Haplotypes of "Candidatus Liberibacter solanacearum" identified in Umbeliferous crops in Spain

[EN] 'Candidatus Liberibacter solanacearum' is a phloem-limited Gram-negative bacterium that causes serious damage to different crops of the botanical families Solanaceae and Apiaceae. Five haplotypes have been described: LsoA and LsoB are present in solanaceous crops in America and vectored by the tomato/potato psyllid Bactericera cockerelli; LsoC affects carrots from Northern and Central Europe, and is transmitted by the carrot psyllid Trioza apicalis; haplotypes LsoD and LsoE are present in Southern Europe and Morocco in carrot and celery, and are associated with the psyllid Bactericera trigonica. Thirty-four 'Ca. L. solanacearum' isolates were collected in six different regions of Spain from distinct Apiaceae hosts (carrot, celery, parsley and parsnip) in eight consecutive years and were analysed. Their haplotypes were determined by a sequence analysis of 16S ribosomal RNA, the 16S-26S ribosomal RNA intergenic spacer, and the 23S ribosomal RNA and rplJ and rplL genes. Both haplotypes LsoD and LsoE were found across Spain, and no host specificity appeared between these two haplotypes. This is the first report of 'Ca. L. solanacearum' associated with parsley and parsnip.This work has been supported by grant INIA (RTA2011-00142). This paper is dedicated to the memory of F.J. Villaescusa (1981-2011). The technical support of S. Sanjuan and J.C. Ferrandiz from Agricola Villena Coop. V. is also acknowledged.Alfaro Fernández, AO.; Hernández-Llopis, D.; Font San Ambrosio, MI. (2017). Haplotypes of "Candidatus Liberibacter solanacearum" identified in Umbeliferous crops in Spain. European Journal of Plant Pathology. 149(1):127-131. https://doi.org/10.1007/s10658-017-1172-21271311491Alfaro-Fernández A., Cebrián M.C., Villaescusa F.J., Hermoso de Mendoza A., Ferrándiz J.C., Sanjuán S., Font M.I. (2012). ‘Candidatus Liberibacter solanacearum’ associated with Bactericera trigonica affected carrots in the Canary Islands. Plant Disease 96, 581.Bertolini, E., Teresani, G. R., Loiseau, M., Tanaka, F. A. O., Barbé, S., Martínez, C., Gentit, P., López, M. M., & Cambra, M. (2014). Transmission of ‘Candidatus Liberibacter solanacearum’ in carrot seeds. Plant Pathology, 64, 276–285.EPPO. (2013). Data sheets on pests recommended for regulation. Candidatus Liberibacter solanacearum. EPPO Bulletin, 43, 197–201.Green, M. J., Thompson, D. A., & Mackenzie, D. J. (1999). Easy and efficient DNA extraction from woody plants for the detection of phytoplasmas by polymerase chain reaction. Plant Disease, 83, 482–485.Hansen, A. K., Trumble, J. T., Stouthamer, R., & Paine, T. D. (2008). A new huanglongbing species, “Candidatus Liberibacter psyllaurous,” found to infect tomato and potato, is vectored by the psyllid Bactericera cockerelli (Sulc). Applied Environmental Microbiology, 74, 5862–5865.Larkin, M. A., Blackshields, G., Brown, N. P., Chenna, R., McGettigan, P. A., McWilliam, H., Valentin, F., Wallace, I. M., Wilm, A., Lopez, R., Thompson, J. D., Gibson, T. J., & Higgins, D. G. (2007). Clustal W and Clustal X version 2.0. Bioinformatics, 23, 2947–2948.Loiseau, M., Garnier, S., Boirin, V., Merieau, M., Leguay, A., Renaudin, I., Renvoisé, J.