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Assessing the impact of nitrogen supplementation in oats across multiple growth locations and years with targeted phenotyping and high-resolution metabolite profiling approaches

ABSTRACT: Oats (Avena sativa L.) are a healthy food, being high in dietary fibre (e.g. β-glucans), antioxidants, minerals, and vitamins. Understanding the effect of variety and crop management on nutritional quality is important. The response of four oat varieties to increased nitrogen levels was investigated across multiple locations and years with respect to yield, grain quality and metabolites (assessed via GC- and LC- MS). A novel high-resolution UHPLC-PDA-MS/MS method was developed, providing improved metabolite enrichment, resolution, and identification. The combined phenotyping approach revealed that, amino acid levels were increased by nitrogen supplementation, as were total protein and nitrogen containing lipid levels, whereas health-beneficial avenanthramides were decreased. Although nitrogen addition significantly increased grain yield and β-glucan content, supporting increasing the total nitrogen levels recommended within agricultural guidelines, oat varietal choice as well as negative impacts upon health beneficial secondary metabolites and the environmental burdens associated with nitrogen fertilisation, require further consideration.

INSTRUMENT(S): Liquid Chromatography MS - negative - reverse phase, Gas Chromatography MS - positive, Liquid Chromatography MS - positive - reverse phase

SUBMITTER: James William Allwood

PROVIDER: MTBLS1801 | MetaboLights | 2021-04-26

REPOSITORIES: MetaboLights

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Assessing the impact of nitrogen supplementation in oats across multiple growth locations and years with targeted phenotyping and high-resolution metabolite profiling approaches.

Allwood J William JW Martinez-Martin Pilar P Xu Yun Y Cowan Alexander A Pont Simon S Griffiths Irene I Sungurtas Julie J Clarke Sarah S Goodacre Royston R Marshall Athole A Stewart Derek D Howarth Catherine C

Food chemistry 20210316

Oats (Avena sativa L.) are a healthy food, being high in dietary fibre (e.g. β-glucans), antioxidants, minerals, and vitamins. Understanding the effect of variety and crop management on nutritional quality is important. The response of four oat varieties to increased nitrogen levels was investigated across multiple locations and years with respect to yield, grain quality and metabolites (assessed via GC- and LC- MS). A novel high-resolution UHPLC-PDA-MS/MS method was developed, providing improve ...[more]

