【背景和目的】中国黄花稔曲叶病毒(Sida yellow mosaic China virus,SiYMCNV)是侵染海南儋州雪茄烟的重要病原,为了分析该病毒的致病性,以侵染海南雪茄烟的SiYMCNV分离物为材料,构建SiYMCNV的侵染性克隆。【方法】以侵染SiYMCNV的雪茄...【背景和目的】中国黄花稔曲叶病毒(Sida yellow mosaic China virus,SiYMCNV)是侵染海南儋州雪茄烟的重要病原,为了分析该病毒的致病性,以侵染海南雪茄烟的SiYMCNV分离物为材料,构建SiYMCNV的侵染性克隆。【方法】以侵染SiYMCNV的雪茄烟植物总基因组为模板,分别构建含有1.5倍串联重复DNA-A全长侵染性克隆pCAMBIA1300-1.5A和含有2倍串联重复DNA-β全长侵染性克隆pCAMBIA1300-2β。采用液氮冻融法转化农杆菌GV3101,通过农杆菌浸润验证侵染性克隆的致病性。【结果】SiYMCNV DNA-A单独侵染本氏烟和雪茄烟H382不表现症状,DNA-A和DNA-β混合侵染本氏烟和雪茄烟H382表现出典型的曲叶症状,且整株植物表现为矮化、皱缩。【结论】构建的中国黄花稔曲叶病毒侵染性克隆可高效侵染本氏烟和雪茄烟H382,为进一步开展该病毒致病性机制研究和抗病烟草种质资源鉴定奠定了坚实基础。展开更多
Using DNA of Chinese Tobacco Leaf Curl Virus Guangxi(TbLCV Guangxi)as the template,a 0.4 kb virus specific fragment induding the total common region of the virus and DNA sequence of 52 aa of N terminal of AV1 was obta...Using DNA of Chinese Tobacco Leaf Curl Virus Guangxi(TbLCV Guangxi)as the template,a 0.4 kb virus specific fragment induding the total common region of the virus and DNA sequence of 52 aa of N terminal of AV1 was obtained by PCR with primers specific to whitefly transmittal gemniviruses,cloned and sequenced.This fragment can basically represent the characteristics of the virus full length genome.The sequence of this fragment is the same as that of chinese tomato yellow leaf curl virus (TYLCV Chi), which indicates that TbLCV Guangxi and TYLCV Chi are the same gemniviruses. According to the sequence of the common region of the 0.4 kb fragment, the virus full length genome DNA was amplified from the infected tobacco plant by PCR with primers back to back.展开更多
文摘Using DNA of Chinese Tobacco Leaf Curl Virus Guangxi(TbLCV Guangxi)as the template,a 0.4 kb virus specific fragment induding the total common region of the virus and DNA sequence of 52 aa of N terminal of AV1 was obtained by PCR with primers specific to whitefly transmittal gemniviruses,cloned and sequenced.This fragment can basically represent the characteristics of the virus full length genome.The sequence of this fragment is the same as that of chinese tomato yellow leaf curl virus (TYLCV Chi), which indicates that TbLCV Guangxi and TYLCV Chi are the same gemniviruses. According to the sequence of the common region of the 0.4 kb fragment, the virus full length genome DNA was amplified from the infected tobacco plant by PCR with primers back to back.
