María Carlota Vaz Patto

Abiotic Stress Responses in Legumes: Strategies Used to Cope with Environmental Challenges

Legumes are well recognized for their nutritional and health benefits as well as for their impact in the sustainability of agricultural systems. The threatening scenario imposed by climate change highlights the need for concerted research approaches in order to develop crops that are able to cope with environmental stresses, while increasing yield and quality. During the last decade, some physiological components and molecular players underlying abiotic stress responses of a broad range of legume species have been elucidated. Plant physiology approaches provided general outlines of plant responses, identifying stress tolerance-related traits or elite cultivars. A thorough identification of candidate genes and quantitative trait loci (QTLs) associated with these traits followed. Model legumes like Medicago truncatula, Lotus japonicus, and more recently, Glycine max provided valuable translational approaches for dissecting legume responses to abiotic stresses. The challenge now focuses on the translation of the information gained in model systems in controlled environments to crops grown under field conditions. In this review, we provide a general overview of the recent achievements on the study of abiotic stress responses in a broad range of model, grain and forage legumes species, highlighting the different approaches used. Major accomplishments, as well as limitations or drawbacks are discussed across the different sections. Some perspectives regarding new approaches for screening, breeding or engineering legumes with desirable abiotic stress resistance traits are anticipated. These advances will support the development of legumes better adapted to environmental constraints, tackling current demands on modern agriculture and food production presently exacerbated by global climate changes.

Achievements and Challenges in Improving the Nutritional Quality of Food Legumes

Quality aspects of food crops have globally important market economic and health repercussions in the current climate of food security. Grain legumes have high potential for the nutritional quality improvement of foods, but limited data on manipulating seed quality is available as the primary focus has been hitherto on phenotypic and agronomic trait improvement. This has resulted in a lack of innovation and low attractiveness of legume food products that, with the emergence of novel food habits, have together contributed to reduced legume food consumption. This trend now needs to be challenged and circumvented. In this review we have assessed the key factors affecting the nutritional quality of legume seeds such as protein, starch, dietary fiber, natural antioxidant compounds and anti-nutritional factors. All have been reviewed with emphasis on how these components might influence consumer acceptance and functional properties of legume based food products. Biofortification approaches and technological processing are discussed as ways in which the nutritional value of legumes and their consumption might be enhanced. In order to increase consumption of grain legumes, we propose that efforts should concentrate on identifying nutritionally enhanced and genetically diverse germplasm, and on linking genetics with sensorial and processing quality. This will assist the development of breeding/selection tools for traits that determine consumer demand, facilitating the implementation of quality breeding objectives in legume breeding programs. Equally important, efforts should focus on developing attractive, convenient ready-to-eat and tasty legume-based food formulations, contributing to the diversification of healthier and more nutritional diets. As a result of such a targeted effort, legume cultivation and consumption could be enhanced leading to a reduction in both the global economic burden caused by malnutrition and associated chronic diseases, and the environmental impact of agriculture.

The impact of CdSe/ZnS Quantum Dots in cells of Medicago sativa in suspension culture

BackgroundNanotechnology has the potential to provide agriculture with new tools that may be used in the rapid detection and molecular treatment of diseases and enhancement of plant ability to absorb nutrients, among others. Data on nanoparticle toxicity in plants is largely heterogeneous with a diversity of physicochemical parameters reported, which difficult generalizations. Here a cell biology approach was used to evaluate the impact of Quantum Dots (QDs) nanocrystals on plant cells, including their effect on cell growth, cell viability, oxidative stress and ROS accumulation, besides their cytomobility.ResultsA plant cell suspension culture of Medicago sativa was settled for the assessment of the impact of the addition of mercaptopropanoic acid coated CdSe/ZnS QDs. Cell growth was significantly reduced when 100 mM of mercaptopropanoic acid -QDs was added during the exponential growth phase, with less than 50% of the cells viable 72 hours after mercaptopropanoic acid -QDs addition. They were up taken by Medicago sativa cells and accumulated in the cytoplasm and nucleus as revealed by optical thin confocal imaging. As part of the cellular response to internalization, Medicago sativa cells were found to increase the production of Reactive Oxygen Species (ROS) in a dose and time dependent manner. Using the fluorescent dye H2DCFDA it was observable that mercaptopropanoic acid-QDs concentrations between 5-180 nM led to a progressive and linear increase of ROS accumulation.ConclusionsOur results showed that the extent of mercaptopropanoic acid coated CdSe/ZnS QDs cytotoxicity in plant cells is dependent upon a number of factors including QDs properties, dose and the environmental conditions of administration and that, for Medicago sativa cells, a safe range of 1-5 nM should not be exceeded for biological applications.

