《Table 2.Segregation of transgene-free rice in T1 generation.》

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《Production of Two Elite Glutinous Rice Varieties by Editing Wx Gene》

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Because inherited Cas9 could also induce new mutations and lead to unpredicted segregation，distortion and chimerism in later generations，it is important to eliminate Cas9 by segregation in the T1 generation and obtain stable mutants（Ishizaki，2016；Yin et al，2017）.Three wx-SK118（wx-SK118-1，wx-SK118-2and wx-SK118-3）and three wx-HD5（wx-HD5-1，wx-HD5-2and wx-HD5-3）T0 mutants were self-pollinated to generate T1 plants.Transgene-free lines were analyzed by PCR using specific primers for the Cas9 and HPTII sequences in the T1 plants（Table 1）.The results were determined using negative PCRs of both Cas9 and HPTII.The 235 wx-HD5 and 118 wx-SK118 T1plants（including selections from all the six lines）were subjected to PCR tests，respectively，and 36 wx-HD5 and 18 wx-SK118transgene-free plants were isolated，respectively（Table 2）.