Production of antibodies against the common epitopes of the
nonstructural NSs proteins of tospoviruses and their applications in tospovirus detection
Nonstructural (NSs) protein of tospoviruses plays the role as a gene- silencing suppressor that overcomes natural defense mechanisms of plants. In the previously study, the produced monoclonal antibodies (MAbs) against the NSs protein of Watermelon silver mottle virus (WSMoV) serologically reacted not only with all members of WSMoV serogroup but also with those of non-WSMoV serological related tospoviruses, such as Melon yellow spot virus (MYSV), Iris yellow spot virus (IYSV) and Tomato yellow ring virus (TYRV). Epitope mapping showed that the MAbs recognized the amino acid (aa) 98 to 120 region of the NSs protein of WSMoV, the sequence is
“VRKPGVKNTGCKFTMHNQIFNPN”. Sequence alignment of the
reported NS proteins revealed that the WSMoV NSs MAb-recognized region is consensus at the same position of those of the
forementioned tospoviruses. In addition, another consensus
sequence “ISVCSNTVNTNGVKHQGHLKVLS” was found at the same position, the aa 98 to 120, among the NSs proteins of Tomato spotted wilt virus (TSWV), Groundnut ringspot virus (GRSV) and Impatiens necrotic spot virus (INSV). These results imply that the homology of NSs proteins is related to the evolution of tospoviruses and these conserved regions of NSs proteins are able to be the important target for detection of tospoviruses. In this project, we will carry out the same approach as expression of the NSs protein of WSMoV that applying the Zucchini yellow mosaic virus (ZYMV) vector to fast express large amounts of NSs proteins of TSWV, INSV, Peanut chlorotic fan-spot virus (PCFV) and Peanut yellow spot virus (PYSV). These expressed NSs proteins will be used as antigens for production of antisera and MAbs which with broad-spectrum serological reactions to establish completely detection system for worldwide tospoviruses. These antibodies are not only able to be used as useful tools for tospovirus detection but also to be used as the detective tools for functional analyses of NSs protein in the future studies.
Keywords: Tospoviruses, gene-silencing suppressor, plant viral
vector, serology, and monoclonal antibody.
