Search

A deficit in protein intake can severely halt children growth, and is associated with both structural and functional pathologies of the brain, as well as increase the risk of contracting infectious diseases. Protein-enriched defatted meals from sacha inchi (Plukenetia volubilis) (sdSI) and chocho (Lupinus mutabilis) (sdCH) were obtained by a solvent defatting procedure. The concentration of protein was higher in sdSI (64%) than in sdCH (60%). Lysine and tryptophan were the limiting amino acid (AA) in each case. However, the AA profile of the protein meals met daily intake FAO requirements for adults, while their blends met the requirements for infants. The sdSI had an in vitro digestibility of 100. Regarding functional properties, sdSI presented higher values for water absorption index (3.5), emulsifying activity (56%), and foaming capacity (167%), while the sdCH had a slightly better oil absorption capacity (2.8). However, when mixing sdSI with sdCH some improvements in functional properties were observed. The nutritional and functional properties of the mixtures of meals can be used to develop protein-enriched foods and beverages.

Andean lupin (Lupinus mutabilis) seeds are appreciated for their high protein and lipid contents and have potential applications as ingredients in food, cosmetic, and pharmaceutical industries. Nevertheless, the information about the seed composition (especially in lipophilic antioxidants) of ecotypes from distinct cropping areas is currently limited. Thus, the aim of the present research was to assess the morphological characteristics, chemical composition, tocopherol and carotenoid contents of the seeds of 33 Andean lupin ecotypes from different Peruvian regions, along with three L. albus, one L. angustifolius and one L. luteus controls. Significant differences were noted among the Andean ecotypes for all analyzed features. The protein, lipid and ash contents were 32.0-46.9, 13.6-18.6 and 2.7-4.4 g/100 g dry matter (DM), respectively. The seeds were rich in tocopherols (172.1-249.8 mg/kg DM; γ-tocopherol was 98% of total tocols) and low in carotenoids (0.69-2.89 mg/kg DM). Debittering increased the tocopherol content (227.0-378.2 mg/kg DM), probably because of the soluble components loss, although the carotenoid concentration remained unchanged. The Andean lupins had higher protein, lipid and tocopherol contents than L. albus and L. angustifolius; the L. luteus values were within the L. mutabilis range. These results suggest that L. mutabilis harbors nutritional characteristics that are well suited to modern food trends.

The evaluation of the level of alkaloids in edible Lupinus species is crucial from a food safety point of view. Debittering of lupin seeds has a long history; however, the control of the level of alkaloids after processing the seeds is typically only evaluated by changes in the bitter taste. The aim of this study was to evaluate the profile and residual levels of quinolizidine alkaloids (QA) in (Lupinus mutabilis Sweet) after aqueous debittering process. Samples from 10 ecotypes from different areas of Peru were analyzed before and after the process. Based on results obtained by gas chromatography and mass spectrometry, from eight alkaloids identified before the debittering process, only small amounts of lupanine (avg. 0.0012 g/100 g DM) and sparteine (avg. 0.0014 g/100 g DM) remained in the seeds after the debittering process, and no other alkaloids were identified. The aqueous debittering process reduced the content of alkaloids to levels far below the maximal level allowed by international regulations (≤ 0.2 g/kg DM). Electronic supplementary material The online version of this article (10.1007/s11130-020-00799-y) contains supplementary material, which is available to authorized users.

Universidad Nacional Agraria La Molina. Facultad de Industrias Alimentarias The aim of this research was to develop gluten-free (GF) and egg-free quinoa pasta with high nutritional value. Extruded and non-extruded quinoa (red and white) four, potato starch, tara gum, and potato, pea and rice protein isolates were investigated in diferent recipes, some of them included egg white as an initial reference point. Results showed that extruded quinoa four, potato starch and tara gum had deteriorating efects on GF and egg-free pasta frmness and cooking quality. Lupine four addition itself was not able to replace egg white when added in the same amounts, but after increasing the concentration to 12%, the frmness and cooking quality decreasing efects could be improved again, especially when tara gum was absent in the formulations. In the fnal recipe, the content of lupine four was increased to 30% because its protein is complementary to the quinoa protein. From the three studied protein isolates, pea protein was superior to potato or rice protein, addition of the oxidizing enzyme POx could even further improve texture frmness. After these trials, the fnal recipe containing lupine four, pea protein and POx showed satisfying GF noodle quality and possessed a valuable nutritional composition with high protein and dietary fbre content.

