RNA干涉培育低木质素杨树_宋恩慧.pdfVIP

安徽林业科技，2012，38 （2）：58~62 安 徽 林 业 科 技 2012 年 Anhui Forestry Science and Technology RNA 干涉培育低木质素杨树 宋恩慧，蔡 诚，魏 国，高 慧，项 艳 （安徽农业大学生物质改良与转化省级重点实验室，合肥230036） 摘 要：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。 关键词：CCR ；RNAi ；木质素；遗传转化 中图分类号：S718.46 文献标识码：A 文章编号：2095-0152 （2012）02-0058-05 Research on RNA- interference Breeding of Low- lignin Poplar Varieties SONG En-hui ；CAI Cheng，WEI Guo ；GAO Hui，XIANG Yan （Key Provincial Laboratory of Biomass Improvement and Conversion ，Anhui Agricultural University ，Hefei 230036，China） Abstract: 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 cocultivation ，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 pla