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Hibiscus is not native to Colombia but well suited to its arid soil and dry climates. A single hibiscus plant from Risaralda, showing black spots on upper and lower sides of its leaves, was collected for virome analysis using meta-transcriptomic high-throughput sequencing technology. Bioinformatic analysis identified 12.5% of the total reads in the Ribo-Zero cDNA library which mapped to viral genomes. BLAST searches revealed the presence of carlavirus, potexvirus, and of known members of the genera Betacarmovirus, Cilevirus, Nepovirus, and Tobamovirus in the sample; confirmed by RT-PCR with virus-specific primers followed by amplicon sequencing. Furthermore, in silico analysis suggested the possibility of a novel soymovirus, and a new hibiscus strain of citrus leprosis virus C2 in the mixed infection. Both RNA dependent RNA polymerase and coat protein gene sequences of the potex and carla viruses shared less than 72% nucleotide and 80% amino acid identities with any alphaflexi- and betaflexi-virus sequences available in GenBank, identifying three novel carlavirus and one potexvirus species in the Hibiscus rosa-sinensis plant. The detection of physalis vein necrosis nepovirus and passion fruit green spot cilevirus in hibiscus are also new reports from Colombia. Overall, the meta-transcriptome analysis identified the complex virome associated with the black spot symptoms on hibiscus leaves and demonstrated the diversity of virus genera tolerated in the mixed infection of a single H. rosa-sinensis plant.

Assuntos

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Viruses are an important disease source for beans. In order to evaluate the impact of virus disease on Phaseolus biodiversity, we determined the identity and distribution of viruses infecting wild and domesticated Phaseolus spp. in the Mesoamerican Center of Domestication (MCD) and the western state of Nayarit, Mexico. We used small RNA sequencing and assembly to identify complete or near-complete sequences of forty-seven genomes belonging to nine viral species of five genera, as well as partial sequences of two putative new endornaviruses and five badnavirus- and pararetrovirus-like sequences. The prevalence of viruses in domesticated beans was significantly higher than in wild beans (97% vs. 19%; p < 0.001), and all samples from domesticated beans were positive for at least one virus species. In contrast, no viruses were detected in 80-83% of the samples from wild beans. The Bean common mosaic virus and Bean common mosaic necrosis virus were the most prevalent viruses in wild and domesticated beans. Nevertheless, Cowpea mild mottle virus, transmitted by the whitefly Bemisia tabaci, has the potential to emerge as an important pathogen because it is both seed-borne and a non-persistently transmitted virus. Our results provide insights into the distribution of viruses in cultivated and wild Phaseolus spp. and will be useful for the identification of emerging viruses and the development of strategies for bean viral disease management in a center of diversity.

Assuntos

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Major themes in pathogen evolution are emergence, evolution of virulence, host adaptation and the processes that underlie them. RNA viruses are of particular interest due to their rapid evolution. The in vivo molecular evolution of an RNA plant virus was demonstrated here using a necrotic isolate of cowpea mild mottle virus (CPMMV) and a susceptible soybean genotype submitted to serial inoculations. We show that the virus lost the capacity to cause necrosis after six passages through the host plant. When a severe bottleneck was imposed, virulence reduction occurred in the second passage. The change to milder symptoms had fitness benefits for the virus (higher RNA accumulation) and for its vector, the whitefly Bemisia tabaci. Genetic polymorphisms were highest in ORF1 (viral replicase) and were independent of the symptom pattern. Recombination was a major contributor to this diversity - even with the strong genetic bottleneck, recombination events and hot spots were detected within ORF1. Virulence reduction was associated with different sites in ORF1 associated to recombination events in both experiments. Overall, the results demonstrate that the reduction in virulence was a consequence of the emergence of new variants, driven by recombination. Besides providing details of the evolutionary mechanisms behind a reduction in virulence and its effect under viral and vector fitness, we propose that this recombination-driven switch in virulence allows the pathogen to rapidly adapt to a new host and, potentially, switch back.

Assuntos

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By having an extensive territory and suitable climate conditions, South America is one of the most important agricultural regions in the world, providing different kinds of vegetable products to different regions of the world. However, such favorable conditions for plant production also allow the development of several pests, increasing production costs. Among them, whiteflies (Hemiptera: Aleyrodidae) stand out for their potential for infesting several crops and for being resistant to insecticides, having high rates of reproduction and dispersal, besides their efficient activity as virus vectors. Currently, the most important species occurring in South America are Bemisia afer, Trialeurodes vaporariorum, and the cryptic species Middle East-Asia Minor 1, Mediterranean, and New World, from Bemisia tabaci complex. In this review, a series of studies performed in South America were compiled in an attempt to unify the advances that have been developed in whitefly management in this continent. At first, a background of the current whitefly distribution in South American countries as well as factors affecting them are shown, followed by a background of the whitefly transmitted viruses in South America, addressing their location and association with whiteflies in each country. Afterwards, a series of management strategies are proposed to be implemented in South American fields, including cultural practices and biological and chemical control, finalizing with a section containing future perspectives and directions for further research.

