GENETICS AND GENOMICS OF CULTIVATED EGGPLANTS AND WILD RELATIVES

Tesis por compendioAlimentar a la futura población en crecimiento en un contexto de cambio climático exige nuevos enfoques y herramientas de mejora genética para desarrollar nuevas variedades de cultivos que sean resilientes y eficientes en el uso de los recursos. Entre las hortalizas, la berenjena (Solanum melongena) es reconocida como un cultivo importante y como tal está incluida en el Anexo 1 del Tratado Internacional sobre los Recursos Fitogenéticos para la Alimentación y la Agricultura, que incluye 34 cultivos considerados como los más relevantes para la humanidad. Antes del inicio de esta tesis, se disponía de pocas herramientas genéticas y genómicas para la mejora genética de la berenjena, las cuales se revisan en un capítulo introductorio. Asimismo, hemos reconocido la importancia de los parientes silvestres de la berenjena, que apenas se han utilizado en la mejora genética de la misma. A este respecto, para hacer un uso más eficiente de los parientes silvestres en la mejora genética, propusimos un enfoque ambicioso, llamado "introgressiomics", que consiste en un desarrollo sistemático y masivo de materiales que lleven introgresiones de parientes silvestres (CWR), los cuales generalmente son una fuente inexplorada e desaprovechada de variación genética. Los trabajos realizados en esta tesis están relacionados con la aplicación de este enfoque a la berenjena. En este marco, los objetivos generales de la tesis son el desarrollo de la información y herramientas genéticas y genómicas en el acervo genético de la berenjena, utilizando un enfoque multidisciplinario y multifacético para la mejora genética en el desarrollo de nuevas variedades mejoradas y resistentes usando especies relacionadas como fuente de variación. Concretamente, en el primer capítulo de esta tesis, secuenciamos el transcriptoma de dos especies relacionadas con la berenjena, la silvestre Solanum incanum y la especie cultivada africana S. aethiopicum, que tienen un gran interés en la mejora genética de la berenjena. Los transcriptomas se ensamblaron en 83,905 y 87,084 unigenes para S. incanum y S. aethiopicum respectivamente, los cuales fueron extensivamente anotados estructuralmente y funcionalmente. La busqueda de variantes alélicas identificó decenas de miles de polimorfismos intraespecíficos e interespecíficos, así como alrededor de un millar de SSRs en cada especie. En el segundo capítulo, un subconjunto de esos marcadores (11 SSRs y 35 SNPs) se analizaron para confirmar su utilidad para la huella genética, la evaluación de la diversidad y el establecimiento de relaciones entre las berenjenas cultivadas (común, escarlata y gboma). Observamos que los SSRs y los SNPs proporcionaron resultados diferentes en el establecimiento de las relaciones, lo que sugiere que cada tipo de marcador muestreó diferentes niveles de variación genética. Sin embargo, aunque ambos marcadores proporcionaron un nivel similar de información, los SNPs parecen proporcionar una mejor resolución que los SSRs para materiales filogenéticamente más distantes. En el tercer capítulo, para ampliar la estimación de la diversidad genética y las relaciones genéticas entre y dentro de las especies silvestres y cultivadas pertenecientes al complejo berenjena, realizamos un genotipado masivo, mediante un enfoque de genotipado por secuenciación, de 76 accesiones pertenecientes a 17 especies del acervo genético primario, secundario y terciario de la berenjena común. De los 75,399 sitios polimórficos identificados, 12,859 se asociaron a regiones CDS y se usaron para establecer una evaluación exhaustiva y detallada de la diversidad alélica natural y las relaciones genéticas en el acervo genético de la berenjena utilizando tres enfoques diferentes (estructura jerárquica de la población, dendrograma basado en UPGMA y análisis de PCoA). En el cuatro capítulo, desarrollamos un mapa genético interespecífico entre S. incanum y S. melongena, vinculado a otrosFeeding the future burgeoning population in a climate change scenario demands new breeding approaches and tools to develop new resource-efficient and resilient crop varieties. Among vegetable crops, eggplant (Solanum melongena) is recognized as an important food crop and as such is included in the Annex 1 of the International Treaty on Plant Genetic Resources for Food and Agriculture, which includes 34 crops considered as most relevant for mankind. Before the start of this thesis, few genetic and genomic tools and resources for eggplant breeding, which are reviewed in an introductory chapter, were available. We have recognized the importance of wild eggplant relatives, which have been barely used in eggplant breeding. In this respect, in order to make a more efficient use of wild relatives in plant breeding, we proposed an ambitious approach, called "introgressiomics", consisting of a systematic and massive development of materials carrying introgressions from crop wild relatives (CWRs), which usually are an unexplored and unexploited source of genetic variation for breeding traits. The works done in this thesis are related to the application of the introgressiomics approach to eggplant. In this framework, the general objectives of the thesis are the development of genetic and genomic information and tools in eggplant genepool, using a multidisciplinary and multi-pronged approach to assist eggplant breeding in the development of new improved and resilient varieties using eggplant relatives as a source of variation. Specifically, in the first chapter of this thesis, we sequenced the transcriptome of two eggplant related species, the wild Solanum incanum and the cultivated S. aethiopicum, that have a great interest in eggplant breeding. The transcriptomes were assembled in 83,905 and 87,084 unigenes for S. incanum and S. aethiopicum respectively, which were extensively structurally and functionally annotated. The variant call analysis identified tens of thousands intraspecific and interspecific