Polarized light microscopy guarantees the use of autochthonous wheat in the production of flour for the Protected Geographical Indication ‘Galician Bread’

‘Galician Bread’ is a traditional baked product and a national benchmark that has recently been granted the European mark of Protected Geographical Indication (PGI), which requires at least 25% of the flour to be produced from autochthonous varieties such as ‘Caaveiro’. The objective of this work was to find a method that guarantees the presence and the percentage of ‘Caaveiro’ wheat in blended flours by using microscopy techniques. Using optical microscopy, including bright-field and polarizing microscopy, autochthonous and foreign flours were analyzed and compared. ‘Caaveiro’ starch presented a different birefringence pattern (associated with a higher amount of amylose) with respect to other cultivars used to produce flours, a feature used to make a computation of the two starch granule types in the mixtures of ‘Caaveiro’ with foreign flours. Repetitions with different mixture percentages allowed us to develop a mathematical model to estimate the percentage of ‘Caaveiro’ flour present in the mixture. Firstly, the most effective method for preparing samples was determined by ensuring the homogeneity of the samples and, subsequently, a validation was carried out with blind samples. Starch birefringence properties allowed the detection of ‘Caaveiro’ wheat flour in mixtures with foreign/Castilian wheat flours and to determine the percentages used in the flour mixtures applying a calibration line (R2 = 0.9577). Deviations were due to the difficulty in obtaining precise mixtures of the blended flours, as happened with other Simple Sequence Repeat (SSR)-based methods used in the same samples. This is a novel method for detecting contraventions/infractions of the percentage of ‘Caaveiro’ used in wheat flours, which is simple, effective and inexpensiveThis research was supported by the Spanish Ministry of Science and Innovation, Proyectos de Generación de Conocimiento 2021-2023 (PID2021-123905OB-I00). We would like to thank Da Cunha Group for the samples and their support of the ‘Cátedra do Pan e do Cereal’. Nerea Fernández-Canto is grateful to Xunta de Galicia for her predoctoral research fellowship (ED481A-2019/263)S

Genetic Diversity of Castanea sativa Mill. Accessions from the Tuscan-Emilian Apennines and Emilia Romagna Region (Italy)

This work investigated the genetic diversity of 134 Castanea sativa Mill. accessions present in the Italian region of Emilia-Romagna. Samples were taken from three collection fields (Granaglione, Zocca and Paloneta) in the Tuscan-Emilian Apennines. The accessions were analyzed by using 16 microsatellite markers (SSR). Genetic distances among accessions, calculated through the DICE coefficient, were used to construct an UPGMA cluster analysis. One major genotype (named “Marroni”) was identified across the three investigated collection fields; this variety corresponds to a sweet chestnut cultivar that has been propagated and widely diffused in the Emilia-Romagna region. Other genotypes were represented by different varieties of Italian chestnuts. The results of this study will be used to define and share guidelines for the characterization and varietal certification of the chestnut varieties in the Emilia-Romagna regionField and laboratory work were developed as the contribution of the regional project PSR ‘BIODIVERSAMENTE CASTAGNO’ and of the National Academy of Agriculture (ANA), in particular for supporting the analyses on the Granaglione’s samples. The Parco Didattico Sperimentale del Castagno di Granaglione and the Collection of Zocca host the main varieties of the Tuscan-Emilian Apennines. Sara Alessandri’s fellowship was funded by the Agricultural, Environmental, and Food Science and Technology (STAAA) PhD program offered by the Department of Agriculture and Food Sciences (DISTAL, University of Bologna)S

Traceability of the local cultivar ‘Caaveiro’ in flour mixtures used to produce Galician bread by simple sequence repeats and droplet digital polymerase chain reaction technology

