Cassava common mosaic disease (CCMD) has been reported in all regions where cassava is grown in the Americas and the causal agent, Cassava common mosaic virus (CsCMV), has been identified as a mechanically transmitted potexvirus (Alphaflexiviridae). In Argentina, cassava is grown mainly in the northeast (NEA) region that shares borders with Brazil and Paraguay. Increasing incidences of CCMD were observed during the years 2014 to 2016 associated with severe leaf mosaic symptoms and yield reductions where the occurrence of CsCMV was confirmed by RT‐PCR and sequencing. In this work, the virus has been successfully purified and a double‐antibody sandwich (DAS‐) ELISA test has been developed from an Argentinean isolate of CsCMV to extend the diagnostics of the disease. A collection of 726 samples was screened and CsCMV was detected with 100% prevalence in the NEA region. Additional co‐infecting viruses were detected in some plants (64.4%); in these, CCMD symptoms correlated with CsCMV only, although more severe symptoms could be observed in mixed infected plants. Sequence analysis of the conserved RdRp domain showed a wider diversity of CsCMV isolates. Interestingly, a separate phylogenetic cluster was formed by isolates from the NEA region that only shared 77.1% to 80.3% nucleotide identity with the other clusters. These results indicate the presence of mixed strains occurring in the NEA region and suggest the presence of geographically distinct strains of CsCMV in South America.
Cassava common mosaic virus (CsCMV) is a potexvirus that causes systemic infections in cassava plants, leading to chlorotic mosaic and producing significant yield losses. To date, the physiological alterations and the mechanism underlying biotic stress during the cassava-CsCMV compatible interaction remains unknown. In this study, we found that CsCMV infection adversely modified chloroplast structure and had functional effects on chloroplasts in source leaves during the course of viral infection. Extrusion of the chloroplast membrane with amoeboid-shaped appearance and disorganized grana stacks were observed in infected mesophyll cells. These alterations were associated with up to 35% reduction of relative chlorophyll content, and a decline of CO 2 fixation (13.5% and 24.2% at 90 and 210 days after planting, respectively). The effects of CsCMV infection on the performance index on absorption basis dropped up to 37%. The analysis of chlorophyll a fluorescence showed a progressive loss of both oxygen evolving complex activity and "connectivity" within the tripartite system (core antenna-LHCII-reaction centre). Here, we report the latter phenomenon for the first time in a viral infection. The oxidative stress process was observed in CsCMV-infected plants (20.8% reduction of antioxidant capacity with respect to noninfected plants). Other effects of the pathogen included reduction of starch and maltose content in source leaves, and a significant increase (24.7%) of the sucrose:starch ratio, which indicates an altered pattern of carbon allocation. Our results suggest that CsCMV induces chloroplast distortion associated with progressive chloroplast function loss and diversion of carbon flux in source leaf tissue, leading to the loss of cassava tuber yield.
RESUMENMal de Río Cuarto virus en trigo provoca una enfermedad resultante de la acción combinada con su principal vector, Delphacodes kuscheli. La severa sintomatología inducida por virus e insecto condujo a determinar la posible existencia de alteraciones fisiológicas en inducida por virus e insecto condujo a determinar la posible existencia de alteraciones fisiológicas en el cultivar de trigo ProINTA Federal, infectado artificialmente con dos aislamientos geográficamente distantes del MRCV y sometido a picadura de insectos no virulíferos, respectivamente, en tres estadios fenológicos. ProINTA Federal fue picado por insectos no virulíferos e inoculado en la primera hoja emergiendo del coleóptilo, en la cuarta y quinta hojas desplegadas, respectivamente. La condición sanitaria de los diferentes tratamientos fue constatada mediante ELISA. Las plantas tratadas manifestaron alteraciones fisiológicas con similares variaciones entre estadios fenológicos. Azúcares solubles totales, almidón y proteínas solubles incrementaron significativamente su contenido en plantas infectadas por MRCV. En plantas PS y con MRCV, el contenido de clorofilas disminuyó marcadamente y lo inverso sucedió con el malondialdehído, lo que indicaría daño oxidativo por estrés en las mismas. No existieron diferencias entre estadios fenológicos y las variaciones de los indicadores de alteraciones fisiológicas considerados, excepto para el incremento de almidón, significativamente superior en plantas inoculadas en la primera hoja emergiendo del coleóptilo. Palabras clave: infección viral, toxicidad por insectos, azúcares solubles, almidón, proteínas, clorofilas, malondialdehído, estadios fenológicos. ABSTRACT Physiological alterations associated to the Mal de Rio Cuarto virus (MRCV) infection and to vector (Delphacodes kuscheli Fennah) phytotoxicity in wheatMal de Río Cuarto virus produces a wheat disease, resulting from the combined action of this pathogen and its main vector, resulting from the combined action of this pathogen and its main vector, from the combined action of this pathogen and its main vector, of this pathogen and its main vector, this pathogen and its main vector, Delphacodes kuscheli. Knowing that viruses and insects cause a severe symptomatology in different genotypes, the objective of this nowing that viruses and insects cause a severe symptomatology in different genotypes, the objective of this severe symptomatology in different genotypes, the objective of this in different genotypes, the objective of this genotypes, the objective of this the objective of this the objective of this of this work was to determine the possible physiological alterations in three phenological stages of wheat (cv ProINTA Federal) inoculated by was to determine the possible physiological alterations in three phenological stages of wheat (cv ProINTA Federal) inoculated by possible physiological alterations in three phenological stages of wheat (cv ProINTA Federal) inoculated by le physiological alterations in three phenological stages of wheat (cv Pro...