-P., & Gentit, P. (2014). First report of ‘Candidatus Liberibacter solanacearum’ in carrot in France. Plant Disease, 98, 839.Munyaneza, J. E. (2012). Zebra chip disease of potato: biology, epidemiology and management. American Journal of Potato Research, 89, 329–350.Munyaneza, J., Buchman, J., Upton, J., Goolsby, J., Crosslin, J., & Bester, G. (2008). Impact of different potato psyllid populations on zebra chip disease incidence, severity, and potato yield. Subtropical Plant Science, 60, 27–37.Munyaneza, J., Sengoda, V., Crosslin, J., de la Rosa-Lorenzo, G., & Sanchez, A. (2009). First report of ‘Candidatus Liberibacter psyllaurous’ in potato tubers with zebra Chip disease in Mexico. Plant Disease, 93, 552.Munyaneza, J. E., Fisher, T. W., Sengoda, V. G., & Garczynski, S. F. (2010a). First report of ‘Candidatus Liberibacter solanacearum’ associated with psyllid-affected carrots in Europe. Plant Disease, 94, 639.Munyaneza, J. E., Fisher, T. W., Sengoda, V. G., Garczynski, S. F., Nissinen, A., & Lemmetty, A. (2010b). Association of ‘Candidatus Liberibacter solanacearum’ with the psyllid Trioza apicalis (hemiptera: Triozidae). Journal of Economic Entomology, 103, 1060–1070.Munyaneza, J. E., Swisher, K. D., Hommes, M., Willhauck, A., Buck, H., & Meadow, R. (2015). First report of ‘Candidatus Liberibacter solanacearum’ associated with psyllid-infested carrots in Germany. Plant Disease, 99, 1269.Nelson, W. R., Fisher, T. W., & Munyaneza, J. E. (2011). Haplotypes of “Candidatus Liberibacter solanacearum” suggest long-standing separation. European Journal of Plant Patholology, 130, 5–12.Nelson, W. R., Sengoda, V. G., Alfaro-Fernández, A., Font, M. I., Crosslin, J. M., & Munyaneza, J. E. (2013). A new haplotype of ‘Candidatus Liberibacter solanacearum’ identified in the Mediterranean region. European Journal of Plant Pathology, 135, 633–639.Tahzima, R., Maes, M., Achbani, E. H., Swisher, K. D., Munyaneza, J. E., & De Jonghe, K. (2014). First report of ‘Candidatus Liberibacter solanacearum’ on carrot in Africa. Plant Disease, 98, 1426.Teresani, G. R., Bertolini, E., Alfaro-Fernández, A., Martínez, C., Tanaka, F. A. O., Kitajima, E. W., Roselló, M., Sanjuán, S., Ferrándiz, J. C., López, M. M., Cambra, M., & Font, M. I. (2014). Association of ‘Candidatus Liberibacter solanacearum’ with a vegetative disorder of celery in Spain and development of a real-time PCR method for its detection. Phytopathology, 104, 804–811.Teresani, G., Hernández, E., Bertolini, E., Siverio, F., Marroquín, C., Molina, J., de Hermoso Mendoza, A., & Cambra, M. (2015). Search for potential vectors of ‘Candidatus Liberibacter solanacearum’: population dynamics in host crops. Spanish Journal of Agricultural Research, 13, e10–002