The aim of this study was to determine thrips abundance, species composition, sex ratio and seasonal dynamics of these insects on two morphological forms of Andean lupin (epigonal and semiepigonal). Thrips were studied in the lupine plants sown at two dates at the Experimental Research Station at Pawlowice, near Wroclaw, Lower Silesia, Poland, during two growing seasons (2011–2012). The experiment was designed as a split-plot with four replicates for each morphological form and time of sowing. For the determination of thrips species’ composition, plants were collected from the central part of each plot when lupin was at the full blooming stage. In the laboratory, plants were shaken over a sheet of white paper. Once in each season thrips were also collected with a sweep net. In the laboratory larvae and adults were identified to species level and separated into males and females. During the whole lupin growing season, insects were also counted weekly on 10 consecutive plants in the middle row of each plot. Frankliniella intonsa, Thrips major and Aeolothrips intermedius were the dominant species in the full blooming stage of both semiepigonal and epigonal forms of Andean lupin. Thrips tabaci also occurred in greater numbers. There were no significant differences between numbers of thrips and number of species occurring on semiepigonal and epigonal plants sown at the same time. In the most numerous species, like F. intonsa and T. major, mainly females occurred in their population. In the population of A. intermedius males were abundant. Thrips occurring in Lupinus mutabilis, irrespective of the used form of lupin or sowing time, had one peak of abundance, i.e., the last ten days of June and the first ten days of July, when plants were at the blooming stage.

A robust, reproducible method of Agrobacterium-mediated transformation was developed for Lupinus mutabilis Sweet (tarwi), a large-seeded Andean legume. Initially, a regeneration and transformation protocol was developed using a plasmid which contained a bifunctional fusion gene conferring both β-glucuronidase (gus) and neomycin phosphotransferase activities, under the control of a constitutive 35S35SAMV promoter. The tissue explants consisted of longitudinal slices from embryonic axes of imbibed, mature seed. Using a series of tissue culture media for cocultivation, shoot initiation, shoot elongation, and rooting, kanamycin-resistant transgenic plants were recovered from approximately 1% of the explants. This transformation protocol was further used with a construct that contained the human adenosine deaminase (hADA) gene under the control of a legumin seed-specific promoter, also with a kanamycin resistance cassette for chemical selection. Changes made during the course of this study, which included adjustments to the antibiotic concentration during the shoot elongation and rooting phases plus the incorporation of techniques to improve ventilation in the tissue culture system, resulted in major improvements in shoot quality and, most significantly, rooting. The outcome was an increased frequency of transgenic plant recovery (7.4%), with a low (9.6%) rate of plants that escaped selection. The inheritance of the hADA gene was documented and showed the expected Mendelian segregation pattern. The produced hADA protein was a fully functional enzyme and localized only in the seed, as expected. Thus, this legume species is an excellent candidate for a nonfood plant host platform for the production of plant-made proteins.

Research of the regulatory function of sucrose in storage lipid breakdown was conducted on isolated embryo axes, excised cotyledons and whole seedlings of three lupine species grown in vitro on medium with 60 mM sucrose or without the sugar. Lack of sucrose in the medium caused significant increase in total lipid content in yellow, white and Andean lupine isolated embryo axes but in Andean lupine seedling cotyledons and excised cotyledons, lipid level was clearly lower in carbohydrates deficient conditions. Sucrose caused no significant effect on fatty acids spectra. The main fatty acid in yellow lupine seeds was linoleic acid, in white lupine oleic acid and in Andean lupine both oleic and linoleic acids. The main phospholipid in organs of three lupine species was phosphatidylcholine. In sugar-deficient conditions, content of phosphatidylcholine and some others phospholipids was decreased. The peculiar features of regulation by sugars of storage lipid breakdown in germinating lupine seeds and induction of autophagy in young carbohydrate starved embryo axes is discussed.

