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Here we present the approach used to develop the INCREASE "Intelligent Chickpea" Collections, from analysis of the information on the life history and population structure of chickpea germplasm, the availability of genomic and genetic resources, the identification of key phenotypic traits and methodologies to characterize chickpea. We present two phenotypic protocols within H2O20 Project INCREASE to characterize, develop, and maintain chickpea single-seed-descent (SSD) line collections. Such protocols and related genetic resource data from the project will be available for the legume community to apply the standardized approaches to develop Chickpea Intelligent Collections further or for multiplication/seed-increase purposes. © 2022 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Characterization of chickpea seeds for seed-trait descriptors Basic Protocol 2: Characterization of chickpea lines for plant-trait descriptors specific for primary seed increase.

RESUMEN La dirofilariosis cardiaca es una enfermedad de distribución mundial, causada por el nematodo Dirofilaria immitis y transmitida por mosquitos culícidos, siendo los caninos los hospederos principales. Las pruebas serológicas proporcionan cierta ayuda diagnóstica, pero no son concluyentes. En el presente reporte se describe un caso clínico de un canino en el cual las pruebas serológicas para detección de filarias y las pruebas para detección de microfilaremias no fueron concluyentes. Sin embargo, el uso de protocolos estandarizados empleando el ultrasonido como parte del examen físico, evidenció la presencia de filarias cardiacas. La importancia de reportar este caso es presentar un protocolo clínico que incluye el ultrasonido cardíaco para una aproximación diagnóstica certera. ABSTRACT Heartworm disease is a parasitic infection with worldwide distribution, caused by the Dirofilaria immitis nematode and transmitted by culicid mosquitoes, with canines being the main hosts. Serologic tests provide some diagnostic help but are not conclusive. This report describes a clinical case of a canine in which the serological tests for filarial detection and the tests for the detection of microfilaraemia were inconclusive. However, the use of standardized protocols using ultrasound as part of the physical examination, evidenced the presence of cardiac filarials. The importance of reporting this case is to present a clinical protocol that includes cardiac ultrasound for an accurate diagnostic approach.

16 Pág. Centro de Recursos Fitogenéticos y Agricultura Sostenible (CRF), Common vetch (Vicia sativa L.) is a legume used for animal feed because of its high protein content and great capacity for nitrogen fixation, making this crop relevant in sustainable agriculture. The Spanish vetch collection, conserved at the Spanish Plant Genetic Resources Center (CRF), is one of the largest collections of this species worldwide, including landraces, wild relatives mainly collected in Spain, and commercial cultivars, but also accessions of international origin. The analysis of the genetic diversity of this material, whose genome has not been sequenced yet, and the assembly of a representative collection could play a pivotal role in conserving and exploiting these genetic resources in breeding programs mainly in those focused on consequences and demands of climate change. In this work, a set of 14 simple sequence repeat (SSR) reference alleles for genetic diversity analysis of the CRF vetch collection has been developed, used for genotyping more than 545 common vetch accessions from all over the world and validated. All the tested markers were polymorphic for the analyzed accessions. Overall, at least 86 different loci were identified with 2-11 alleles per locus with an average of 6.1 alleles per locus. Also, the analyses of the generated SSR database support that most of these SSR markers are transferable across closely related species of Vicia genus. Analysis of molecular variance revealed that wild relatives have a higher genetic diversity than landraces. However, cultivars have similar diversity than landraces, indicating that genetic variability has been barely lost due to the breeding of this legume. Low differences of genetic variations between Spanish and non-Spanish accessions have been observed, suggesting a high degree of diversity within Spanish genotypes, which provide 95% of the total genetic variation, so we have focused our efforts on characterizing genotypes of Spanish origin that were further studied using storage protein profiles. Based on SSR, seed protein profiles, and agromorphological and passport data, a vetch core collection (VCC) containing 47 V. sativa accessions of Spanish origin has been established. In this collection, the characterization has been expanded using ISSR markers, and it has been reevaluated with new agromorphological data, including drought tolerance characters. This VCC presents a minimum loss of genetic diversity concerning the total collection and constitutes an invaluable material that can be used in future breeding programs for direct use in a resilient agricultural system., This work was funded by the Spanish Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA) (Project RF2007-00005-00-00 and Project AT2016-009), by the Spanish Government MICIU (RTI2018-094037-R-I00), and by the Grant Severo Ochoa Program for Center of Excellence in R&D (CBGP)