polymorphisms, as well as around a thousand of SSRs in each species. In the second chapter, a subset of those markers (11 SSRs and 35 SNPs) was tested for confirming their usefulness for genetic fingerprinting, diversity evaluation and the establishment of relationships in cultivated eggplant (common, scarlet and gboma) genepools. We observed that SSRs and SNPs provided different results in the establishment of the relationships, suggesting that each marker type sampled different levels of genetic variation. However, although both markers provided a similar level of information, SNPs seem to provide a better resolution than SSRs for materials phylogenetically more distant. In the third chapter, in order to broaden the estimation of the genetic diversity and genetic relationships among and within wild and cultivated species belonging to eggplant complexes, we performed a massive genotyping, by a genotype-by-sequencing approach, of 76 accessions belonging to 17 species from the primary, secondary and tertiary genepool of common eggplant. Out of 75,399 polymorphic sites identified, 12,859 were associated to CDS regions and used to establish an exhaustive and detailed evaluation of the natural allelic diversity and genetic relationships in eggplant genepool using three different approaches (hierarchical population structure, UPGMA-based dendrogram, and PCoA analysis). In the four chapter, we developed an interspecific genetic map between S. incanum and S. melongena, linked to four previous eggplant maps and to one tomato map. A total of 243 molecular markers were successfully mapped consisting of 42 COSII, 99 SSRs, 88 AFLPs, 9 CAPS, 4 SNPs and one morphological polymorphic markers encompassed 1085 cM distributed in 12 linkage groups. Based on the syntheny with tomato, the candidate genes involved in the core chlorogenic acid synthesis pathway in eggplant (PAL, C4H, 4CL, HCT, C3¿H, HQT), five polyphenol oxidase genes (PPO1, PPO2, PPO3, PPAlimentar la futura població en creixement en un context de canvi climàtic exigeix nous enfocaments i eines de millora genètica per desenvolupar noves varietats de cultius que siguin resilients i eficients en l'ús dels recursos. Entre les hortalisses, l'albergínia (Solanum melongena) és reconeguda com un cultiu important i com a tal està inclosa en l'Annex 1 del Tractat Internacional sobre els Recursos Fitogenètics per a l'Alimentació i l'Agricultura, que inclou 34 cultius considerats com els més rellevants per a la humanitat. Abans de l'inici d'aquesta tesi, es disposava de poques eines genètiques i genòmiques per a la millora genètica de l'albergínia, les quals es revisen en un capítol introductori. Així mateix, hem reconegut la importància dels parents silvestres de l'albergínia, que tot just s'han utilitzat en la millora genètica de la mateixa. Referent a això, per fer un ús més eficient dels parents silvestres a la millora genètica, vam proposar un enfocament ambiciós, anomenat "introgressiomics", que consisteix en un desenvolupament sistemàtic i massiu de materials que portin introgresions de parents silvestres (CWR), els quals generalment són una font inexplorada i desaprofitada de variació genètica. Els treballs realitzats en aquesta tesi estan relacionats amb l'aplicació d'aquest enfocament a l'albergínia. En aquest marc, els objectius generals de la tesi són el desenvolupament de la informació i eines genètiques i genòmiques en el patrimoni genètic de l'albergínia, utilitzant un enfocament multidisciplinari i multifacètic per a la millora genètica en el desenvolupament de noves varietats millorades i resistents emprent spècies relacionades com a font de variació. Concretament, en el primer capítol d'aquesta tesi, seqüenciem el transcriptoma de dues espècies relacionades amb l'albergínia, la silvestre Solanum incanum i l'espècie cultivada africana S. aethiopicum, que tenen un gran interès en la millora genètica de l'albergínia. Els transcriptomes s'ensamblaren en 83,905 i 87,084 unigenes per S. incanum i S. aethiopicum respectivament, els quals van ser extensivament anotats estructuralment i funcionalment. La recerca de variants al·lèliques va identificar desenes de milers de polimorfismes intraespecífics i interespecífics, així com al voltant d'un miler de SSRs en cada espècie. En el segon capítol, un subconjunt d'aquests marcadors (11 SSRs i 35 SNPs) es van analitzar per confirmar la seva utilitat per l'empremta genètica, l'avaluació de la diversitat i l'establiment de relacions entre les albergínies conreades (comú, escarlata i gboma). Observem que els SSRs i els SNPs van proporcionar resultats diferents en l'establiment de les relacions, el que suggereix que cada tipus de marcador va mostrejar diferents nivells de variació genètica. No obstant això, encara que tots dos marcadors van proporcionar un nivell similar d'informació, els SNPs semblen proporcionar una millor resolució que els SSRs per materials filogenèticament més distants. En el tercer capítol, per a ampliar l'estimació de la diversitat genètica i les relacions genètiques entre i dins de les espècies silvestres i conreades pertanyents al complex albergínia, vam realitzar un genotipat massiu, mitjançant un enfocament de genotipat per seqüenciació, de 76 accessions pertanyents a 17 espècies del patrimoni genètic primari, secundari i terciari de l'albergínia comuna. Dels 75,399 llocs polimòrfics identificats, 12,859 es van associar a regions CDS i es van usar per a establir una avaluació exhaustiva i detallada de la diversitat al·lèlica natural i les relacions genètiques en el patrimoni genètic de l'albergínia utilitzant tres enfocaments diferents (estructura jeràrquica de la població, dendrograma basat en UPGMA i anàlisi de PCoA). En el quart capítol, desenvolupem un mapa genètic interespecífic entre S. incanum i S. melongena, vinculat a altres quatre mapes anteriorGramazio, P. (2018). GENETICS AND GENOMICS OF CULTIVATED EGGPLANTS AND WILD RELATIVES [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/104605TESISCompendi