The analysis of simple sequence repeats (SSRs) and of bulk ground samples by droplet digital polymerase chain reaction (ddPCR) was investigated as an alternative to individual kernel testing for assessing the presence of local cultivars in common wheat (Triticum aestivum L.) varietal blends. The recent Protected Geographical Indication (PGI) of ‘Pan Galego’ (Galician bread) requires that the flour comprise a minimum 25% local wheat cultivars. As a test for compliance with this minimum level, wheat flours were prepared by mixing commercial flours with 0%, 5%, 20%, 25% and 30% ‘Caaveiro’ and 100% ‘Caaveiro’ and 100% commercial flours were used as controls. A second analysis was performed with a second set of wheat flours with 5% and 25% ‘Caaveiro’. These were mixed with two different commercial flours to assess the potential ability of five SSRs to identify the percentage of ‘Caaveiro’, constituting the first reference of the use of SSRs in the traceability of specific autochthonous cultivars in flour blends. ddPCR using the QX200 system platform was used to the targeted proportions across the simulated range with two out of five SSRs, indicating that they can be used in the traceability of ‘Caaveiro’ in mixed flours and breadsS

Genetic diversity of the Spanish apple genetic resources using SSRs

The Spanish Program of Plant Genetic Resources integrates, among others, the collections located at Public University of Navarre, Centro de Investigaciones Agrarias de Mabegondo, Cabildos (Tenerife, La Palma and Gran Canaria), University of Lleida, Estación Experimental de Aula Dei-CSIC and CITA of Aragon. Those collections include mainly local cultivars from their respective regions, covering most of the Spanish apple-growing areas. Though some previous studies about the genetic variability of apple genetics resources from Spain were already performed, a complete analysis is needed in order to evaluate the complete diversity of Malus spp. in Spain. For doing that, the Spanish Government funded the project ¿Harmonization of the methodology of characterization, assessment of genetic diversity and definition of the core collection of the apple germplasm conserved in Spanish genebanks¿. In total, we have evaluated 1206 accessions using standardized methodologies, with SSR markers and morphological descriptors. SSR fingerprinting was performed with 13 SSR markers. SSR profiles were obtained independently and allele sizes were compared using a common set of cultivars selected as references. Results showed 601 genotypes for 1206 accessions. Most of the genotypes (438) were identified only in one accession. The other 163 genotypes were repeated in two to 81 accessions (involving 767 accessions in total). The harmonization of morphological descriptors will allow us to determine if the accessions with the same genotype are synonymies or closely related individuals. Results of this study highlight the interest of coordinated actions in order to optimize the management of germplasm collections and to evaluate the complete genetic diversity of Malus spp. in Spain.Peer Reviewe

Definición del núcleo optimizado de la colección de conservación del manzano español

5 Pags.- 1 Tabl.Este trabajo tiene como objetivo determinar la estrategia más adecuada para la selección del conjunto mínimo de accesiones (núcleo optimizado) que represente eficientemente la variación genética del manzano conservado en las colecciones españolas. Este núcleo optimizado mediante criterios genéticos constituirá la base de la colección nuclear de conservación, que podrá ser complementado con accesiones seleccionadas por otros criterios (morfo-fisiológicos, agronómicos, valor histórico, etc.). Se ha evaluado la eficiencia de estrategias de selección por búsqueda local estocástica avanzada (ASLS) que diferían tanto por el tamaño final del núcleo como por la combinación (y peso relativo) de las medidas de distancia genética y riqueza alélica a optimizar. Las estrategias empleadas han proporcionado núcleos optimizados con grandes diferencias en la diversidad conservada, así como en el nivel de representación de la estructura genética general. Teniendo en cuenta el uso principal de la colección nuclear, la estrategia que ofrece un mejor equilibrio entre representatividad y adecuación al uso es la que combina la optimización de la distancia media entre cada accesión de la colección y la entrada en el núcleo más cercana con el índice de Shannon y la recuperación de alelos.Este trabajo ha sido financiado por los proyectos INIA RF2011-00017-C05-00 y RTA2015-00052-C02-00Peer reviewe

Genetic diversity of the Spanish apple genetic resources using SSRs: abstract

PublishedPoster number 6

In vivo multimodal imaging of adenosine A1 receptors in neuroinflammation after experimental stroke

