转抗菌肽基因毛白杨的培育及其对溃疡病的抗性  

Transformation of an Antimicrobial Peptide Gene LJAMP2 into Populus tomentosa and Resistance of the Transgenic Poplar to Canker Disease

作　　者：贾之春[1,2] 孙一铭[1,2] 秦志超[1,2] 肖训焰[1,2] 齐代华[1,2] 罗克明[1,2] 

机构地区：[1]西南大学生命科学学院三峡库区生态环境教育部重点实验室 [2]西南大学资源植物研究所,重庆400715

出　　处：《林业科学》2011年第7期123-127,共5页Scientia Silvae Sinicae

基　　金：国家自然科学基金(3081576);中央高校基本科研业务费专项资金(XDJK2009B018);重庆市自然科学基金(2009BA1004,2009BB0004)

摘　　要：以毛白杨为转基因受体材料,利用根癌农杆菌介导法转化来源于益母草的阳离子抗菌肽基因LJAMP2。经卡那霉素筛选,共获得50株抗性植株。GUS组织染色和PCR检测显示有30株抗性植株呈阳性,初步证明外源目的基因已整合到毛白杨基因组中。RT-PCR证实抗菌肽基因LJAMP2在转基因植株中能大量表达。离体抗病性试验表明:转基因毛白杨细胞粗提液的抑菌能力明显强于非转基因植株。进一步将溃疡病菌接种在转基因和野生型毛白杨茎段上培养30天,转基因植株的病级指数均低于非转化植株。上述抗性试验结果表明:在毛白杨中超量表达益母草抗菌肽基因LJAMP2能显著提高其溃疡病抗病性。An nsLTPs-like antimicrobial protein gene（LJAMP2）from the seeds of motherwort （Leonurus japonicus）was introduced into Chinese white poplar（Populus tomentosa）via Agrobacterium-mediated transformation using nptII gene conferring kanamycin resistance as a selectable marker. Thirty GUS-positive poplar lines were obtained and stable integration of transgenes in the plant genome was identified by PCR analysis. LJAMP2 expression was detected in transgenic plants by reverse transcription-PCR（RT-PCR）. Transgenic poplar lines were tested for resistance to the fungal pathogen Cytospora chrysosperma crude extracts from the transformants showed in vitro inhibitory activity against the fungal pathogen. in vivo assays demonstrated that the transgenic poplar plants displayed a significant reduction in their disease symptoms after infection with C. chrysosperma, compared to the control plants. These results suggested that constitutive expression of the LJAMP2 gene from motherwort could improve resistance to canker disease in Chinese white poplar.

关 键 词：毛白杨 LJAMP2基因 遗传转化 溃疡病 

分 类 号：S763.1[农业科学—森林保护学]