Challenges and prospects of new plant breeding techniques for GABA improvement in crops: tomato as an example

[EN] Over the last seven decades, gamma-aminobutyric acid (GABA) has attracted great attention from scientists for its ubiquity in plants, animals and microorganisms and for its physiological implications as a signaling molecule involved in multiple pathways and processes. Recently, the food and pharmaceutical industries have also shown significantly increased interest in GABA, because of its great potential benefits for human health and the consumer demand for health-promoting functional compounds, resulting in the release of a plethora of GABA-enriched products. Nevertheless, many crop species accumulate appreciable GABA levels in their edible parts and could help to meet the daily recommended intake of GABA for promoting positive health effects. Therefore, plant breeders are devoting much effort into breeding elite varieties with improved GABA contents. In this regard, tomato (Solanum lycopersicum), the most produced and consumed vegetable worldwide and a fruit-bearing model crop, has received much consideration for its accumulation of remarkable GABA levels. Although many different strategies have been implemented, from classical crossbreeding to induced mutagenesis, new plant breeding techniques (NPBTs) have achieved the best GABA accumulation results in red ripe tomato fruits along with shedding light on GABA metabolism and gene functions. In this review, we summarize, analyze and compare all the studies that have substantially contributed to tomato GABA breeding with further discussion and proposals regarding the most recent NPBTs that could bring this process to the next level of precision and efficiency. This document also provides guidelines with which researchers of other crops might take advantage of the progress achieved in tomato for more efficient GABA breeding programsPG is grateful to the Japanese Society for the Promotion of Science for the JSPS postdoctoral grant FY2019-P19105Gramazio, P.; Takayama, M.; Ezura, H. (2020). Challenges and prospects of new plant breeding techniques for GABA improvement in crops: tomato as an example. Frontiers in Plant Science. 11:1-16. https://doi.org/10.3389/fpls.2020.577980S1161

