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1.
Food Chem ; 367: 130722, 2022 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-34375893

RESUMO

Lupin is slated as a potential contributor towards future food security. Lupin possesses several nutritional and nutraceutical attributes, many linked to seed proteins. For in-depth characterisation of the lupin proteome, liquid chromatography-tandem mass spectrometry was used to evaluate four protein extraction procedures. The proteomes of three narrow-leafed lupin were qualitatively evaluated using protein/peptide identifications and further quantitatively assessed by data-independent proteome measurement. Each extraction buffer led to unique protein identifications; altogether yielding 2,760 protein identifications from lupin varieties. The analysis of protein abundance data highlighted distinct differences between Tris-HCl and urea extracted proteomes, while also revealing variation amongst the cultivar proteomes with the wild accession (P27255) distinctly different from the domesticated cultivars (Tanjil, Unicrop). The extraction buffer used influenced the proteome coverage, downstream functional annotation results and consequently the biological interpretation demonstrating the need to optimise and understand the impact of protein extraction conditions.


Assuntos
Lupinus , Lupinus/genética , Espectrometria de Massas , Folhas de Planta , Proteoma , Sementes
2.
BMC Genomics ; 22(1): 723, 2021 Oct 06.
Artigo em Inglês | MEDLINE | ID: mdl-34615466

RESUMO

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.


Assuntos
Lupinus , Transportadores de Cassetes de Ligação de ATP/genética , Biologia Computacional , Lupinus/genética , Fósforo , Filogenia
3.
Molecules ; 26(14)2021 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-34299594

RESUMO

Modifying hen fodder is a common way of changing eggs composition today. However, there is no information on the effect of the source of protein in the fodder replacement on egg allergenicity. This research aimed to detect potential differences in the immunoreactivity and protein composition of eggs from hens fed with fodder containing legume. The aim of the first step of the study was to select the proper solvent for extracting allergenic proteins from hen eggs. Two of them (containing Tween 20 and Triton 100) were selected, based on protein profile and concentration analysis. Egg-white- and egg-yolk-proteins extracts prepared with them were checked for potential differences, using SDS-PAGE electrophoresis, and then the Western-blot method, using sera from children allergic to eggs and soy. Preliminary studies on the influence of fodder composition on the composition of egg proteins suggest that the addition of soy and lupine to fodder modifies the expression of egg proteins. The observed differences in the immunoreactivity of proteins contained in hen egg-white samples do not seem to be as significant as the appearance of protein with a molecular weight of ~13 kDa in the yolk of eggs obtained from soybean-fed hens. This protein may increase the immunoreactivity of eggs for children allergic solely to soy.


Assuntos
Alérgenos/imunologia , Ração Animal , Galinhas/imunologia , Proteínas do Ovo/imunologia , Ovos , Lupinus , Soja , Animais , Hipersensibilidade a Ovo/prevenção & controle , Feminino , Humanos
4.
Int J Food Microbiol ; 354: 109316, 2021 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-34247020

RESUMO

Fermented soy sauces are used as food seasonings in Eastern countries and all over the world. Depending on their cultural origins, their production differs in parameters such as wheat addition, temperature, and salt concentration. The fermentation of lupine seeds presents an alternative to the use of soybeans; however, the microbiota and influencing factors are currently unknown. In this study, we analyse the microbiota of lupine Moromi (mash) fermentations for a period of six months and determine the influence of different salt concentrations on the microbiota dynamics and the volatile compound composition. Cultured microorganisms were identified by protein profiling using matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS), and 16S rRNA gene amplicon sequencing provided an overview of the microbiota including non-cultured bacteria. The volatile compounds were determined by gas chromatography-mass spectrometry (GC-MS). At all salt concentrations, we found that Tetragenococcus halophilus (up to 1.4 × 109 colony forming units (CFU)/mL on day 21) and Chromohalobacter japonicus (1.9 × 109 CFU/mL, day 28) were the dominating bacteria during Moromi fermentation. Debaryomyces hansenii (3.6 × 108 CFU/mL, day 42) and Candida guilliermondii (2.2 × 108 CFU/mL, day 2) were found to be the most prevalent yeast species. Interestingly, Zygosaccharomyces rouxii and other yeasts described as typical for soy Moromi were not found. With increasing salinity, we found lower diversity in the microbiota, the prevalence-gain of typical species was delayed, and ratios differed depending on their halo- or acid tolerance. GC-MS analysis revealed aroma-active compounds, such as pyrazines, acids, and some furanones, which were mostly different from the aroma compounds found in soy sauce. The absence of wheat may have caused a change in yeast microbiota, and the use of lupine seeds may have led to the differing aromatic composition. Salt reduction resulted in a more complex microbiome, higher cell counts, and did not show any spoiling organisms. With these findings, we show that seasoning sauce that uses lupine seeds as the sole substrate is a suitable gluten-free, soy-free and salt reduced alternative to common soy sauces with a unique flavour.


