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Cross-allergenicity in the legume botanical family in children with food hypersensitivity. II. Laboratory correlates.

Only two of 41 legume-allergic patients diagnosed by double-blind, placebo-controlled oral food challenge or "convincing history" of anaphylaxis had an IgE-mediated hypersensitivity reaction to more than one member of the legume family. However, extensive immunologic cross-reactivity was demonstrated among legume antigens on Immunoblot and Immunodot-blot analyses and prick skin tests. The proteins of six legumes (peanut, soybean, lima bean, pea, garbanzo bean, and green beans) were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to nitrocellulose, and probed with sera from six legume-allergic patients. Multiple IgE-binding bands were identified in each legume lane by the sera from each of these legume-allergic patients. In vitro cross-reactivity did not correlate with clinical hypersensitivity. All the legumes studied (except green bean) had a prominent band at 20 kd. Numerous proteins and protein subunits can be identified in each of the legumes (16 peanut, 21 soybean, 23 lima bean, 25 pea, 22 garbanzo bean, and 11 green bean protein bands) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and it appears that legume-allergic patients' sera may recognize multiple similar fractions from each legume. A second in vitro test was performed in which the six legume extracts were bound directly onto nitrocellulose paper. These "legume" Immunodot blots were probed for specific IgE-binding activity with sera from 62 patients with positive legume prick skin tests. The legume Immunodot blots again demonstrated extensive clinically irrelevant cross-reactivity. However, this test may prove useful as a simple technique for screening food-specific IgE with minimal quantities of sera.

Antigens

Compositional evaluation of some cowpea varieties and some under-utilized edible legumes in Nigeria.

The nutritive potentials of some cowpea varieties such as Ife Brown, Ife Bimpe, IT84E-124, K59 and TVX716 and some under-utilized edible legumes grown in Nigeria such as pigeon pea (Cajanus cajan), lima bean (Phaseolus lunatus), lablab bean (Dolichos lablab), mucuna bean (Mucuna sp.) and Sphenostilis sternocarpa have been evaluated with respect to their proximate chemical composition, mineral content and some endogenous toxic constituents. The cowpea varieties contained on the average 22.5 g crude protein (CP), 2.60 g crude fibre (CF), 5.89 g either extract (EE) and 3.36 g ash/100 g DM while the under-utilized legumes contained 21.7, 6.10, 2.86, and 3.56 g/100 g DM for CP, CF, EE and ash respectively. Distinct varietal differences were observed for EE values as indicated by the coefficients of variation (CV) of 102% for cowpea and 60.8% for the under-utilized legumes. The CF content of the under-utilized legumes were generally higher than those of the cowpea varieties. Potassium was the most abundant mineral in both the cowpea varieties and the under-utilized legumes with mean values of 1.45 and 1.66% respectively, while P was the least abundant with 13.1 and 8.50 ppm, respectively. There were marked intra-varietal differences in the P content as shown by the high CV of 84.0 and 73.9% for the cowpea varieties and the other legumes. The cowpea varieties generally had higher levels of thioglucosides, trypsin inhibitor activity (TIA) and lower haemagglutinating activity (mean values of thioglucosides: 3.86%, of TIA: 13.9 mg/g protein and of haemagglutinating activity: 13.0 HU/mg N respectively), than the under-utilized legumes with mean respective values of 1.22%, 9.84 mg/g protein and 22.7 HU/mg N. The nutritional implications of these anti-nutritional components were discussed and some reasons adduced for the under-utilization of some of these legumes inspite of their apparent similarity in nutritional quality to the more commonly consumed grain legumes.

Calcium

Responses of yearling steers to different stocking rates on a subtropical grass-legume pasture.

Despite potential benefits, limitations of individual tropical legumes have restricted development of sustainable grass-legume pastures in tropical and subtropical regions. Sowing mixtures of complementary legumes may overcome limitations of individual species. Responses of yearling steers grazing a mixture of three tropical legumes with bahiagrass (Paspalum notatum Flugge) were evaluated at three stocking rates under continuous grazing. Carpon desmodium (Desmodium heterocarpon [L.] DC.), which is persistent under grazing but often difficult to establish, was combined with the short-lived legumes aeschynomene (Aeschynomene americana L.) and phasey bean (Macroptilium lathyroides [L.] Urb.). Diet composition, as determined by microhistological analysis of fecal samples, and animal performance were evaluated in three grazing periods: summer 1987 and spring 1988 (2.0, 3.5, and 5.0 steers/ha) and summer 1988 (3.0, 5.3, and 7.5 steers/ha). Stocking rate did not affect percentage of the selectively grazed legumes, aeschynomene and phasey bean, in the diet. Average daily gain decreased linearly (P less than .05) with increased stocking rate, as is typical for grass pastures. Aeschynomene and phasey bean contributed to diets during the first summer, and carpon desmodium contribution was greater in the second summer. These results indicate that this pasture mixture can provide legume herbage from aeschynomene and phasey bean in the year of sowing and from carpon desmodium thereafter. Over the range of grazing pressures obtained, legume responses were generally consistent; thus, optimizing stocking rate for gain per hectare or for economic returns can be targeted without additional constraints to maintain the contribution of these legumes to grazing livestock.

