KLU基因转化亚麻荠的初步研究  

Preliminary Study on Transforming KLUinto Camelina sativa (L.) Crantz

作　　者：高立虎[1] 蔡永智[1] 王爱英[1] 沈海涛[1] 祝建波[1] 

机构地区：[1]石河子大学生命科学学院农业生物技术重点实验室,新疆石河子832003

出　　处：《河南农业科学》2013年第10期25-30,共6页Journal of Henan Agricultural Sciences

基　　金：国家转基因专项(2011ZX005-004);兵团博士基金项目(2009jc01)

摘　　要：为进一步提高油料作物亚麻荠的含油量,克隆拟南芥胚珠组织特异型基因的启动子pINO和拟南芥细胞色素P450 KLUH基因(KLU),以植物表达载体pCAMBIA2301为基础载体,构建组织特异性启动子pINO调控KLU基因的植物表达载体pCAMBIA2301-pINO-KLU,用根癌农杆菌介导的floral dip法转化亚麻荠。结果表明,农杆菌培养至OD600值为0.8时,用等体积渗入培养基(1/2MS、5%蔗糖、200μL/L Silwet L-77)重悬菌体转化亚麻荠,50mg/L卡那霉素检测转基因种子的阳性率为15%,PCR检测初步证明,KLU基因已整合到部分抗性植株的基因组中,转化率为1.8%。For further improving the oil content of an alternative oil crop species Camelina sativa （L.） Crantz,we cloned the Arabidopsis ovule organ-specific promoter （pINO） and the Arabidopsis cytochrome P450 KLUH gene （KLU）. Next,the plant expression vector pCAMBIA2301 was digested and fused KLU with pINO to construct the plant express vector pCAMBIA 2301 pINO-KLU,which was introduced into Camelina sativa （L.） Crantz with Agrobacterium-mediated floral dip transformation. The results showed that through using the Agrobacterium pellet to transform Camelina sativa （L.） Crantz, which was resuspended by the same volume of infiltration medium （1/2MS, 5% sucrose,200 μL/L Silwet L-77） when Agrobacterium culture was cul tured to OD600 = 0.8, positive rate of screening transgenic seeds by Kan （50 mg/L） was 15 %. Further PCR analysis confirmed that KLU gene had been incorporated into the genome of parts of resistant plant, and the transformation efficiency was 1.8 %.

关 键 词：亚麻荠 KLU 根癌农杆菌 滴花法 

分 类 号：S565.9[农业科学—作物学]