Assuntos
Alimentos e Bebidas Fermentados , Lupinus , Microbiota , Sementes , Chromohalobacter/metabolismo , Enterococcaceae/metabolismo , Alimentos e Bebidas Fermentados/microbiologia , Microbiologia de Alimentos , Lupinus/química , Microbiota/efeitos dos fármacos , Microbiota/genética , RNA Ribossômico 16S/genética , Saccharomycetales/metabolismo , Sementes/microbiologia , Cloreto de Sódio/farmacologia
5.
Theor Appl Genet ; 134(10): 3411-3426, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34258645

RESUMO

KEY MESSAGE: A plant-specific Trimethylguanosine Synthase1-like homologue was identified as a candidate gene for the efl mutation in narrow-leafed lupin, which alters phenology by reducing vernalisation requirement. The vernalisation pathway is a key component of flowering time control in plants from temperate regions but is not well understood in the legume family. Here we examined vernalisation control in the temperate grain legume species, narrow-leafed lupin (Lupinus angustifolius L.), and discovered a candidate gene for an ethylene imine mutation (efl). The efl mutation changes phenology from late to mid-season flowering and additionally causes transformation from obligate to facultative vernalisation requirement. The efl locus was mapped to pseudochromosome NLL-10 in a recombinant inbred line (RIL) mapping population developed by accelerated single seed descent. Candidate genes were identified in the reference genome, and a diverse panel of narrow-leafed lupins was screened to validate mutations specific to accessions with efl. A non-synonymous SNP mutation within an S-adenosyl-L-methionine-dependent methyltransferase protein domain of a Trimethylguanosine Synthase1-like (TGS1) orthologue was identified as the candidate mutation giving rise to efl. This mutation caused substitution of an amino acid within an established motif at a position that is otherwise highly conserved in several plant families and was perfectly correlated with the efl phenotype in F2 and F6 genetic population and a panel of diverse accessions, including the original efl mutant. Expression of the TGS1 homologue did not differ between wild-type and efl genotypes, supporting altered functional activity of the gene product. This is the first time a TGS1 orthologue has been associated with vernalisation response and flowering time control in any plant species.


Assuntos
Flores/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Genética Populacional , Lupinus/crescimento & desenvolvimento , Metiltransferases/metabolismo , Folhas de Planta/crescimento & desenvolvimento , Proteínas de Plantas/metabolismo , Flores/genética , Lupinus/genética , Metiltransferases/genética , Mutação , Fenótipo , Filogenia , Folhas de Planta/genética , Proteínas de Plantas/genética
6.
BMC Plant Biol ; 21(1): 314, 2021 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-34215194

RESUMO

BACKGROUND: Anther dehiscence resulting in the release of pollen grains is tightly regulated in a spatiotemporal manner by various factors. In yellow lupine (Lupinus luteus L.), a species that shows cleistogamy, the anthers split before the flowers open, but the course and regulation of this process are unknown. The specific control of anther development takes place via hormonal pathways, the wide action of which ensures reproductive success. In our previous research concerning flower and early pod development in yellow lupine, we showed that the lowest transcript level of LlDELLA1, a main repressor of gibberellin (GA) signalling, occurs approximately at the time of anther opening; therefore, the main purpose of this study was to precisely investigate the gibberellic acid (GA3)-dependent regulation of the anther dehiscence in this species. RESULTS: In this paper, we showed the specific changes in the yellow lupine anther structure during dehiscence, including secondary thickening in the endothecium by lignocellulosic deposition, enzymatic cell wall breakdown at the septum/stomium and cell degeneration via programmed cell death (PCD), and identified several genes widely associated with this process. The expression profile of genes varied over time, with the most intense mRNA accumulation in the phases prior to or at the time of anther opening. The transcriptional activity also revealed that these genes are highly coexpressed and regulated in a GA-dependent manner. The cellular and tissue localization of GA3 showed that these molecules are present before anther opening, mainly in septum cells, near the vascular bundle and in the endothecium, and that they are subsequently undetectable. GA3 localization strongly correlates with the transcriptional activity of genes related to GA biosynthesis and deactivation. The results also suggest that GA3 controls LlGAMYB expression via an LlMIR159-dependent pathway. CONCLUSIONS: The presented results show a clear contribution of GA3 in the control of the extensive anther dehiscence process in yellow lupine. Understanding the processes underlying pollen release at the hormonal and molecular levels is a significant aspect of controlling fertility in this economically important legume crop species and is of increasing interest to breeders.


