Experimental study of corona discharge in the production of transgenic silkworms from diapause strains

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  • Yamada Nobuto
    Division of Silk-Producing Insect Biotechnology, Institute of Agrobiological Sciences, NARO
  • Kojima Katsura
    Division of Silk-Producing Insect Biotechnology, Institute of Agrobiological Sciences, NARO
  • Mise Yoshiko
    Division of Silk-Producing Insect Biotechnology, Institute of Agrobiological Sciences, NARO
  • Yonemura Naoyuki
    Division of Silk-Producing Insect Biotechnology, Institute of Agrobiological Sciences, NARO
  • Sakai Hiroki
    Division of Silk-Producing Insect Biotechnology, Institute of Agrobiological Sciences, NARO
  • Sezutsu Hideki
    Division of Silk-Producing Insect Biotechnology, Institute of Agrobiological Sciences, NARO
  • Uchino Keiro
    Division of Silk-Producing Insect Biotechnology, Institute of Agrobiological Sciences, NARO
  • Tamura Toshiki
    Silk Sciences and Technology Research Institute
  • Iizuka Tetsuya
    Division of Silk-Producing Insect Biotechnology, Institute of Agrobiological Sciences, NARO

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<p> The effect of corona discharge on the silkworm’s egg diapause, Bombyx mori, has been studied for more than 150 years ago and showed that it can effectively prevent embryonic diapause. In this study, we evaluated the applicability of the corona discharge to the transgenesis of the diapause silkworm strains. We developed our corona discharge equipment to optimize the treatment parameters for the transgenesis of the diapause strain. In the first experiment, we treated the diapause eggs with a corona discharge 2 h after oviposition, then microinjected the plasmid DNAs to produce transgenic silkworms and reared the hatched silkworms. In G1 embryos, when we screened the transgenic silkworms, we discovered that they were present in one-fourth of the broods. In the second experiment, we injected the genetic materials into the eggs <6 h after oviposition, incubated them for one night at 25°C in the second experiment, and then treated them with corona discharge. Consequently, the hatched larvae were reared, G1 eggs were obtained, and the G1 embryo was screened. In both experiments, we obtained numerous transgenic silkworms. Our results depict that both diapause eggs, when treated with corona discharge before and after the injection, could be convenient and effective in producing transgenic silkworms. The first experiment results concurred with the report by Zhang et al. (2022). Moreover, we included new findings in the second experiment depicting that the corona discharge after the microinjection benefits transgenesis.</p>

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