Target Lines for in Planta Gene Stacking in Japonica Rice

作者：Li, Ruyu^*; Han, Zhiguo; Yin, Qian; Li, Meiru; Zhang, Mingyong; Li, Zhenzhen; Wang, Ping; Jiang, Li; Ow, David W.^*

来源：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 2022, 23(16): 9385.

DOI：10.3390/ijms23169385

• transgenic
• Bxb1
• integrase
• recombinase
• GMO

The clustering of transgenes at a chromosome location minimizes the number of segregating loci that needs to be introgressed to field cultivars. Transgenes could be efficiently stacked through site-specific recombination and a recombinase-mediated in planta gene stacking process was described previously in tobacco based on the Mycobacteriophage Bxb1 site-specific integration system. Since this process requires a recombination site in the genome, this work describes the generation of target sites in the Japonica rice genome. Agrobacterium-mediated gene transfer yielded similar to 4000 random-insertion lines. Seven lines met the criteria of being single copy, not close to a centromere, not inserted within or close to a known gene or repetitive DNA, having precise recombination site sequences on both ends, and able to express the reporter gene. Each target line tested was able to accept the site-specific integration of a new gfp-containing plasmid and in three of those lines, we regenerated fertile plants. These target lines could be used as foundation lines for stacking new traits into Japonica rice.

• 单位
  中国科学院; 中国科学院研究生院