Assuntos
Flores/fisiologia , Giberelinas/farmacologia , Lupinus/fisiologia , Apoptose/efeitos dos fármacos , Apoptose/genética , Sequência de Bases , Parede Celular/efeitos dos fármacos , Parede Celular/genética , Simulação por Computador , Flores/efeitos dos fármacos , Flores/genética , Flores/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Giberelinas/metabolismo , Lupinus/efeitos dos fármacos , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
7.
Curr Protoc ; 1(7): e191, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34242495

RESUMO

Well-characterized genetic resources are fundamental to maintain and provide the various genotypes for pre-breeding programs for the production of new cultivars (e.g., wild relatives, unimproved material, landraces). The aim of the current article is to provide protocols for the characterization of the genetic resources of two lupin crop species: the European Lupinus albus and the American Lupinus mutabilis. Intelligent nested collections of lupins derived from homozygous lines (single-seed descent) are being developed, established, and exploited using cutting-edge approaches for genotyping, phenotyping, data management, and data analysis within the INCREASE project (EU Horizon 2020). This will allow us to predict the phenotypic performance of genotyped lines, and will further boost research and development in lupins. Lupins stand out due to their high-quality seed protein (∼40% of seed dry weight) and other primary components in the seeds, which include fatty acids, dietary fiber, and minerals. The potential of lupins as a crop is highlighted by the multiple benefits of plant-based food in terms of food security, nutrition, human health, and sustainable production. The use of lupins in foods, along with other well-studied and widely used food legumes, will also provide a greatly diversified plant-based food palette to meet the Global Goals for Sustainable Development to improve people's lives by 2030. © 2021 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Lupin seed phenotypic descriptors Basic Protocol 2: Lupin seed imaging Basic Protocol 3: Standardized phenotypic characterization of lupin genetic resources grown towards primary seed increase (development of single-seed descent genetic resources).


Assuntos
Lupinus , Fibras na Dieta , Genótipo , Humanos , Lupinus/genética , Melhoramento Vegetal , Sementes/genética
8.
Molecules ; 26(11)2021 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-34200152

RESUMO

Lupin seeds can represent a valuable source of phenolics and other antioxidant compounds. In this work, a comprehensive analysis of the phytochemical profile was performed on seeds from three Lupinus species, including one cultivar (Lupinus albus) and two wild accessions (Lupinus cossentinii and Lupinus luteus), collected from the northern region of Tunisia. Untargeted metabolomic profiling allowed to identify 249 compounds, with a great abundance of phenolics and alkaloids. In this regard, the species L. cossentinii showed the highest phenolic content, being 6.54 mg/g DW, followed by L. luteus (1.60 mg/g DW) and L. albus (1.14 mg/g DW). The in vitro antioxidant capacity measured by the ABTS assay on seed extracts ranged from 4.67 to 17.58 mg trolox equivalents (TE)/g, recording the highest values for L. albus and the lowest for L. luteus. The DPPH radical scavenging activity ranged from 0.39 to 3.50 mg TE/g. FRAP values varied between 4.11 and 5.75 mg TE/g. CUPRAC values for lupin seeds ranged from 7.20 to 8.95 mg TE/g, recording the highest for L. cossentinii. The results of phosphomolybdenum assay and metal chelation showed similarity between the three species of Lupinus. The acetylcholinesterase (AChE) inhibition activity was detected in each methanolic extract analyzed with similar results. Regarding the butyrylcholinesterase (BChE) enzyme, it was weakly inhibited by the Lupinus extracts; in particular, the highest activity values were recorded for L. albus (1.74 mg GALAE/g). Overall, our results showed that L. cossentinii was the most abundant source of polyphenols, consisting mainly in tyrosol equivalents (5.82 mg/g DW). Finally, significant correlations were outlined between the phenolic compounds and the in vitro biological activity measured, particularly when considering flavones, phenolic acids and lower-molecular-weight phenolics.


