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利用基因枪转化法,以优质粳稻保持系早花二B为受体,获得了含有GNA和Hyg基因的转基因植株。对T2产生的6个纯合株系进行多代潮霉素筛选、PCR分子检测及稻飞虱接种鉴定,表明标记基因在受体中稳定遗传,并高效表达,且一经纯合不再分离;而目的基因表达效果不一样,只有T-10(克隆号A5-32)表达效果最好,其抗虫性达到中抗(MR)以上水平,比受体抗性(S)明显提高。将转基因保持系与不育系早花二A回交,获得的转基因不育系高抗潮霉素,且抗性也完全纯合,表明转基因保持系的抗性基因已转化到不育系中,并得到高效表达。
The transgenic plants containing GNA and Hyg genes were obtained by the gene gun transformation method and the high-quality japonica rice maintainer Bachem B as receptor. Six generations of T2 homozygous lines were screened by hygromycin selection, PCR and DNA inoculation. The results showed that the tagged genes were stably inherited in the recipient and highly expressed, and were no longer separated after being homozygous. The expression of target gene was not the same, only T-10 (clone No.5-32) had the best expression and its insect resistance reached the level of medium resistance (MR), which was significantly higher than that of receptor (S). The transgenic maintainer line was backcrossed back to the CMS line early. The resulting transgenic sterile lines were highly hygromycin resistant and also completely homozygous, indicating that the resistance gene from the transgenic maintainer line had been transformed into a CMS line and Efficient expression.