Adenosine A(l) receptors (A(l)ARs) are promising imaging biomarkers and targets for the treatment of stroke. Nevertheless, the role of A(l)ARs on ischemic damage and its subsequent neuroinflammatory response has been scarcely explored so far. Methods: In this study, the expression of A(1)ARs after transient middle cerebral artery occlusion (MCAO) was evaluated by positron emission tomography (PET) with [F-18]CPFPX and immunohistochemistry (IHC). In addition, the role of AIARs on stroke inflammation using pharmacological modulation was assessed with magnetic resonance imaging (MRI), PET imaging with [F-18]DPA-714 (TSPO) and [F-18]FLT (cellular proliferation), as well as IHC and neurofunctional studies. Results: In the ischemic territory, [F-18]CPFPX signal and IHC showed the overexpression of A(l)ARs in microglia and infiltrated leukocytes after cerebral ischemia. Ischemic rats treated with the AAR agonist ENBA showed a significant decrease in both [F-18]DPA-714 and [F-18]FLT signal intensities at day 7 after cerebral ischemia, a feature that was confirmed by IHC results. Besides, the activation of A(l)AR promoted the reduction of the brain lesion, as measured with T2W-MRI, and the improvement of neurological outcome including motor, sensory and reflex responses. These results show for the first time the in vivo PET imaging of A(l)AR expression after cerebral ischemia in rats and the application of [F-18]FLT to evaluate glial proliferation in response to treatment. Conclusion: Notably, these data provide evidence for A(l)AR playing a key role in the control of both the activation of resident glia and the de novo proliferation of microglia and macrophages after experimental stroke in rats.The authors would like to thank A. Leukona and V. Salinas for technical support in the radiosynthesis. This study was funded by grants from the Spanish Ministry of Education and Science/FEDER RYC-201722412, SAF2016-75292-R, SAF2017-87670-R and PID2019-107989RB-I00, the Basque Government (IT1203/19, BIO18/IC/006) and CIBERNED. Maria Ardaya holds a fellowship from the University of Pais Vasco. Ana Joya acknowledges funding from Fundacio La Marato de TV3 (17/C/2017). Juan Jose Gutierrez acknowledges funding from Euskampus Fundazioa. Jordi Llop also acknowledges The Spanish Ministry of Economy and Competitiveness (Grant CTQ2017-87637-R). Part of the work has been performed under the Maria de Maeztu Units of Excellence Program from the Spanish State Research Agency (Grant No. MDM-2017-0720)

Development of a standardized methodology for phenotypical characterizations in apple

4 Pags.- 2 Tabls. Articles derived from XIV EUCARPIA Symposium on Fruit Breeding and Genetics (Bologna,Italy. June 14-18 2015) . The definitive version is available at: http://www.actahort.org/index.htmThe description of phenotypic traits in apple cultivars is generally performed using internationally agreed descriptors such as UPOV guidelines, which defines for each trait several states of expression. However, it is not always possible to classify a cultivar unambiguously using those guidelines, because in practice the states are not clearly enough defined or the example cultivars are not always available in the collections. This work presents the results of a harmonization project performed by the teams responsible of the main apple germplasm collections in Spain. The objective was to develop a standardized method for the 57 traits included in the TG/14/9 UPOV guidelines for apple characterization, defining their states of expression in a clear and unambiguous way for Spanish germplasm. Phenotypic data collected for more than 1,600 accessions from Spanish collections were used and the method to define each state depended on the type of expression. For quantitative traits the number of states and their limits were defined according to the variability that exists within and between accessions. For qualitative traits, high-resolution images clearly depicting each state were selected. A standardized characterization protocol for the 57 traits of apple germplasm has been provided, enabling to comparing properly the phenotypes of Spanish genetic resources.This Project has been funded by the Spanish Ministry of Science and Innovation/National Institute for Agricultural and Food Research and Technology (RF2011- 00017-C05-00).Peer reviewe

Development of a standardized methodology for phenotypical characterizations in apple

1 pag.This work presents the results of a harmonization project performed by the teams responsible of the main apple germplasm collections in Spain. The objective was to develop a standardized method for the 57 traits included in the TG/14/9 UPOV guidelines for apple characterization, defining their states of expression in a clear and unambiguous way for Spanish germplasm.Peer reviewe