The presence of acrylamide in popular foods has become one of the most difficult problems facing the food industry and its supply chain. Acrylamide is a contaminant that forms from an amino acid called asparagine in its free (non-protein) form and sugars such as glucose, fructose and maltose. The reaction occurs during frying, baking, roasting, toasting and high-temperature processing. Acrylamide is classified as probably cancer-causing in humans and affects development and fertility at high doses. Products made from wheat and other cereals are major sources of dietary acrylamide. Acrylamide in food is covered by regulation (EU) 2017/2158 (2018), which set Benchmark Levels for the presence of acrylamide in food, described compulsory mitigation measures and required all food businesses to monitor the levels of acrylamide in their products. Methods developed to limit acrylamide formation during processing do not work for all products, may adversely affect product quality and are expensive to implement. Our approach has been to lower the acrylamide-forming potential of crop products: for wheat and other cereals that means reducing the concentration of free asparagine in the grain. The project arises from previous BBSRC-funded work in which we have demonstrated large varietal differences in free asparagine concentration in wheat grain and shown it to be highly responsive to sulphur deficiency and other nutritional and environmental stress factors, as well as disease. Asparagine is made by an enzyme called asparagine synthetase and wheat has five genes that encode this enzyme. One of these genes, TaASN2, has emerged as a genetic target because it is highly active specifically in the grain. In our current work, involving teams at Rothamsted Research, Univ. of Bristol and partners AHDB, KWS, Limagrain, RAGT, Saaten Union and Syngenta, we have used the genome editing technique, CRISPR-Cas9, to knock out TaASN2 genes. In addition, we have screened a wheat population in which mutations have been introduced by chemical mutagenesis (a much older technique that has been used in plant breeding since the 1950s) and identified lines with mutations in each of the six versions of TaASN2 in the wheat genome. The TaASN2 mutations in these lines are being 'stacked' by one of our partners and partial knockouts will be available by Year 2 of the project. The concentration of free asparagine in one of the CRISPR lines is reduced by approximately 90%. The aim of this project, which will continue the partnership with Univ. of Bristol, AHDB, Limagrain, RAGT, Saaten Union and Syngenta, is to undertake field trials of these ultra-low asparagine wheat lines with a view to the development of low acrylamide wheat varieties for the UK market. In year 1 we will bulk up seed for the trial. We will also investigate the germination rate of the CRISPR lines: they have shown poor germination so far but this can be reversed by treatment with low concentrations of asparagine. We will check the efficacy and practicality of the asparagine and other potential treatments. We will also develop genetic markers/tools to enable breeders to integrate the ultra-low asparagine trait into breeding lines. Lastly, we will prepare a risk assessment and submit an application for permission to hold the field trial. In year 2 we will conduct the field trial, analysing the low asparagine lines for emergence, physical characteristics, developmental differences, composition, yield and other agronomic characteristics. We will also conduct glasshouse experiments to ascertain whether the low asparagine lines respond to sulphur deficiency in the same way as normal wheat. The project would represent a landmark for crop gene editing, with the field trial, to our knowledge, being the first for gene edited wheat in the UK and Europe, while the involvement of AHDB and wheat breeders will provide a direct pathway to commercialisation if the plants perform well in the field trial.

Bio-lubricants have both environmental and technical advantages over their counterparts derived from mineral oils. In addition to being renewable, they are biodegradable, have lower volatile emissions and low environmental toxicity. They provide superior anti-wear protection and exhibit reduced combustibility. In addition, bio-lubricants have lower coefficients of friction, which results in reduced energy costs for equipment in which bio-lubricants as used. Although vegetable oils are used in blending some less stressed lubricants, their thermal stability is inadequate for the majority of applications as a consequence of the presence of excessive polyunsaturation of their constituent fatty acids. In view of the poor stability of conventional refined rapeseed oil, lubricant blenders currently favour the use of synthetic esters with a high renewables content of the production of the more stressed lubricant types; this more expensive base oil currently inhibits uptake of bio-lubricants by end users. Rapeseed oil has many physical and chemical properties that are advantageous for base oil for the lubricants industry. However, the total content of polyunsaturated fatty acids remains too high and the resulting instability is the principal barrier to its widespread use. The target set by the industry is reduction to less than 5% total PUFAs, whilst retaining the other desirable physical and chemical properties of rapeseed oil. To be economically competitive, some yield penalty in the crop and increased processing costs can be tolerated, as its principal competitor in the market place, low PUFA sunflower oil, is presently priced at up to $120/tonne more on the commodity markets. Nevertheless, the approaches we propose should result in little, if any, yield loss from fully developed varieties. The purpose of the project is to underpin the development of oilseed rape varieties for the production of oil for use in the lubricants industry. A key knowledge gap is an understanding of how to substantially reduce the content of polyunsaturated fatty acids in rapeseed oil without reducing the oil yield of the crop. We will address this knowledge gap and enable establishment of a closed supply chain. This involves: (a) The genetic improvement of oilseed rape by mutagenesis of specific genes in order to produce, from a high-yielding winter crop, oil very low in polyunsaturated fatty acids. (b) Assessment of the physical properties of the oil produced in order to validate its utility. (c) Provision of characterised oilseed rape lines to the breeding industry for the development of cultivars. (d) Catalysing assembly of a supply chain. The strategy is non-GM, so we anticipate no barriers to the widespread utilization of the resultant varieties in the UK.

