Tomato fruit blotch virus cytopathology strengthens evolutionary links between plant blunerviruses and insect negeviruses

Tomato fruit blotch virus (ToFBV) is a blunervirus that causes blotches on mature tomato (Solanum lycopersicon L.) fruits in Italy and Australia in 2020, and was newly detected in Brazil. A cytological study on pericarp tissues from the blotched areas of infected fruits collected in Brasília, Brazil, revealed characteristic cell alterations. Small and slender bacilliform particles (ca. 25 nm wide × 100 nm long) were found accumulating in the perinuclear space and the lumen of the endoplasmic reticulum of the epidermis, peri- and mesocarp cells. No viroplasm-like inclusion was observed either in the nuclei or in the cytoplasm. Such cell alterations are reminiscent of those described in cultured mosquito cells infected by negeviruses, an unofficial group of insect viruses. Negeviruses and some other arthropod-borne viruses shared a common ancestor in the RdRp gene with kitavirids, including blunerviruses. Although additional detailed studies are required, we show evidence that ToFBV particles are enveloped and bacilliform, and that such similarity in cytopathology seems to support the evolutionary relationship between plant kitavirids and insect negeviruses.(AU)

First report of sunflower chlorotic mottle virus infecting sunflower plants in Brazil

Sunflower (Helianthus annuus) plants showing symptoms of chlorosis, mosaic, chlorotic ringspot, and necrosis on younger leaves were found in a small experimental plot in Piracicaba, in the state of São Paulo, Brazil. Preliminary examinations by transmission electron microscopy of symptomatic leaf tissue revealed flexuous filamentous particles 13-15 nm wide and 700-750 nm long, and cytoplasmatic cylindrical inclusions typical of those found in plant cells infected by members of the Potyvirus genus. Total RNA extracted from symptomatic leaves and subjected to RT-PCR followed by partial nucleotide sequencing confirmed the presence of a potyvirus in the affected plants, which was identified as sunflower chlorotic mottle virus (SuCMoV), a member of the Sunflower chlorotic mottle virus (genus Potyvirus, family Potyviridae) species. Mechanical transmission assays with extracts of symptomatic sunflower leaves reproduced the original symptoms in sunflowers, mosaic symptoms in Zinnia elegans, and chlorotic local lesions in Chenopodium amaranticolor and C. quinoa. Sunflower and zinnia plants became infected after aphid transmission experiments with Myzus persicae. RT-PCR tests using specific primers for SuCMoV confirmed the presence of this virus in experimentally infected plants, meeting the criteria of Koch's postulate. This is the first report of SuCMoV infecting sunflower plants in Brazil.

Phytoplasma of 16SrVII-B subgroup associated to shoot proliferation in Physalis peruviana plants

Physalis is an herbaceous species native to the Andes region. Currently, it is cultivated in various Brazilian states due to the economic interest of growers for this new fruit. Physalis plants grown in the field showed symptoms of shoot proliferation, leaf malformation, and chlorosis. Since these symptoms are commonly induced by phytoplasmas, this study investigated to confirm the presence of these prokaryotes in symptomatic plants. After DNA extraction from symptomatic and asymptomatic plants, phytoplasmas were found in all affected plants through the nested PCR. Examination by transmission electron microscopy (TEM) using appropriately prepared segments of leaf veins allowed the visualization of typical pleomorphic cells of phytoplasmas in the phloem of symptomatic plants. The computer-simulated RFLP patterns and the phylogenetic analysis allowed identifying the detected phytoplasmas as a 'Candidatus Phytoplasma fraxini'-related strain belonging to the 16SrVII-B subgroup. Moreover, physalis was identified as an additional host species for phytoplasmas in the 16SrVII group, expanding the current knowledge on the host range of phytoplasmas in this group.

Sesame phyllody associated with a 16SrI-B phytoplasma, a Candidatus Phytoplasma asteris-related strain, in Paraguay

Sesame (Sesamum indicum L.) plants exhibiting symptoms of phyllody disease were observed in commercial fields in Paraguay. The symptoms were indicative of infection by phytoplasmas. Thus, the present study investigated the association between affected plants and phytoplasma, which was later analyzed using molecular and phylogenetic methods. Total DNA was extracted from symptomatic and asymptomatic plants and used in nested PCR assays using primers R16SN910601/R16SN011119 and R16F2n/16R2. Amplified products of 1.2 Kb revealed the presence of phytoplasma in all diseased plants, and electron microscopy confirmed the presence of phytoplasmas within phloem vessels. Nucleotide sequences from sesame phytoplasma shared 99 % similarity with phytoplasmas belonging to group 16SrI. Computer-simulated RFLP indicated that the detected phytoplasma is representative of the 16SrI-B, therefore, a Candidatus Phytoplasma asteris-related strain. Phylogenetic analysis was in agreement with virtual RFLP. Our findings expand the current knowledge regarding distribution of representatives of the aster yellows group in a new agroecosystem and implicate sesame as a new host of 16SrI-B phytoplasma in Latin America.

