ABSTRACT
Transcriptome sequencing analysis of a symptomatic Rehmannia glutinosa plant revealed a virome containing two known RNA viruses and one novel virus. In this study, we examined the molecular and biological characteristics of the novel virus. The complete genome of the novel virus is composed of monopartite single-stranded RNA of 15,322 nucleotides with 69% nucleotide sequence identity (with 68% coverage) to tobacco virus 1. Its genome organization is typical of the members of the genus Closterovirus, containing nine putative open reading frames. Molecular and phylogenetic analyses of the genome and encoded protein sequences strongly support that the identified virus is a new species of the genus Closterovirus in the family Closteroviridae. The name rehmannia virus 1 (ReV1) is proposed for this novel virus.
Subject(s)
Closterovirus/isolation & purification , Plant Diseases/virology , Rehmannia/virology , Closterovirus/classification , Closterovirus/genetics , Genome, Viral , Open Reading Frames , Phylogeny , Viral Proteins/geneticsABSTRACT
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.
Subject(s)
Rehmannia/virology , Reverse Transcriptase Polymerase Chain Reaction/methods , Tobacco Mosaic Virus/isolation & purification , Amino Acid Sequence , Base Sequence , Molecular Sequence Data , Tobacco Mosaic Virus/genetics , Tobacco Mosaic Virus/immunologyABSTRACT
The stem tip from germ-free stem segment of Rehmannia glutinosa cultured in test tube can be induced into virus-free seedling. The experiment showed that the proper disinfectant for stem segments of Rehmannia glutinosa was 0.05% HgCl2. The seedings from stem segments grew better with MS in the concentration of agar 0.7% and pH7. The stem tips could be directly induced to seedlings by using MS + 6-BA(0.05 mg/L). The MS media for seedlings virus-free culture are 1/4 macro-elements + 1/2 micro-elements and using edible sugar instead of sucrose, so the cost of media could be decreased to 48.7%.