Transcriptome Analysis of Stigmas of Vicia faba L. Flowers.

Pollination in angiosperms depends on complex communication between pollen grains and stigmas, classified as wet or dry, depending on the presence or absence of secretions at the stigma surface, respectively. In species with wet stigma, the cuticle is disrupted and the presence of exudates is indicative of their receptivity. Most stigma studies are focused on a few species and families, many of them with self-incompatibility systems. However, there is scarce knowledge about the stigma composition in Fabaceae, the third angiosperm family, whose stigmas have been classified as semidry. Here we report the first transcriptome profiling and DEGs of Vicia faba L. styles and stigmas from autofertile (flowers able to self-fertilize in the absence of manipulation, whose exudate is released spontaneously) and autosterile (flowers that need to be manipulated to break the cuticle and release the exudates to be receptive) inbred lines. From the 76,269 contigs obtained from the de novo assembly, only 45.1% of the sequences were annotated with at least one GO term. A total of 115,920, 75,489, and 70,801 annotations were assigned to Biological Process (BP), Cellular Component (CC), and Molecular Function (MF) categories, respectively, and 5918 differentially expressed genes (DEGs) were identified between the autofertile and the autosterile lines. Among the most enriched metabolic pathways in the DEGs subset were those related with amino acid biosynthesis, terpenoid metabolism, or signal transduction. Some DEGs have been related with previous QTLs identified for autofertility traits, and their putative functions are discussed. The results derived from this work provide an important transcriptomic reference for style-stigma processes to aid our understanding of the molecular mechanisms involved in faba bean fertilization.

Genome-wide association study for yield-related traits in faba bean (Vicia faba L.).

Yield is the most complex trait to improve crop production, and identifying the genetic determinants for high yield is a major issue in breeding new varieties. In faba bean (Vicia faba L.), quantitative trait loci (QTLs) have previously been detected in studies of biparental mapping populations, but the genes controlling the main trait components remain largely unknown. In this study, we investigated for the first time the genetic control of six faba bean yield-related traits: shattering (SH), pods per plant (PP), seeds per pod (SP), seeds per plant (SPL), 100-seed weight (HSW), and plot yield (PY), using a genome-wide association study (GWAS) on a worldwide collection of 352 homozygous faba bean accessions with the aim of identifying markers associated with them. Phenotyping was carried out in field trials at three locations (Spain, United Kingdom, and Serbia) over 2 years. The faba bean panel was genotyped with the Affymetrix faba bean SNP-chip yielding 22,867 SNP markers. The GWAS analysis identified 112 marker-trait associations (MTAs) in 97 candidate genes, distributed over the six faba bean chromosomes. Eight MTAs were detected in at least two environments, and five were associated with multiple traits. The next step will be to validate these candidates in different genetic backgrounds to provide resources for marker-assisted breeding of faba bean yield.

A Clear Cell Sarcoma Case: A Diagnostic and Treatment Challenge, with a Promising Response to Trabectedin.

Introduction: Clear cell sarcoma (CCS) is a rare and aggressive soft tissue sarcoma. CCS is characterized by the translocation t(12;22) (q13;q12), involving the fusion of EWSR1 and ATF1 genes, and less frequently the fusion gene EWSR1-CREB1. Usually, CCSs are considered poorly responsive to conventional chemotherapy. However, trabectedin has shown activity against translocation-related sarcomas. Furthermore, preclinical results suggest that trabectedin is a promising antitumor agent for CCS, potentially inducing melanocytic differentiation. Case Presentation: We report the case of a challenging anatomopathological diagnosis in a patient with an aggressive metastatic CCS. Following the diagnosis of CCS, the patient experienced a clinical and radiological tumor response to trabectedin after four lines of treatment. Conclusion: This is a novel report of CCS treated with trabectedin that resulted in a partial response and suggests the need for further research on trabectedin as a therapeutic option for CCS.

Variability in drought stress response in a panel of 100 faba bean genotypes.