Animal Feed

Influence of legume intake on biliary lipids and cholesterol saturation in young Chilean men. Identification of a dietary risk factor for cholesterol gallstone formation in a highly prevalent area.

Chileans and North American Indians have one of the highest prevalence rates of cholesterol gallstones in the world. The most common theory to explain this has been the operation of some as yet undefined genetic risk factor in these populations. Searching for some common environmental factor for gallstones in Chileans and North American Indians, we found that beans and other legumes are common foods consumed by both populations. In this study we tested the hypothesis that legume intake may favor the production of biliary cholesterol supersaturation. We studied 20 young men subjected to a diet containing 120 g/day of legumes and a control diet without legumes for a period of 1 mo each. Both diets supplied identical quantities of energy, carbohydrates, protein, total fat, fiber, and cholesterol. Low-density lipoprotein cholesterol concentration decreased by 16% (p less than 0.001) after the legume diet. Biliary cholesterol saturation increased in 19 of the 20 subjects; the mean of the group markedly increased from 110% to 169% (p less than 0.001) after the legume diet. These results are consistent with the hypothesis that legume intake is a potential risk factor for cholesterol gallstone disease.

Adult

[Aflatoxins produced by Aspergillus flavus in soya and other legumes].

There is no doubt that our programme of applied nutrition must include soya on account of the high nutritive value of the legume. This underlines research undertaken here regarding the risks of contamination by A. flavus and the subsequent formation of aflatoxins on and in soya beans. We have studied on parallel lines soya beans and other legumes important in the local dietary habits. These legumes are either obtained locally or imported. On every specimen of legume we have tried to confirm, or otherwise, the presence of A. flavus and the aflatoxins. This was followed by experimenting on the conditions for growth and formation of aflatoxins on every one of the legumes. During subsequent experiments we have studied mixtures of legumes containing soya. Research on these lines has shown the inhibitory effects of legumes such as lentils, dried garden peas, Bengal gram, green peas, red peas, and broad beans on the growth of A. flavus and the formation of aflatoxins. Groundnut, on the other hand, seems to encourage such growth and such formation. The results of the experiments carried out, as above, lead us to conclude that soya must form part of our national food monitoring programme.

Aflatoxins

Food legumes in human nutrition: a personal perspective.

Perhaps with the notable exception, and that only in recent years, of red meat, which contributes dietary saturated fats and cholesterol, two well-known reasons in the etiology of heart-related disorders, no single group of foods has been portrayed in such negative terms as the food legumes traditionally have been during the last 50 years of research in food science and human nutrition. Even more alarming are the trends of continued research on such aspects as the deficiency of sulfur-amino acids (both by amino acid analyses as well as rat feeding studies), and the heat lability/stability of proteinase inhibitors and phytohemagglutinins in various legume species. A survey of literature indicates that over 100 research papers were published during the 1981 to 1990 period alone, in just three journals (Journal of Food Science, Journal of Agriculture and Food Chemistry, and Journal of the Science of Food and Agriculture) having the highest citation ratings in food sciences on these three topics, with a general consensus about the facts that were well established as early as the late 1950s. Considering the proliferation of journals publishing food science and human nutrition related work, especially in the Third World countries, the actual number probably would be much higher. This trend also indicates that we are repeating certain aspects of research on the importance of food legumes in human nutrition. Are we really any closer today in our understanding and appreciation of why the nomadic human made such a choice for their very existence during the transition to a more civilized society? This is a high time to project the image of legumes in human nutrition in proper perspective. The validity of our continued research on certain aspects of legumes in human nutrition, at a time when worldwide the research dollars are becoming increasingly harder to come by, is challenged in this review. Essentially, it is a journey through the author's personal diary that raises several questions in justifying the continued research support for at least some nutrition-related work on legumes and an account of what research areas perhaps need to be targeted in the 21st century.

