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LET dependence on killing effect and mutagenicity in the model filamentous fungus Neurospora crassa
Authors:Liqiu Ma  Yusuke Kazama  Tomonari Hirano  Ryouhei Morita  Shuuitsu Tanaka  Tomoko Abe
Affiliation:1. Laboratory of Genetics, Department of Regulatory Biology, Faculty of Science, Saitama University, Saitama, Japan;2. RIKEN Nishina Center, Saitama, Japan;3. Gunma University Heavy Ion Medical Center, Maebashi, Japan;4. RIKEN Nishina Center, Saitama, Japan;5. Faculty of Agriculture, University of Miyazaki, Miyazaki, Japan
Abstract:Purpose: To assess the unique biological effects of different forms of ionizing radiation causing DNA double-strand breaks (DSBs), we compared the killing effect, mutagenesis frequency, and mutation type spectrum using the model filamentous fungus Neurospora.

Materials and methods: Asexual spores of wild-type Neurospora and two DSB repair-deficient strains [one homologous recombination- and the other non-homologous end-joining (NHEJ) pathway-deficient] were irradiated with argon (Ar)-ion beams, ferrous (Fe)-ion beams, or X-rays. Relative biological effectiveness (RBE), forward mutation frequencies at the ad-3 loci, and mutation spectra at the ad-3B gene were determined.

Results: The canonical NHEJ (cNHEJ)-deficient strain showed resistance to higher X-ray doses, while other strains showed dose-dependent sensitivity. In contrast, the killing effects of Ar-ion and Fe-ion beam irradiation were dose-dependent in all strains tested. The rank order of RBE was Ar-ion?>?Fe-ion?>?C-ion. Deletion mutations were the most common, but deletion size incremented with the increasing value of linear energy transfer (LET).

Conclusions: We found marked differences in killing effect of a cNHEJ-deficient mutant between X-ray and high-LET ion beam irradiations (Ar and Fe). The mutation spectra also differed between irradiation types. These differences may be due to the physical properties of each radiation and the repair mechanism of induced damage in Neurospora crassa. These results may guide the choice of irradiation beam to kill or mutagenize fungi for agricultural applications or further research.
Keywords:High-LET ion beam  homologous recombination repair  non-homologous end-joining  large deletion  multiple mutations
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