Publication: Interacción cucurbitáceas-virus-microbioma de pulgón en el desarrollo de enfermedades virales
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Date
2025-10-28
Authors
Moya Ruiz, Celia de
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Escuelas::Escuela Internacional de Doctorado
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Gómez López, Pedro
Publisher
Universidad de Murca
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DOI
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info:eu-repo/semantics/doctoralThesis
Description
Abstract
En primer lugar, se realizó una revisión bibliográfica profunda del estado actual y distribución de los virus transmitidos por pulgón en las principales regiones productoras de cucurbitáceas en España. Tras esto se estudió (I) la distribución de los principales virus transmitidos por pulgón incluyendo aquellos que son transmitidos por mosca blanca en muestras sintomáticas de dos cultivos de cucurbitáceas de tres zonas productoras de España durante los años 2021 a 2023. CABYV y WMV fueron los virus más ampliamente distribuidos en los tres años y cultivos. Además, identificamos un nuevo polerovirus, Polerovirus PABYV, que no había sido descrito anteriormente en los cultivos de cucurbitáceas de España. Este estudio mostró que PABYV emergió en 2018 y que había permanecido oculto bajo los síntomas de amarilleo causados por CABYV. Además, con esta metodología se detectó el virus críptico Alphaendornavirus cucumis por primera en muestras de calabaza.
En segundo lugar, se estudió (II) cómo el orden temporal de las infecciones mixtas podría afectar la dinámica de las poblaciones virales. Para ello, se hizo un ensayo experimental en el que se evaluó la carga viral de CABYV y WMV en plantas de melón bajo diferente orden de infección (secuencial) y de forma paralela bajo infección simultánea (co-infección). La co-infección de CABYV y WMV resultó en una mayor acumulación de ambos virus, lo que sugiere un efecto doble sinérgico. En el caso de las infecciones secuenciales, se observó que el orden temporal de la infección afecta el tipo de interacción entre estos dos virus en melón, con una interacción sinérgica cuando CABYV precede a WMV y una interacción antagónica cuando WMV precede a CABYV. Con este estudio, se destaca la importancia de las interacciones bióticas en la configuración de la epidemiología viral y la dinámica de las enfermedades de las plantas en los agro-ecosistemas.
En tercer lugar, se estudió (III) cómo la combinación de factores abióticos (temperatura) y bióticos (infección viral con WMV) tienen un efecto en la respuesta de la planta (melón y calabacín con alta y baja tolerancia a estrés por temperatura) y por lo tanto en la interacción virus-planta mediante un estudio de transcriptómica por 3’ mRNA-seq. Además, se cuantificó la carga viral de WMV bajo las diferentes condiciones mencionadas. Los resultados mostraron que la carga viral de WMV fue mucho mayor en las plantas de calabacín que de melón y que además fue dependiente de temperatura y el grado de tolerancia de las plantas a esta. En cuanto a la respuesta génica de las plantas a ambos estreses, se observó que el porcentaje de genes diferencialmente expresados (DEGs) fue mayor en las plantas termo-susceptibles para ambas especies de cucurbitáceas y se encontraron DEGs exclusivos en la respuesta a la combinación de ambos estreses para las variedades termo-tolerantes y en concreto, dos genes ortólogos ligados al estrés por temperatura y por infección viral en melón y calabacín.
Por último, se evaluó (IV) cómo la estructura y composición del microbioma de pulgón (A. gossypii) es alterado por su alimentación en diferentes especies de plantas y cuando están infectadas con CABYV y/o WMV, usando análisis de secuenciación del gen 16S rRNA. En este estudio, se observó que la diversidad de la comunidad de bacterias fue mayor en los pulgones alimentados en melón que en pepino por lo que el microbioma se ve fuertemente afectado por el huésped del que se alimenta. También se observó que la diversidad microbiana fue mayor en los pulgones alimentados en plantas sanas que en plantas infectadas. Los géneros más abundantes fueron Buchnera y Arsenophonus, cuyas abundancias se vieron modificadas en función de la alimentación del pulgón.
