Direct Gene Introduction into Plant Cells Using an N2-Laser Microbeam.

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  • Ｎ２レーザーマイクロビームによる植物細胞への直接遺伝子導入
  • N2 レーザー マイクロビーム ニ ヨル ショクブツ サイボウ エ ノ チョク

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Abstract

An N2-laser with a 337.1nm wavelength and 3ns pulse width burned a localized hole ca. 0.5μm in diameter on the cell wall and cell membrane of cultured cells of melon under selected irradiation conditions (1.5-6μJ/spot). No visible damage due to the irradiation was detected under these conditions. Efficiency of gene delivery into plant cells was then tested using this laser treatment. Mature pollen of Impatiens balsamina, I. walleriana and Lycoris albiflora was irradiated with a laser at 3μJ/spot in medium containing 0.3M mannitol and plasmid PBI221 harboring β-glucuronidase. The maximum frequency of transient gene expression was 24% in these treatments. When the melon cotyledons were treated with a laser in the presence of cucumber mosaic virus-RNA, virus multiplication was observed in the epidermal and around the cells 24hr after irradiation. These results suggest that the present laser method is useful for the transformation of plant cells.

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