Amino Acids

Nissolia brasiliensis as a nonnodulating model legume.

The nitrogen-fixing root nodule symbiosis is specifically formed by 4 orders of angiosperms. The largest of these 4 orders includes the legume family, the Fabaceae. Among legumes, historical model species have emerged, such as the root nodule symbiosis-forming Medicago truncatula and Lotus japonicus or, more recently, Aeschynomene evenia. By contrast, legume species that have lost root nodule symbiosis have been largely ignored. Here, we describe the first near chromosome-level assembly for a non-root nodule symbiosis-forming legume, the tropical Papilionoideae Nissolia brasiliensis. We compared its genome to closely related legumes and identified genes associated with root nodule symbiosis. Finally, we developed a stable transformation protocol that can be deployed in the future to reevolve root nodule symbiosis in legumes, a first step toward the goal of engineering root nodule symbiosis in nonlegume crops.

Fabaceae

Beyond the CO-FT regulatory module: E1 and PHYA emerge as players in photoperiodic regulation of flowering in legumes.

The legume family (Fabaceae) is the third largest in plants and includes several crop species that are able to fix nitrogen, promote soil health, and contribute to food security worldwide. Recent progress in legume genetics and genomics allowed the identification of photoperiod-dependent flowering loci, which were incorporated into specific signalling networks. Functional characterization of these regulators revealed new roles for known photoreceptors such as phytochrome A, and it also identified legume-specific B3 domain transcriptional factors (E1 and E1-like proteins). This suggests some diversification from the traditional CONSTANS-FLOWERING LOCUS T module present in other angiosperms. Although most of the findings discussed herein pertain to species from the two main legume clades, the Galegoids (e.g. alfalfa, clover, and pea) and the Phaseoloids (e.g. common bean, soybean, cowpea, and pigeon pea), research on flowering regulation in the basal Genistoid clade (e.g. lupins) will also be addressed. We propose that functional diversification of photoperiod-dependent flowering strategies in the different legume species could have contributed to their environmental adaptation and allowed their geographical expansion and success worldwide.

Photoperiod

Control through breeding methods of factors affecting nutritional quality of cereals and grain legumes.

The comparison of nutritional quality parameters of proteins from cultivated cereal and legume species with animal proteins indicate the poor nutritional value of these plant products. The nutritive value of different cereal and legume species is very variable and large differences have also been observed coming from cultivars belonging to the same species. Many interpreting factors, such as protein content, essential amino acid composition and availability, protein digestibility and others are involved in such a highly variability. In addition, cereals as well as legumes may contain large amounts of antinutritional factors which can have serious effects under particular circumstances (e.e. tannins in a low-protein diet or phytates in a metal-deficient diet). Some legume species also contain chemicals of a different nature (i. e. lathyrogens, cyanogenetic glycosides, and others) which may be extremely toxic when ingested in significant amounts. The plant breeder attempting to develop higher-yielding, disease-resistant and nutritionally-improved crop varieties should be aware of such a complex of factors and alert to the possible production or increase of undesirable products or deleterious changes in chemical composition. Available data, although rather limited, indicate valuable breeding approaches to the improvement of nutritive value of cereal and legume grains for humans.

Amino Acids

[Examination and composition of some legume seeds (author's transl)].

The chemical composition of some legume seeds is reviewed. It was found that soy (Glycine soja), field beans (Vicia faba), Vicia sativa and Lens culingris (red seeds) have the highest protein content. There were only minor differences in amino acid composition except for glutamic acid. The lysine content of legume seeds was high. Lysine is the most limiting essential amino acid in cereal proteins, therefore cereal products can be improved nutritionally by addition of legume seed flour. Examination of polar lipids by thin layer chromatography revealed that lecithin, phosphatidyl ethanolamine and phosphatidyl inositol were the major components. However, Lupinus angustifolius contained two unknown polar lipid classes.--In contrary to previous literature the ripe legume seeds contained only a very small amount of galactosyl diglycerides; however legume seeds contain higher amounts of the compounds. The conection between biosynthesis of linolenic acid and the presence of galactosyl diglycerides in higher plants is discussed.

Amino Acids

Condensed tannins in some forage legumes: their role in the prevention of ruminant pasture bloat.