The capacity of plant roots to increase their carboxylate exudation at a low plant phosphorus (P) status is an adaptation to acquire sufficient P at low soil P availability. Our objective was to compare crop species in their adaptive response to a low-P availability, in order to gain knowledge to be used for improving crop P-acquisition efficiency from soils that are low in P or that have a high capacity to retain P. In the present screening study we compared 13 crop species, grown in sand at either 3 or 300 μM of P, and measured root mass ratio, cluster-root development, rhizosphere pH and carboxylate composition of root exudates. Root mass ratio decreased with increasing P supply for Triticum aestivum L., Brassica napus L., Cicer arietinum L. and Lens culinaris Medik., and increased only for Pisum sativum L., while the Lupinus species and Vicia faba L. were not responsive. Lupinus species that had the potential to produce root clusters either increased or decreased biomass allocation to clusters at 300 μM of P compared with allocation at 3 μM of P. All Lupinus species acidified their rhizosphere more than other species did, with average pH decreasing from 6.7 (control) to 4.3 for Lupinus pilosus L. and 5.9 for Lupinus atlanticus L.; B. napus maintained the most alkaline rhizosphere, averaging 7.4 at 300 μM of P. Rhizosphere carboxylate concentrations were lowest for T. aestivum, B. napus, V. faba, and L. culinaris than for the other species. Exuded carboxylates were mainly citrate and malate for all species, with the exception of L. culinaris and C. arietinum, which produced mainly citrate and malonate. Considerable variation in the concentration of exuded carboxylates and protons was found, even with a genus. Cluster-root forming species did not invariably have the highest concentrations of rhizosphere carboxylates. Lupinus species varied both in P-uptake and in the sensitivity of their cluster-root development to external P supply. Given the carbon cost of cluster roots, a greater plasticity in their formation and exudation (i.e. reduced investment in cluster roots and exudation at higher soil P, a negative feedback response) is a desirable trait for agricultural species that may have variable access to readily available P.

Transformed roots of Lupinus mutabilis cv. Potosi induced by Agrobacterium rhizogenes strain R1601 were cultured on Murashige and Skoog-based medium lacking kanamycin sulphate, or with this antibiotic at 40 mg l−1. The neomycin phosphotransferase gene in the genome of transformed roots was confirmed by non-radioactive Southern hybridisation. Neomycin phosphotransferase protein was detected by ELISA. Transformed roots synthesised isoflavones, but not quinolizidine alkaloids; the latter are typical secondary metabolites of lupin normally produced in aerial parts of the plant. Genistein and 2′-hydroxygenistein, were the main secondary metabolites in cultured, transformed roots, whereas the glycoside genistin was more abundant in roots of non-transformed plants. Wighteone concentrations in transgenic roots were higher than those of non-transformed roots. Transformed roots produced twice the concentration of isoflavones compared with roots from non-transformed plants, indicating that Ri plasmid T-DNA genes modified isoflavone concentration and pattern of biosynthesis.

The complete protocols for long-term micropropagation of some cultivars of four lupin species: Lupinus luteus, L. albus, L. angustifolius and L. mutabilis were elaborated. The shoots were regenerated in vitro via induction of axillary buds development. Plantlets were multiplicated on lowered salts MS-derived media containing BAP in diverse and generally low concentrations. Significant differences in regeneration capacity between species and cultivars were observed. The highest multiplication ratio revealed L. mutabilis and L. luteus. Regenerated shoots were rooted in vitro on low-salts MS-derived media with B5 vitamins. Media were supplemented with different auxins that affected roots formation of particular species and cultivars. Rooting ability of regenerated shoots decreased rapidly through in vitro culture. For that reason, grafting was applied as an alternative method of transfer of shoots to in vivo conditions. This method turned out to be successful for the majority of studied species and cultivars. Complete rooted or grafted plantlets were cultivated in pots with perlit in greenhouse.

Seed triacylglycerols (TAGs) are stored as energy reserves and extracted for various end-product uses. In lupins, seed oil content varies from 16% in Lupinus mutabilis to 8% in L. angustifolius. We have shown that TAGs rapidly accumulate during mid-stages of seed development in L. mutabilis compared to the lower seed oil species, L. angustifolius. In this study, we have targeted the key enzymes of the lipid biosynthetic pathway, acetyl-CoA carboxylase (ACCase) and diacylglycerol acyltransferase (DAGAT), to determine factors regulating TAG accumulation between two lupin species. A twofold increase in ACCase activity was observed in L. mutabilis relative to L. angustifolius and correlated with rapid TAG accumulation. No difference in DAGAT activity was detected. We have identified, cloned and partially characterised a novel gene differentially expressed during TAG accumulation between L. angustifolius and L. mutabilis. The gene has some identity to the glucose dehydrogenase family previously described in barley and bacteria and the significance of its expression levels during seed development in relation to TAG accumulation is discussed. DNA sequence analysis of the promoter in both L. angustifolius and L. mutabilis identified putative matrix attachment regions and recognition sequences for transcription binding sites similar to those found in the Adh1 gene from Arabidopsis. The identical promoter regions between species indicate that differential gene expression is controlled by alternative transcription factors, accessibility to binding sites or a combination of both.