A novel and rapid method for Agrobacterium-mediated production of stably transformed Cannabis sativa L. plants

[EN] The development of genetically transformed plants is an elusive landmark in Cannabis sativa L. breeding. Despite its economic interest, at present, protocols for producing transgenic C. sativa plants are scarce. We studied the ability of hypocotyl, cotyledon and meristem explants from six C. sativa hemp varieties for transgenic plant regeneration. For this, we firstly evaluated in vitro regeneration rates of hypocotyls cultured in medium without plant growth regulators, and cotyledons cultured in medium supplemented with 0.4 mg L- 1 of thidiazuron (TDZ) and 0.2 mg L- 1 of ¿-naphthaleneacetic (NAA). Subsequently, the effect of different kanamycin concentrations (50, 100, 200, 500 and 750 mg L- 1) on hypocotyl regeneration rate was determined. Finally, we assessed transformation rates after hypocotyl, cotyledon and meristem co-culture with Agrobacterium tumefaciens strain LBA4404 carrying the binary plasmid pBIN19 containing the ß-glucuronidase (uidA) reporter gene and the kanamycin resistance neomycin phosphotransferase (nptII) genes. Plant transformation was validated through in vitro culture of regenerating shoots in kanamycin-containing selective regeneration medium, by GUS histochemical assay for uidA expression, and by PCR amplification of uidA and nptII genes. Our results showed that hypocotyls reached a higher regeneration rate (53.3 %) than cotyledons (18.1 %) without Agrobacterium coculture. On the other hand, 100 mg L- 1 kanamycin proved to be the best concentration in terms of regeneration rate (63.3 %) and spontaneous rooting rate of hypocotyl regenerating shoots (12.2 %), which displayed a 7.1 % of albinism rate. After co-culture with A. tumefaciens and subsequent culture in antibiotic-containing selective regeneration medium, hypocotyl was the best explant type achieving 23.1 % of regeneration rate, which contrasts with the 1.0 % regeneration rate detected for cotyledons. Transgenic plants were obtained from all explant types evaluated. Although there were significant differences among varieties evaluated, hypocotyls proved to be superior to already-developed meristems, reaching a transformation rate of 5.0 % and 0.8 % respectively. Despite the extremely low regeneration rate of cotyledons after A. tumefaciens co-culture, all cotyledon-derived regenerating shoots analyzed were successfully transformed. Our hormone-free protocol doubles the transformation rate of regenerating shoots, also producing transgenic plants three times faster than other already published protocols. This has relevant implications for C. sativa breeding, enabling not only genetic transformation, but also the use of new plant breeding techniques such as targeted genome editing by using CRISPR/Cas systems. This may foster the development of C. sativa varieties with specific biochemical profiles, or tolerant to biotic and abiotic stresses among others.The authors received no specific funding for this work. Pietro Gramazio is grateful to Japan Society for the Promotion of Science (JSPS) for a post-doctoral grant (P19105, FY2019 [Standard])Galán-Ávila, A.; Gramazio, P.; Ron, M.; Prohens Tomás, J.; Herraiz García, FJ. (2021). A novel and rapid method for Agrobacterium-mediated production of stably transformed Cannabis sativa L. plants. Industrial Crops and Products. 170:1-15. https://doi.org/10.1016/j.indcrop.2021.113691S11517

spectroscopic ellipsometry meets afm nanolithography about hydration of bio inert oligo ethylene glycol terminated self assembled monolayers on gold

An accurate thickness determination provides insight into the complex vertical morphology of OEG-terminated SAMs

De novo Transcriptome Assembly and Comprehensive Annotation of Two Tree Tomato Cultivars (Solanum betaceum Cav.) with Different Fruit Color