Virosis en tomate transmitidas por semilla y su control

[ES] Las virosis transmitidas por semilla en el cultivo del tomate crean gran preocupación entre los productores, y son de especial atención en aquellos que se dedican al cultivo de variedades locales donde las semillas se extraen durante la campaña y son empleadas para cultivos posteriores con lo que la infección y dispersión de estos virus es mucho más frecuente. Entre los virus transmitidos por semilla en tomate destacan el virus del mosaico del tomate (ToMV) y el virus del mosaico del pepino dulce (PepMV). Ambos virus se caracterizan por transmitirse, además de por semilla, de manera mecánica fácilmente y son muy estables manteniéndose en los restos del cultivo anterior y en las infraestructuras empleadas durante el manejo del cultivo. Sin embargo, la localización de estos virus en las semillas contaminadas difiere, mientras que PepMV se localiza únicamente de manera superficial, ToMV puede encontrarse además en zonas más internas como en el endospermo. Esto hace que los tratamientos empleados para la desinfección de semillas infectadas con cada uno de estos virus sea distinto: mientras que PepMV puede ser inactivado con tratamientos químicos superficiales, el tratamiento para descontaminar semillas con ToMV debe ser térmico a elevadas temperaturas.[EN] Viral diseases transmitted through seed create a great concern among the tomato producers, especially those who use local varieties that harvest their own seeds from the previous growing season fruits. In this case the infection and spread of seed-transmitted viruses is more usual. ToMV and PepMV are the two main seed-transmitted viruses which affect tomato crops. Both viruses are easily mechanically and seed transmitted, and remain infective in the plant debris of the previous crop and in the crop structures. However, the location of the virus in the contaminated seed is different. PepMV is present only externally in the seed coat, but ToMV could be also found in the endosperm. Therefore seed treatments to inactivate these two viruses are different; while PepMV could be inactivated by external chemical treatments, ToMV infected seeds should be thermal treated in order to eliminate further seedling infections.Alfaro Fernández, AO.; Font San Ambrosio, MI. (2020). Virosis en tomate transmitidas por semilla y su control. En I Congrés de la Tomaca Valenciana: La Tomaca Valenciana d'El Perelló. Editorial Universitat Politècnica de València. 97-114. https://doi.org/10.4995/TOMAVAL2017.2017.6524OCS9711

First report of cucurbit chlorotic yellows virus infecting watermelon and zucchini in the Canary Islands, Spain

This work was funded by grants from Spanish Ministerio de Econolnia, industria y competitividad (RTA2017-00061-C03-02) and from Instituto Valenciano de Investigaciones Agrarias (IVlA) (51912), both co-funded by the European Regional Development Fund (ERDF).Alfaro Fernández, AO.; Espino De Paz, AI.; Botella-Guillen, M.; Font San Ambrosio, MI.; Sanauja, E.; Galipienso, L.; Rubio, L. (2022). First report of cucurbit chlorotic yellows virus infecting watermelon and zucchini in the Canary Islands, Spain. Plant Disease. 106(7):1-1. https://doi.org/10.1094/PDIS-10-21-2296-PDN11106

Search for reservoirs of `Candidatus Liberibacter solanacearum¿ and mollicutes in weeds associated with carrot and celery crops

