The experiment was performed on induction tetraploid of Haliotis discus hannai at two-cell stage through cell fusion with PEG treatment. In this paper, the orthogonal experiment of three factors and three levels [L9 (34)] was used. Three factors and three levels were molecular weight of PEG: 8 000, 6 000, 4 000 mol·g-1; PEG concentration: 45%, 50%, 55%; treatment duration time: 1, 2, 3 min, respectively. The results showed that the optimal pattern of three factors and three levels on inducing tetraploid of Haliotis discus hannai at two-cell stage through using PEG treatment were: molecular weight 4 000, concentration 55%, treatment duration time: 2 min. The highest tetraploid induction rate was 10.8% at embryo period. The three factors treatment sequence was treatment duration time→concentration→molecular weight.
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