Assuntos
Antioxidantes/química , Lupinus/química , Compostos Fitoquímicos/química , Sementes/química , Alcaloides/química , Alcaloides/metabolismo , Antioxidantes/metabolismo , Lupinus/metabolismo , Metabolômica/métodos , Fenóis/química , Fenóis/metabolismo , Compostos Fitoquímicos/metabolismo , Extratos Vegetais/química , Extratos Vegetais/metabolismo , Sementes/metabolismo , Tunísia
9.
Plant Physiol Biochem ; 166: 531-539, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34174658

RESUMO

Phosphorus (P) deficiency largely restricts plant growth and lead to severe yield losses. Therefore, identification of novel root traits to improve P uptake is needed to circumvent yield losses. White lupin (Lupinus albus) is a legume crop that develops cluster roots and has the high phosphorus use efficiency in low P soils. We aimed to investigate the association between cluster roots (CR) rhizosheath formation and P uptake in white lupin. Rhizosheath formation and P concentration were evaluated under four soil treatments. CR increased up to 2.5-fold of overall plant dry weight under SD-P compared to WW + P (control), partly attributable to variations in CR development. Our data showed that SD-P significantly increase rhizosheath weight in white lupin. Among the root segments, MCR showed improved P accumulation in the root which is associated with increased MCR rhizosheath weight. Additionally, a positive correlation was observed between MCR rhizosheath weight and P uptake. Moreover, high sucrose content was recorded in MCR, which may contribute in CR growth under SD-P. Expression analysis of genes related to sucrose accumulation (LaSUC1, LaSUC5, and LaSUC9) and phosphorus uptake (LaSPX3, LaPHO1, and LaPHT1) exhibited peaked expression in MCR under SD-P. This indicate that root sucrose status may facilitate P uptake under P starvation. Together, the ability to enhance P uptake of white lupin is largely associated with MCR rhizosheath under SD-P. Our results showed that gene expression modulation of CR forming plant species, demonstrating that these novel root structures may play crucial role in P acquisition from the soil. Our findings could be implicated for developing P and water efficient crop via CR development in sustainable agriculture.


Assuntos
Lupinus , Transporte Biológico , Lupinus/genética , Fósforo , Raízes de Plantas , Solo
10.
Sci Total Environ ; 788: 147622, 2021 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-34034171

RESUMO

Mine tailings pose physical and chemical challenges for plant establishment. Our aim was to learn from natural processes in long-term soil and ecosystem development to use tailings as novel parent materials and pioneer ecological-engineering plant species to ameliorate extreme conditions of tailings, and facilitate the establishment of subsequent native plants. A glasshouse trial was conducted using magnetite tailings containing various amendments, investigating the potential of the nitrogen (N)-fixing, non-native pioneer species Lupinus angustifolius (Fabaceae), narrow-leaf lupin, as a potential eco-engineer to promote soil formation processes, and whether amendment type or the presence of pioneer vegetation improved the subsequent establishment and growth of 40 species of native plants. We found that L. angustifolius eco-engineered the mine tailings, by enhancing the N status of tailings and mobilising primary mineral P into organic P via a carboxylate-exudation strategy, thereby enabling subsequent growth of native species. The substantial increases of the soil organic P (from ca. 10 to 150 mg kg-1) pool and organo-bound Al minerals (from 0 to 2 mg kg-1) were particularly evident, indicating the initiation of pedogenesis in mine tailings. Our findings suggest that the annual legume L. angustifolius has eco-engineering potential on mine tailings through N-fixation and P-mobilisation, promoting the subsequent growth of native plants. We proposed Daviesia (Fabaceae) species as native species alternatives for the non-native L. angustifolius in the Western Australian context. Our findings are important for restoration practitioners tasked with mine site restoration in terms of screening pioneer eco-engineering plant species, where native plants are required to restore after mine operations.


Assuntos
Lupinus , Poluentes do Solo , Austrália , Ecossistema , Óxido Ferroso-Férrico , Folhas de Planta/química , Solo , Poluentes do Solo/análise
11.
Int J Mol Sci ; 22(8)2021 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-33917799

RESUMO

White lupin (Lupinus albus L.) is a pulse annual plant cultivated from the tropics to temperate regions for its high-protein grain as well as a cover crop or green manure. Wild populations are typically late flowering and have high vernalization requirements. Nevertheless, some early flowering and thermoneutral accessions were found in the Mediterranean basin. Recently, quantitative trait loci (QTLs) explaining flowering time variance were identified in bi-parental population mapping, however, phenotypic and genotypic diversity in the world collection has not been addressed yet. In this study, a diverse set of white lupin accessions (n = 160) was phenotyped for time to flowering in a controlled environment and genotyped with PCR-based markers (n = 50) tagging major QTLs and selected homologs of photoperiod and vernalization pathway genes. This survey highlighted quantitative control of flowering time in white lupin, providing statistically significant associations for all major QTLs and numerous regulatory genes, including white lupin homologs of CONSTANS, FLOWERING LOCUS T, FY, MOTHER OF FT AND TFL1, PHYTOCHROME INTERACTING FACTOR 4, SKI-INTERACTING PROTEIN 1, and VERNALIZATION INDEPENDENCE 3. This revealed the complexity of flowering control in white lupin, dispersed among numerous loci localized on several chromosomes, provided economic justification for future genome-wide association studies or genomic selection rather than relying on simple marker-assisted selection.