Faba bean is an important protein crop for food and feed worldwide and provides a range of advantages in crop rotations. Its limited use in modern agriculture is mainly due to the high fluctuations in yield. A well known limiting factor in most legumes, and particularly in faba bean, is the high sensitivity to water shortage, which is further aggravated by climate change. The present study was undertaken to exploit the genetic variation in drought stress response in a faba bean collection of 100 accessions with diverse origins and to assess selection criteria for identifying drought tolerant genotypes. Physiological, phenological and yield related traits evaluated under drought or water-sufficient conditions responded significantly to the end-terminated drought stress. Comparison of yield relations showed the advantage of using a stress tolerance index (STI) to identify genotypes combining high yield potential with high stress yield. With regard to physiological traits, SPAD (chlorophyll content) values were significantly related to yield as well as to STI, while the other traits also contributed to different extents to variation in yield formation. Among the yield related traits, seeds per plant proved to be the most important trait followed by pods per plant. Interestingly, the eight genotypes with the best STI performance use different strategies to cope with drought stress.

Genetic analysis of global faba bean diversity, agronomic traits and selection signatures.

KEY MESSAGE: We identified marker-trait associations for key faba bean agronomic traits and genomic signatures of selection within a global germplasm collection. Faba bean (Vicia faba L.) is a high-protein grain legume crop with great potential for sustainable protein production. However, little is known about the genetics underlying trait diversity. In this study, we used 21,345 high-quality SNP markers to genetically characterize 2678 faba bean genotypes. We performed genome-wide association studies of key agronomic traits using a seven-parent-MAGIC population and detected 238 significant marker-trait associations linked to 12 traits of agronomic importance. Sixty-five of these were stable across multiple environments. Using a non-redundant diversity panel of 685 accessions from 52 countries, we identified three subpopulations differentiated by geographical origin and 33 genomic regions subjected to strong diversifying selection between subpopulations. We found that SNP markers associated with the differentiation of northern and southern accessions explained a significant proportion of agronomic trait variance in the seven-parent-MAGIC population, suggesting that some of these traits were targets of selection during breeding. Our findings point to genomic regions associated with important agronomic traits and selection, facilitating faba bean genomics-based breeding.

Genome-wide association analysis for drought tolerance and associated traits in faba bean (Vicia faba L.).

Faba bean (Vicia faba L.) is an important high protein legume adapted to diverse climatic conditions with multiple benefits for the overall sustainability of the cropping systems. Plant-based protein demand is being expanded and faba bean is a good candidate to cover this need. However, the crop is very sensitive to abiotic stresses, especially drought, which severely affects faba bean yield and development worldwide. Therefore, identifying genes associated with drought stress tolerance is a major challenge in faba bean breeding. Although the faba bean response to drought stress has been widely studied, the molecular approaches to improve drought tolerance in this crop are still limited. Here we built on recent genomic advances such as the development of the first high-density SNP genotyping array, to conduct a genome-wide association study (GWAS) using thousands of genetic polymorphisms throughout the entire faba bean genome. A worldwide collection of 100 faba bean accessions was grown under control and drought conditions and 10 morphological, phenological and physiological traits were evaluated to identify single nucleotide polymorphism (SNP) markers associated with drought tolerance. We identified 29 SNP markers significantly correlated with these traits under drought stress conditions. The flanking sequences were blasted to the Medicago truncatula reference genomes in order to annotate potential candidate genes underlying the causal variants. Three of the SNPs for chlorophyll content after the stress, correspond to uncharacterized proteins indicating the presence of novel genes associated with drought tolerance in faba bean. The significance of stress-inducible signal transducers provides valuable information on the possible mechanisms underlying the faba bean response to drought stress, thus providing a foundation for future marker-assisted breeding in the crop.

Antioxidant and Chemopreventive Activity of Protein Hydrolysates from Raw and Germinated Flour of Legumes with Commercial Interest in Colorectal Cancer.

Legumes are a highly nutritious source of plant protein, fiber, minerals and vitamins. However, they also contain several bioactive compounds with significant potential benefits for human health. The objectives of this study were to evaluate the antioxidant, antitumor and chemopreventive activity of functional extracts from legumes using raw and germinated flours of six legume species of commercial interest. The methodology carried out consisted on the development of protein hydrolysates, assessment of their antioxidant capacity and in vitro tests on T84, HCT15 and SW480 colorectal cancer (CRC) cell lines. Our results showed a high antitumor activity of protein hydrolysate from M. sativa. Likewise, when combined with 5-Fluorouracile (5-Fu), there was a synergistic effect using extract concentrations from 50 to 175 µg/mL and 5-Fu concentrations from 1.5 to 5 µM. Similarly, the induction effect on detoxifying enzymes by the extracts of M. sativa, germinated V. faba Baraca × LVzt1 and V. narbonensis, which produced a higher induction rate than the positive control sulforaphane (10 µM), should be highlighted. Therefore, incorporating these enzymes into the diet could provide nutritional effects, as well as play an effective role in cancer chemoprevention and therapy.