[EN] Currently, the main arthropod vectored pathogens associated with carrot and celery crop diseases are EiCandidatus Liberibacter solanacearumA ', Spiroplasma citri and different phytoplasma species. Mitigation strategies require elucidating whether these pathogens survive in the weeds of these Apiaceae crops, which can act as reservoirs. Weed surveys were conducted in a vegetative cycle (April to October 2012) in the spontaneous vegetation that surrounded crops affected by EiCa. L. solanacearumA ', S. citri and/or phytoplasmas. Sixty-three species of 53 genera that belong to 23 botanical families were collected in the main carrot and celery Spanish production area. Species were identified, estimating coverage and abundance, and conserved in herbarium. Samples were analysed by nested-PCR with universal primers for phytoplasmas detection, and were sequenced for identification purposes; by conventional PCR for S. citri and real-time PCR for EiCa. L. solanacearumA '. The only detected pathogens were EiCa. Phytoplasma trifoliiA ' (clover proliferation group 16Sr VI-A) in Amaranthus blitoides and Setaria adhaerens and EiCa. P. solaniA ' (stolbur group 16Sr XII-A) in Convolvulus arvensis. These pathogens were also sporadically detected in celery or carrot crops. Unexpectedly, neither EiCa. L. solanacearumA ' nor S. citri was detected in the weed samples, despite the relatively high prevalence of these pathogens (less than 66 % and 25 %, respectively) in the surveyed plots. This suggests that weeds do not play an epidemiological role as reservoirs in the spread of such organisms in the studied region. The use of pathogen-free seed lots and the control of vectors are crucial for preventing the introduction and spread of these economical important pathogens to new areas.This work has been supported by grant INIA (RTA2011-00142). G.R. Teresani was the recipient of a PhD grant from Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES), Ministerio de Educacao, Brazil. This paper is dedicated to the memory of F.J. Villaescusa (1981-2011). The technical support of S. Sanjuan and J.C. Ferrandiz from Agricola Villena Coop. V. is acknowledged.Alfaro Fernández, AO.; Verdeguer Sancho, MM.; Rodríguez-León, F.; Ibañez, I.; Hernández, D.; Teresani, GR.; Bertolini, E.... (2017). Search for reservoirs of `Candidatus Liberibacter solanacearum¿ and mollicutes in weeds associated with carrot and celery crops. European Journal of Plant Pathology. 147(1):15-20. https://doi.org/10.1007/s10658-016-0984-915201471Alfaro-Fernández, A., Cebrián, M. C., Villaescusa, F. J., Hermoso de Mendoza, A., Ferrándiz, J. C., Sanjuán, S., & Font, M. I. (2012). First report of ˋCandidatus Liberibacter solanacearum´ in carrots in mainland Spain. Plant Disease, 96, 582.Bertaccini, A., & Duduk, B. (2009). Phytoplasma and phytoplasma disease: a review of recent research. Phytopathologia Mediterranea, 48, 355–378.Bertolini, E., Teresani, G. R., Loiseau, M., Tanaka, F. A. O., Barbé, S., Martínez, C., Gentit, P., López, M. M., & Cambrá, M. (2015). Transmission of Candidatus Liberibacter solanacearum in carrot seeds. Plant Pathology, 64, 276–285.Bové, J. M. (1986). Stubborn and its natural transmission in the Mediterranean area and the near east. FAO Plant Protection Bulletin, 34, 15–23.Bové J. M., Fos A., Lallemand J., Raie A., Ali Y., Ahmed N. (1988). Epidemiology of Spiroplasma citri in the old world. In: L. W. Timmer, S.M. Garnsey, L. Navarro, eds. Proceedings of the 10th International Organization of Citrus Virologist Conference, (295–299). Riverside, USA. (www.iocv.org/proceedings).Braun-Blanquet, J. (1932). Plant sociology: the study of plant communities (439 pp). New York: McGraw-Hill.Carretero, J. L. (2004). Flora arvense española (754 pp). Valencia: Las malas hierbas de los cultivos españoles. Phytoma Ed.Cebrián, M. C., Villaescusa, F. J., Alfaro-Fernández, A., Hermoso De Mendoza, A., Córdoba- Sellés, M. C., Jordá, C., Ferrándiz, J. C., Sanjuán, S., & Font, M. I. (2010). First report of Spiroplasma citri in carrot in Europe. Plant Disease, 94, 1264.Davis, R. M., & Raid, R. N. (2002). Compendium of Umbelliferous crop disease (110 pp).American Phytopathological SocietyEmber, I., Acs, Z., Munyaneza, J. E., Crosslin, J. M., & Kolber, M. (2011). Survey and molecular detection of