[EN] The tree tomato (Solanum betaceum Cav.) is an underutilized fruit crop native to the Andean region and phylogenetically related to the tomato and potato. Tree tomato fruits have a high amount of nutrients and bioactive compounds. However, so far there are no studies at the genome or transcriptome level for this species. We performed a de novo assembly and transcriptome annotation for purple-fruited (A21) and an orange-fruited (A23) accessions. A total of 174,252 (A21) and 194,417 (A23) transcripts were assembled with an average length of 851 and 849 bp. A total of 34,636 (A21) and 36,224 (A23) transcripts showed a significant similarity to known proteins. Among the annotated unigenes, 22,096 (A21) and 23,095 (A23) were assigned to the Gene Ontology (GO) term and 14,035 (A21) and 14,540 (A23) were found to have Clusters of Orthologous Group (COG) term classifications. Furthermore, 22,096 (A21) and 23,095 (A23) transcripts were assigned to 155 and 161 (A23) KEGG pathways. The carotenoid biosynthetic process GO terms were significantly enriched in the purple-fruited accession A21. Finally, 68,647 intraspecific single-nucleotide variations (SNVs) and almost 2 million interspecific SNVs were identified. The results of this study provide a wealth of genomic data for the genetic improvement of the tree tomato.Pietro Gramazio is grateful to the Spanish Ministerio de Ciencia e Innovacion for a Juan de laCierva-Formacion post-doctoral grant FJC2019-038921-I funded by MCIN/AEI/10.13039/501100011033)Pacheco, J.; Vilanova Navarro, S.; Grillo-Risco, R.; García-García, F.; Prohens Tomás, J.; Gramazio, P. (2021). De novo Transcriptome Assembly and Comprehensive Annotation of Two Tree Tomato Cultivars (Solanum betaceum Cav.) with Different Fruit Color. Horticulturae. 7(11):1-18. https://doi.org/10.3390/horticulturae7110431S11871

Performance of a Set of Eggplant (Solanum melongena) Lines With Introgressions From Its Wild Relative S. incanum Under Open Field and Screenhouse Conditions and Detection of QTLs

[EN] Introgression lines (ILs) of eggplant (Solanum melongena) represent a resource of high value for breeding and the genetic analysis of important traits. We have conducted a phenotypic evaluation in two environments (open field and screenhouse) of 16 ILs from the first set of eggplant ILs developed so far. Each of the ILs carries a single marker-defined chromosomal segment from the wild eggplant relative S. incanum (accession MM577) in the genetic background of S. melongena (accession AN-S-26). Seventeen agronomic traits were scored to test the performance of ILs compared to the recurrent parent and of identifying QTLs for the investigated traits. Significant morphological differences were found between parents, and the hybrid was heterotic for vigour-related traits. Despite the presence of large introgressed fragments from a wild exotic parent, individual ILs did not display differences with respect to the recipient parent for most traits, although significant genotype x environment interaction (G x E) was detected for most traits. Heritability values for the agronomic traits were generally low to moderate. A total of ten stable QTLs scattered across seven chromosomes was detected. For five QTLs, the S. incanum introgression was associated with higher mean values for plant- and flower-related traits, including vigour prickliness and stigma length. For one flower- and four fruit-related-trait QTLs, including flower peduncle and fruit pedicel lengths and fruit weight, the S. incanum introgression was associated with lower mean values for fruit-related traits. Evidence of synteny to other previously reported in eggplant populations was found for three of the fruit-related QTLs. The other seven stable QTLs are new, demonstrating that eggplant ILs are of great interest for eggplant breeding under different environments.This work was undertaken as part of the initiative "Adapting Agriculture to Climate Change: Collecting, Protecting, and Preparing Crop Wild Relatives", which is supported by the Government of Norway. The project is managed by the Global Crop Diversity Trust with the Millennium Seed Bank of the Royal Botanic Gardens, Kew and implemented in partnership with national and international gene banks and plant breeding institutes around the world. For further information, see the project website: http://www.cwrdiversity.org/.Funding was also received from Spanish Ministerio de Economia, Industria y Competitividad and Fondo Europeo de Desarrollo Regional (grant AGL2015-64755-R from MINECO/FEDER); from Ministerio de Ciencia, Innovacion y Universidades, Agencia Estatal de Investigacion and Fondo Europeo de Desarrollo Regional (grant RTI-2018-094592-B-100 from MCIU/AEI/FEDER, UE); from European Union's Horizon 2020 Research and Innovation Programme under grant agreement No. 677379 (G2P-SOL project: Linking genetic resources, genomes and phenotypes of Solanaceous crops); and from Vicerrectorado de Investigacion, Innovacion y Transferencia de la Universitat Politecnica de Valencia (Ayuda a Primeros Proyectos de Investigacion; PAID-06-18). Giulio Mangino is grateful to Generalitat Valenciana for a predoctoral grant within the Santiago Grisolia programme (GRISOLIAP/2016/012). Pietro Gramazio is grateful to Japan Society for the Promotion of Science for a postdoctoral grant (P19105, FY2019 JSPS Postdoctoral Fellowship for Research in Japan (Standard)).Mangino, G.; Plazas Ávila, MDLO.; Vilanova Navarro, S.; Prohens Tomás, J.; Gramazio, P. (2020). Performance of a Set of Eggplant (Solanum melongena) Lines With Introgressions From Its Wild Relative S. incanum Under Open Field and Screenhouse Conditions and Detection of QTLs. Agronomy. 10(4):1-15. https://doi.org/10.3390/agronomy10040467S115104FAOSTAThttp://www.fao.org/faostat/Gebhardt, C. (2016). The historical role of species from the Solanaceae plant family in genetic research. Theoretical and Applied Genetics, 129(12), 2281-2294. doi:10.1007/s00122-016-2804-1Hirakawa, H., Shirasawa, K., Miyatake, K., Nunome, T., Negoro, S., Ohyama, A., … Fukuoka, H. (2014). Draft Genome Sequence of Eggplant (Solanum melongena L.): the Representative Solanum Species Indigenous to the Old World. DNA Research, 21(6), 649-660. doi:10.1093/dnares/dsu027Barchi, L., Pietrella, M., Venturini, L., Minio, A., Toppino, L., Acquadro, A., … Rotino, G. L. (2019). A chromosome-anchored eggplant genome sequence reveals key events in Solanaceae evolution. Scientific Reports, 9(1). doi:10.1038/s41598-019-47985-wGramazio, P., Yan, H., Hasing, T., Vilanova, S., Prohens, J., & Bombarely, A. (2019). Whole-Genome Resequencing of Seven Eggplant (Solanum melongena) and One Wild Relative (S. incanum) Accessions Provides New Insights and Breeding Tools for Eggplant Enhancement. Frontiers in Plant Science, 10. doi:10.3389/fpls.2019.01220GRAMAZIO, P., PROHENS, J., PLAZAS, M., MANGINO, G., HERRAIZ, F. J., GARCÍA-FORTEA, E., & VILANOVA, S. (2018). Genomic Tools for the Enhancement of Vegetable Crops: A Case in Eggplant. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 46(1), 1-13. doi:10.15835/nbha46110936Frary, A., Frary, A., Daunay, M.-C., Huvenaars, K., Mank, R., & Doğanlar, S. (2014). 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Screening of pepino (Solanum muricatum) and wild relatives against four major tomato diseases threatening its expansion in the Mediterranean region

