Breeding for intercropping: the case of red clover persistence in grasslands

Legumes are important in sustainable agriculture and particularly so when they are intercropped with other species. In breeding programs, little attention is paid to their agronomic performance in species mixtures. In red clover, improved persistence is an important breeding goal. We identified traits associated with survival of red clover cultivated in pure stands (PS 3) or in mixtures with grasses (MS 3) and managed under a 3-cut system (two locations), as well as in pure stands in a 5-cut system (PS 5, one location). Survivors from replicate plots were collected and a new generation made from each plot. The new generations were characterized in a growth experiment with light or simulated shade, and in a freezing experiment. We show that the traits related to red clover persistence depend on both plant community composition and cutting frequency. MS 3 had more leaves with larger leaf blades and longer petioles during the vegetative stage, followed by earlier stem elongation, higher number of elongating stems, higher biomass (also when accounting for earlier stem elongation) and more leaves in the regrowth after cutting than PS 3. MS 3 also had better freezing tolerance. PS 5 was similar to MS 3 and different from PS 3 in the number of leaves, leaf blade size, petiole length and number of elongating stems. These results show that breeding and cultivar evaluation, which is currently almost exclusively considering performance in pure stands, may miss some variation which provides persistence of red clover in mixtures with grasses.Breeding for intercropping: the case of red clover persistence in grasslandspublishedVersio

Cultivar development of kelps for commercial cultivation - Past lessons and future prospects

Cultivated kelps and other macroalgae have great potential in future provision of food, feed, bioenergy, fertilizer, and raw material for a range of chemical products including pharmaceuticals, food and feed additives, and cosmetics. Only a few species are currently cultivated, almost exclusively in Asia. There is a range of species that could be utilized in different parts of the world, providing that protocols for reproduction, propagation, and cultivation are developed. Domestication of species involves selection of traits that are desirable in cultivation and in the utilization of the harvested biomass. Genetic improvement of cultivated species through recombination of alleles and selection (breeding) has ensured high productivity and product quality in both agri- and aquaculture and will likely do so for macroalgae cultivation and use as well. According to the published literature, genetic improvement of kelps in Asia has so far largely relied on utilization of heterosis expressed in certain combinations of parental material, sometimes species hybrids. Here, we explore and evaluate the various methods that could be used in kelp breeding and propose an initial simple and low-cost breeding strategy based on recurrent mixed hybridization and phenotypic selection within local populations. We also discuss the genetic diversity in wild populations, and how this diversity can be protected against genetic pollution, either by breeding and cultivating local populations, or by developing cultivars that are not able to establish in, or hybridize with, wild populations.publishedVersio

Allele frequency changes provide evidence for selection and identification of candidate loci for survival in red clover (Trifolium pratense L)

Survivor populations of red clover (Trifolium pratense L.) from plots in a field experiment in southern Norway were genetically characterized using genotyping by sequencing, and compared with the original population and each other. Genetic differentiation between populations was characterized on the basis of allele frequencies of single nucleotide polymorphisms (SNPs), using principal component analysis. SNPs that had been under selection, i.e., SNPs with significantly different allele frequencies in survivor populations relative to the original population, or between survivor populations that had received different treatments, were identified by analysis of FST values, using BayeScan and a simple and stringent FST-based test utilizing replicate populations from the field experiment. In addition, we tested the possibility of pooling DNA samples prior to sequencing, and pooling leaf samples prior to DNA extraction and sequencing, followed by allele frequency estimation on the basis of number of variant reads. Overall, survivor populations were more different from each other than from the original population, indicating random changes in allele frequency, selection in response to local variation in conditions between plots in the field experiment, or sampling error. However, some differentiation was observed between plots sown as pure stands or species mixtures, plots sown at different densities, and plots subjected to different harvesting regimes. Allele frequencies could be accurately estimated from pooled DNA, and SNPs under selection could be identified when leaf samples were pooled prior to DNA extraction. However, substantial sampling error required replicate populations and/or a high number of sampled individuals. We identified a number of chromosomal loci that had been under selection in pure stand plots relative to the original sown population, and loci that had been under differential selection in pure stands of red clover vs. red clover grown in species mixtures. These are all candidate loci for establishment success or persistence in red clover

Species interactions in a grassland mixture under low nitrogen fertilization and two cutting frequencies II:Nutritional quality

Mixtures and pure stands of perennial ryegrass, tall fescue, white clover and red clover were grown in a three-cut and a five-cut system in southern Norway, at a low fertilization rate (100 kg N ha−1 year−1). The nutritional quality (annual weighted averages) of the dried forage from the two-first harvesting years was analysed. There was no significant effect of species diversity on crude protein (CP) concentration. In the three-cut system, we found a significant species diversity effect leading to 10% higher concentrations of acid detergent fibre (ADF), 20–22% lower concentrations of water-soluble carbohydrate (WSC) and 4% lower net energy for lactation (NEL) concentrations in mixtures compared with pure stands (averaged across the two-first years). In the five-cut system, similar effects were seen in the first year only. This diversity effect was associated with a reduction in WSC and NEL concentrations and an increase in ADF, NDF and CP concentrations in the grass species, and not in red clover, when grown in mixtures. This is thought to be a combined result of better N availability and more shading in the mixtures. Species diversity reduced the intra-annual variability in nutritional quality in both cutting systems.acceptedVersio

Assessment of Quarantine Pest Dispersal from Norwegian Potato and Root Vegetable Packing Plants with Evaluation of Risk Reducing Options. Opinion of the Panel on Plant Health of the Norwegian Scientific Committee for Food and Environment