International audience; Food legumes are crucial for all agriculture-related societal challenges, including climate change mitigation, agrobiodiversity conservation, sustainable agriculture, food security and human health. The transition to plant-based diets, largely based on food legumes, could present major opportunities for adaptation and mitigation, generating significant co-benefits for human health. The characterization, maintenance and exploitation of food-legume genetic resources, to date largely unexploited, form the core development of both sustainable agriculture and a healthy food system. INCREASE will implement, on chickpea (Cicer arietinum), common bean (Phaseolus vulgaris), lentil (Lens culinaris) and lupin (Lupinus albus and L. mutabilis), a new approach to conserve, manage and characterize genetic resources. Intelligent Collections, consisting of nested core collections composed of single-seed descent-purified accessions (i.e., inbred lines), will be developed, exploiting germplasm available both from genebanks and on-farm and subjected to different levels of genotypic and phenotypic characterization. Phenotyping and gene discovery activities will meet, via a participatory approach, the needs of various actors, including breeders, scientists, farmers and agri-food and non-food industries, exploiting also the power of massive metabolomics and transcriptomics and of artificial intelligence and smart tools. Moreover, INCREASE will test, with a citizen science experiment, an innovative system of conservation and use of genetic resources based on a decentralized approach for data management and dynamic conservation. By promoting the use of food legumes, improving their quality, adaptation and yield and boosting the competitiveness of the agriculture and food sector, the INCREASE strategy will have a major impact on economy and society and represents a case study of integrative and participatory approaches towards conservation and exploitation of crop genetic resources.

Crop wild relatives (CWR) have recently received significant attention due to their value as plant genetic resources and their contribution to world food security. We present a prioritized checklist of CWR in Spain in which the criteria of crossability with crops of economic importance, endemicity and threat status have been taken into account. First, we selected a list of genera corresponding to the most relevant crops for Spain and at the international level. These crops were classified into use categories (Food, Forage & Fodder, Ornamental, and Industrial & Other uses) depending on their main use. The wild plant species native to Spain belonging to these genera were then listed. After evaluation by national experts in plant breeding, the resulting checklist contained 929 species. Further selection based on crossability, endemicity and threat status led to the generation of the Prioritized Spanish Checklist of crop wild relatives containing 578 species. Thirty-two percent of these species belong to the Forage & Fodder use category, 28% to the Ornamental category, 24% to the Food category and 16% to the Industrial & Other uses category. Thirty-five percent of the prioritized species are endemic to Spain, and over one-fourth are classified under some category of threat according to the International Union for Conservation of Nature. Endemicity and threat status rates in the Prioritized Spanish Checklist of CWR were higher than those found in the prioritized CWR inventories of other countries. A ex situ assessment reporting number of accessions showed that 70% of the prioritized Spanish CWR have accessions preserved in genebanks.

Background Common vetch (Vicia sativa L.) is a forage grain legume of high protein content and high nitrogen fixation, relevant in sustainable agriculture systems. Drought is the main limiting factor of this crop yield. Genetic resources collections are essential to provide genetic variability for breeding. The analysis of drought associated parameters has allowed us to identify drought tolerant and sensitive ecotypes in a vetch core collection. Results To understand the mechanisms involved in drought response we analysed transcriptomic differences between tolerant and sensitive accessions. Polymorphic variants (SNPs and SSRs) in these differential expressed genes (DEGs) have also been analysed for the design of drought-associated markers. A total of 1332 transcripts were commonly deregulated in both genotypes under drought. To know the drought adaptive response, we also analysed DEGs between accessions. A total of 2646 transcripts are DEG between sensitive and tolerant ecotypes, in watered and drought conditions, including important genes involved in redox homeostasis, cell wall modifications and stress-response. The integration of this functional and genetic information will contribute to understand the molecular mechanisms of drought response and the adaptive mechanisms of drought tolerance in common vetch. The identification of polymorphic variants in these DEGs has also been screened for the design of drought-associated markers that could be used in future breeding program strategies. Conclusions Our studies shed light for the first time in common vetch about the genes and pathways associated with drought tolerance. In addition, we identify over 100 potential drought associated polymorphism, as SNPs or SSRs, which are differently present in drought and tolerant genotypes. The use of these molecular markers for trait prediction would enable the development of genomic tools for future engineering strategies by screening of germplasm crop collections for traits related with crop drought resilience, adaptability or yield in vetch.