Bio-lubricants have both environmental and technical advantages over their counterparts derived from mineral oils. In addition to being renewable, they are biodegradable, have lower volatile emissions and low environmental toxicity. They provide superior anti-wear protection and exhibit reduced combustibility. In addition, bio-lubricants have lower coefficients of friction, which results in reduced energy costs for equipment in which bio-lubricants as used. Although vegetable oils are used in blending some less stressed lubricants, their thermal stability is inadequate for the majority of applications as a consequence of the presence of excessive polyunsaturation of their constituent fatty acids. In view of the poor stability of conventional refined rapeseed oil, lubricant blenders currently favour the use of synthetic esters with a high renewables content of the production of the more stressed lubricant types; this more expensive base oil currently inhibits uptake of bio-lubricants by end users. Rapeseed oil has many physical and chemical properties that are advantageous for base oil for the lubricants industry. However, the total content of polyunsaturated fatty acids remains too high and the resulting instability is the principal barrier to its widespread use. The target set by the industry is reduction to less than 5% total PUFAs, whilst retaining the other desirable physical and chemical properties of rapeseed oil. To be economically competitive, some yield penalty in the crop and increased processing costs can be tolerated, as its principal competitor in the market place, low PUFA sunflower oil, is presently priced at up to $120/tonne more on the commodity markets. Nevertheless, the approaches we propose should result in little, if any, yield loss from fully developed varieties. The purpose of the project is to underpin the development of oilseed rape varieties for the production of oil for use in the lubricants industry. A key knowledge gap is an understanding of how to substantially reduce the content of polyunsaturated fatty acids in rapeseed oil without reducing the oil yield of the crop. We will address this knowledge gap and enable establishment of a closed supply chain. This involves: (a) The genetic improvement of oilseed rape by mutagenesis of specific genes in order to produce, from a high-yielding winter crop, oil very low in polyunsaturated fatty acids. (b) Assessment of the physical properties of the oil produced in order to validate its utility. (c) Provision of characterised oilseed rape lines to the breeding industry for the development of cultivars. (d) Catalysing assembly of a supply chain. The strategy is non-GM, so we anticipate no barriers to the widespread utilization of the resultant varieties in the UK.

Dietary fibre (DF) is essential for human health, improving gastro-intestinal function and reducing the risk of a range of chronic diseases (including type 2 diabetes, cardio-vascular disease and types of cancer). However, most UK consumers do not eat DF, with the average daily intake being 17.2 g for women and 20.1g for men, compared with a target of 30g. Cereal products are the major source of DF in the UK diet, with bread alone contributing about 20%. However, the contribution from wheat is limited by the fact that most wheat products are made from white flour, which contains about 3.5% DF compared with 11.5-15.5% in wholemeal. We have therefore identified wheat lines with high DF in white flour, which can be used to develop high fibre wheat lines for UK farmers and products for UK consumers. The proposal will remove the constraints to the development of high fibre lines and products in the UK, by collaborating with four wheat breeders, 2 milling and baking companies and the organisations representing the milling (NABIM) and baking (Federation of Bakers) sectors. This will be achieved in two ways: by providing high fibre pre-breeding lines and molecular makers to wheat breeders, and high fibre lines to millers and bakers to optimise their processes. These advances will be disseminated by the BBSRC Designing Future Wheat programmes and by NABIM and FoB. It will therefore have a fundamental impact on the diet and health of UK consumers.

Maltsters, brewers and distillers are concerned about the long-term sustainability of the barley crop. Seasonal problems in many parts of Europe resulted in a restricted malting barley supply that has only just been alleviated by an above average harvest in Argentina. Within the UK, drought conditions resulted in reduced barley crop quality, i.e. higher protein samples, particularly in Eastern England, where much English malting barley is sourced. Under predicted climate change scenarios, such drought conditions are likely to become more frequent and will affect the spring crop much more than the winter crop, which can escape the worst effects of summer drought through a much earlier maturity. Whilst winter barley might therefore provide a more consistent supply, the proportion bought by English maltsters has declined by over 25% over the past 20 years. This decline is due to the reduced quality level of the winter crop compared to the spring so that distillers can produce 16 more litres of raw spirit per tonne of malt on average from the latter. For an industry predicted to use 600,000t of barley from the 2012 harvest, this is a highly significant difference in production efficiency. All current UK winter barley malting varieties have been derived from Maris Otter, first recommended in 1965. Maris Otter combined the spring malting quality attributes of an older variety, Proctor, with the winter habit of Pioneer. Proctor was the major spring malting variety in the UK for many years but the introduction of Triumph was a quantum leap forward for the spring crop in terms of both quality and yield. In a previous project, we have analysed DNA fingerprints of UK spring and winter barley malting cultivars to identify genetic differences between the two crops that are associated with malting quality. Whilst plant breeders have previously tried to introgress spring quality attributes into winter barley, they have relied on chance events to assemble the right genes, which is an impossible task when the crops differ at thousands of genes. But we now have the knowledge and tools to conduct the introgression of spring attributes into winter barley in a highly targeted manner to test the hypothesis that their introduction will improve winter malting quality. The germplasm emerging from this proposal will then be used by the plant breeding partners of the project in further rounds of crossing and selection to develop improved winter malting quality cultivars that approached the spring quality levels but in a suitable agronomic background for contemporary farming practise and would thus re-generate interest in using winter barley for malting for use in brewing and distilling. As indicated in the previous paragraph, greater use of the winter crop is likely to provide a more consistent supply of malting barley in the future. As malting supplies are becoming tighter due to a variety of market factors, a switch to the higher yielding winter crop would also mean that the effects of competition for land for more profitable crops would have a less pronounced effect upon malting barley supply. As six row barley varieties tend to have a higher yield than two row, a longer term aim is to develop six row malting types that would further decrease the land area required to secure a malting barley supply.