phytoplasmas associated with potato in Romania and southern Rusia. European Journal of Plant Pathology, 130, 367–377.Fialová, R., Valová, P., Balakishiyevá, G., Danet, J. L., Safarová, D., Foissac, X., & Navratil, M. (2009). Genetic variability of stolbur phytoplasma in anual crop and wild plant species in South Moravia. Journal of Plant Pathology, 91, 411–416.Fujiwara, K. (1987). Aims and methods of phytosociology or "vegetation science", Papers on plant ecology and taxonomy to the memory of Dr. Satoshi Nakanishi. pp. 607–628.Green, M. J., Thompson, D. A., & MacKenzie, D. J. (1999). Easy and efficient DNA extraction from Woody plants for the detection of Phytoplasmas by polymerase chain reaction. Plant Disease, 83, 482–485.Gundersen, D. E., & Lee, I. M. (1996). Ultrasensitive detection of phytoplasmas by nested- PCR assays using two universal primer pairs. Phytopathologia Mediterranea, 35, 144–151.Haapalanien, M. (2014). Biology and epidemics of Candidatus Liberibacter species, psyllid-transmitted plant-pathogenic bacteria. Annals of Applied Biology, 165, 172–198.Herbario de la Universidad Pública de Navarra. (2012). http://www.unavarra.es/herbario. Accessed 2012.Herbario Virtual del Mediterráneo Occidental. (2012). http://herbarivirtual.uib.es/. Accessed 2012.Flora Ibérica. (2012). http://www.floraiberica.org/. Accessed 2012.Jomantiene, R., Maas, J. L., Dally, E. L., Davis, R. E., & Postman, J. D. (1999). First report of clover proliferation Phytoplasma in strawberry. Plant Disease, 83, 967.Jomantiene, R., Postman, J. D., Montano, H. G., Maas, J. L., Davis, R. E., & Johnson, K. B. (2000). First report of clover yellow edge Phytoplasma in Corylus (hazelnut). Plant Disease, 84, 102.Lee, I. M., Gundersen-Rindal, D. E., Davis, R. E., & Bartoszyk, I. M. (1998). Revised classification scheme of phytoplasmas based on RFLP analyses of 16S rRNA and ribosomal protein gene sequences. International Journal of Systematic and Evolutionary Microbiology, 48, 1153–1169.Lee, I. M., Dane, R. A., & Black, M. C. (2001). First report of a member of Aster yellows Phytoplasma group and of clover proliferation Phytoplasma group associated with onion in Texas. Plant Disease, 85, 448.Lee, I. M., Bottner, K. D., Miklas, P. N., & Pastor-Corrales, M. A. (2004). Clover proliferation group (16SrVI) subgroup a (16SrVI-A) Phytoplasma is a probable causal agent of dry bean Phyllody disease in Washington. Plant Disease, 88, 429–429.Mateo, G., & Crespo, M. (2009). Manual Para la determinación de la flora valenciana (4ª ed.507 pp). Alicante: Librería Compás.Ed.Murphy, A. F., Cating, R. A., Goyer, A., Hamm, P. B., & Rondon, S. I. (2014). First report of natural infection by ‘Candidatus Liberibacter solanacearum’ in bittersweet nightshade (Solanum dulcamara) in the Columbia Basin of eastern Oregon. Plant Disease, 94, 1425.Nejat, N., Vadamalai, G., & Dickinson, M. (2011). Spiroplasma citri: a wide host range phytopathogen. Plant Pathology Journal, 10, 46–56.Schneider, B., Seemüller, E., Smart, C. D., & Kirkpatrick, B. C. (1995). Phylogenetic classification of plant pathogenic mycoplasma-like organisms or phytoplasmas. In S. Razin & J. G. Tully (Eds.), Molecular and diagnostic procedures in mycoplasmology (Vol. Vol.I, pp. 369–380). San Diego: Academic Press.Teresani, G., Bertolini, E., Alfaro-Fernandez, A., Martínez, C., Tanaka, F. A., Kitajima, E., Rosello, M., Sanjuan, S., Ferrandiz, J. C., López, M. M., Cambra, M., & Font-San-Ambrosio, M. I. (2014). Association of ‘Candidatus Liberibacter solanacearum’ with a vegetative disorder of celery in Spain and development of a real-time PCR method for its detection. Phytopahology, 104, 804–811.Teresani, G., Hernández, E., Bertolini, E., Siverio, F., Marroquín, C., Molina, J., Hermoso de Mendoza, A., & Cambra, M. (2015). Search for potencial vectors of ‘Candidatus Liberibacter solanacearum’: population dynamics in host crops. Spanish Journal of Agricultural Research, 13, e10–002.Flora Vascular. (2012). http://www.floravascular.com/. Accessed 2012.Weed Science Society of America. (2012). http://wssa.net/weed/weed-identification/. Accessed 2012.Yokomi, R. K., Mello, A. F. S., Saponari, M., & Fletcher, J. (2008). Polymerase chain reactionbased detection of Spiroplasma citri associated with citrus stubborn disease. Plant Disease, 92, 253–260

