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Mutation rate and novel ${it tt}$ mutants of ${it Arabidopsis thaliana}$ induced by carbon ions

シロイヌナズナにおけるカーボンイオン誘発突然変異率と新規${it tt}$突然変異体の単離

鹿園 直哉; 横田 幸彦*; 北村 智; 鈴木 智広*; 渡辺 宏; 田野 茂光; 田中 淳

Shikazono, Naoya; Yokota, Yukihiko*; Kitamura, Satoshi; Suzuki, Chihiro*; Watanabe, Hiroshi; Tano, Shigemitsu; Tanaka, Atsushi

高等植物におけるイオンビームの突然変異誘発効果を調べるため、シロイヌナズナにカーボンイオンを照射した。カーボンイオンは電子線に比べ胚発生致死及び葉緑素欠損突然変異体をそれぞれ高頻度(11倍,7.8倍)に誘発した。カーボンイオン照射後の突然変異体選抜の過程で2つの新しいフラボノイド突然変異体(${it tt18, tt19}$)を単離した。3系統の${it tt18}$のうちの2系統では${it LDOX}$遺伝子内に小さな欠失をもつこと、残りの1系統では${it LDOX}$遺伝子を含む断片に大きな構造変化が起こっていることがPCR及び塩基配列解析からわかった。分離頻度を用いて突然変異率を算出したところ、カーボンイオンは電子線に比べ17倍突然変異率が高いことが明らかになった。新しい突然変異体が単離できたこと、及び突然変異率が高いことは、イオンビームが植物遺伝学において有用な変異原として利用できることを示唆する。

Irradiation of ${it Arabidopsis thaliana}$ by carbon ions was carried out to investigate the mutational effect of ion particles in higher plants. Frequencies of embryonic lethals and chlorophyll deficient mutants were found to be significantly higher after carbon ion-irradiation than after electron-irradiation (11-fold and 7.8-fold per unit dose, respectively). During the isolation of mutants, two new classes of flavonoid mutants (${it tt18, tt19}$) were isolated from carbon ion-mutagenized M2 plants. From PCR and sequence analysis, two of the three ${it tt18}$ mutant alleles were found to have a small deletion within the ${it LDOX}$ gene and the other was revealed to contain a rearrangement. Using the segregation rates, the mutation rate of carbon ions was estimated to be 17-fold higher than that of electrons. The isolation of novel mutants and the high mutation rate suggest that ion particles can be used as a valuable mutagen for plant genetics.

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