Symptomatic sweet potato cv Arapey INIA samples were collected from a commercial production field in Colonia Molina, Guaymallén department, Mendoza province, Argentina. They showed dark rounded lesions, sometimes coalescing with white granular mycelium. Fungus was obtained from symptomatic sweet potatoes, which represented the generalized infection that affected the crop. They were seeded in PDA with streptomycin sulfate and incubated for seven days at 21°C, alternating white/black (UV400nm) light. Observations with an optical microscope revealed the presence of hyaline, not septated, cylindrical endoconidia with rounded ends. They were 8-16 μm length and 4–6 μm width. Phialides were 43-46 μm length, rounded bases (7-9 μm width) and tapering to the neck´s tip (4-6 μm width). Brown chlamydospores (aleuriospores), 9-13 μm length and 8-12 μm width, in chains of 2-8 spores were observed. For molecular identification, total genomic DNA was extracted. ITS fragment of 565 pb was amplified using ITS5/ITS4 primers and sequenced. The sequence indicated 99% identity with Berkeleyomyces basicola (synonymous: Thielaviopsis basicola). This was deposited in GenBank as (KX580957) (CBS: C430.74, Gen Bank accession number AF275482.1). This is the first report of B. basicola in sweet potato in Argentina, a potential threat to storage root yields. Highlights: Sweet potato black root rot, new disease in Argentina. First report of Berkeleyomyces basicola causing black root rot on sweet potato in Mendoza, Argentina.
Sweet potato yellow curling (YC), the most severe disease of sweet potato detected in Argentina, causes symptoms and damage to sweet potato crops in all cultivated regions. Since 2010/11, the presence of four viruses has been detected in symptomatic cv. Arapey INIA: two potyviruses non-persistently transmitted by Myzus persicae (sweet potato feathery mottle virus, SPFMV and sweet potato virus G, SPVG); a closterovirus, sweet potato chlorotic stunt virus (SPCSV) and a geminivirus, sweet potato leaf curl virus (SPLCV), both transmitted by Bemisia tabaci in a semi-persistent and persistent manner, respectively. All the plants were collected from fields in Colonia Caroya, Córdoba province, Argentina. The objectives of the present work are to isolate and identify the virus or viruses involved in YC disease of sweet potato, and to elucidate the viral combination that reproduces YC symptoms. The most severe YC symptoms for this genotype in the field were only reproduced by a combination of the four viruses. The symptoms include chlorosis, stunting, mosaic, blistering, leaf curling, chlorotic spots, chlorotic patterns, leaf area reduction and distortion, and upward curling of leaf edges. The presence of each virus was detected by serological (DAS, NCM and TAS-ELISA) and molecular (PCR) tests. It is concluded that the interaction of SPFMV, SPVG, SPCSV and SPLCV is needed for the development of YC symptoms. Highlights Four viruses involved in yellow curling disease, SPFMV, SPVG, SPCSV and SPLCV were isolated and identified. The first antiserum produced in Argentina allows the fast, safe and efficient diagnosis of SPVG. The interaction of SPFMV, SPVG, SPCSV and SPLCV is needed for the manifestation of yellow curling in Argentina.
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