Development of a Real-Time Loop-Mediated Isothermal Amplification Assay for the Rapid Detection of Olea Europaea Geminivirus

A real-time loop-mediated isothermal amplification (LAMP) assay was developed for simple, rapid and efficient detection of the Olea europaea geminivirus (OEGV), a virus recently reported in different olive cultivation areas worldwide. A preliminary screening by end-point PCR for OEGV detection was conducted to ascertain the presence of OEGV in Sicily. A set of six real-time LAMP primers, targeting a 209-nucleotide sequence elapsing the region encoding the coat protein (AV1) gene of OEGV, was designed for specific OEGV detection. The specificity, sensitivity, and accuracy of the diagnostic assay were determined. The LAMP assay showed no cross-reactivity with other geminiviruses and was allowed to detect OEGV with a 10-fold higher sensitivity than conventional end-point PCR. To enhance the potential of the LAMP assay for field diagnosis, a simplified sample preparation procedure was set up and used to monitor OEGV spread in different olive cultivars in Sicily. As a result of this survey, we observed that 30 out of 70 cultivars analyzed were positive to OEGV, demonstrating a relatively high OEGV incidence. The real-time LAMP assay developed in this study is suitable for phytopathological laboratories with limited facilities and resources, as well as for direct OEGV detection in the field, representing a reliable method for rapid screening of olive plant material

Inoculation of cucumber, melón and zucchini varieties with Tomato leaf curl New Delhi virus (ToLCNDV) and evaluation of infection using different methods

This is the peer reviewed version of the following article: Figás-Moreno, MDR.; Alfaro Fernández, AO.; Font San Ambrosio, MI.; Borràs Palomares, D.; Casanova-Calancha, C.; Hurtado Ricart, M.; Plazas Ávila, MDLO.... (2017). Inoculation of cucumber, melón and zucchini varieties with Tomato leaf curl New Delhi virus (ToLCNDV) and evaluation of infection using different methods. Annals of Applied Biology. 170(3):405-414. doi:10.1111/aab.12344, which has been published in final form at http://doi.org/10.1111/aab.12344. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.[EN] The disease caused by Tomato leaf curl New Delhi virus (ToLCNDV), which is naturally transmitted by the whitefly Bemisia tabaci, causes important economic losses in cucurbit crops. The availability of simple and efficient inoculation protocols and detection methods is necessary for screening varieties and germplasm collections as well as for breeding populations. We evaluated the infectivity of ToLCNDV inocula prepared using three different buffers for mechanical sap inoculation in a susceptible variety of zucchini. We found that inoculum prepared with buffer III, which contains polyvinylpyrrolidone, is highly efficient for mechanical inoculation, with 100% of plants displaying severe symptoms 21 days post-inoculation. Using this buffer, we mechanically inoculated 19 commercial varieties of cucurbit crops (six of cucumber, six of melon and seven of zucchini), evaluated the evolution of symptoms and diagnosed infection using nine different ToLCNDV detection methods (four based on serology, four based on molecular hybridization and one based on PCR detection). The results revealed that all varieties are susceptible, although cucumber varieties display less severe symptoms than those of melon or zucchini. All detection methods were highly efficient (more than 85% of plants testing positive) in melon and zucchini, but in cucumber, the percentage of positive plants detected with serology and molecular hybridization methods ranged from 20.4% with Squash leaf curl virus (SLCV) antiserum, to 78.5% with DNA extract hybridization. Overall, the best detection results were obtained with PCR, with 92.6%, 92.4% and 98.4% cucumber, melon and zucchini plants, respectively, testing positive. When considering the overall results in the three crops, the best serology and molecular hybridization methods were those using Watermelon chlorotic stunt virus (WmCSV) antiserum and DNA extract, respectively. The inoculation methodology developed and the information on detection methods are of great relevance for the selection and breeding of varieties of cucurbit crops that are tolerant or resistant to ToLCNDV.Figás-Moreno, MDR.; Alfaro Fernández, AO.; Font San Ambrosio, MI.; Borràs Palomares, D.; Casanova-Calancha, C.; Hurtado Ricart, M.; Plazas Ávila, MDLO.... (2017). Inoculation of cucumber, melón and zucchini varieties with Tomato leaf curl New Delhi virus (ToLCNDV) and evaluation of infection using different methods. Annals of Applied Biology. 170(3):405-414. doi:10.1111/aab.12344S405414170