Assuntos
Flores/fisiologia , Lupinus/fisiologia , Desenvolvimento Vegetal , Locos de Características Quantitativas , Característica Quantitativa Herdável , Fatores de Tempo
12.
Int J Mol Sci ; 22(6)2021 Mar 16.
Artigo em Inglês | MEDLINE | ID: mdl-33809409

RESUMO

Yellow lupine is a great model for abscission-related research given that excessive flower abortion reduces its yield. It has been previously shown that the EPIP peptide, a fragment of LlIDL (INFLORESCENCE DEFICIENT IN ABSCISSION) amino-acid sequence, is a sufficient molecule to induce flower abortion, however, the question remains: What are the exact changes evoked by this peptide locally in abscission zone (AZ) cells? Therefore, we used EPIP peptide to monitor specific modifications accompanied by early steps of flower abscission directly in the AZ. EPIP stimulates the downstream elements of the pathway-HAESA and MITOGEN-ACTIVATED PROTEIN KINASE6 and induces cellular symptoms indicating AZ activation. The EPIP treatment disrupts redox homeostasis, involving the accumulation of H2O2 and upregulation of the enzymatic antioxidant system including superoxide dismutase, catalase, and ascorbate peroxidase. A weakening of the cell wall structure in response to EPIP is reflected by pectin demethylation, while a changing pattern of fatty acids and acyl lipids composition suggests a modification of lipid metabolism. Notably, the formation of a signaling molecule-phosphatidic acid is induced locally in EPIP-treated AZ. Collectively, all these changes indicate the switching of several metabolic and signaling pathways directly in the AZ in response to EPIP, which inevitably leads to flower abscission.


Assuntos
Flores/crescimento & desenvolvimento , Homeostase , Lipídeos/química , Lupinus/crescimento & desenvolvimento , Pectinas/metabolismo , Peptídeos/farmacologia , Parede Celular/efeitos dos fármacos , Parede Celular/metabolismo , Flores/efeitos dos fármacos , Homeostase/efeitos dos fármacos , Peróxido de Hidrogênio/metabolismo , Lupinus/efeitos dos fármacos , Oxirredução , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Transdução de Sinais/efeitos dos fármacos , Superóxido Dismutase/metabolismo
13.
Nutrients ; 13(3)2021 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-33808034

RESUMO

P5 (LILPKHSDAD) is a hypocholesterolemic peptide from lupin protein with a multi-target activity, since it inhibits both 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCoAR) and proprotein convertase subtilisin/kexin type-9 (PCSK9). This work shows that, during epithelial transport experiments, the metabolic transformation mediated by intestinal peptidases produces two main detected peptides, ILPKHSDAD (P5-frag) and LPKHSDAD (P5-met), and that both P5 and P5-met are linearly absorbed by differentiated human intestinal Caco-2 cells. Extensive comparative structural, biochemical, and cellular characterizations of P5-met and the parent peptide P5 demonstrate that both peptides have unique characteristics and share the same mechanisms of action. In fact, they exert an intrinsically multi-target behavior being able to regulate cholesterol metabolism by modulating different pathways. The results of this study also highlight the dynamic nature of bioactive peptides that may be modulated by the biological systems they get in contact with.


Assuntos
Transporte Biológico/efeitos dos fármacos , Metabolismo dos Lipídeos/efeitos dos fármacos , Lupinus/química , Peptídeos/farmacocinética , Proteínas de Plantas/farmacocinética , Células CACO-2 , Células Hep G2 , Humanos , Hidroximetilglutaril-CoA Redutases/metabolismo , Mucosa Intestinal/metabolismo , Pró-Proteína Convertase 9/metabolismo
14.
Int J Mol Sci ; 22(5)2021 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-33800929