First, a comprehensive literature review was conducted to examine the current state and distribution of aphid-transmitted viruses in the main cucurbit-producing regions of Spain. This was followed (I) by a study of the distribution of the main aphid-transmitted viruses, including some whitefly-transmitted viruses, in symptomatic cucurbit crop samples (melon and watermelon) showing yellowing and mosaic symptoms, collected from three major production regions of Spain between 2021 and 2023. CABYV and WMV were the most widely distributed viruses across all three years and both crops. Moreover, a novel polerovirus was identified, Polerovirus PABYV, not previously reported in cucurbit crops in Spain. This study showed that this virus emerged in 2018 and had remained undetected, masked by symptoms attributed to CABYV, as it was frequently found in mixed infections with CABYV. Additionally, Alphaendornavirus cucumis, a cryptic Alphaendornavirus, was detected for the first time in pumpkin samples. Second, it was studied (II) how the temporal order of mixed infections could influence viral population dynamics. An experimental assay was conducted to assess the viral load of CABYV and WMV in melon plants under different infection sequences (sequential) and in co-infection scenarios (simultaneous). Co-infection led to a greater accumulation of both viruses, indicating a dual synergistic effect. In sequential infections, the order of infection significantly influenced virus interaction: synergy occurred when CABYV preceded WMV, while antagonism was observed when WMV preceded CABYV. Third, it was investigated (III) how the combination of abiotic (temperature) and biotic (WMV viral infection) factors affects plant responses and, consequently, virus–plant interactions. It was examined the effects of WMV infection and heat stress on the gene expression profiles of two plant species (melon and zucchini) with high and low heat stress tolerance using 3’ mRNA-seq transcriptomic analysis. WMV viral load was also quantified under the different conditions. Results showed that WMV accumulation was significantly higher in zucchini than in melon and was dependent on both temperature and the plant’s tolerance level. Regarding gene expression responses, a higher percentage of differentially expressed genes (DEGs) was found in heat-susceptible plants of both species under combined virus and low-temperature stress. Moreover, specific DEGs were identified in thermotolerant varieties exposed to both stresses, including two orthologous genes linked to temperature and viral stress responses in melon and zucchini. Finally, it was evaluated (IV) how the structure and composition of the aphid (A. gossypii) microbiome is altered by feeding on different plant species and in the presence of CABYV and/or WMV infections. Using 16S rRNA gene sequencing, it was compared the bacterial communities of aphids feeding on melon, followed by transitions to cucumber and back to melon plants. It was assessed the microbiome of aphids feeding on melon plants infected with CABYV and WMV, in both single and mixed infections. This study revealed that bacterial diversity was higher in aphids feeding on melon compared to cucumber, indicating that the host plant strongly influences the aphid microbiome. Additionally, microbial diversity was greater in aphids feeding on healthy plants than on infected ones. The most abundant bacterial genera were Buchnera and Arsenophonus, with their relative abundances affected by feeding conditions.
First, a comprehensive literature review was conducted to examine the current state and distribution of aphid-transmitted viruses in the main cucurbit-producing regions of Spain. This was followed (I) by a study of the distribution of the main aphid-transmitted viruses, including some whitefly-transmitted viruses, in symptomatic cucurbit crop samples (melon and watermelon) showing yellowing and mosaic symptoms, collected from three major production regions of Spain between 2021 and 2023. CABYV and WMV were the most widely distributed viruses across all three years and both crops. Moreover, a novel polerovirus was identified, Polerovirus PABYV, not previously reported in cucurbit crops in Spain. This study showed that this virus emerged in 2018 and had remained undetected, masked by symptoms attributed to CABYV, as it was frequently found in mixed infections with CABYV. Additionally, Alphaendornavirus cucumis, a cryptic Alphaendornavirus, was detected for the first time in pumpkin samples. Second, it was studied (II) how the temporal order of mixed infections could influence viral population dynamics. An experimental assay was conducted to assess the viral load of CABYV and WMV in melon plants under different infection sequences (sequential) and in co-infection scenarios (simultaneous). Co-infection led to a greater accumulation of both viruses, indicating a dual synergistic effect. In sequential infections, the order of infection significantly influenced virus interaction: synergy occurred when CABYV preceded WMV, while antagonism was observed when WMV preceded CABYV. Third, it was investigated (III) how the combination of abiotic (temperature) and biotic (WMV viral infection) factors affects plant responses and, consequently, virus–plant interactions. It was examined the effects of WMV infection and heat stress on the gene expression profiles of two plant species (melon and zucchini) with high and low heat stress tolerance using 3’ mRNA-seq transcriptomic analysis. WMV viral load was also quantified under the different conditions. Results showed that WMV accumulation was significantly higher in zucchini than in melon and was dependent on both temperature and the plant’s tolerance level. Regarding gene expression responses, a higher percentage of differentially expressed genes (DEGs) was found in heat-susceptible plants of both species under combined virus and low-temperature stress. Moreover, specific DEGs were identified in thermotolerant varieties exposed to both stresses, including two orthologous genes linked to temperature and viral stress responses in melon and zucchini. Finally, it was evaluated (IV) how the structure and composition of the aphid (A. gossypii) microbiome is altered by feeding on different plant species and in the presence of CABYV and/or WMV infections. Using 16S rRNA gene sequencing, it was compared the bacterial communities of aphids feeding on melon, followed by transitions to cucumber and back to melon plants. It was assessed the microbiome of aphids feeding on melon plants infected with CABYV and WMV, in both single and mixed infections. This study revealed that bacterial diversity was higher in aphids feeding on melon compared to cucumber, indicating that the host plant strongly influences the aphid microbiome. Additionally, microbial diversity was greater in aphids feeding on healthy plants than on infected ones. The most abundant bacterial genera were Buchnera and Arsenophonus, with their relative abundances affected by feeding conditions.
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