文献类型：期刊文献

中文题名：通过RNAi技术抑制杨树c3h基因表达提高糖转化效率

英文题名：Downregulation of Coumaroyl Shikimate 3-hydroxylase in Poplar by RNAi Technique

作者：杨少寨[1,2] 柳新红[2] 赵树堂[1] 王敏杰[1] 卢孟柱[1]

第一作者：杨少寨

机构：[1]中国林业科学研究院林业研究所国家林业局林木培育重点实验室,北京100091;[2]浙江省林业科学研究院,杭州310023

年份：2012

卷号：32

期号：3

起止页码：1-8

中文期刊名：浙江林业科技

外文期刊名：Journal of Zhejiang Forestry Science and Technology

收录：北大核心:【北大核心2011】；

基金：863项目“高产优质多抗杨树分子与细胞高效育种技术及品种创制”（2006AA100109-1）,863项目“特色值物高效转化技术的建立”（2007AA10Z182）,863项目“杨树木材发育的基因调控研究”（2006AA10Z122）

语种：中文

中文关键词：RNA干扰;杨树;c3h;木质素;糖转化效率

外文关键词：RNAi; poplar; c3h; lignin; sacchrification

分类号：S718.4

摘要：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84K，RealfirnePCR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％．
Efficient hydrolysis of lignocellulose is the biggest technical challenges on forest bioenergy development due to lignin hindrance to bioconversion from lignocellulose to ethanol in poplar. Experiments were conducted to genetically modify poplar to decrease lignin content in order to enhance fermentable sugars which can be converted to ethanol, c3hl gene in Populus tomentosa was cloned and its RNAi vector expressing ds-RNA was constructed. 8 transgenic lines harboring the RNAi constructs were obtained via the leaf-disc method and propagated by cutting for each lines in the greenhouse. The transcription level of c3hl in RNAi inhibition transgenic lines analyzed by real-time PCR decreased 89.04%, 82.22% and 68.38% in the RNAi inhibition line 323, 325 and 322 compared with the control. Stem cross-section staining and microsa＇ucture observations showed that the xylem development and lignin deposition pattern＇in transgenic plant changed. Transgenic plants with lowest lignin content generally matched with the highest content of cellulose and soluble total sugars and the highest sacchri？cation el？. ciency. The results indicated that lignin is probably the major factor in recalcitrance of cell walls to sacchari？cation. Moreover, it demonstrated that genetic reduction of lignin content effectively overcame cell wall recalcitrance to bioconversion.