Identification of Leukemia-Associated Immunophenotypes by Databaseguided Flow Cytometry Provides a Highly Sensitive and Reproducible Strategy for the Study of Measurable Residual Disease in Acute Myeloblastic Leukemia.

Background: Multiparametric Flow Cytometry (MFC) is an essential tool to study the involved cell lineages, the aberrant differentiation/maturation patterns and the expression of aberrant antigens in acute myeloid leukemia (AML). The characterization of leukemia-associated immunophenotypes (LAIPs) at the moment of diagnosis is critical to establish reproducible strategies for the study of measurable residual disease using MFC (MFC-MRD). Methods: In this study, we identify and characterize LAIPs by comparing the leukemic populations of 145 AML patients, using the EuroFlow AML/ MDS MFC panel, with six databases of normal myeloid progenitors (MPCs). Principal component analysis was used to identify and characterize the LAIPs, which were then used to generate individual profiles for MFC-MRD monitoring. Furthermore, we investigated the relationship between the expression patterns of LAIPs and the different subtypes of AML. The MFC-MRD study was performed by identifying residual AML populations that matched with the LAIPs at diagnosis. To further validate this approach, the presence of MRD was also assessed by qPCR (qPCR-MRD). Finally, we studied the association between MFC-MRD and progression-free survival (PFS). Results: The strategy used in this study allowed us to describe more than 300 different LAIPs and facilitated the association of specific phenotypes with certain subtypes of AML. The MFC-MRD monitoring based on LAIPs with good/strong specificity was applicable to virtually all patients and showed a good correlation with qPCR-MRD and PFS. Conclusions: The described methodology provides an objective method to identify and characterize LAIPs. Furthermore, it provides a theoretical basis to develop highly sensitive MFC-MRD strategies.

Asociación entre estado nutricional y la cantidad de dientes permanentes en niños escolares en Costa Rica / Associação entre o estado nutricional e o número de dentes permanentes em crianças em idade escolar na Costa Rica / Association between nutritional status and the number of permanent teeth in a school child in Costa Rica

Resumen: El sobrepeso y la obesidad infantil se han convertido en uno de los desafíos de la salud pública a nivel mundial. Estas condiciones pueden afectar varios sistemas, asociándose con un inicio temprano de la pubertad y erupción dental. Objetivo: evaluar si existe una asociación entre el estado nutricional y la cantidad de dientes permanentes erupcionados en una población de niños escolares en Costa Rica. Material y métodos: Estudio transversal con una muestra de 753 niños entre los 6 y 12 años. Se realizó un examen clínico donde se registró la presencia o ausencia de dientes permanentes y se tomaron medidas de peso y talla. El estado antropométrico fue evaluado según los criterios de la Organización Mundial de la Salud. Se efectuó análisis descriptivo para todas las variables, comparación de medias entre los grupos (sobrepeso/obesidad vs peso normal) con la prueba U de Mann-Whitney. Se utilizó un análisis de regresión lineal con la edad y el sexo como covariantes. Resultados: Un 69.20% niños presentaba un peso normal y 30.80 % sobrepeso/obesidad. El promedio de dientes erupcionados era mayor en los niños con sobrepreso/obesidad que en aquellos con un peso normal (p=0.001). Al realizar la comparación según el sexo, la diferencia se mantiene (hombres, p=0.001; mujeres, p=0.018). El análisis de regresión lineal demostró que el estado nutricional (p=0.001) y la edad (p=0.001) están asociadas con el número de dientes permanentes erupcionados. Conclusión: los niños en edad escolar con sobrepeso/obesidad presentan mayor cantidad de dientes erupcionados que sus pares con un peso saludable.

