Genome-wide association study reveals white lupin candidate gene involved in anthracnose resistance

Key message GWAS identifies candidate gene controlling resistance to anthracnose disease in white lupin. Abstract White lupin (Lupinus albus L.) is a promising grain legume to meet the growing demand for plant-based protein. Its cultivation, however, is severely threatened by anthracnose disease caused by the fungal pathogen Colletotrichum lupini. To dissect the genetic architecture for anthracnose resistance, genotyping by sequencing was performed on white lupin accessions collected from the center of domestication and traditional cultivation regions. GBS resulted in 4611 high-quality single-nucleotide polymorphisms (SNPs) for 181 accessions, which were combined with resistance data observed under controlled conditions to perform a genome-wide association study (GWAS). Obtained disease phenotypes were shown to highly correlate with overall three-year disease assessments under Swiss field conditions (r > 0.8). GWAS results identified two significant SNPs associated with anthracnose resistance on gene Lalb_Chr05_g0216161 encoding a RING zinc-finger E3 ubiquitin ligase which is potentially involved in plant immunity. Population analysis showed a remarkably fast linkage disequilibrium decay, weak population structure and grouping of commercial varieties with landraces, corresponding to the slow domestication history and scarcity of modern breeding efforts in white lupin. Together with 15 highly resistant accessions identified in the resistance assay, our findings show promise for further crop improvement. This study provides the basis for marker-assisted selection, genomic prediction and studies aimed at understanding anthracnose resistance mechanisms in white lupin and contributes to improving breeding programs worldwide.

Adaptive Mechanisms Make Lupin a Choice Crop for Acidic Soils Affected by Aluminum Toxicity

Almost half of the world’s agricultural soils are acidic, and most of them present significant levels of aluminum (Al) contamination, with Al3+ as the prevailing phytotoxic species. Lupin is a protein crop that is considered as an optimal alternative to soybean cultivation in cold climates. Lupins establish symbiosis with certain soil bacteria, collectively known as rhizobia, which are capable of fixing atmospheric nitrogen. Moreover, some lupin species, especially white lupin, form cluster roots, bottlebrush-like structures specialized in the mobilization and uptake of nutrients in poor soils. Cluster roots are also induced by Al toxicity. They exude phenolic compounds and organic acids that chelate Al to form non-phytotoxic complexes in the rhizosphere and inside the root cells, where Al complexes are accumulated in the vacuole. Lupins flourish in highly acidic soils where most crops, including other legumes, are unable to grow. Some lupin response mechanisms to Al toxicity are common to other plants, but lupin presents specific tolerance mechanisms, partly as a result of the formation of cluster roots. Al-induced lupin organic acid secretion differs from P-induced secretion, and organic acid transporters functions differ from those in other legumes. Additionally, symbiotic rhizobia can contribute to Al detoxification. After revising the existing knowledge on lupin distinct Al tolerance mechanisms, we conclude that further research is required to elucidate the specific organic acid secretion and Al accumulation mechanisms in this unique legume, but definitely, white lupin arises as a choice crop for cultivation in Al-rich acidic soils in temperate climate regions.

Development and evaluation of potential functional food biscuits made from White Lupin

Potential functional food bakery products were developed and characterized based on White Lupin (Lupinus albus cv. Nelly) flour. Analytical properties of the seeds resemble to previously described Lupinus species, with significantly high protein content (45%). The high protein and dietetic fiber content of the seeds makes Lupin flour suitable to develop potential functional food products with high nutritional values. Results of the development of sweet biscuits and salty crackers enriched with Lupin flour are presented. Sensory evaluation of the bakery products was carried out by 15 panelists using the nine points hedonic scale. Heat stability of White Lupin proteins were investigated by gel-electrophoretic analysis, White Lupin proteins are quite stable at 140°C, after 35 min heating the biscuits still contain 69% of the original amount of proteins. Baking conditions were optimized also based on gel-electrophoretic experiments, the optimal baking time was 30 min at 140°C. Gluten-free Lupin-based biscuits and crackers were produced by completely omitting wheat flour from the recipes.

