应用IC—RT—PCR法检测怀地黄中烟草花叶病毒(TMV)
[9] 杜琳，钟先锋，陈剑泓，等.植物病毒检测技术研究进展[J].农产品加工学刊，2007(7)：48.
[10] 李志勇，郭京泽，司贺龙，等.应用IC-RT-PCR方法检测齿兰环斑病毒和建兰花叶病毒[J].河北农业大学学报，2008，31(6)：40.
[11] 贾丽霞，李志勇，马继芳.应用IC-RT-PCR方法检测水仙中百合斑驳病毒[J].北方园艺，2011，(14)：123.
[12] Kilic H C，Yardimci N.Nested RT-PCR and immunocapture RT-PCR for detection of beet necrotic yellow vein virus on sugar beet in lake district of turkey[J].Rom Agr Res，2012，29：333.
Detection of tobacco mosaic virus(TMV) in Rehmannia glutinosa f.
hueichingensis by IC-RT-PCR
DU Lin*， XIANG Jin-le， FAN Jin-ling， LI Xin， LUO Lei
(Food and Bioengineering College， Henan University of Science and Technology， Luoyang 471023， China)
[Abstract] Objective： To establish a rapid，sensitive and efficient detection method for tobacco mosaic virus (TMV)， and provide technical support of TMV detection of Rehmannia glutinosa f. hueichingensis. The virus-free plantlets could be produced on a large scale to ameliorate breed degeneration caused by viral disease. Method： Specific primers were designed based on the conserved region of coat protein(CP)gene of TMV. Immunocapture RT-PCR (IC-RT-PCR) was employed to detect TMV and the sequence of the products was detected. Result： The expected nucleotide acid fragments were amplified by IC-RT-PCR. The homology of nucleotide acid sequence and amino acid sequence were 95.29% and 96.7% between the PCR products and the CP gene of TMV(accession number AY555269). Conclusion： The method was established for the detection of TMV in R. glutinosa f. hueichingensis by IC-RT-PCR. This detection combined molecular biology technology with immunology，was convenient for a quick， sensitive and simple detection of TMV.
[Key words] Rehmannia glutinosa f. hueichingensis； TMV； IC-RT-PCR； sequence analysis； virus detection
doi：10.4268/cjcmm20131311
[责任编辑 吕冬梅], 百拇医药(杜琳，向进乐，樊金玲，李欣，罗磊)
[10] 李志勇，郭京泽，司贺龙，等.应用IC-RT-PCR方法检测齿兰环斑病毒和建兰花叶病毒[J].河北农业大学学报，2008，31(6)：40.
[11] 贾丽霞，李志勇，马继芳.应用IC-RT-PCR方法检测水仙中百合斑驳病毒[J].北方园艺，2011，(14)：123.
[12] Kilic H C，Yardimci N.Nested RT-PCR and immunocapture RT-PCR for detection of beet necrotic yellow vein virus on sugar beet in lake district of turkey[J].Rom Agr Res，2012，29：333.
Detection of tobacco mosaic virus(TMV) in Rehmannia glutinosa f.
hueichingensis by IC-RT-PCR
DU Lin*， XIANG Jin-le， FAN Jin-ling， LI Xin， LUO Lei
(Food and Bioengineering College， Henan University of Science and Technology， Luoyang 471023， China)
[Abstract] Objective： To establish a rapid，sensitive and efficient detection method for tobacco mosaic virus (TMV)， and provide technical support of TMV detection of Rehmannia glutinosa f. hueichingensis. The virus-free plantlets could be produced on a large scale to ameliorate breed degeneration caused by viral disease. Method： Specific primers were designed based on the conserved region of coat protein(CP)gene of TMV. Immunocapture RT-PCR (IC-RT-PCR) was employed to detect TMV and the sequence of the products was detected. Result： The expected nucleotide acid fragments were amplified by IC-RT-PCR. The homology of nucleotide acid sequence and amino acid sequence were 95.29% and 96.7% between the PCR products and the CP gene of TMV(accession number AY555269). Conclusion： The method was established for the detection of TMV in R. glutinosa f. hueichingensis by IC-RT-PCR. This detection combined molecular biology technology with immunology，was convenient for a quick， sensitive and simple detection of TMV.
[Key words] Rehmannia glutinosa f. hueichingensis； TMV； IC-RT-PCR； sequence analysis； virus detection
doi：10.4268/cjcmm20131311
[责任编辑 吕冬梅], 百拇医药(杜琳，向进乐，樊金玲，李欣，罗磊)