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The effect of harvest time of forage on carbohydrate digestion in horses quantified by in vitro and mobile bag techniques

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Global transcriptome changes in perennial ryegrass during early infection by pink snow mould

Lack of resistance to pink snow mould (Microdochium nivale) is a major constraint for adaptation of perennial ryegrass (Lolium perenne L.) to continental regions with long-lasting snow cover at higher latitudes. Almost all investigations of genetic variation in resistance have been performed using cold acclimated plants. However, there may be variation in resistance mechanisms that are functioning independently of cold acclimation. In this study our aim was to identify candidate genes involved in such resistance mechanisms. We first characterized variation in resistance to M. nivale among non-acclimated genotypes from the Norwegian cultivar ‘Fagerlin’ based on relative regrowth and fungal quantification by real-time qPCR. One resistant and one susceptible genotype were selected for transcriptome analysis using paired-end sequencing by Illumina Hiseq 2000. Transcriptome profiles, GO enrichment and KEGG pathway analysis indicate that defense response related genes are differentially expressed between the resistant and the susceptible genotype. A significant up-regulation of defense related genes, as well as genes involved in cell wall cellulose metabolic processes and aryl-alcohol dehydrogenase (NADP+) activity, was observed in the resistant genotype. The candidate genes identified in this study might be potential molecular marker resources for breeding perennial ryegrass cultivars with improved resistance to pink snow mould.publishedVersio

Comparative assessment of food safety regulations and standards for arsenic, cadmium, lead, mercury and iodine in macroalgae used as food and feed in China and Europe

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Red clover (Trifolium pratense L.) draft genome provides a platform for trait improvement

Red clover (Trifolium pratense L.) is a globally significant forage legume in pastoral livestock farming systems. It is an attractive component of grassland farming, because of its high yield and protein content, nutritional value and ability to fix atmospheric nitrogen. Enhancing its role further in sustainable agriculture requires genetic improvement of persistency, disease resistance, and tolerance to grazing. To help address these challenges, we have assembled a chromosome-scale reference genome for red clover. We observed large blocks of conserved synteny with Medicago truncatula and estimated that the two species diverged ~23 million years ago. Among the 40,868 annotated genes, we identified gene clusters involved in biochemical pathways of importance for forage quality and livestock nutrition. Genotyping by sequencing of a synthetic population of 86 genotypes show that the number of markers required for genomics-based breeding approaches is tractable, making red clover a suitable candidate for association studies and genomic selection

Genome editing in food and feed production – implications for risk assessment. Scientific Opinion of the Scientific Steering Committee of the Norwegian Scientific Committee for Food and Environment

The Norwegian Scientific Committee for Food and Environment (VKM) initiated this work to examine the extent to which organisms developed by genome-editing technologies pose new challenges in terms of risk assessment. This report considers whether the risk assessment guidance on genetically modified organisms, developed by the European Food Safety Authority (EFSA), can be applied to evaluate potential risks of organisms developed by genome editing. Background Gene technology has allowed for the transfer of genes between organisms and species, and thereby to design altered genotypes with novel traits, i.e. GMOs. A new paradigm started in the early 2000s with the development of genome-editing techniques. Unlike traditional genetic modification techniques resulting in insertion of foreign DNA fragments at random locations in the genome, the new genome-editing techniques additionally open for a few single nucleotide edits or short insertions/deletions at a targeted site in an organism’s genome. These new techniques can be applied to most types of organisms, including plants, animals and microorganisms of commercial interest. An important question is how the novel, genome-edited organisms should be evaluated with respect to risks to health and the environment. The European Court of Justice decided in 2018 to include genome-edited organisms in the GMO definition and hence in the regulatory system already in place. This implies that all products developed by genome-editing techniques must be risk-assessed within the existing regulatory framework for GMOs. The European and Norwegian regulatory frameworks regulate the production, import and placing on the market of food and feed containing, consisting of or produced from GMOs, as well as the release of GMOs into the environment. The assessment draws on guidance documents originally developed by EFSA for risk assessment of GMOs, which were drawn up mainly to address risks regarding insertion of transgenes. The new genome-editing techniques, however, provide a new continuum of organisms ranging from those only containing a minor genetic alteration to organisms containing insertion or deletion of larger genomic regions. Risk assessment of organisms developed by genome editing The present discourse on how new genome-editing techniques should be regulated lacks an analysis of whether risk assessment methodologies for GMOs are adequate for risk assessment of organisms developed through the use of the new genome-editing techniques. Therefore, this report describes the use of genome-editing techniques in food and feed production and discusses challenges in risk assessment with the regulatory framework. Specifically, this report poses the question as to whether the EFSA guidance documents are sufficient for evaluating risks to health and environment posed by genome-edited plants, animals and microorganisms. To address these questions, the report makes use of case examples relevant for Norway. These examples, intended for food and feed, include oilseed rape with a modified fatty acid profile, herbicide-tolerant and pest-resistant crops, sterile salmon, virus-resistant pigs and hornless cattle. The report considers all aspects of the stepwise approach as described in the EFSA guidance documents. Conclusions The inherent flexibility of the EFSA guidance makes it suitable to cover health and environmental risk assessments of a wide range of organisms with various traits and intended uses. Combined with the embedded case-by-case approach the guidance is applicable to genome-edited organisms. The evaluation of the guidance demonstrates that the parts of the health and environmental risk assessment concerned with novel traits (i.e. the phenotype of the organism) may be fully applied to all categories of genome-edited organisms. ............publishedVersio