFR2) was absent from the genome of the xwf1 mutant. The other copy of the DFR gene (NtDFR1) contained a single-base deletion resulting in a frameshift mutation, which is a spontaneous mutation in cv. Xanthi. Introduction of NtDFR2 cDNA into the petal limbs of xwf1 by particle bombardment resulted in production of the pink-colored cells, whereas introduction of NtDFR1 cDNA did not. These results indicate that xwf1 is a DFR-deficient mutant. One copy of NtDFR1 harbors a spontaneous frameshift mutation, while the other copy of NtDFR2 was deleted by heavy-ion beam irradiation.

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Molecular nature of mutations induced by high-LET irradiation with argon and carbon ions in Arabidopsis thaliana

Cited by 55 publications

References 46 publications

Effect of high-LET Fe-ion beam irradiation on mutation induction in <i>Arabidopsis thaliana</i>

Effect of high-LET Fe-ion beam irradiation on mutation induction in <i>Arabidopsis thaliana</i>

Mutants as a Genetic Resource for Future Crop Improvement

Characterization of a heavy-ion induced white flower mutant of allotetraploid Nicotiana tabacum

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