Identification of common genomic regions controlling resistance to Mycosphaerella pinodes, earliness and architectural traits in different pea genetic backgrounds

In a previous study a genetic map had been developed using a RIL population derived from a cross between the Pisum sativum ssp. syriacum accession P665 and the P. sativum ssp. sativum cv. Messire. This population segregated for several agricultural important traits and was successfully used to identify QTLs (Quantitative Trait Loci) controlling resistance to Mycosphaerella pinodes and Orobanche crenata, earliness, root length and aerial biomass. However, this map contained only a few markers in common with the international pea consensus map, hampering comparison with other pea maps. The objective of this study was to incorporate a set of common transferable and reproducible markers into the P665 × Messire map to favour comparative mapping and QTL validation. Seventy-eight out of the 248 SSRs assayed resulted polymorphic in the parental lines. Thirty-eight of them, uniformly distributed all over the genome, were genotyped in the whole population and included in the map. This SSR enriched map allowed identification of six new QTLs (three for resistance to M. pinodes, two for resistance to broomrape and one for root length). Inclusion of the SSRs confirmed the homology of some of the QTLs identified in the population P665 × Messire with other QTLs associated with related traits in different pea genetic backgrounds.

Genetic Architecture of Ear Fasciation in Maize (Zea mays) under QTL Scrutiny.

Maize ear fasciation Knowledge of the genes affecting maize ear inflorescence may lead to better grain yield modeling. Maize ear fasciation, defined as abnormal flattened ears with high kernel row number, is a quantitative trait widely present in Portuguese maize landraces. Material and Methods Using a segregating population derived from an ear fasciation contrasting cross (consisting of 149 F2:3 families) we established a two location field trial using a complete randomized block design. Correlations and heritabilities for several ear fasciation-related traits and yield were determined. Quantitative Trait Loci (QTL) involved in the inheritance of those traits were identified and candidate genes for these QTL proposed. Results and Discussion Ear fasciation broad-sense heritability was 0.73. Highly significant correlations were found between ear fasciation and some ear and cob diameters and row number traits. For the 23 yield and ear fasciation-related traits, 65 QTL were identified, out of which 11 were detected in both environments, while for the three principal components, five to six QTL were detected per environment. Detected QTL were distributed across 17 genomic regions and explained individually, 8.7% to 22.4% of the individual traits or principal components phenotypic variance. Several candidate genes for these QTL regions were proposed, such as bearded-ear1, branched silkless1, compact plant1, ramosa2, ramosa3, tasselseed4 and terminal ear1. However, many QTL mapped to regions without known candidate genes, indicating potential chromosomal regions not yet targeted for maize ear traits selection. Conclusions Portuguese maize germplasm represents a valuable source of genes or allelic variants for yield improvement and elucidation of the genetic basis of ear fasciation traits. Future studies should focus on fine mapping of the identified genomic regions with the aim of map-based cloning.

Lathyrus sativus transcriptome resistance response to Ascochyta lathyri investigated by deepSuperSAGE analysis

Lathyrus sativus (grass pea) is a temperate grain legume crop with a great potential for expansion in dry areas or zones that are becoming more drought-prone. It is also recognized as a potential source of resistance to several important diseases in legumes, such as ascochyta blight. Nevertheless, the lack of detailed genomic and/or transcriptomic information hampers further exploitation of grass pea resistance-related genes in precision breeding. To elucidate the pathways differentially regulated during ascochyta-grass pea interaction and to identify resistance candidate genes, we compared the early response of the leaf gene expression profile of a resistant L. sativus genotype to Ascochyta lathyri infection with a non-inoculated control sample from the same genotype employing deepSuperSAGE. This analysis generated 14.387 UniTags of which 95.7% mapped to a reference grass pea/rust interaction transcriptome. From the total mapped UniTags, 738 were significantly differentially expressed between control and inoculated leaves. The results indicate that several gene classes acting in different phases of the plant/pathogen interaction are involved in the L. sativus response to A. lathyri infection. Most notably a clear up-regulation of defense-related genes involved in and/or regulated by the ethylene pathway was observed. There was also evidence of alterations in cell wall metabolism indicated by overexpression of cellulose synthase and lignin biosynthesis genes. This first genome-wide overview of the gene expression profile of the L. sativus response to ascochyta infection delivered a valuable set of candidate resistance genes for future use in precision breeding.

