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利用克隆得到的毛白杨c3h1基因构建其RNAi抑制表达载体,通过根癌农杆菌介导的叶盘法转化银腺杨无性系84 K,Realtime PCR检测表明其转基因株系323、325和322中c3h1基因表达量较野生型植株分别下调89.04%、82.22%和68.38%;茎横切片组化染色和显微结构观察表明转基因植株木质部发育和木质素沉积方式发生了改变;木质素、纤维素含量测定及苯酚—硫酸法总糖含量与HPLC法可溶性总糖和单糖含量检测结果表明:转基因植株木质素含量平均降低23.00%,最高可达39.71%;酸前处理效率最高提高了41.39%;未经酸处理直接酶解的糖化效率是对照植株的2.34～2.72倍,322株系和323株系比对照植株经酸前处理后再酶解的糖化效率高出81.18%和375.53%。
The cloned Populus tomentosa c3h1 gene was used to construct its RNAi expression vector. Agrobacterium tumefaciens-mediated leaf disk method was used to transform 84 G clones of A. gossypii. Realtime PCR results showed that the c3h1 gene of transgenic lines 323, 325 and 322 The gene expression was 89.04%, 82.22% and 68.38% lower than that of the wild-type plants respectively. The histochemical staining and microscopic observation of stem cross-sections showed that the development of xylem and the deposition of lignin in transgenic plants changed. The content of lignin and cellulose The results showed that the average lignin content of transgenic plants was reduced by 23.00% and the highest was 39.71%, the highest efficiency of acid pretreatment was increased by 41.39% Saccharification efficiency was 2.34 ~ 2.72 times higher than that of the control plants by acid treatment. The efficiency of saccharification of 322 plants and 323 plants was 81.18% and 375.53% higher than that of the control plant after acid treatment.