RESUMO

The main restraint obstructing the wider adoption of lupins as protein crops is the presence of bitter and toxic quinolizidine alkaloids (QAs), whose contents might increase under exposure to stressful environmental conditions. A poor understanding of how QAs accumulate hinders the breeding of sweet varieties. Here, we characterize the expression profiles of QA-related genes, along with the alkaloid content, in various organs of sweet and bitter narrow-leafed lupin (NLL, Lupinus angustifolius L.). Special attention is paid to the RAP2-7 transcription factor, a candidate regulator of the QA pathway. We demonstrate the upregulation of RAP2-7 and other QA-related genes, across the aerial organs of a bitter cultivar and the significant correlations between their expression levels, thus supporting the role of RAP2-7 as an important regulatory gene in NLL. Moreover, we showed that the initial steps of QA synthesis might occur independently in all aerial plant organs sharing common regulatory mechanisms. Nonetheless, other regulatory steps might be involved in RAP2-7-triggered QA accumulation, given its expression pattern in leaves. Finally, the examination of QA-related gene expression in plants infected with Colletotrichum lupini evidenced no connection between QA synthesis and anthracnose resistance, in contrast to the important role of polyamines during plant-pathogen interactions.


Assuntos
Colletotrichum/fisiologia , Regulação da Expressão Gênica de Plantas , Lupinus/genética , Doenças das Plantas/genética , Quinolizidinas/metabolismo , Cromatografia Gasosa , Lupinus/metabolismo , Lupinus/microbiologia , Especificidade de Órgãos , Melhoramento Vegetal , Doenças das Plantas/microbiologia , Proteínas de Plantas/biossíntese , Proteínas de Plantas/genética , Estruturas Vegetais/metabolismo , Estruturas Vegetais/microbiologia , Poliaminas/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Transcrição/biossíntese , Fatores de Transcrição/genética
15.
Molecules ; 26(6)2021 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-33808718

RESUMO

This study addresses an innovative approach to generate aerated foods with appealing texture through the utilization of lupin protein isolate (LPI) in combination with edible fats. We show the impact of transglutaminases (TGs; SB6 and commercial), glycerol (Gly), soy lecithin (Lec) and linoleic acid (LA) on the micro- and nanostructure of health promoting solid foods created from LPI and fats blends. 3-D tomographic images of LPI with TG revealed that SB6 contributed to an exceptional bubble spatial organization. The inclusion of Gly and Lec decreased protein polymerization and also induced the formation of a porous layered material. LA promoted protein polymerization and formation of homogeneous thick layers in the LPI matrix. Thus, the LPI is a promising protein resource which when in blend with additives is able to create diverse food structures. Much focus has been placed on the great foamability of LPI and here we show the resulting microstructure of LPI foams, and how these were improved with addition of TGs. New food applications for LPI can arise with the addition of food grade dispersant Lec and essential fatty-acid LA, by improved puffiness, and their contributing as replacer of chemical leavening additives in gluten-free products.


Assuntos
Gorduras na Dieta , Congelamento , Lupinus/química , Proteínas de Plantas/química , Transglutaminases/química
16.
Food Res Int ; 140: 110046, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33648271

RESUMO

Lupinus albus or white lupine has recently received increase attention for its medicinal values. Several studies have described the hypoglycemic effect of the white lupine, which is known as a food plant with potential value for treatment of diabetes. This study provides useful information for the identification and quantification of compounds in L. albus fractions by proton nuclear magnetic resonance (1H NMR) spectroscopy. In total, 35 metabolites were identified from L. albus fractions.Principal component analysis (PCA) was used as a multivariate projection method for visualizing the different composition of four different fractions. The bioactivities of fractions with different polarity obtained from the extract of L. albus seeds are reported. Among the fractions studied, the chloroform fraction (CF) exhibits a high free radical scavenging (DPPH) and α-glucosidase inhibitory activities with IC50 values of 24.08 and 20.08 µg/mL, respectively. A partial least-squares analyses (PLS) model had been successfully performed to correlate the potential active metabolites with the corresponding biological activities. Metabolites containing proline, caprate, asparagine, lupinoisolone C, hydroxyiso lupalbigenin and some unknown compounds show high correlation with the bioactivities studied. Moreover, the structural identification in the active fraction was supported by ultrahigh-performance-liquid chromatography-electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS) analysis. A total of 21 metabolites were tentatively identified from MS/MS data by comparison with previously reported data. Most of these compounds are isoflavonoids without known biological activity. This information may be useful for developing functional food from L. albus with potential application in the management of diabetes.