Allelic diversity in the transcriptomes of contrasting rust-infected genotypes of Lathyrus sativus, a lasting resource for smart breeding

BackgroundGrass pea (Lathyrus sativus L.) is a valuable resource for potentially durable partial resistance to rust. To gain insight into the resistance mechanism and identify potential resistance genes, we generated the first comprehensive transcriptome assemblies from control and Uromyces pisi inoculated leafs of a susceptible and a partially rust-resistant grass pea genotype by RNA-seq.Results134,914 contigs, shared by both libraries, were used to analyse their differential expression in response to rust infection. Functional annotation grouped 60.4% of the contigs present in plant databases (37.8% of total) to 33 main functional categories, being “protein”, “RNA”, “signalling”, “transport” and “stress” the most represented. Transcription profiles revealed considerable differences in regulation of major phytohormone signalling pathways: whereas Salicylic and Abscisic Acid pathways were up-regulated in the resistant genotype, Jasmonate and Ethylene pathways were down-regulated in the susceptible one. As potential Resistance-genes we identified a mildew resistance locus O (MLO)-like gene, and MLO-related transcripts. Also, several pathogenesis-related genes were up-regulated in the resistant and exclusively down regulated in the susceptible genotype. Pathogen effectors identified in both inoculated libraries, as e.g. the rust Rtp1 transcript, may be responsible for the down-regulation of defence-related transcripts. The two genotypes contained 4,892 polymorphic contigs with SNPs unevenly distributed between different functional categories. Protein degradation (29.7%) and signalling receptor kinases (8.2%) were the most diverged, illustrating evolutionary adaptation of grass pea to the host/pathogens arms race.ConclusionsThe vast array of novel, resistance-related genomic information we present here provides a highly valuable resource for future smart breeding approaches in this hitherto under-researched, valuable legume crop.

Relevance, structure and analysis of ferulic acid in maize cell walls

Phenolic compounds in foods have been widely studied due to their health benefits. In cereals, phenolic compounds are extensively linked to cell wall polysaccharides, mainly arabinoxylans, which cross-link with each other and with other cell wall components. In maize, ferulic acid is the phenolic acid present in the highest concentration, forming ferulic acid dehydrodimers, trimers and tetramers. The cross-linking of polysaccharides is important for the cell wall structure and growth, and may protect against pathogen invasion. In addition to the importance for maize physiology, ferulic acid has been recognized as an important chemical structure with a wide range of health benefits when consumed in a diet rich in fibre. This review paper presents the different ways ferulic acid can be present in maize, the importance of ferulic acid derivatives and the methodologies that can be used for their analysis.

Unveiling common responses of Medicago truncatula to appropriate and inappropriate rust species.

Little is known about the nature of effective defense mechanisms in legumes to pathogens of remotely related plant species. Some rust species are among pathogens with broad host range causing dramatic losses in various crop plants. To understand and compare the different host and nonhost resistance (NHR) responses of legume species against rusts, we characterized the reaction of the model legume Medicago truncatula to one appropriate (Uromyces striatus) and two inappropriate (U. viciae-fabae and U. lupinicolus) rusts. We found that similar pre and post-haustorial mechanisms of resistance appear to be operative in M. truncatula against appropriate and inappropriate rust fungus. The appropriate U. striatus germinated better on M. truncatula accessions then the inappropriate U. viciae-fabae and U. lupinicolus, but once germinated, germ tubes of the three rusts had a similar level of success in finding stomata and forming an appressoria over a stoma. However, responses to different inappropriate rust species also showed some specificity, suggesting a combination of non-specific and specific responses underlying this legume NHR to rust fungi. Further genetic and expression analysis studies will contribute to the development of the necessary molecular tools to use the present information on host and NHR mechanisms to breed for broad-spectrum resistance to rust in legume species.

Participatory Plant Quality Breeding: An Ancient Art Revisited by Knowledge Sharing. The Portuguese Experience

Since the first domestications of wild plants about 12.000 years ago, farmers have been re‐ sponsible for the development and conservation of thousands of crop landraces in hundreds of species [1]. Farmers put aside, for the next generation, a part of the harvested seed. De‐ pending on the crop and the farmer, selection is carried out to obtain a crop answering bet‐ ter to the wishes of the growers and communities [2].