Mediterranean White Lupin Landraces as a Valuable Genetic Reserve for Breeding

Legumes crops are important for sustainable agriculture and global food security. Among them white lupin (Lupinus albus L.), is characterized by exceptional protein content of high nutritional value, competitive to that of soybean (Glycine max) and is well adapted to rainfed agriculture. However, its high seed-quinolizidine alkaloid (QA) content impedes its direct integration to human diet and animal feed. Additionally, its cultivation is not yet intensive, remains confined to local communities and marginal lands in Mediterranean agriculture, while adaptation to local microclimates restrains its cultivation from expanding globally. Hence, modern white lupin breeding aims to exploit genetic resources for the development of “sweet” elite cultivars, resilient to biotic adversities and well adapted for cultivation on a global level. Towards this aim, we evaluated white lupin local landrace germplasm from Greece, since the country is considered a center of white lupin diversity, along with cultivars and breeding lines for comparison. Seed morphological diversity and molecular genetic relationships were investigated. Most of the landraces were distinct from cultivars, indicating the uniqueness of their genetic make-up. The presence of pauper “sweet” marker allele linked to low seed QA content in some varieties was detected in one landrace, two breeding lines, and the cultivars. However, QA content in the examined genotypes did not relate with the marker profile, indicating that the marker’s predictive power is limited in this material. Marker alleles for vernalization unresponsiveness were detected in eight landraces and alleles for anthracnose resistance were found in two landraces, pointing to the presence of promising germplasm for utilization in white lupin breeding. The rich lupin local germplasm genetic diversity and the distinct genotypic composition compared to elite cultivars, highlights its potential use as a source of important agronomic traits to support current breeding efforts and assist its integration to modern sustainable agriculture.

Identification of an Isoflavonoid Transporter Required for the Nodule Establishment of the Rhizobium-Fabaceae Symbiotic Interaction

Nitrogen (N) as well as Phosphorus (P) are key nutrients determining crop productivity. Legumes have developed strategies to overcome nutrient limitation by, for example, forming a symbiotic relationship with N-fixing rhizobia and the release of P-mobilizing exudates and are thus able to grow without supply of N or P fertilizers. The legume-rhizobial symbiosis starts with root release of isoflavonoids that act as signaling molecules perceived by compatible bacteria. Subsequently, bacteria release nod factors, which induce signaling cascades allowing the formation of functional N-fixing nodules. We report here the identification and functional characterization of a plasma membrane-localized MATE-type transporter (LaMATE2) involved in the release of genistein from white lupin roots. The LaMATE2 expression in the root is upregulated under N deficiency as well as low phosphate availability, two nutritional deficiencies that induce the release of this isoflavonoid. LaMATE2 silencing reduced genistein efflux and even more the formation of symbiotic nodules, supporting the crucial role of LaMATE2 in isoflavonoid release and nodulation. Furthermore, silencing of LaMATE2 limited the P-solubilization activity of lupin root exudates. Transport assays in yeast vesicles demonstrated that LaMATE2 acts as a proton-driven isoflavonoid transporter.

Identification of ABC transporter G subfamily in white lupin and functional characterization of L.albABGC29 in phosphorus use

Background White lupin (Lupinus albus) is a leguminous crop with elite adaptive ability in phosphorus-deficient soil and used as a model plant for studying phosphorus (P) use. However, the genetic basis of its adaptation to low P (LP) remains unclear. ATPase binding cassette (ABC) transports G subfamily play a crucial role in the transportation of biological molecules across the membrane. To date, identification of this subfamily has been analyzed in some plants, but no systematic analysis of these transporters in phosphorus acquisition is available for white lupin. Results This study identified 66 ABCG gene family members in the white lupin genome using comprehensive approaches. Phylogenetic analysis of white lupin ABCG transporters revealed six subclades based on their counterparts in Arabidopsis, displaying distinct gene structure and motif distribution in each cluster. Influences of the whole genome duplication on the evolution of L.albABCGs were investigated in detail. Segmental duplications appear to be the major driving force for the expansion of ABCGs in white lupin. Analysis of the Ka/Ks ratios indicated that the paralogs of the L.albABCG subfamily members principally underwent purifying selection. However, it was found that L.albABCG29 was a result of both tandem and segmental duplications. Overexpression of L.albABCG29 in white lupin hairy root enhanced P accumulation in cluster root under LP and improved plant growth. Histochemical GUS staining indicated that L.albABCG29 expression increased under LP in white lupin roots. Further, overexpression of L.albABCG29 in rice significantly improved P use under combined soil drying and LP by improving root growth associated with increased rhizosheath formation. Conclusion Through systematic and comprehensive genome-wide bioinformatics analysis, including conserved domain, gene structures, chromosomal distribution, phylogenetic relationships, and gene duplication analysis, the L.albABCG subfamily was identified in white lupin, and L.albABCG29 characterized in detail. In summary, our results provide deep insight into the characterization of the L.albABCG subfamily and the role of L.albABCG29 in improving P use.

