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

Abstract

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.

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

“…To take the advantage of plant production system, we adopted the plant viral vector developed previously in our group, BaMV-based vector, pCB-GFP2a-CP [ 18 ] to express recombinant proteins consisting of superfolder GFP (sGFP) [ 34 ] and the above SpaED (GfED) as a universal labeling agent for antibodies used in serological detection methods. It was expected that sGFP domain could bind to IgG through the SpaED domain, and the target recognized by IgG could be visualized by fluorescence of sGFP.…”

“…In this study, we constructed the GfED fusion protein as the antibody-labeling agent for the fluorescent detection technique. The BaMV-based vector [ 18 ] was employed for the practical production of GfED in C . quinoa plants.…”

“…In this regard, the usage of plants as “bio-reactors” for target protein production [ 9 , 10 ] in combination of the expression vectors based on plant viruses [ 11 – 13 ] offers an applicable alternative for the production of fluorescent antibody labeling proteins. In this study, we have adopted the BaMV-based vector previously developed in our group [ 18 ] for the practical production of GfED in C . quinoa .…”

“…Isopropylthio-β-galactoside (IPTG, 1 mM) induction was performed at 37°C for 3 hr, and the bacterial cells were collected by centrifugation and lysed for further protein analysis. To express the fluorescent tags for antibodies (GfED) in plants, a BaMV-based vector, pCB-GFP2a-CP [ 18 ] was used. The spaED segments flanked either by artificially designed Age I or Not I restriction enzyme sites were amplified by PCR using pET21d-spaED as the template with primers listed in Table 1 .…”

“…We have previously developed a plant viral vector based on Bamboo mosaic virus (BaMV) and demonstrated the application of such a vector in the expression of vaccine candidates as chimeric virus particles (CVPs) in plants [ 14 – 17 ]. In addition, we have constructed an infectious clone of BaMV, designated pCB-GFP2a-CP, which led to the production of GFP-tagged BaMV CVPs in the inoculated plants [ 18 ]. The CVP strategy offers an advantage of displaying multiple copies of target proteins on the surface of CVPs.…”