Assuntos
Lupinus , Espectrometria de Massas em Tandem , Cromatografia Líquida de Alta Pressão , Espectroscopia de Ressonância Magnética , Metabolômica , Extratos Vegetais , Espectroscopia de Prótons por Ressonância Magnética
17.
Food Chem ; 354: 129473, 2021 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-33743449

RESUMO

Vegetable proteins are appearing as a sustainable source for human consumption. Food-derived peptides are an important field of research in terms of bioactive molecules. In this study, seven vegetable proteins were enzymatically hydrolysed following an optimised treatment (sequential hydrolysis with subtilisin-trypsin-flavourzyme) to obtain dipeptidyl peptidase IV (DPP-IV) inhibitory peptides. Hydrolysates were fractionated by size exclusion chromatography and, from the most bioactive fractions (corresponding to Glycine max, Chenopodium quinoa and Lupinus albus proteins); peptides responsible for this bioactivity were identified by mass spectrometry. Peptides with adequate molecular features and based on in silico analysis were proposed as DPP-IV inhibitors from soy (EPAAV) lupine (NPLL), and quinoa (APFTVV). These vegetable protein sources are adequate to obtain protein hydrolysates for functional food.


Assuntos
Dipeptidil Peptidase 4/química , Inibidores da Dipeptidil Peptidase IV/química , Peptídeos/química , Proteínas de Vegetais Comestíveis/metabolismo , Animais , Chenopodium quinoa/metabolismo , Cromatografia em Gel , Dipeptidil Peptidase 4/metabolismo , Inibidores da Dipeptidil Peptidase IV/metabolismo , Humanos , Hidrólise , Lupinus/metabolismo , Espectrometria de Massas , Peptídeos/metabolismo , Proteínas de Vegetais Comestíveis/química , Soja/metabolismo
18.
Food Chem ; 354: 129480, 2021 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-33765465

RESUMO

A multidimensional analysis aimed to determine the thermal impact on γ-conglutin at the two oligomeric states was carried out. A wide range of biophysical and bioinformatic methods allowed to get insight into a thermal unfolding mechanism. The determined midpoint transition temperature (Tm) values were remarkably different, being 56.5 °C and 71.1 °C for γ-conglutin monomer and hexamer, respectively. The unfolding pattern for hexamer molecules included aggregation/precipitation, while monomers tended to form soluble aggregates after heat exposure. Interestingly, differences in the aromatic amino acid residues movements indicate that during thermal treatment of γ-conglutin hexamer red-shift occurred contrary to the monomer in the case of which blue-shift was noted. The obtained results provide an essential contribution to expand our knowledge about the molecular characterization of this intriguing lupin seed protein.


Assuntos
Lupinus/metabolismo , Proteínas de Plantas/química , Varredura Diferencial de Calorimetria , Dicroísmo Circular , Concentração de Íons de Hidrogênio , Proteínas de Plantas/metabolismo , Estabilidade Proteica , Sais/química , Sementes/metabolismo , Temperatura
19.
J Chromatogr A ; 1642: 462027, 2021 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-33714772

RESUMO

Phytotoxins are plant secondary metabolites. They have recently been considered as chemicals of emerging concern (CECs) and there is a growing interest in their environmental fate and potential threat to public health. Dedicated target and non-target screening (NTS) analysis of phytotoxins in environmental samples are sparse, meanwhile phytotoxins are rarely detected in NTS-based analysis due to lack of an efficient methodology. Development of new analytical measurement methods is therefore highly needed. In this study, we for the first time investigated key parameters of reversed phase liquid chromatography-high resolution mass spectrometry (RPLC-HRMS) for five major classes of phytotoxins (alkaloids, steroids, terpenoids, flavonoids and aromatic polyketides) in environmental matrices; the investigation included analytical conditions which have not yet been explored by others, e.g. ionization at alkaline pH above 9. As the outcome we established a new analytical method for target/suspect screening and NTS of phytotoxins in the environment, which significantly improved the detection sensitivity with up to 40 times compared to previous methods, and enabled the discovery of over 30 phytotoxins in a NTS-based environmental study. We also observed that the negative ionization of phenols could be facilitated by the number of hydroxyl groups on the ring rather than their position of substitution. This study is of interest for a better fundamental understanding of the behavior of phytotoxins in LC-MS. Dedicated target/suspect screening and NTS methods will facilitate a better risk characterization of phytotoxins in the environment and stimulate implementation of new public regulation on phytotoxins.


