Proceedings of the National Academy of Sciences of the United States of America, Oct 29, 2021
Significance Plant cell surface receptors perceive carbohydrate signaling molecules and hereby es... more Significance Plant cell surface receptors perceive carbohydrate signaling molecules and hereby establish communication with surrounding microbes. Genetic studies have identified two different classes of lysin motif receptor kinases as gatekeepers that together trigger the symbiotic pathway in plants; however, no structural or functional data of the perception mechanisms switching these receptors from resting state into activation is known. In this study, we use structural biology, biochemical, and genetic approaches to demonstrate how the NFP/NFR5 class of lipochitooligosaccharide (LCO) receptors discriminate bacterial symbionts based on a kinetic proofreading mechanism that controls receptor activation and signaling specificity. We show that the LCO binding site can be engineered to support symbiotic functions, which greatly advance future opportunities for receptor engineering in legumes and nonlegumes.
Increasing the proportion of locally produced plant protein in currently meat-rich diets could su... more Increasing the proportion of locally produced plant protein in currently meat-rich diets could substantially reduce greenhouse gas emission and loss of biodiversity. However, plant protein production is hampered by the lack of a cool-season legume equivalent to soybean in agronomic value. Faba bean (Vicia faba L.) has a high yield potential and is well-suited for cultivation in temperate regions, but genomic resources are scarce. Here, we report a high-quality chromosome-scale assembly of the faba bean genome and show that it has grown to a massive 13 Gb in size through an imbalance between the rates of amplification and elimination of retrotransposons and satellite repeats. Genes and recombination events are evenly dispersed across chromosomes and the gene space is remarkably compact considering the genome size, though with significant copy number variation driven by tandem duplication. Demonstrating practical application of the genome sequence, we develop a targeted genotyping ass...
Faba bean is a widely adapted and high-yielding legume cultivated for its protein-rich seeds1. Ho... more Faba bean is a widely adapted and high-yielding legume cultivated for its protein-rich seeds1. However, the seeds accumulate the anti-nutritional pyrimidine glucosides vicine and convicine, which can cause haemolytic anaemia—favism—in the 400 million individuals genetically predisposed by a deficiency in glucose-6-phosphate dehydrogenase2. Here, we identify the first enzyme associated with vicine and convicine biosynthesis, which we name VC1. We show that VC1 co-locates with the major QTL for vicine and convicine content and that the expression of VC1 correlates highly with vicine content across tissues. We also show that low-vicine varieties express a version of VC1 carrying a small, frame-shift insertion, and that overexpression of wild-type VC1 leads to an increase in vicine levels. VC1 encodes a functional GTP cyclohydrolase II, an enzyme normally involved in riboflavin biosynthesis from the purine GTP. Through feeding studies, we demonstrate that GTP is a precursor of vicine bo...
Switching perception of friend and foe Lysine motif receptors in plants perceive glycans that sig... more Switching perception of friend and foe Lysine motif receptors in plants perceive glycans that signal the presence of pathogenic or symbiotic nitrogen-fixing microbes. Bozsoki et al. now define the portions of these receptors that create the discriminatory binding pocket (see the Perspective by Bisseling and Geurts). The motifs were conserved in receptors that initiate immune responses, reflecting the invariable nature of the chitin fragments that they sense. Conversely, the motifs in receptors that respond to symbiotic signals were more varied, reflecting the greater diversity of the lipochitooligosaccharides (Nod factors) that they sense. With domain swapping, the authors switched the Nod factor specificity of receptors from two legume species and also enabled a chitin receptor that was otherwise dedicated to the detection of pathogenic microbes to instead recognize Nod factors. Science , this issue p. 663 ; see also p. 620
Publisher Summary This chapter discusses the use of Agrobacterium for the transformation of the d... more Publisher Summary This chapter discusses the use of Agrobacterium for the transformation of the diploid legume Lotus japonicas . Using the transformation–regeneration method, transgenic Lotus japonicus plants can be obtained after selection for two different resistance genes expressing hygromycin phosphotransferase and neomycin phosphotransferase. Selection for hygromycin and geneticin resistance is very effective and up to 90% of the hypocotyl explants cocultivated with Agrobacterium give rise to one or more transgenic calli. Transgenic calli are easily distinguished from the explants and are moved to regeneration media. Rooted transformed plants in pots are available after 4 months. Longer time is required to discern and separate transgenic calli from explant tissue.
Current plant science and biotechnology in agriculture, 1998
... A LOTUS F1LICAULIS X LOTUS JAPONICUS F2 POPULATION FOR GENETIC AND PHYSICAL MAPPING NN Sandal... more ... A LOTUS F1LICAULIS X LOTUS JAPONICUS F2 POPULATION FOR GENETIC AND PHYSICAL MAPPING NN Sandal and J. Stougaard Laboratory of Gene Expression, Department of Molecular and Structural Biology, University of Aarhus, Gustav Wieds Vej 10, DK-8000 ...