Molecular analysis of a Spanish isolate of chili pepper mild mottle virus and evaluation of seed transmission and resistance genes

[EN] An isolate of chili pepper mild mottle virus (CPMMV-Sp; GenBank OQ920979) with a 99% identity to CPMMV (GenBank MN164455.1) was found in symptomatic pepper plants in Spain. RACE analysis, performed using a stem-loop primer developed in this study to prime at the end of the introduced poly(A)/(U) tail, revealed the presence of an extra 22 nt at the 5' end, starting with a cytosine, which were essential to generate infectious clones. However, the 5' terminal cytosine was dispensable for initiating the infection. The design of two specific digoxigenin riboprobes targeting the more divergent area of CPMMV-Sp, compared to the closely related bell pepper mottle virus (BPeMV) (identity percentage of 80.6% and 75.8%, respectively), showed that both probes specifically detected CPMMV-Sp when the hybridization was performed at 68oC and 60oC, respectively. However, the BPeMV probe, targeting a region with an 89.4% identity percentage to CPMMV-Sp, showed cross-hybridization at 60oC but not at 68oC. The comparison of the detection limits between molecular hybridization and RT-PCR techniques revealed that the former was 125 times less sensitive than RT-PCR. The analysis of the vertical transmission of CPMMV-Sp using seeds from naturally or mechanically infected pepper plants revealed a transmission percentage ranging from 0.9% to 8.5%. Finally, the analysis of the resistance of capsicum species carrying different alleles of the L gene (L1, L2, L3, and L4) revealed that varieties with the L1 gene were infected by CPMMV-Sp (20-40% of inoculated plants), while varieties with the L2, L3, and L4 genes were resistant.This work was supported by grants PID2020-115571RB-100 and TED2021-131949B-I00 from the Spanish Agencia Estatal de Investigacion (AEI) and Fondo Europeo de Desarrollo Regional (FEDER). Project 20-00032-VIRUSPIM from Dept. of Environment, Territorial Planning, Agriculture and Fisheries (Basque Government). Mikel Ojinaga was the recipient of a PhD contract "Introduction of Resistance to Tobamovirus and other Viruses in Landraces of Gernika Pepper and Ibarra Chili Pepper" (Order of 24 October 2018 of the Minister of Economic Development and Competitiveness of the Basque Government). Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature.Ontañon, C.; Ojinaga, M.; Larregla, S.; Zabala, JA.; Reva, A.; Losa, A.; Heribia, R.... (2023). Molecular analysis of a Spanish isolate of chili pepper mild mottle virus and evaluation of seed transmission and resistance genes. European Journal of Plant Pathology. 1-18. https://doi.org/10.1007/s10658-023-02765-1118Al-Tamimi, N., Kawas, H., & Mansour, A. (2010). Seed Transmission Viruses in Squash Seeds (Cucurbita pepo) in Southern Syria and Jordan Valley. 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Seed Transmission of Tobamoviruses: Aspects of Global Disease Distribution. Advances in Seed Biology. https://doi.org/10.5772/INTECHOPEN.70244Gallois, J. L., Moury, B., & German-Retana, S. (2018). Role of the genetic background in resistance to plant viruses. International Journal of Molecular Sciences, 19(10), 2856. https://doi.org/10.3390/ijms19102856Genda, Y., Kanda, A., Hamada, H., Sato, K., Ohnishi, J., & Tsuda, S. (2007). Two amino acid substitutions in the coat protein of Pepper mild mottle virus are responsible for overcoming the L4 gene-mediated resistance in Capsicum spp. Phytopathology, 97(7), 787–793.Genda, Y., Sato, K., Nunomura, O., Hirabayashi, T., & Tsuda, S. (2011). Immunolocalization of Pepper mild mottle virus in developing seeds and seedlings of Capsicum annuum. Journal of General Plant Pathology, 77(3), 201–208. https://doi.org/10.1007/s10327-011-0307-0Genda, Y., Sato, K., Nunomura, O., Hirabayashi, T., Ohnishi, J., & Tsuda, S. (2005). 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Use of tomato crop residues into soil for control of bacterial wilt caused by Ralstonia solanacearum