Assuntos
Poluentes Ambientais/química , Espectrometria de Massas/métodos , Toxinas Biológicas/química , Cromatografia Líquida , Cromatografia de Fase Reversa , Concentração de Íons de Hidrogênio , Lupinus/química , Fenóis/análise , Padrões de Referência , Solo/química , Espectrometria de Massas por Ionização por Electrospray , Temperatura , Fatores de Tempo , Toxinas Biológicas/análise , Água/química
20.
Ars pharm ; 62(1): 52-65, ene.-mar. 2021. tab, graf
Artigo em Inglês | IBECS | ID: ibc-199700

RESUMO

INTRODUCTION: The prevalence of diabetes type 2 is increasing worldwide, thus the search of novel alternative ther¬apies is needed. According to their traditional use, we selected five Bolivian plants Chenopodium quinoa (CQ) Ama¬ranthus caudatus (AC), Chenopodium pallidicaule (CP), Lupinus mutabilis (LM) and Smallanthus sonchifolius (SS) that are traditionally used to control glycemia. METHODS: The effect of a single oral administration of Ethanolic (EtOH), hydro-ethanolic (EtOH70) and aqueous (Aq) extracts from all plant species were tested for their effect on blood glucose in non-fasted mice and during the oral glucose tolerance test (OGTT). The effect on insulin secretion was evaluated in mice pancreatic islets. RESULTS: EtOH70 extracts of all the plants showed glucose-reducing effect at the highest dose evaluated (2000 mg/ kg b.w.). EtOH70 extracts improved the glucose tolerance evaluated by the OGTT in mice fasted for 12 hours. The extracts have different effects on glucose homeostasis since just extracts of AC, LM and CQ but not CP and SS in¬creased insulin secretion as shown on mice pancreatic islets. The phytochemical qualitative characterization of EtOH70 extracts detected phenolic acids and flavonoids in AC, CP and CQ; alkaloids in LM and anthocyanidins in SS. None of EtOH70 extracts tested showed in vitro or in vivo acute toxicity at concentrations where they exhibit glucose lowering effects. CONCLUSIONS: We report here that extracts from AC, CQ, CP, LM and SS exhibit glucose lowering effect while just AC, CQ and LM stimulate directly the insulin secretion


INTRODUCCIÓN: La prevalencia de diabetes tipo 2 está aumentando en todo el mundo, por lo que se necesita la búsqueda de nuevas terapias alternativas. Según su uso tradicional, seleccionamos cinco plantas bolivianas Chenopodium quinoa (CQ) Amaranthus caudatus (AC), Chenopodium pallidicaule (CP), Lupinus mutabilis (LM) y Smallanthus sonchifolius (SS) que se usan tradicionalmente para controlar la glucemia. MÉTODOS: Se evaluó el efecto de la administración oral única de extractos etanólicos (EtOH), hidroetanólicos (EtOH70) y acuosos (Aq) de las plantas mencionadas para determinar su efecto sobre la glucosa en sangre en ratones en o sin ayunas y durante la prueba de tolerancia a la glucosa oral (PTGO). El efecto sobre la secreción de insulina se evaluó en islotes pancreáticos de ratones. RESULTADOS: Los extractos de EtOH70 de todas las plantas disminuyeron la glucemia a la dosis más alta evaluada (2000 mg / kg b.w.). Los extractos de EtOH70 mejoraron la tolerancia a la glucosa evaluada mediante la PTGO en ratones con ayuno de 12 horas. Los extractos tienen diferentes efectos sobre la homeostasis de la glucosa, ya que solo los extractos de AC, LM y CQ pero no CP y SS aumentaron la secreción de insulina como se muestra en los islotes pancreáticos de los ratones. La caracterización cualitativa fitoquímica de extractos de EtOH70 detectó ácidos fenólicos y flavonoides en AC, CP y CQ, alcaloides en LM y antocianidinas en SS. Ninguno de los extractos de EtOH70 probados mostró toxicidad aguda in vitro o in vivo a concentraciones en las que exhiben efectos reductores de glucosa. CONCLUSIÓN: Los extractos de AC, CQ, CP, LM y SS exhiben un efecto reductor de la glucosa, mientras que solo AC, CQ y LM estimulan directamente la secreción de insulina


Assuntos
Animais , Masculino , Camundongos , Glicemia/efeitos dos fármacos , Diabetes Mellitus Tipo 2/tratamento farmacológico , Diabetes Mellitus Experimental/tratamento farmacológico , Secreção de Insulina/efeitos dos fármacos , Extratos Vegetais/farmacologia , Suplementos Nutricionais , Chenopodium quinoa/química , Amaranthus/química , Chenopodium/química , Lupinus/química , Ilhotas Pancreáticas/efeitos dos fármacos , Fatores de Tempo , Valores de Referência , Reprodutibilidade dos Testes , Bolívia , Teste de Tolerância a Glucose
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