PMID: 33799237

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Project description:<div><b>INTRODUCTION:</b> Oats (Avena sativa L.) are a whole grain cereal recognised for their health benefits and which are cultivated largely in temperate regions providing both a source of food for humans and animals, as well as being used in cosmetics and as a potential treatment for a number of diseases. Oats are known as being a cereal source high in dietary fibre (e.g. β-glucans), as well as being high in antioxidants, minerals and vitamins. Recently, oats have been gaining increased global attention due to their large number of beneficial health effects. Consumption of oats has been proven to lower blood LDL cholesterol levels and blood pressure, thus reducing the risk of heart disease, as well as reducing blood-sugar and insulin levels.</div><div><b>OBJECTIVES:</b> Oats are seen as a low input cereal. Current agricultural guidelines on nitrogen application are believed to be suboptimal and only consider the effect of nitrogen on grain yield. It is important to understand the role of both variety and of crop management in determining nutritional quality of oats. In this study the response of yield, grain quality and grain metabolites to increasing nitrogen application to levels greater than current guidelines were investigated.</div><div><b>METHODS:</b> Four winter oat varieties (Mascani, Tardis, Balado and Gerald) were grown in a replicated nitrogen response trial consisting of a no added nitrogen control and four added nitrogen treatments between 50 and 200 kg N ha-1 in a randomised split-plot design. Grain yield, milling quality traits, β-glucan, total protein and oil content were assessed. The de-hulled oats (groats) were also subjected to a rapid Ultra High Performance Liquid Chromatography-Mass Spectrometry (UHPLC-MS) metabolomic screening approach.</div><div><b>RESULTS:</b> Application of nitrogen had a significant effect on grain yield but there was no significant difference between the response of the four varieties. Grain quality traits however displayed significant differences both between varieties and nitrogen application level. β-glucan content significantly increased with nitrogen application. The UHPLC-MS approach has provided a rapid, sub 15 min per sample, metabolite profiling method that is repeatable and appropriate for the screening of large numbers of cereal samples. The method captured a wide range of compounds, inclusive of primary metabolites such as the amino acids, organic acids, vitamins and lipids, as well as a number of key secondary metabolites, including the avenanthramides, caffeic acid, and sinapic acid and its derivatives and was able to identify distinct metabolic phenotypes for the varieties studied. Amino acid metabolism was massively upregulated by nitrogen supplementation as were total protein levels, whilst the levels of organic acids were decreased, likely due to them acting as a carbon skeleton source. Several TCA cycle intermediates were also impacted, potentially indicating increased TCA cycle turn over, thus providing the plant with a source of energy and reductant power to aid elevated nitrogen assimilation. Elevated nitrogen availability was also directed towards the increased production of nitrogen containing phospholipids. A number of both positive and negative impacts on the metabolism of phenolic compounds that have influence upon the health beneficial value of oats and their products were also observed.</div><div><b>CONCLUSIONS:</b> Although the developed method has broad applicability as a rapid screening method or a rapid metabolite profiling method and in this study has provided valuable metabolic insights, it still must be considered that much greater confidence in metabolite identification, as well as quantitative precision, will be gained by the application of higher resolution chromatography methods, although at a large expense to sample throughput. Follow up studies will apply higher resolution GC (gas chromatography) and LC (reversed phase and HILIC) approaches, oats will be also analysed from across multiple growth locations and growth seasons, effectively providing a cross validation for the results obtained within this preliminary study. It will also be fascinating to perform more controlled experiments with sampling of green tissues, as well as oat grains, throughout the plants and grains development, to reveal greater insight of carbon and nitrogen metabolism balance, as well as resource partitioning into lipid and secondary metabolism.</div>

Project description:Durum wheat requires high nitrogen inputs to obtain the high protein concentration necessary to satisfy pasta and semolina quality criteria. Optimizing plant nitrogen use efficiency is therefore of major importance for wheat grain quality. Here, we studied the impact on grain yield, protein concentration, and for the first time on protein composition of a marine (DPI4913) and a fungal (AF086) biostimulants applied to plant leaves. Plants were grown in a greenhouse and sampled at maturity for grain analysis. The protein concentration and quantity in grains per plant were determined, as well as water-use efficiency. A large-scale quantitative proteomics study of wheat flour samples was performed to determine the effect of biostimulant treatment on grain protein composition. Grain dry biomass and protein quantity were increased by 23.9% and 24.8% respectively with DPI4913, and by 27.4% and 25.9%, respectively, with AF086. A total of 1471 proteins were identified and 1391 were quantified. Quantitative proteomics analysis revealed 26 and 38 proteins with a significantly varying abundance after DPI4913 and AF086 treatment, respectively. Among these, 14 proteins were affected by both DPI4913 and AF086 treatments. The highest variations in proteins abundances were found for proteins involved in grain technological properties such as grain hardness, in storage functions with the substantial over-representation of the gluten protein gamma-gliadin, in regulation processes with the over-representation of proteins that play a transcription regulator role, and in stress responses with the over-representation of proteins implied in biotic and abiotic stress defense. The involvement of biostimulants in the abiotic stress response is suggested by the increase in water-use efficiency for DPI4913 treatment (15.4%) and for AF086 treatment (9.9%). In conclusion, our work, performed in greenhouse, has shown that DPI4913 and AF086 treatments promote grain yield while maintaining protein concentration in grains, and positively affect protein composition in term of grain quality. This study suggests that these biostimulants could be used to optimize durum wheat production and quality in field conditions.

Dataset Information

Assessing the impact of nitrogen supplementation in oats across multiple growth locations and years with targeted phenotyping and high-resolution metabolite profiling approaches

Publications

Assessing the impact of nitrogen supplementation in oats across multiple growth locations and years with targeted phenotyping and high-resolution metabolite profiling approaches.

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