摘要
研究了半夏凝集素(Pinellia ternataagglutinin,PTA)基因对桃蚜(Myzus persicae)的抗性。从半夏幼叶中提取总RNA,采用RT-PCR法分离克隆出PTA1基因cDNA片段,该基因在GenBank中登录号为DQ092435。序列分析表明:该cDNA片段全长为810bp,编码269个氨基酸组成的蛋白,具有单子叶植物甘露糖凝集素基因家族的特征,含有3个由4个氨基酸(QDNY)组成的甘露糖专一结合位点,与报道的天南星科凝集素基因氨基酸序列的同源性在72.7%~92.9%。构建了35S启动子控制下的PTA1基因的植物表达载体pBI121PTA1,该载体含有抗卡那霉素的选择标记基因(nptⅡ),利用根癌农杆菌介导法转化烟草获得抗卡那霉素的植株。PCR和半定量RT-PCR分析表明:PTA1基因已经整合到植物基因组中并在转录水平上得到了有效表达。抗蚜虫鉴定表明:转基因植株有效地抑制了蚜虫的群体增长率,平均抑制率达77.02%,表明该基因对同翅目害虫的抗虫基因工程有重要的应用价值。
The effects of PTA gene encoding Pinellia ternata agglutinin(PTA)on the Myzus persicae were studied.Using total RNA isolated from Pinellia ternata young leaves,a cDNA containing PTA-1 gene was amplified by RT-PCR and cloned,and the gene sequence was accepted and released by GenBank with accession number DQ092435.Sequence analysis results showed that this gene is of 810 base pairs,and encoding a lectin protein of 269 anmino acids,it was found that the PTA-1 gene had many common characters of mannose-binding ...
出处
《南京农业大学学报》
CAS
CSCD
北大核心
2010年第2期45-50,共6页
Journal of Nanjing Agricultural University
基金
河北省农林科学院青年基金项目(A06030101)
河北省自然科学基金项目(C2009001301)
国家转基因重大专项资助项目(2009ZX08002-005B)
关键词
半夏凝集素
基因克隆
转基因烟草
抗性
桃蚜
Pinellia ternata agglutinin（PTA）
gene cloning
transgenic tobacco plants
resistance
Myzus persicae
