The glycoprotein gene of Chrysanthemum stem necrosis virus and Zucchini lethal chlorosis virus and molecular relationship with other tospoviruses

Abstract

Two tospoviruses, Chrysanthemum stem necrosis virus (CSNV) and Zucchini lethal chlorosis virus (ZLCV), cause economical losses in several ornamental and vegetable crops in Brazil. The nucleocapsid gene and movement protein sequences had already been reported for both viruses, though the glycoprotein precursor gene sequence was not available. In this study, cDNA fragments (ca. 4 kb) of the M RNA 3' portion of CSNV (isolate Chry-1) and ZLCV (isolate 1038), including the complete glycoprotein precursor gene, partial NSm gene, and the entire intergenic and 3' untranslated regions, were cloned and sequenced. The sequences were assembled with the corresponding 5' region sequence (NSm gene and 5'UTR) of the same isolates to build up the complete sequence of the M RNA segment of both species. The M RNA of CSNV was 4,828 nucleotide-long, while of ZLCV 4,836 nucleotides. Both M RNA molecules comprised two ORFs in an ambisense arrangement. The vcRNA coded for the viral glycoprotein (Gn/Gc) precursor gene of CSNV and ZLCV (both with 127.5 kDa). Comparison of deduced amino acids of the CSNV and ZLCV glycoprotein precursor genes with those of other tospoviruses showed the highest identity with that of Tomato spotted wilt virus (86%) and with that of CSNV (82%), respectively. However, the nucleotide sequence of the intergenic and 3' untranslated regions of CSNV and ZLCV shared lower identities with other tospoviruses. The glycoprotein precursor gene is thought to be a good candidate as additional classification parameter for Tospovirus taxonomy. The presence of the RGD motif in both Gc proteins indicated that they are typical American tospoviruses, which was confirmed by phylogenetic analysis. The membrane topology of both glycoproteins is discussed.