Molecular and serological detection of Parietaria mottle virus in Phytolacca americana, a new host of the virus

Parietaria mottle virus (PMoV) is an emerging virus in Mediterranean countries, responsible for severe disease in tomato and pepper crops in the field and protected cultivation. The principal wild reservoir of PMoV is Parietaria officinalis, and only few additional wild plants have been described as natural reservoirs of the virus. During field survey in southern Italy, several plants of Phytolacca americana showing virus-like symptoms were collected. Serological and molecular assays showed that these plants were infected by PMoV. Sequence comparison of the movement protein gene of the PMoV isolate from P. americana showed the greatest similarity to the corresponding sequence from tomato plants growing nearby. These results indicate that P. ameriacana is a new natural host of PMoV, and further investigation is warranted to establish the potential of this host as reservoir of the virus in the field.

Detection and identification of TMV infecting tomato under protected cultivation in Paraná State

During an inspection in plastic houses in Sapopema, Paraná, 90% of tomato plants showed leaf abnormalities, probably associated with herbicide toxity. However, virus like symptoms developed in selected hosts after mechanical inoculatation. RT-PCR reactions using primers for an internal region within the movement protein gene of TMV and ToMV resulted in the amplification of a 409 bp cDNA fragment only by TMV primers. Deduced amino acids showed 100% identity when compared to TMV movement protein and 94% with ToMV. The RT-PCR protocol was efficient for quick and conclusive determination of virus species. The virus was purified and a polyclonal antiserum was raised for future surveys in tomato crops of Paraná. The partial genomic sequence obtained for TMV-Sapopema has been deposited under the accession number DQ173945, which is the first partial genomic sequence of an isolate of TMV from Brazil in the GenBank, and the first tomato virus isolate from Paraná to have some of its biological and molecular properties determined.

Packaging of Brome Mosaic Virus RNA3 Is Mediated through a Bipartite Signal

ABSTRACT The three genomic and a single subgenomic RNA of brome mosaic virus (BMV), an RNA virus infecting plants, are packaged by a single-coat protein (CP) into three morphologically indistinguishable icosahedral virions with T = 3 quasi-symmetry. Genomic RNAs 1 and 2 are packaged individually into separate particles whereas genomic RNA3 and subgenomic RNA4 (coat protein mRNA) are copackaged into a single particle. We report here that packaging of dicistronic RNA3 requires a bipartite signal. A highly conserved 3′ tRNA-like structure postulated to function as a nucleating element (NE) for CP subunits (Y. G. Choi, T. W. Dreher, and A. L. N. Rao, Proc. Natl. Acad. Sci. USA 99:655-660, 2002) and a cis-acting, position-dependent packaging element (PE) of 187 nt present in the nonstructural movement protein gene are the integral components of the packaging core. Efficient incorporation into BMV virions of nonviral RNA chimeras containing NE and the PE provides confirmatory evidence that these two elements are sufficient to direct packaging. Analysis of virion RNA profiles obtained from barley protoplasts transfected with a RNA3 variant lacking the PE provides the first genetic evidence that de novo synthesized RNA4 is incompetent for autonomous assembly whereas prior packaging of RNA3 is a prerequisite for RNA4 to copackage.

Analysis of local and systemic spread of the crucifer-infecting TMV-Cg virus in tobacco and several Arabidopsis thaliana ecotypes

The crucifer-infecting tobacco mosaic virus, TMV-Cg, infects Arabidopsis thaliana (L.) Heynh. efficiently without causing severe symptoms. The systemic spread of TMV-Cg in Arabidopsis was evaluated in 14�ecotypes. Five days after inoculation, TMV-Cg was detected in apical leaves of 8 out of 14 ecotypes. As expected, the spread of TMV-Cg in the ecotypes tested was considerably faster than that of tobacco mosaic virus (TMV-U1). To study the participation of viral proteins in the TMV-Cg-induced infection, a complete genomic cDNA of TMV-Cg was cloned. The role of TMV-Cg movement protein in systemic spread was tested with a hybrid virus, constructed from the TMV-U1 genome and the TMV-Cg movement protein gene. Contrary to expectations, the systemic spread of this hybrid in Arabidopsis was similar to that of TMV-U1. The failure of the hybrid virus to spread at rates similar to those of TMV-Cg was not due to restrictions in local movement. In tobacco (Nicotiana tabacum L.), the hybrid virus spread efficiently and induced systemic mosaic symptoms characteristic of TMV-U1. The TMV-Cg cDNA clone provides an attractive tool to study virus–host interactions.