A transgenic plant cell‐suspension system for expression of epitopes on chimeric <i><scp>B</scp>amboo mosaic virus</i> particles

Muthamilselvan, Thangarasu; Lee, Chin‐Wei; Cho, Yu-Hsin; Wu, Feng-Chao; Hu, Chung-Chi; Liang, Y.; Lin, Na-Sheng; Hsu, Yau‐Heiu

doi:10.1111/pbi.12377

Cited by 17 publications

(24 citation statements)

References 55 publications

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“…By quantification of specific protein bands using the Multi Gauge version 3.0 software (Fujifilm Life Sciences), the yield of GfED was estimated to be about 0.13 and 0.17 mg/g fresh leaf in N6 and C31 inoculated plants, respectively. The yields were comparable to, or better than, those of our previous studies [ 15 , 16 ] and the other plant virus based expression system, e.g., 2.4 mg/g fresh leaf tissue for the expression of hepatitis B virus core antigen (HBcAg) virus-like particles (VLPs) or 0.8 mg/g fresh leaf tissue for the expression of Norwalk virus VLPs using TMV based expression systems in Nicotiana benthamiana plant [ 35 , 36 ].…”

Section: Resultssupporting

confidence: 85%

Production of fluorescent antibody-labeling proteins in plants using a viral vector and the application in the detection of Acidovorax citrulli and Bamboo mosaic virus

et al. 2018

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Serological methods are relatively convenient and simple for the detection of pathogens for front-line workers. On-site visualization of the test results plays a pivotal role in the process. However, an efficient, universal labeling agent for antibodies is needed for the development of efficient serological detection tools. In this study, a Bamboo mosaic virus (BaMV)-based viral vector was employed to express recombinant proteins, collectively designated GfED, consisting of Staphylococcus aureus Protein A domain ED (SpaED) fused to either the N- or C-terminal of an improved green florescent protein (GFP) with or without the coat protein (CP) of BaMV, efficiently in Chenopodium quinoa. The GfED in crude leaf extracts could specifically attach to IgG molecules of rabbits and mice, effectively labeling IgG with GFP, emitting green light at 506 nm when excited at 450 nm using simple, handheld equipment. To demonstrate the applicability of GfED in serological assays, we have developed a fluorescent dot blot assay for the rapid detection of Acidovorax citrulli (Ac), a bacterial pathogen of cucurbits, and BaMV, a viral pathogen of bamboos. By using the crude extracts of inoculated C. quinoa leaves expressing GfED as an IgG-labeling agent, the pathogens were easily and quickly detected through uncomplicated operations using simple equipment, with results observable by the naked eye. Examination using fluorescent microscopy and transmission electron microscopy revealed that the GfED subunits may assemble into virus-like particles, which were further involved in the formation of aggregates of GfED-antibody-antigen complexes with the potential for fluorescence signal enhancement. The results suggested that plant-expressed GfED may serve as a promising alternative of IgG-labeling agent for current serological assays.

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Section: Resultssupporting

confidence: 85%

Production of fluorescent antibody-labeling proteins in plants using a viral vector and the application in the detection of Acidovorax citrulli and Bamboo mosaic virus

et al. 2018

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“…For the FMD vaccine, N. benthamiana , tomato, and rice have been used for both transient and stable transformation. Most recently, Muthamilselvan et al [99] have presented an important advance with 2.1 mg/20 g fresh weight of plant callus-based biomass in the cost-effective large-scale production of immunopeptide vaccines against FMD. They established transgenic cell-suspension cultures from N. benthamiana leaves expressing FMDV VP1 and gene silencing suppressor p19 and then validated the efficacy using immunized guinea pigs that produced humoral antibodies.…”

Section: Studies On Plant Vaccinesmentioning

confidence: 99%

The Last Ten Years of Advancements in Plant-Derived Recombinant Vaccines against Hepatitis B

Joung

Park

Moon

et al. 2016

IJMS

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Disease prevention through vaccination is considered to be the greatest contribution to public health over the past century. Every year more than 100 million children are vaccinated with the standard World Health Organization (WHO)-recommended vaccines including hepatitis B (HepB). HepB is the most serious type of liver infection caused by the hepatitis B virus (HBV), however, it can be prevented by currently available recombinant vaccine, which has an excellent record of safety and effectiveness. To date, recombinant vaccines are produced in many systems of bacteria, yeast, insect, and mammalian and plant cells. Among these platforms, the use of plant cells has received considerable attention in terms of intrinsic safety, scalability, and appropriate modification of target proteins. Research groups worldwide have attempted to develop more efficacious plant-derived vaccines for over 30 diseases, most frequently HepB and influenza. More inspiring, approximately 12 plant-made antigens have already been tested in clinical trials, with successful outcomes. In this study, the latest information from the last 10 years on plant-derived antigens, especially hepatitis B surface antigen, approaches are reviewed and breakthroughs regarding the weak points are also discussed.

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“…Purified BaMV virions were dissolved in BE buffer (50 mM Borate, pH 8.0, 1 mM EDTA), then stored at -20°C until used. It should be noted that the purification procedure include an ultracentrifugation through a 5-ml 20% sucrose cushion, which precludes most of the unassembled viral proteins in the pellets (Muthamilselvan et al, 2016). However, the possibility of the presence of some unassembled BaMV CP, GFP-tagged BaMV CP, and GFP-2a fusion proteins in the virion preparations could not be ruled out.…”

Section: Methodsmentioning

confidence: 99%

Transmission of Bamboo mosaic virus in Bamboos Mediated by Insects in the Order Diptera

Chang

Huang

et al. 2017

Front. Microbiol.

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Bamboo mosaic virus (BaMV), a member of the genus Potexvirus, is the major threat to bamboo cultivation. Similar to most potexviruses, the transmission of BaMV by insect vectors has not been documented previously. However, field observations of BaMV disease incidences suggested that insect vectors might be involved. In this study, we aimed to investigate the possibility of insect-mediated transmission of BaMV among bamboo clumps, in order to provide further insights into the infection cycles of BaMV for the development of effective disease management measures. From the major insects collected from infected bamboo plantations, BaMV genomic RNAs were detected inside the bodies of two dipteran insects, Gastrozona fasciventris and Atherigona orientalis, but not in thrips (Scirtothrips dorsalis). Artificial feeding assays using green fluorescent protein-tagged BaMV virions revealed that BaMV could enter the digestive systems and survive in the regurgitant and excretion of the dipterans. BaMV RNA could be retained in the dipterans for up to 4 weeks. Insect-mediated transmission assays indicated that both dipterans could transmit BaMV to bamboo seedlings through artificially created wounds with low infection efficiency (14 – 41%), suggesting that the dipterans may mediate the transmission in a mechanical-like manner. These results demonstrated that dipterans with sponge-like mouthparts may also serve as vectors for at least one potexvirus, BaMV, among bamboo plants. The finding suggested that dipteran insect control should be integrated into the disease management measures against BaMV infections.

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A transgenic plant cell‐suspension system for expression of epitopes on chimeric Bamboo mosaic virus particles

Cited by 17 publications

References 55 publications

Production of fluorescent antibody-labeling proteins in plants using a viral vector and the application in the detection of Acidovorax citrulli and Bamboo mosaic virus

Production of fluorescent antibody-labeling proteins in plants using a viral vector and the application in the detection of Acidovorax citrulli and Bamboo mosaic virus

The Last Ten Years of Advancements in Plant-Derived Recombinant Vaccines against Hepatitis B

Transmission of Bamboo mosaic virus in Bamboos Mediated by Insects in the Order Diptera

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