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为了解百脉根细胞壁形成中纤维素含量的变化,以其成熟叶片为材料,筛选优化原生质体分离制备的酶解液组成和制备培养原生质体,并测定原生质体再生壁形成过程中的纤维素含量。结果表明,优化后的酶解液配比为:1.00%纤维素酶+1.00%果胶酶+0.50%半纤维素酶,该酶液制备的原生质体产量和活力分别为1.12×106个/g·FW和68.7%。纤维素含量随着细胞壁的形成而逐渐增加,原生质体在培养1 d、3 d、5 d和7 d时,其纤维素含量分别占再生壁干重的1.13%、4.17%、8.08%和12.79%。
In order to understand the changes of cellulose content in the cell walls of Lotus japonicus, the mature leaves were used as material to screen and optimize the composition of enzymolysis solution prepared from protoplasts and to prepare protoplasts for protoplast regeneration. The cellulose content. The results showed that the optimized ratio of the hydrolyzate was 1.00% cellulase + 1.00% pectinase + 0.50% hemicellulase. The yield and viability of the prepared protoplasts were 1.12 × 106 / g FW and 68.7%. The content of cellulose increased with the formation of cell wall. The protoplasts accounted for 1.13%, 4.17%, 8.08% and 12.79% of the regenerated wall dry weight at 1 d, 3 d, 5 d and 7 d respectively %.