RNA干涉培育低木质素杨树

Research on RNA-interference Breeding of Low-lignin Poplar Varieties

采用改良的CTAB法提取南林95杨的基因组DNA,经PCR扩增得到肉桂酰辅酶A还原酶CCR基因的第4个外显子部分序列,通过中间载体pUCCRNAi,构建含正反向干涉片段的pBI121表达载体,导入农杆菌LBA4404。利用叶盘法侵染南林95杨,获得3株转基因植株,经分子鉴定证实干涉片段已导入南林95杨。测定Klason木质素及综纤维素含量的结果显示:转基因植株Klason木质素含量与对照相比平均降低了9.86%,综纤维素含量与对照相比平均增加了3.17%,纤维长宽比明显增加,均表明转基因植株更有利于造纸。

Total genome DNA of＇Nanlin 95＇poplar was extracted with improved CTAB method, and the 4th exon sequence segment of CCR genome from Cinnamoyl coenzyme A was obtained with PCR. The pBI121 expression vector of the forward and reverse short interfering RNA segments was structured with the intermediate vector pUCCRNAi and led to Agrobacterium tumefaciens LBA4404. 3 genetically-modified plants were obtained with ＇Nanlin 95＇ leaf disc cocuhivation, and the plants were attested to have led-in genome DNA of ＇Nanlin 95＇. Determination of the Klason lignin and holocellulose contents showed that the Klason lignin content of the genetically-modified plants was lower than that of the control group by 9.86%, and the holocellulose content of the genetically-modified plants was higher than that of the control group by 3.17% with significantly increased length-width ratio. The results all showed that the genetically-modified plants are more suitable for making paper.