Cloning of cDNA Encoding CCoAOMT from Populus tomentosa and Down-regulation of Lignin Content in Transgenic Plant Expressing Antisense Gene  

毛白杨CCoAOMT cDNA片段的克隆与转基因杨木质素含量的调控(英文)

作　　者：魏建华[1] 赵华燕[1] 张景昱[1] 刘惠荣[1] 宋艳茹[1] 

机构地区：[1]中国科学院植物研究所,北京100093

出　　处：《Acta Botanica Sinica》2001年第11期1179-1183,共5页Acta Botanica Sinica（植物学报：英文版）

摘　　要：cDNA encoding caffeoyl CoA O-methyltransferase (CCoAOMT) from Chinese white poplar ( Populus tomentosa Carr.) was cloned by RT-PCR and sequenced. Northern analysis displayed that the CCoAOMT was expressed specifically in the developing secondary xylem and its expression was coincident with lignification. The antisense CCoAOMT cDNA was transformed into P. tremula x P. alba mediated by Agrobacterium tumefaciens ( Smith et Townsend) Conn. Transgenic plants were identified with PCR, PCR-Southern and Southern analysis. Lignin content in 5- to 6-month-old transgenic plants was measured. One of the transgenic lines had significant reduction of 17.9% in Klason lignin content as compared with that of untransformed poplar. The results demonstrate that antisense repression of CCoAOMT is an efficient way to reduce lignin content for improving pulping property in engineered trees.用RT_PCR方法从毛白杨 (PopulustomentosaCarr.)中克隆了CCoAOMT基因的cDNA片段 ,并对其序列进行了分析。Northern杂交表明该基因在毛白杨正在木质化的次生木质部高水平表达 ,且与木质化进程同步。构建了该基因反义表达载体 ,根癌土壤杆菌 (Agrobacteriumtumefaciens (SmithetTownsend)Conn)介导转化杂交杨 (欧洲山杨×银白杨 ,P .tremula×P .alba)。采用PCR、PCR_Southern及Southern杂交对获得的转基因植株进行分子检测 ,并测定移栽 5～ 6个月的转基因植株的Klason木质素含量 ,其中一个转基因株系的Klason木质素含量比未转基因对照杨树下降 17.9%。结果表明利用反义RNA技术对杨树CCoAOMT基因的表达可以降低转基因植株的木质素含量 。

关 键 词：RT-PCR Populus tomentosa CCOAOMT antisense RNA LIGNIN 

分 类 号：Q943.2[生物学—植物学]