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The effects of 2 mM silicon (Si) and 10 mM KNO3 (N)-prime signals for plant resistance to pathogens-were analyzed in healthy and Cowpea chlorotic mottle virus (CCMV) or Cowpea mild mottle virus (CMMV)-infected Bradyrhizobium-nodulated cowpea, yardlong bean and mung bean plants. In healthy plants of the three Vigna taxa, nodulation and growth were promoted in the order of Si + N > N > Si > controls. In the case of healthy cowpea and yardlong bean, the addition of Si and N decreased ureide and α-amino acids (AA) contents in the nodules and leaves in the order of Si + N> N > Si > controls. On the other hand, the addition of N arrested the deleterious effects of CCMV or CMMV infections on growth and nodulation in the three Vigna taxa. However, the addition of Si or Si + N hindered growth and nodulation in the CCMV- or CMMV-infected cowpea and yardlong bean, causing a massive accumulation of ureides in the leaves and nodules. Nevertheless, the AA content in leaves and nodules of CCMV- or CMMV-infected cowpea and yardlong bean was promoted by Si but reduced to minimum by Si + N. These results contrasted to the counteracting effects of Si or Si + N in the CCMV- and CMMV-infected mung bean via enhanced growth, nodulation and levels of ureide and AA in the leaves and nodules. Together, these observations suggest the fertilization with Si + N exclusively in virus-free cowpea and yardlong bean crops. However, Si + N fertilization must be encouraged in virus-endangered mung bean crops to enhance growth, nodulation and N-metabolism. It is noteworthy to see the enhanced nodulation of the three Vigna taxa in the presence of 10 mM KNO3.

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Infectious cDNA clones are an important tool to study the molecular and cellular process of RNA virus infection. In vitro and in vivo transcription systems are the two main strategies used in the generation of infectious cDNA clones for RNA viruses. This study describes the first generation of a full-length infectious cDNA clone of Cowpea mild mottle virus (CPMMV), a Carlavirus. The full-length genome was synthesized by Overlap Extension PCR of two overlapping fragments and cloned in a pUC-based vector under control of the SP6 RNA polymerase promoter. After in vitro run-off transcription, the produced RNA was mechanically inoculated into soybean plants cv. CD206. The systemic infection was confirmed by RT-PCR and further sequencing of amplified cDNA fragments. To simplify the transfection process, the complete genome was subcloned into a binary vector under control of the 35S promoter of cauliflower mosaic virus by the Gibson Assembly protocol. The resulting clones were inoculated by particle bombardment onto soybean seedlings and the recovery of the virus was confirmed 2 weeks later by RT-PCR. Our results indicate the constructs of the full-length cDNA of CPMMV are fully infectious in both in vitro and in vivo transcription strategies.

Assuntos

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Garlic (Allium sativum L.) is a host to several viruses, most commonly those belonging to theAllexivirus,Carlavirus, orPotyvirusgenera. Nine species distributed among these three genera have been reported in Brazil: two species within carlaviruses, two within potyviruses, and five within allexiviruses. To quantify the prevalence of these viruses, young leaves from 520 plants (plants either symptomatic or asymptomatic) were collected from commercial fields grown in four Brazilian states and analyzed using universal and species-specific primers via the reverse transcription polymerase chain reaction (RT-PCR). Potyvirus presence was positive in 306 samples (81 %), 151 of them (38 %) in mixed infections with other viruses. The most frequent potyviruses wereOnion yellow dwarf virus(OYDV, 56 %) andLeek yellow stripe virus(LYSV, 55 %). 187 samples (49 %) were positive for allexivirus, with 33 (9 %) showing single infections and 154 (41 %) showing mixed infections withGarlic virus A (GarV-A),Garlic virus B(GarV-B),Garlic virus C(GarV-C),Garlic virus D(GarV-D), and species belonging to theCarlavirus andPotyvirusgenera. The predominant species in which allexiviruses were found were GarV-A and GarV-D. Only 15 samples (4 %) were infected solely by a carlavirus, and 63 (17 %) showed mixed infections with viruses from different genera. The dominant species of carlavirus wasGarlic commom latent virus(GarCLV). Carlaviruses and allexiviruses are frequently associated with mixed infections with potyviruses, whereas mixed infections with carlaviruses and allexiviruses are rare. About 70 % of the plants collected were positive for at least one species of virus.(AU)

Assuntos

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Garlic (Allium sativum L.) is a host to several viruses, most commonly those belonging to theAllexivirus,Carlavirus, orPotyvirusgenera. Nine species distributed among these three genera have been reported in Brazil: two species within carlaviruses, two within potyviruses, and five within allexiviruses. To quantify the prevalence of these viruses, young leaves from 520 plants (plants either symptomatic or asymptomatic) were collected from commercial fields grown in four Brazilian states and analyzed using universal and species-specific primers via the reverse transcription polymerase chain reaction (RT-PCR). Potyvirus presence was positive in 306 samples (81 %), 151 of them (38 %) in mixed infections with other viruses. The most frequent potyviruses wereOnion yellow dwarf virus(OYDV, 56 %) andLeek yellow stripe virus(LYSV, 55 %). 187 samples (49 %) were positive for allexivirus, with 33 (9 %) showing single infections and 154 (41 %) showing mixed infections withGarlic virus A (GarV-A),Garlic virus B(GarV-B),Garlic virus C(GarV-C),Garlic virus D(GarV-D), and species belonging to theCarlavirus andPotyvirusgenera. The predominant species in which allexiviruses were found were GarV-A and GarV-D. Only 15 samples (4 %) were infected solely by a carlavirus, and 63 (17 %) showed mixed infections with viruses from different genera. The dominant species of carlavirus wasGarlic commom latent virus(GarCLV). Carlaviruses and allexiviruses are frequently associated with mixed infections with potyviruses, whereas mixed infections with carlaviruses and allexiviruses are rare. About 70 % of the plants collected were positive for at least one species of virus.

Assuntos