A procedure for regenerating plants of Lupinus mutabilis from shoot apices, from which the leaf primordia and initial cell layer(s) of the apical meristem were removed, has been used to generate transgenic plants following Agrobacterium tumefaciens-mediated gene delivery. Transformation competent cells, from which buds developed, were located at the periphery of the apical meristem. Kanamycin resistant plants were obtained which expressed β-glucuronidase activity. Integration of the neomycin phosphotransferase II and β-glucuronidase genes into the genomes of transgenic plants was confirmed by non-radioactive DNA-DNA hybridisation. This is the first report of the generation of transgenic plants in L. mutabilis.

When thin cell layers (TCLn), isolated from the nodalregion of the hypocotyl of 5 day old seedlings, werecultured in medium M1 with IAA and BA, entire plantswere obtained from L. mutabilis explants andprolific shoots from L. albus explants.Histological studies carried out on L. mutabilisTCLn showed that meristematic cells, present in theexplants at the time of inoculation, gave rise to theformation of a primary meristem resulting inorganogenesis. In TCLn where apical or axillarymeristematic cells were absent, we observed theformation of a nodular structure with a cambium-likelayer at its periphery which divided to form a primarymeristem leading to organogenesis. Plant regenerationfrom TCLn may be useful in genetic transformationassays.

2S albumins were isolated from seeds of Andean lupin (Lupinus mutabilis Sweet) by buffer extraction, ammonium sulphate precipitation and ultrafiltration followed ion-exchange and reversed-phase HPLC. The 2S albumin preparation (LM2S) contained eight albumins. The complete amino acid sequences of small and large subunits of three major albumins (LM2S-4, -5 and -6) were determined by automated Edman degradation of S-pyridylethylated polypeptides and peptides obtained from them by enzymatic digestions. The small subunit of the dominant 2S albumin (LM2S-4) contains 39 amino acid residues and has a molecular mass of 4731 Da. The large subunit of LM2S-4 contains 74 amino acid residues (molecular mass=8708 Da). Two 2S albumin isoforms (LM2S-4 and -6) are due to the expression of two distinct genes; LM2S-6 isoform has eight amino acid replacements when its sequence is compared with the sequence of LM2S-4. The LM2S-5 isoform contains an identical small subunit to LM2S-4, and has in comparison with LM2S-4 two additional amino acid residues at the N-terminus of the large subunit. The amino acid sequences of 2S isoforms from L. mutabilis showed high homology (78–83% identity) with 2S albumins from different Old World Lupinus species.

Severe blight of stems, leaves and pods caused by Botrytis cinerea was found on pearl lupine (Lupinus mutabilis), a legume crop, grown in Kagawa Prefecture, Japan, in March–June 1996–2002. This disease was named “gray mold of pearl lupine” as a new disease.

Somatic embryos were obtained from immature cotyledons of Lupinus angustifolius, L. albus and L. mutabilis but not from L. luteus. Different kinds of basal media and plant growth regulators in primary and secondary culture were tested. The best induction media were based on B5 and were supplemented with 5 mg I-1 2,4-D alone or with 0.25 mg I-1 kinetin. Mature stage somatic embryos were obtained on media containing ABA (0.1–0.5 mg I-1) and a high NH4/NO3 ratio. Embryo germination and plantlet development occurred on MS media supplemented with glutamine or GA3.

