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A combined physiological and biophysical approach to understand the ligand-dependent efficiency of 3-hydroxy-4-pyridinone Fe-chelates
Publication . Santos, Carla S.; Leite, Andreia; Vinhas, Sílvia; Ferreira, Sofia; Moniz, Tânia; Vasconcelos, Marta W.; Rangel, Maria da Conceição
Ligands of the 3‐hydroxy‐4‐pyridinone (3,4‐HPO) class were considered eligible to formulate new Fe fertilizers for Iron Deficiency Chlorosis (IDC). Soybean (Glycine max L.) plants grown in hydroponic conditions and supplemented with Fe‐chelate [Fe(mpp)3] were significantly greener, had increased biomass, and were able to translocate more iron from the roots to the shoots than those supplemented with an equal amount of the commercially available chelate [FeEDDHA]. To understand the influence of the structure of 3,4‐HPO ligand on the role of the Fe‐chelate to improve Fe‐uptake, we investigated and report here the effect of Fe‐chelates ([Fe(mpp)3], [Fe(dmpp)3], and [Fe(etpp)3]) in addressing IDC. Chlorosis development was assessed by measurement of morphological parameters, quantification of chlorophyll and Fe, and other micronutrient contents, as well as measurement of enzymatic activity (FCR) and gene expression (FRO2, IRT1, and Ferritin). All [Fe(3,4‐HPO)3] chelates were able to provide Fe to plants and prevent IDC but with a different efficiency depending on the ligand. We hypothesize that this may be related with the distinct physicochemical characteristics of ligands and complexes, namely, the diverse hydrophilic–lipophilic balance of the three chelates. To test the hypothesis, we performed an EPR biophysical study using liposomes prepared from a soybean (Glycine3 max L.) lipid extract and spin probes. The results showed that the most effective chelate [Fe(mpp)3] shows a preferential location close to the surface while the others prefer the hydrophobic region inside the bilayer.
Iron partitioning at an early growth stage impacts iron deficiency responses in soybean plants (Glycine max L.)
Publication . Santos, Carla S.; Roriz, Mariana; Carvalho, Susana M. P.; Vasconcelos, Marta
Iron (Fe) deficiency chlorosis (IDC) leads to leaf yellowing, stunted growth and drastic yield losses. Plants have been differentiated into ‘Fe-efficient’ (EF) if they resist to IDC and ‘Fe-inefficient’ (IN) if they do not, but the reasons for this contrasting efficiency remain elusive. We grew EF and IN soybean plants under Fe deficient and Fe sufficient conditions and evaluated if gene expression and the ability to partition Fe could be related to IDC efficiency. At an early growth stage, Fe-efficiency was associated with higher chlorophyll content, but Fe reductase activity was low under Fe-deficiency for EF and IN plants. The removal of the unifoliate leaves alleviated IDC symptoms, increased shoot:root ratio, and trifoliate leaf area. EF plants were able to translocate Fe to the aboveground plant organs, whereas the IN plants accumulated more Fe in the roots. FRO2-like gene expression was low in the roots; IRT1-like expression was higher in the shoots; and ferritin was highly expressed in the roots of the IN plants. The efficiency trait is linked to Fe partitioning and the up-regulation of Fe-storage related genes could interfere with this key process. This work provides new insights into the importance of mineral partitioning among different plant organs at an early growth stage.
Nuclear magnetic resonance metabolomics of iron deficiency in soybean leaves
Publication . Lima, Marta R. M.; Diaz, Sílvia O.; Lamego, Inês; Grusak, Michael A.; Vasconcelos, Marta; Gil, Ana M.
Iron (Fe) deficiency is an important agricultural concern that leads to lower yields and crop quality. A better understanding of the condition at the metabolome level could contribute to the design of strategies to ameliorate Fe-deficiency problems. Fe-sufficient and Fe-deficient soybean leaf extracts and whole leaves were analyzed by liquid 1H nuclear magnetic resonance (NMR) and high-resolution magic-angle spinning NMR spectroscopy, respectively. Overall, 30 compounds were measurable and identifiable (comprising amino and organic acids, fatty acids, carbohydrates, alcohols, polyphenols, and others), along with 22 additional spin systems (still unassigned). Thus, metabolite differences between treatment conditions could be evaluated for different