This is the peer reviewed version of the following article: Pacheco, J, Soler, S, Figàs, MR, San Bautista, A, Prohens, J, Gramazio, P. Screening of pepino (Solanum muricatum) and wild relatives against four major tomato diseases threatening its expansion in the Mediterranean region. Ann Appl Biol. 2021; 179: 288¿ 301, which has been published in final form at https://doi.org/10.1111/aab.12698. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.[EN] The pepino (Solanum muricatum) is an Andean vegetable crop closely related to tomato. In the last decades, it has been introduced in the Mediterranean region and other parts of the world as a potential new crop. However, several tomato major pathogens may threaten the expansion of pepino cultivation. We identified Fusarium oxysporum f. sp. lycopersici (FOL), Verticillium dahliae (VE), pepino mosaic virus (PepMV) and tomato mosaic virus (ToMV) as four of the most likely pathogens to cause damage to pepino crops in Mediterranean climates. In order to evaluate the response of the pepino genepool against these pathogens, as well as to identify sources of tolerance, we inoculated six accessions of cultivated pepino, nine accessions of seven pepino wild relatives, and one interspecific hybrid with FOL, VE, PepMV and ToMV and followed its symptomatology for 30 days (FOL and VE) or 60 days (PepMV and ToMV). ELISA tests were also performed for PepMV and ToMV. Susceptible tomato materials were used as controls. The pepino genepool displayed fewer symptoms than susceptible tomato controls after inoculation with FOL, with most accessions being tolerant or resistant. Regarding VE, a wide variation of values for the symptoms index (SI) was observed, with three cultivated pepino accessions displaying tolerance. For PepMV a wide variation for SI was also observed, with one accession of S. caripense being resistant, and several accessions of pepino and other wild relatives displaying different degrees of tolerance. PepMV absorbance values obtained by ELISA tests followed a pattern similar to that of SI. For ToMV no resistances were found, although two wild accessions and the interspecific hybrid displayed low values for the SI and were considered as moderately tolerant. ELISA tests against ToMV revealed that the virus replicated well in all materials. None of the accessions evaluated displayed resistance or high levels of tolerance to the four pathogens, but some of them were complementary for resistance or high levels of toler- ance. Although the interspecific hybrid tested was not resistant to any of the pathogens, it was tolerant to FOL and PepMV and moderately tolerant to VE and ToMV. A multivariate hierarchical clustering revealed similar patterns among accessions in the response to the two fungal diseases (FOL and VE) on one side and to the two viral ones (PepMV and ToMV) on the other. The information generated in this study has allowed identifying materials within the pepino genepool for the development of multi-resistant pepino cultivars to major diseases threatening its expansion in the Mediterranean region.Japan Society for the Promotion of Science, Grant/Award Numbers: FY2019, P19105; Universitat Politecnica de Valencia, Grant/Award Number: 20200268Pacheco, J.; Soler Aleixandre, S.; Figás-Moreno, MDR.; San Bautista Primo, A.; Prohens Tomás, J.; Gramazio, P. (2021). Screening of pepino (Solanum muricatum) and wild relatives against four major tomato diseases threatening its expansion in the Mediterranean region. Annals of Applied Biology. 179(3):288-301. https://doi.org/10.1111/aab.12698S288301179