One hundred and one Spanish landraces of lentil maintained in the Plant Genetic Resources Centre (CRF) and coming from different micro-climatic areas were evaluated for 16 morphological characters. Origin zones were classified following Papadakis agro-climatic parameters and the relationships between climatic data zones and morphological traits were analysed. The results indicated that a great part of the landraces came from temperate areas with dry Mediterranean moisture regime. Most of the varieties had plant pigmentation, slight leaf pubescence, white flowers with blue or violet veins, green testa without pattern and yellow cotyledons. Two clear morphological groups based on seed characters could be established coinciding with Barulina's description for the macrosperma and microspermatypes. Other traits of high agronomic importance as phenology and plant height showed less usefulness in discriminating accessions. Characters related to seed and productivity had most correlation with climate of the origin sites, while phenology and plant height did not show significant correlations. The climatic variables related with extreme temperatures were more effective than average temperatures or rainfall to explain the morphological variability. The agro-climatic patterns detected seems to be due to the geographical location of the two morphological lentil groups in zones with different climatic conditions, indicating that lentil landraces must be migrated following criteria related with seed characters preferred by farmers.

In this article, the importance and situation of the conservation of plant genetic resources for food and agriculture is reviewed, taking into consideration not only the aspects specifically related to the conservation activity but also their role as components of biodiversity and providers of ecosystem services. Special attention is drawn to ex situ collections as providers of germplasm for direct use and for plant breeding, two elements that make an essential contribution to a greater resilience of agriculture to environmental changes. In particular, the state of conservation at national and global levels of genetic resources of Fabaceae species cultivated for human food and animal feed, is examined., En el artículo se revisa la importancia y la situación de la conservación de recursos fitogenéticos para la alimentación y la agricultura, tanto en los aspectos asociados específicamente a esta actividad como en aquellos otros derivados de su faceta como componente de la biodiversidad y proveedor de servicios ecosistémicos. Se incide en la función de las colecciones ex situ como fuente de materiales tanto para su utilización directa como en programas de mejora genética, ambos elementos imprescindibles para una agricultura resiliente frente al cambio ambiental. De forma específica, se describe el estado de la conservación de germoplasma de las especies de la familia de Fabaceae tanto para utilización en alimentación humana como en alimentación animal, en el ámbito nacional e internacional.

This study evaluated the phenotypic representativeness of core collections generated independently with ecogeographical data or with morphological data. The Spanish Phaseolus vulgaris L. collection was the target germplasm collection, and one map of ecogeographical categories and 15 morphological variables were used to create core collections. Three ecogeographical and 12 morphological core collections were produced combining different grouping methods (categories from ecogeographical land characterization map [CEM], two-step clustering [TSC], Ward-modified location model clustering [WM], and unweighted pair-group method using arithmetic average [UPGMA]) and allocation strategies (constant [C], proportional [P], logarithmic [L], and Gower's distance [D]). An additional morphological core collection was created using a modified least distance stepwise sampling (LDSSm) method. Core collections were evaluated with morphological data using parameters for qualitative and quantitative variables that were summarized in a new synthetic evaluation parameter (SEP). Synthetic evaluation parameter results showed that the ecogeographical core collections had high phenotypic representativeness, similar to that of morphological core collections generated by the combination of WM and TSC grouping methods and P allocation strategy. A graphical evaluation of core collections supported these results. In conclusion, ecogeographical information was confirmed as a good alternative or complement to other types of data to create reliable and representative core collections. © Crop Science Society of America.

The characterization of plant genetic resources is the first step towards improving their use. The Spanish Plant Genetic Resources Centre, which belongs to the National Institute for Agricultural and Food Technology Research (CRF-INIA), conserves accessions of wild underground vetches collected in Spain. In the present work, 26 underground vetch accessions were characterized in terms of their seed storage proteins (separated by SDS-PAGE) as a means of assessing the genetic variation of these plants and their agronomic value. Vicia sativa cv. Vereda was used as control. A total of 54 bands were detected, of which 49 were polymorphic. Fifty eight different electrophoretic patterns were observed in total. Protein bands were scored in terms of their presence (1) or absence (0) for all the seeds studied, and two matrices constructed, one with all the bands present in each accession, the other with the different patterns for each accession. Dendrograms based on the Jaccard similarity index and the UPGMA clustering method were produced from these matrices, and the degree of genetic variation between and within accessions was calculated. The groups obtained were compared with the chromosome number for each accession. The results reflect the great diversity of underground vetch seed storage proteins. The aerial and underground seeds of 16 accessions were then analysed separately. In some cases, the aerial and underground flowers of the same plant produced different proteins. © 2009 Springer Science+Business Media B.V.