compound families simultaneously. Statistically relevant metabolite changes upon Fe deficiency included higher levels of alanine, asparagine/aspartate, threonine, valine, GABA, acetate, choline, ethanolamine, hypoxanthine, trigonelline, and polyphenols and lower levels of citrate, malate, ethanol, methanol, chlorogenate, and 3-methyl-2-oxovalerate. The data indicate that the main metabolic impacts of Fe deficiency in soybean include enhanced tricarboxylic acid cycle activity, enhanced activation of oxidative stress protection mechanisms and enhanced amino acid accumulation. Metabolites showing accumulation differences in Fe-starved but visually asymptomatic leaves could serve as biomarkers for early detection of Fe-deficiency stress.
Cultivar variability of iron uptake mechanisms in rice (Oryza sativa L.)
Publication . Pereira, Margarida P.; Santos, Carla; Gomes, Ana M. P.; Vasconcelos, Marta W.
Rice (Oryza sativa L.) is the most important staple food in the world. It is rich in genetic diversity and can grow in a wide range of environments. Iron (Fe) deficiency is a major abiotic stress in crop production and in aerobic soils, where Fe forms insoluble complexes, and is not readily available for uptake. To cope with Fe deficiency, plants developed mechanisms for Fe uptake, and although rice was described as a Strategy II plant, recent evidence suggests that it is capable of utilizing mechanisms from both Strategies. The main objective of this work was to compare two cultivars, Bico Branco (japonica) and Nipponbare (tropical japonica), to understand if the regulation of Fe uptake mechanisms could be cultivar (cv.) dependent. Plants of both cultivars were grown under Fe-deficient and -sufficient conditions and physiological and molecular responses to Fe deficiency were evaluated. Bico Branco cv. developed more leaf chlorosis and was more susceptible to Fe deficiency, retaining more nutrients in roots, than Nipponbare cv., which translocated more nutrients to shoots. Nipponbare cv. presented higher levels of Fe reductase activity, which was significantly up-regulated by Fe deficiency, and had higher expression levels of the Strategy I-OsFRO2 gene in roots, while Bico Branco cv. induced more genes involved in Strategy II. These new findings show that rice cultivars have different responses to Fe deficiency and that the induction of Strategy I or II may be rice cultivar-dependent, although the utilization of the reduction mechanisms seems to be an ubiquitous advantage.
High relative air humidity influences mineral accumulation and growth in iron deficient soybean plants
Publication . Roriz, Mariana; Carvalho, Susana M. P.; Vasconcelos, Marta W.
Iron (Fe) deficiency chlorosis (IDC) in soybean results in severe yield losses. Cultivar selection is the most commonly used strategy to avoid IDC but there is a clear interaction between genotype and the environment; therefore, the search for quick and reliable tools to control this nutrient deficiency is essential. Several studies showed that relative humidity (RH) may influence the long distance transport of mineral elements and the nutrient status of plants. Thus, we decided to analyze the response of an “Fe-efficient” (EF) and an “Fe-inefficient” (INF) soybean accession grown under Fe-sufficient and deficient conditions under low (60%) and high (90%) RH, evaluating morphological, and physiological parameters. Furthermore, the mineral content of different plant organs was analyzed. Our results showed beneficial effects of high RH in alleviating IDC symptoms as seen by increased SPAD values, higher plant dry weight (DW), increased plant height, root length, and leaf area. This positive effect of RH in reducing IDC symptoms was more pronounced in the EF accession. Also, Fe content in the different plant organs of the EF accession grown under deficient conditions increased with RH. The lower partitioning of Fe to roots and stems of the EF accessions relative to dry matter also supported our hypothesis, suggesting a greater capacity of this accession in Fe translocation to the aerial parts under Fe deficient conditions, when grown under high RH.

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Funding agency

Fundação para a Ciência e a Tecnologia

Funding programme

3599-PPCDT

Funding Award Number

PTDC/AGR-GPL/118772/2010

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