Fostering Conservation via an Integrated Use of Conventional Approaches and High-Throughput SPET Genotyping: A Case Study Using the Endangered Canarian EndemicsSolanum lidiiandS. vespertilio (Solanaceae)

[EN] Islands provide unique opportunities to integrated research approaches to study evolution and conservation because boundaries are circumscribed, geological ages are often precise, and many taxa are greatly imperiled. We combined morphological and hybridization studies with high-throughput genotyping platforms to streamline relationships in the endangered monophyletic and highly diverse lineage ofSolanumin the Canarian archipelago, where three endemic taxa are currently recognized. Inter-taxa hybridizations were performed, and morphological expression was assessed with a common-garden approach. Using the eggplant Single Primer Enrichment Technology (SPET) platform with 5,093 probes, 74 individuals of three endemic taxa (Solanum lidii,S. vespertiliosubsp.vespertilio, andS. vespertiliosubsp.doramae) were sampled for SNPs. While morphological and breeding studies showed clear distinctions and some continuous variation, inter-taxon hybrids were fertile and heterotic for vigor traits. SPET genotyping revealed 1,421 high-quality SNPs and supported four, not three, distinct taxonomic entities associated with post-emergence geological, ecological and geographic factors of the islands. Given the lack of barriers to hybridization among all the taxa and their molecular differences, great care must be taken in population management. Conservation strategies must take account of the sexual and breeding systems and genotypic distribution among populations to successfully conserve and restore threatened/endangered island taxa, as exemplified bySolanumon the Canary Islands.This work was supported by funds from project DEMIURGO to the Jardin Botanico Canario "Viera y Clavijo" -Unidad Asociada al CSIC of the Cabildo de Gran Canaria. Consumables and genotyping were funded by the Jardin Botanico Canario "Viera y Clavijo" -Unidad Asociada al CSIC. Participation of authors from Universitat Politecnica de Valencia has been partially funded by the European Union's Horizon 2020 Research and Innovation Programme under the Grant Agreement No. 677379 (G2P-SOL project: Linking genetic resources, genomes, and phenotypes of solanaceous crops). PG was grateful to Universitat Politecnica de Valencia and to Japan Society for the Promotion of Science for their respective Postdoctoral Grants [PAID-10-18 and FY2019-P19105 JSPS Postdoctoral Fellowship for Research in Japan (Standard)].Gramazio, P.; Jaén-Molina, R.; Vilanova Navarro, S.; Prohens Tomás, J.; Marrero, Á.; Caujapé-Castells, J.; Anderson, GJ. (2020). Fostering Conservation via an Integrated Use of Conventional Approaches and High-Throughput SPET Genotyping: A Case Study Using the Endangered Canarian EndemicsSolanum lidiiandS. vespertilio (Solanaceae). Frontiers in Plant Science. 11:1-17. https://doi.org/10.3389/fpls.2020.00757S11711Ablay, G., & Hürlimann, M. (2000). 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