[Detection of tobacco mosaic virus (TMV) in Rehmannia glutinosa f. hueichingensis by IC-RT-PCR].
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.


METHODS
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.


RESULTS
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).


CONCLUSIONS
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.
Food and Bioengineering College, Henan University of Science and Technology, Luoyang 471023, China. dulin1980@sohu.com
Lin
L
Du
Jin-Le
JL
Xiang
Jin-Ling
JL
Fan
Xin
X
Li
Lei
L
Luo
D000595:Amino Acid Sequence; D001483:Base Sequence; D008969:Molecular Sequence Data; D032267:Rehmannia / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014027:Tobacco Mosaic Virus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*