Development of polyclonal antibodies-based serological methods for detection of the rehmannia mosaic virus in field plants

Abstract

Rehmannia glutinosa is a top-grade traditional Chinese medicine, and also is an important planting medicinal material for Chinese poor farmers shaking off poverty. Rehmannia mosaic virus (ReMV) causes big economic loss of R. glutinosa in planting area. However, there is no effective methods for quick, accurate, and high-throughput detection for ReMV in Chinese production area. The preserved R. glutinosa samples carrying ReMV was taken for research material. The coat protein coding sequences (CPReMV) was cloned and sequenced. The target sequence was further placed into a prokaryotic expression vector to express the N-terminal-tagged recombinant CPReMV protein (His-CPReMV). Purified His-CPReMV was used as an antigen to immunize New Zealand white rabbits, and antiserum was obtained. The titers and sensitivities of the antisera were analyzed and evaluated. Polyclonal antibodies were purified from the antiserum, and the titers and sensitivity to the target His-CPReMV protein were evaluated. The results demonstrate that the obtained polyclonal antibodies against His-CPReMV could be successfully used for rapid, accurate, and high-throughput detection of ReMV from R. glutinosa planted in the wild. Our investigation established serological-based detection methods for ReMV for the first time, and provides a foundation for future exploration of the pathogenic mechanisms of ReMV in R. glutinosa.

Generation of a Triple-Shuttling Vector and the Application in Plant Plus-Strand RNA Virus Infectious cDNA Clone Construction

“…Rehmannia glutinosa is an important traditional Chinese medicinal plant, and its roots can be used to treat hemoptysis, anemia, and gynecological disease [ 41 ]. Planting R. glutinosa is key for the government and farmers to eliminate poverty in middle China under the current background of rural revitalization [ 15 ]. Raphanus sativus L., commonly named radish, is an important species cultivated in most of the temperate regions of the world and severed as an ancient kitchen garden vegetable.…”

“…Tobacco mosaic virus (TMV), a member of a large group of viruses within the genus Tobamovirus , has single-stranded genomic RNA (~6400 nucleotides) and rod-shaped viral particles (virions) [ 14 ]. Youcai mosaic virus (YoMV) and Rehmannia mosaic virus (ReMV), which were previously isolated from Raphanus sativus and Rehmannia glutinosa , respectively [ 15 , 16 ], also belong to the genus Tobamovirus . Tobamoviruses are the most important viruses in crops that are not transmitted by invertebrate vectors.…”

“…Rape virus disease occurred in rape-producing areas all over the country, among which the winter rape areas in North China, Southwest China, and Central China were the most serious [ 16 ]. In the long-term asexual reproduction process, ReMV and other viruses will accumulate and aggravate the damage, causing a serious impact on the yield and quality of groundnut [ 15 ].…”

Development of Polyclonal Antibodies and a Serological-Based Reverse-Transcription Loop-Mediated Isothermal Amplification (S-RT-LAMP) Assay for Rice Black-Streaked Dwarf Virus Detection in Both Rice and Small Brown Planthopper

“…PAb-CP RBSDV was utilized for the serological diagnosis of RBSDV-infected rice seedlings and SBPHs. On the basis of the PAb-CP RBSDV , the detection methods of Western blot, dot-blot and ELISA were established by following their conventional protocols with slight adjustments [ 18 , 19 , 20 ].…”