CHEMICAL COMPOSITION OF SEEDS OF PLANTS LUPINUS ANGUSTIFOLIUS L. AND LUPINUS ALBUS L. OF THE KALININGRAD REGION

The aim of the work was to study the chemical composition of seeds of plants Lupinus albus L., Lupinus angustifolius L., growing in the Kaliningrad region. Lupin is a promising crop due to the high content of biologically valuable protein in its seeds, as well as minerals, unsaturated fatty acids, water - and fat-soluble biologically active substances. The paper considers the amino acid, mineral, and fatty acid composition of seeds, presents data on the content of water-soluble vitamins in them, and shows the component composition of chloroform extracts of seeds. The total number of amino acids (47.72±0.37 and 38.655±0.32%, respectively) was found in the seeds of narrow-leaved and white lupine, and a high content of glutamic acid and asparagine was found. The macronutrient composition, which is dominated by potassium, phosphorus and calcium, was studied. Among the microelements of seeds, manganese, iron, zinc and silicon predominate. A particularly high content of manganese in white lupin seeds (192.14±1.67 mg per 100 g) was noted. Analysis of the fatty acid composition of lupin seed oils showed a high degree of unsaturation with a high content of oleic and linoleic acids. Both types of lupine contain thiamine, pyridoxine, and niacin, while white lupine seeds also contain Pantothenic and folic acids. Сhromatography-mass spectrometry analysis of the fraction isolated by chloroform from lupine seeds showed the presence of substances of the terpenes class (farnesene, neofitadiene, γ-tocopherol) and steriodic alcohols (campesterol, β-sitosterol).

Synthesis and Characterization of Nano Fe and Mn (hydr)oxides to Be Used as Natural Sorbents and Micronutrient Fertilizers

Fe and Mn (hydr)oxides are widely used as contaminant sorbents in water/wastewater systems but their potential use as micronutrient fertilizers is still poorly known. In this research, four nano-metal (hydr)oxides (amorphous Mn oxide (AMO), Fe-Mn binary oxide (FMBO), two-line ferrihydrite (2L-Fh) and goethite) were successfully synthesized and completely characterized (infrared and Mössbauer spectroscopy, X-ray diffraction particle size, specific surface area, point of zero charge). AMO, FMBO and 2L-Fh were introduced to interact with AgNO3 (20.0 µM) and TlNO3 (100.0 µM) diluted solutions for three days to check their potential capability as potential Ag+ and Tl+ adsorbents. AMO and FMBO (4% w/w) were tested as nanofertilizers by arranging a hydroponic bioassay for 35 days on white lupin culture as a Mn-hyperaccumulator plant model. AMO structure was identified as an amorphous mixture of Mn oxides while FMBO was an Fe dopped birnessite. Both materials were efficient in extracting Ag+ and Tl+ although large Mn concentration was released from FMBO to the solutions. AMO and FMBO promoted Fe and Mn nutrition in plants. Synthetic iron chelate (Fe-EDDHA), present in the nutrient dissolution, could be adsorbed onto AMO surface by producing Fe and Mn accumulation in roots and increasing Mn uptake rate without toxicity symptoms. Therefore, AMO and FMBO not only demonstrated their efficiency as adsorbents, but also displayed they would be promising nanomaterials as micronutrient fertilizers.

Nodulated White Lupin Plants Growing in Contaminated Soils Accumulate Unusually High Mercury Concentrations in Their Nodules, Roots and Especially Cluster Roots

Two white lupin (Lupinus albus L.) cultivars were tested for their capacity to accumulate mercury when grown in Hg-contaminated soils. Plants inoculated with a Bradyrhizobium canariense Hg-tolerant strain or non-inoculated were grown in two highly Hg-contaminated soils. All plants were nodulated and presented a large number of cluster roots. They accumulated up to 600 μg Hg g−1 DW in nodules, 1400 μg Hg g−1 DW in roots and 2550 μg Hg g−1 DW in cluster roots. Soil, and not cultivar or inoculation, was accountable for statistically significant differences. No Hg translocation to leaves or seeds took place. Inoculated L. albus cv. G1 plants were grown hydroponically under cluster root-promoting conditions in the presence of Hg. They accumulated about 500 μg Hg g−1 DW in nodules and roots and up to 1300 μg Hg g−1 DW in cluster roots. No translocation to the aerial parts occurred. Bioaccumulation factors were also extremely high, especially in soils and particularly in cluster roots. To our knowledge, Hg accumulation in cluster roots has not been reported to date. Our results suggest that inoculated white lupin might represent a powerful phytoremediation tool through rhizosequestration of Hg in contaminated soils. Potential uptake and immobilization mechanisms are discussed.