Proceedings of the National Academy of Sciences of the United States of America, Oct 29, 2021
Significance Plant cell surface receptors perceive carbohydrate signaling molecules and hereby es... more Significance Plant cell surface receptors perceive carbohydrate signaling molecules and hereby establish communication with surrounding microbes. Genetic studies have identified two different classes of lysin motif receptor kinases as gatekeepers that together trigger the symbiotic pathway in plants; however, no structural or functional data of the perception mechanisms switching these receptors from resting state into activation is known. In this study, we use structural biology, biochemical, and genetic approaches to demonstrate how the NFP/NFR5 class of lipochitooligosaccharide (LCO) receptors discriminate bacterial symbionts based on a kinetic proofreading mechanism that controls receptor activation and signaling specificity. We show that the LCO binding site can be engineered to support symbiotic functions, which greatly advance future opportunities for receptor engineering in legumes and nonlegumes.
Increasing the proportion of locally produced plant protein in currently meat-rich diets could su... more Increasing the proportion of locally produced plant protein in currently meat-rich diets could substantially reduce greenhouse gas emission and loss of biodiversity. However, plant protein production is hampered by the lack of a cool-season legume equivalent to soybean in agronomic value. Faba bean (Vicia faba L.) has a high yield potential and is well-suited for cultivation in temperate regions, but genomic resources are scarce. Here, we report a high-quality chromosome-scale assembly of the faba bean genome and show that it has grown to a massive 13 Gb in size through an imbalance between the rates of amplification and elimination of retrotransposons and satellite repeats. Genes and recombination events are evenly dispersed across chromosomes and the gene space is remarkably compact considering the genome size, though with significant copy number variation driven by tandem duplication. Demonstrating practical application of the genome sequence, we develop a targeted genotyping ass...
Faba bean is a widely adapted and high-yielding legume cultivated for its protein-rich seeds1. Ho... more Faba bean is a widely adapted and high-yielding legume cultivated for its protein-rich seeds1. However, the seeds accumulate the anti-nutritional pyrimidine glucosides vicine and convicine, which can cause haemolytic anaemia—favism—in the 400 million individuals genetically predisposed by a deficiency in glucose-6-phosphate dehydrogenase2. Here, we identify the first enzyme associated with vicine and convicine biosynthesis, which we name VC1. We show that VC1 co-locates with the major QTL for vicine and convicine content and that the expression of VC1 correlates highly with vicine content across tissues. We also show that low-vicine varieties express a version of VC1 carrying a small, frame-shift insertion, and that overexpression of wild-type VC1 leads to an increase in vicine levels. VC1 encodes a functional GTP cyclohydrolase II, an enzyme normally involved in riboflavin biosynthesis from the purine GTP. Through feeding studies, we demonstrate that GTP is a precursor of vicine bo...
Switching perception of friend and foe Lysine motif receptors in plants perceive glycans that sig... more Switching perception of friend and foe Lysine motif receptors in plants perceive glycans that signal the presence of pathogenic or symbiotic nitrogen-fixing microbes. Bozsoki et al. now define the portions of these receptors that create the discriminatory binding pocket (see the Perspective by Bisseling and Geurts). The motifs were conserved in receptors that initiate immune responses, reflecting the invariable nature of the chitin fragments that they sense. Conversely, the motifs in receptors that respond to symbiotic signals were more varied, reflecting the greater diversity of the lipochitooligosaccharides (Nod factors) that they sense. With domain swapping, the authors switched the Nod factor specificity of receptors from two legume species and also enabled a chitin receptor that was otherwise dedicated to the detection of pathogenic microbes to instead recognize Nod factors. Science , this issue p. 663 ; see also p. 620
Publisher Summary This chapter discusses the use of Agrobacterium for the transformation of the d... more Publisher Summary This chapter discusses the use of Agrobacterium for the transformation of the diploid legume Lotus japonicas . Using the transformation–regeneration method, transgenic Lotus japonicus plants can be obtained after selection for two different resistance genes expressing hygromycin phosphotransferase and neomycin phosphotransferase. Selection for hygromycin and geneticin resistance is very effective and up to 90% of the hypocotyl explants cocultivated with Agrobacterium give rise to one or more transgenic calli. Transgenic calli are easily distinguished from the explants and are moved to regeneration media. Rooted transformed plants in pots are available after 4 months. Longer time is required to discern and separate transgenic calli from explant tissue.
Current plant science and biotechnology in agriculture, 1998
... A LOTUS F1LICAULIS X LOTUS JAPONICUS F2 POPULATION FOR GENETIC AND PHYSICAL MAPPING NN Sandal... more ... A LOTUS F1LICAULIS X LOTUS JAPONICUS F2 POPULATION FOR GENETIC AND PHYSICAL MAPPING NN Sandal and J. Stougaard Laboratory of Gene Expression, Department of Molecular and Structural Biology, University of Aarhus, Gustav Wieds Vej 10, DK-8000 ...
Uploads
Papers by Jens Stougaard