Sesame phyllody associated with a 16SrI-B phytoplasma, a Candidatus Phytoplasma asteris-related strain, in Paraguay

Sesame (Sesamum indicum L.) plants exhibiting symptoms of phyllody disease were observed in commercial fields in Paraguay. The symptoms were indicative of infection by phytoplasmas. Thus, the present study investigated the association between affected plants and phytoplasma, which was later analyzed using molecular and phylogenetic methods. Total DNA was extracted from symptomatic and asymptomatic plants and used in nested PCR assays using primers R16SN910601/R16SN011119 and R16F2n/16R2. Amplified products of 1.2 Kb revealed the presence of phytoplasma in all diseased plants, and electron microscopy confirmed the presence of phytoplasmas within phloem vessels. Nucleotide sequences from sesame phytoplasma shared 99 % similarity with phytoplasmas belonging to group 16SrI. Computer-simulated RFLP indicated that the detected phytoplasma is representative of the 16SrI-B, therefore, a Candidatus Phytoplasma asteris-related strain. Phylogenetic analysis was in agreement with virtual RFLP. Our findings expand the current knowledge regarding distribution of representatives of the aster yellows group in a new agroecosystem and implicate sesame as a new host of 16SrI-B phytoplasma in Latin America.(AU)

Arabidopsis thaliana as a model host for Brevipalpus mite-transmitted viruses

Brevipalpus-transmitted viruses (BTV) are a taxonomically diverse group of plant viruses which severely affect a number of major crops. Members of the group can be sub-classified into cytoplasmic (BTV-C) or nuclear type (BTV-N) according to the accumulation sites of virions in the infected plant cells. Both types of BTV produce only local infections near the point of inoculation by viruliferous mites. Features of BTV-plant interactions such as the failure of systemic spread in their natural hosts are poorly understood. In this study we evaluated Arabidopsis thaliana, a model plant commonly used for the study of plant-virus interactions, as an alternative host for BTV. Infection of Arabidopsis with the BTV-N Coffee ringspot virus and Clerodendrum chlorotic spot virus, and the BTV-C Solanum violaefolium ringspot virus, were mediated by viruliferous Brevipalpus mites collected in the wild. Upon infestation, local lesions appeared in 7 to 10 days on leaves of, at least, 80 % of the assayed plants. Presence of viral particles and characteristic cytopathic effects were detected by transmission electron microscopy (TEM) and the viral identities confirmed by specific reverse-transcriptase polymerase chain reaction (RT-PCR) and further amplicon sequencing. The high infection rate and reproducibility of symptoms of the three different viruses assayed validate A. thaliana as a feasible alternative experimental host for BTV.

Arabidopsis thaliana as a model host for Brevipalpus mite-transmitted viruses

Brevipalpus-transmitted viruses (BTV) are a taxonomically diverse group of plant viruses which severely affect a number of major crops. Members of the group can be sub-classified into cytoplasmic (BTV-C) or nuclear type (BTV-N) according to the accumulation sites of virions in the infected plant cells. Both types of BTV produce only local infections near the point of inoculation by viruliferous mites. Features of BTV-plant interactions such as the failure of systemic spread in their natural hosts are poorly understood. In this study we evaluated Arabidopsis thaliana, a model plant commonly used for the study of plant-virus interactions, as an alternative host for BTV. Infection of Arabidopsis with the BTV-N Coffee ringspot virus and Clerodendrum chlorotic spot virus, and the BTV-C Solanum violaefolium ringspot virus, were mediated by viruliferous Brevipalpus mites collected in the wild. Upon infestation, local lesions appeared in 7 to 10 days on leaves of, at least, 80 % of the assayed plants. Presence of viral particles and characteristic cytopathic effects were detected by transmission electron microscopy (TEM) and the viral identities confirmed by specific reverse-transcriptase polymerase chain reaction (RT-PCR) and further amplicon sequencing. The high infection rate and reproducibility of symptoms of the three different viruses assayed validate A. thaliana as a feasible alternative experimental host for BTV.(AU)