Ri质粒介导fps基因对烟草的转化及表达  

Expression of farnesyl diphosphate synthase gene(fps) in Nicotinan tabacum hairy root via Ri-mediated transformation

作　　者：崔红[1] 慕平利[1] 李雪君[1] 刘梦林[1] 王哲[1] 

机构地区：[1]河南农业大学国家烟草栽培生理生化研究基地,河南郑州450002

出　　处：《浙江大学学报（农业与生命科学版）》2007年第4期355-359,共5页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：国家烟草专卖局科技资助项目(110200101005)

摘　　要：为了探讨外源fps基因表达对发状根合成抗菌物质能力的影响,将含有薄荷(Mentha spicataL.)fps基因的双元表达载体质粒,通过冻融法转化到发根农杆菌菌株Ar1334中,转化菌株与烟草叶圆片共培养诱导发状根产生,共获得18个稳定的发状根无性系,在附加20 mg·L-1卡那霉素的MS培养基上,筛选到4个生长旺盛的无性系.经PCR扩增得到大小为1 kb的目的片段,PCR-Southern检测表明外源fps基因已整合到烟草发状根的基因组中,Northern blot检测证明外源fps基因在转录水平进行了表达;酶活分析显示转基因发状根株系中FPS酶活性明显高于对照,抑菌试验也发现转基因发状根汁液对赤星病菌具有较高水平的抗性,表明外源fps基因在发状根中的表达有利于抗菌物质的合成与积累.To improve phytoalexin synthesis capability of hairy roots, farnesyl diphosphate synthase gene （fps） of Mentha spicata L. was transferred into tobacco via Ri-mediated transformation, The engineering Agrobacterium rhizogenes strain Ar1334 harboured the binary vector pBinARfps, in which the fps cDNA was placed under 35S promoter, The leaf explants of K326 co-cultured with Ar1334 were transformed on MS medium to induce hairy roots. Among the 18 lines of hairy root induced, 4 lines showed some kanamycin （20 mg· L^-1 ） resistance. All the transgenic hairy root lines showed typical hairy root characteristics on MS medium without any plant hormone. Polymerase chain reaction and Southern blot revealed the foreign gene fps was intergrated into these 4 lines. Northern blot analysis indicated that the foreign fps gene was expressed at the transcriptional level. Transgenic hairy root lines showed higher FPS activity than the control and disease challenge also proved that the juice of transgenic hairy roots could inhibit the growth of Alternaria alternata. This result proved that foreign fps gene expression could improve the accumulation of resistance substance in hairy roots in vitro culture,

关 键 词：烟草 法呢基焦磷酸合酶 发状根 赤星病菌 

分 类 号：Q943[生物学—植物学]