毛白杨TIR-NBS-LRR基因转化烟草的研究  

Transformation of tobaccos with a TIR-NBS-LRR gene isolated from Populus tomentosa Carr

作　　者：李海霞[1] 张志毅[1] 张谦[1] 安新民[1] 李琰[1] 刘婷婷[1] 

机构地区：[1]北京林业大学林木育种国家工程实验室、林木花卉遗传育种教育部重点实验室

出　　处：《北京林业大学学报》2009年第1期73-78,共6页Journal of Beijing Forestry University

基　　金：国家自然科学基金项目(30872043);高等学校博士学科点专项科研基金项目(20070022003)

摘　　要：该文采用农杆菌介导法将从毛白杨中克隆得到的TIR-NBS-LRR抗病基因(PtDRG01)导入烟草中,经抗生素筛选和PCR分子检测,获得了一批阳性转化植株。进而对这些阳性转化植株进行烟草花叶病毒接种实验,从表型观察结果发现,在接种病毒1周和6周后,转基因烟草株系TG-11的发病程度明显低于非转基因烟草。荧光定量分析结果显示,转基因烟草株系TG-11叶片中的烟草花叶病毒(TMV)数量显著低于非转基因植株,且该转基因株系中的PtDRG01基因转录水平显著高于非转基因植株。这些结果表明,毛白杨PtDRG01基因具有明显的抗病功能,其高效表达能够显著提高烟草的抗TMV能力。该文通过对烟草的遗传转化与病毒接种实验,鉴定了PtDRG01基因的功能,可为其他TIR-NBS-LRR基因的功能鉴定提供参考。A TIR-NBS-LRR gene（ PtDRG01 ） isolated from Populus tomentosa Can. was transferred into tobacco＇s via the Agrobacterium-mediated method. Antibiotics screening and PCR analysis showed that the gene of interest was integrated into tobacco genome. Resistance test with tobacco mosaic virus （TMV） suggested that the disease severity of transgenic tobacco TG-11 was significantly lower than that of non-transgenic tobaccos one week and six weeks after inoculation. By real-time PCR, the authors found that the number of virus in TG-11 was much lower than that of non-transgenic tobaccos and the transcription level of gene PtDRG01 in TG-11 was significantly higher than that of non-transgenic tobaccos. These results revealed that the PtDRG01 gene had an obvious resistance function and its efficient expression could enhance tobacco＇s resistance to TMV. This research, through tobacco transformation and TMV inoculation test, identifies the function of PtDRG01 gene, thus provides references for functional identification of other TIR-NBS-LRR type of genes.

关 键 词：毛白杨 TIR-NBS-LRR基因 转基因烟草 烟草花叶病毒 抗病性 

分 类 号：S792.117[农业科学—林木遗传育种] S718.46[农业科学—林学]