Plants of nine different species of lupin (Lupinus spp.) were challenged with Alfalfa mosaic virus (AMV) in glasshouse experiments. No infection of Lupinus cosentinii was detected following sap or graft inoculation. Lupinus albus and Lupinus pilosus were not infected by sap inoculation. However, on graft inoculation, L. albus sometimes responded with localised necrosis just below the graft union without further systemic movement, and L. pilosus responded once with severe systemic necrosis. In Lupinus atlanticus, Lupinus digitatus and Lupinus mutabilis, sap inoculation induced necrotic local lesions in inoculated leaves without subsequent systemic infection, while graft inoculation often caused necrosis just below the graft union without further systemic movement. Lupinus angustifolius, Lupinus hispanicus and Lupinus luteus developed systemic susceptible responses when sap-inoculated. AMV caused milder symptoms than Cucumber mosaic virus in L. angustifolius, while mixed infection with both viruses caused more severe symptoms than either virus alone. In addition, AMV was transmitted through seed to seedlings (0.8%) in a seed sample of L. angustifolius. Australasian Plant Pathology Society 2008

Lupinus mutabilis (pearl lupin) P26956 remained uninfected after repeated sap inoculations with Cucumber mosaic virus (CMV) isolates LW, SN-1, CP, LE, SL and BA. In contrast, plants of L. mutabilis P26961 were readily infected by isolates LW, SN-1 and CP. Isolates LE, SL and BA infected most or all plants of P26961 only after repeated inoculation. When scions of L. angustifolius (narrow-leafed lupin) infected with isolate LW were top-grafted onto plants of L. mutabilis P26956, necrosis developed in the shoots produced just below the graft union whereas other axillary shoots grew vigorously with no virus being detected. The responses to sap inoculation with diverse isolates and to graft inoculation suggest that L. mutabilis P26956 has ‘extreme resistance’ to CMV. Australasian Plant Pathology Society 2004

To ensure the cultivation of the lupine and improve the nutritional state of the Andean population, products have been developed which are enriched with lupine. The high content of bitter alkaloids in the seed of theLupinus mutabilis may be considerably reduced by washing with water (traditional method, ‘Cuzco method’). Drying the grains makes it possible to store them. The lupine with a moisture content of 4 to 8%, can be milled; the flour obtained is slightly yellowish and suitable for panification, where 10 to 40% of the wheat flour may be substituted by lupin. The Protein Efficiency Ratio (PER) of bread was found to be 28% (casein 100%) and was much higher after the enrichment with 10% lupine flour (76%). Lupine based dishes have been well accepted according to a test of acceptability performed with children from 8 to 15 years of age over a three year period.

In a 12-week feeding study, water debittered grains of Andean lupins (Lupinus mutabilis) were tested as unique protein source in growing male and female rats. Before the phsyiological study, the lupin flour was evaluated with regard to its possible content of anti-nutritive substances, such as alkaloids, cyanogenetic glucosides, trypsin inhibitor activity and hemagutanin activity. During the feeding study, parameters such as body weight, food consumption, hemoglobin and blood cells, glucose, urea, total protein, total lipid, cholesterol, uric acid, as well as weight and histopathological examinations of some organs (liver, kidneys, heart, spleen and brain) were controlled. The lupin flour did not show physiologically significant amounts of anti-nutritive substances. Rats fed on lupin diet did not show any statistically different growth performances as compared to the casein fed control animals. Food consumption as well showed no difference between the two diets, although food utilization of casein was slightly better during the first two months. All other observed parameters gave no indication of physiological differences of the performance of the diet. Results of the 12-week feeding test did not give any indication against the use of the water extracted grains of Andean lupins in human and animal nutrition.

A new mutant allele, described here as mutal, which reduces the alkaloid content in dry matter of lupinus mutabilis has been identified following seed treatment with ethyl methane sulphanate. The allele, when homozygous, reduces the alkaloid content from levels of > 2.0 per cent found in seed dry matter of existing populations to 0.2–0.3 per cent and produces plants with vegetative and seed tissues that are organoleptically ‘sweet’. Component alkaloids in plants homozygous for mutal differ in respect of the percentage composition of sparteine and of lupanine, as well as possibly of oxa-sparteine and 4-hydroxylupanine, but none is eliminated in genotypes homozygous for the mutant allele. Alkaloid concentration, in so far as not under the control of mutal, has low heritability but lines were isolated following two generations of successive selection which possessed reduced alkaloid levels. These were interpreted to have arisen as a result of recombination of alleles affecting reduction in alkaloid level at several loci. the mutant, mutal, has been established in a pure breeding line which represents a crucial additional step in the evolution of Lupinus mutabilis towards full status as a crop plant.