Resumo: O sobrepeso e a obesidade na infância tornaram-se um dos desafios da saúde pública no mundo. Essas condições podem afetar vários sistemas, estando associadas ao início precoce da puberdade e a erupção dentária. Objetivo: Avaliar se há associação entre o estado nutricional e o número de dentes permanentes erupcionados em uma população de escolares da Costa Rica. Materiais e método: Estudo transversal com amostra de 753 crianças entre 6 e 12 anos. Foi realizado un exame clínico onde foi registrada a presença ou ausência dos dentes permanentes e as medidas do peso e altura. O estado antropométrico foi avaliado de acordo com os critérios da Organização Mundial da Saúde. Fio realizada una análise descritiva para todas as variáveis, comparação das médias entre os grupos (sobrepeso/obesidade vs peso normal) com o teste U de Mann-Whitney. Uma análise de regressão linear foi utilizada como a ideade e o sexo como covariáveis. Resultados: 69,20% das crianças apresentavam peso normal e 30,80% sobrepeso/obesidade. O número médio de dentes erupcionados foi maior em crianças com sobrepeso/obesidades (p=0.001). Ao fazer a comparação pelo sexo, a diferença permanece (homens, p=0.001; mulheres, p=0.018). A análise de regressão linear demostrou que o estado nutricional (p=0.001) e a idade (p=0.001) estão associados ao número de dentes permanentes erupcionados. Conclusão: Crianças em idade escolar com sobrepeso/obesas tam mais dentes erupcionados do que seus pares com peso saudável.

Abstract: Childhood overweight and obesity have become one of the public health challenges worldwide. These conditions can affect various systems, they are associated with an early onset of puberty and tooth eruption. Objective: to evaluate whether there is an association between nutritional status and the number of permanent teeth erupted in a population of school children in Costa Rica. Materials and methods: Cross-sectional study with a sample of 753 children between 6 and 12 years old. A clinical examination was performed the presence or absence of permanent teeth weight and height measurements were recorded. The anthropometric status was evaluated according to the criteria of the World Health Organization. Descriptive analysis was carried out for all variables, comparison of means between groups (overweight/obesity versus normal weight) with the Mann-Whitney U test. A linear regression analysis was used with age and sex as covariates. Results: 69,20% children had a normal weight and 30,80% were overweight/obese. The average number or erupted teeth as higher in children with overweight/obesity than in those with normal weight (p=0.001). This difference remains in the analysis y sex (men, p=0.001; women, p=0.018). Linear regression analysis demonstrated that nutritional status (p=0.001) and age (p=0.001) are associated with the number of permanent teeth erupted. Conclusion: Overweight/obese school-age children have more erupted teeth than their healthy-weight peers.

QTL dissection and mining of candidate genes for Ascochyta fabae and Orobanche crenata resistance in faba bean (Vicia faba L.).

BACKGROUND: Ascochyta blight caused by Ascochyta fabae Speg. and broomrape (Orobanche crenata) are among the economically most significant pathogens of faba bean. Several QTLs conferring resistance against the two pathogens have been identified and validated in different genetic backgrounds. The aim of this study was to saturate the most stable QTLs for ascochyta and broomrape resistance in two Recombinant Inbred Line (RIL) populations, 29H x Vf136 and Vf6 x Vf136, to identify candidate genes conferring resistance against these two pathogens. RESULTS: We exploited the synteny between faba bean and the model species Medicago truncatula by selecting a set of 219 genes encoding putative WRKY transcription factors and defense related proteins falling within the target QTL intervals, for genotyping and marker saturation in the two RIL populations. Seventy and 50 of the candidate genes could be mapped in 29H x Vf136 and Vf6 x Vf136, respectively. Besides the strong reduction of the QTL intervals, the mapping process allowed replacing previous dominant and pedigree-specific RAPD flanking markers with robust and transferrable SNP markers, revealing promising candidates for resistance against the two pathogens. CONCLUSIONS: Although further efforts in association mapping and expression studies will be required to corroborate the candidate genes for resistance, the fine-mapping approach proposed here increases the genetic resolution of relevant QTL regions and paves the way for an efficient deployment of useful alleles for faba bean ascochyta and broomrape resistance through marker-assisted breeding.