[EN] Tomato debris can be used as the soil amendment to reduce agricultural residue accumulation problems and increase soil fertility. However, pathogens present in the debris may form a risk for subsequent crops. In this study, tomato growth substrate was amended with tomato debris artificially inoculated with Ralstonia solanacearum and the effect of heat treatments on the survival of the pathogen was measured. Experiments were carried out in the laboratory and in greenhouses, using peat moss and sand mix in pots as substrates. Pots were enclosed in plastic bags or left open. Then 0, 5, 10 and 15 g of tomato debris were applied to 500 g growing medium, with four replicates per treatment. Treatments at 45 °C lowered tomato wilt indices in tomato cv. Money-Maker and that the pathogen was not eradicated after pot treatments at 25 °C. R. solanacearum remained pathogenic on the assayed growing media after a six-week treatment at 25 °C, but was eradicated after treatments at 45 °C. The lower infectivity of infected debris tomato plants when buried with high doses of organic matter and at temperatures above 45 °C suggests that adverse effects on the soil inoculum would be exerted through increased soil temperatures. This study demonstrates that tomato crop residues, usually considered waste material, could be used as soil amendments to reduce their effect as a source of contamination as they offer additional advantages. © 2011 Elsevier Ltd.This study has been supported by the Spanish Ministry of Science and Technology (AGL2002-04040-C05-05). M.J. Zanon received a fellowship from the "Universidad Politecnica de Valencia. Programa de Formacion de Personal Investigador".Zanón, MJ.; Font San Ambrosio, MI.; Jordá, C. (2011). Use of tomato crop residues into soil for control of bacterial wilt caused by Ralstonia solanacearum. Crop Protection. 30(9):1138-1143. https://doi.org/10.1016/j.cropo.2011.03.025S1138114330

First report of Eggplant mottled dwarf virus in Pittosporum tobira in Spain

Alfaro Fernández, AO.; Córdoba-Sellés, MDC.; Tornos, T.; Cebrián, M.; Font San Ambrosio, MI. (2011). First report of Eggplant mottled dwarf virus in Pittosporum tobira in Spain. Plant Disease. 95(1):75-75. doi:10.1094/PDIS-07-10-0491S757595

Detection and identification of aster yellows and stolbur phytoplasmas in various crops in Spain

Copyright of the bulletin (©Bulletin of Insectology). Full text articles are available at the publisher site freely two years after publication.In this study 'Candidatus Phytoplasma asteris' (subgroups 16SrI-B) and stolbur phytoplasma (subgroup 16SrXII-A) were sporadically identified in several horticultural crops in Spain by nested-PCR and RFLP analyses. One parsnip sample was infected with stolbur, and 'Ca. P. asteris' was detected in lettuce and chicory plants. However, these two phytoplasmas were able to infect also carrot, celery and radish. This work extends the knowledge of phytoplasma diversity affecting horticultural crops in Spain.Alfaro Fernández, AO.; Cebrian Mico, MC.; Villaescusa Sánchez, FJ.; Font San Ambrosio, MI. (2011). Detection and identification of aster yellows and stolbur phytoplasmas in various crops in Spain. Bulletin of insectology. 64:63-64. http://hdl.handle.net/10251/63856S63646