For the past 20 years, the focus in our laboratory has been on finding the causes of ruminant pasture bloat and eventually breeding a bloat-safe alfalfa (Medicago sativa L.); i.e., with bloat potential reduced to the economic threshold. In the mid-seventies, the mechanisms of bloat were explored and found to be more physical than chemical. Characteristic of all bloating legumes after ingestion was a very rapid initial rate of ingestion by rumen microbes. Through the study of bloating and non-bloating legumes, factors were elucidated in the plant that would slow this process. One of these factors was the presence of condensed tannins in the herbage. Some of the non-bloating legumes contained these secondary metabolites, but no condensed tannins were found in any of the bloating legumes. Therefore, species containing an appreciable amount of condensed tannins in their leaves and stems are considered to be non-bloating. Conventional breeding methods have not been successful in producing an alfalfa with condensed tannins in its herbage. New approaches using tissue culture techniques are being attempted, but genetic engineering has the greatest potential for success.

Animals

Effect of processing on some chemical and nutritional characteristics of pre-cooked and dehydrated legumes.

The effect of processing on chemical composition and protein quality of three legumes was studied. The species analyzed were beans (Phaseolus vulgaris), Tórtola and Coscorrón varieties; lentils (Lens esculenta), Laird variety; and chick peas (Cicer arietinum), California-INIA variety. The legumes were soaked in a solution of Na2 EDTA at 0.03% for 16 h and cooked for a predetermined period for each species. They were dried in a tray drier with cross currents of air flowing at a speed of 10 m/min until the residual moisture content was 8%. The water activity in the resulting products ranged from 0.574 to 0.587, thus completely assuring no microbial activity. No important changes were observed in the legumes protein, fat, or fibre contents after processing. The soaking solution was effective in reducing the phytic acid content from 2.99 to 1.64 mg/100 g in the Tórtola beans, which had the highest acid value prior processing. In all the species the heat treatment reduced the activity of the trypsin inhibitors by at least 50%. Cooking and drying significantly increased protein digestibility in all the legumes studied, with chick peas showing the most dramatic increase: form 54.7% pre-processing to 94.9% post-processing. With regard to Net Protein Ratio (NPR), chick peas had a value of 4.03, followed by Tórtola beans (3.29), Coscorrón (3.09) and lentils (2.61). The NPR value for the casein diet was 4.20.

Desiccation

Legume intake and gallstone risk: results from a case-control study.

As legume intake was recently shown to increase biliary cholesterol saturation, it may be a risk factor for gallstone disease. Data from a case-control study in The Netherlands were analysed to confirm this hypothesis. A negative association was found between legume intake (including green beans) and gallstone risk. This appeared not to be explained by diminished legume intake in gallstone cases under the influence of gastrointestinal symptoms. We hypothesize that components from legume pulses (seeds) and pods may have opposite effects on the risk of gallstone development.

Acute Disease

Digestibility of raw and cooked starches from legume seeds using the laboratory rat.

The in vivo digestibility and growth responses to diets containing raw and cooked legume starches were determined using male Wistar rats. These starches were obtained from seven legumes including smooth- and wrinkled-seeded field peas, navy beans, kidney beans, garbanzo beans, mung beans and lentils. Wheat and potato starch diets were fed for comparative purposes. With the exception of potato and the field pea starches, rats fed uncooked starch diets had higher weight gains than those fed cooked starch diets. The starches from wheat and all legumes, excluding the high-amylose wrinkled pea, were nearly 100% digestible but the legume starches reduced the digestibility of casein protein by 3 to 4%. Raw potato starch was shown to be only 57% digestible. The starches from potato and wrinkled pea reduced the digestibility of casein protein to 87% and gave high cecum weights. These characteristics were not altered by cooking the starch of wrinkled pea. The starches of smooth-seeded peas, lentils and all beans were highly digestible, showed no adverse effects on growth and were comparable to wheat starch in this evaluation.

Animals

Iron retention by rats from casein-legume test meals: effect of tannin level and previous diet.

The effect of tannin content on iron (Fe) bioavailability from several legumes was evaluated. Absorption of Fe from a casein (C), isolated soy protein (ISP), chickpea (CP) or red kidney bean (RKB) test meal was tested in marginally Fe-deficient rats [9.9 +/- 0.2 g Hb/100 mL (mean +/- SD)] using the extrinsic tag technique. Also, the effects of a casein habitual diet or of one of three casein-legume habitual diets fed before and after the test meal were investigated. Weanling male rats were fed the habitual diets containing 23 +/- 4 mg Fe/kg of diet (range 17-28) for 13 d. On d 14, after an overnight fast, rats were fed the test meal (1.5 g + 0.1 microCi 59Fe), and whole-body radioactivity was determined. The same habitual diet was refed for seven additional days, and whole-body radioactivity was determined again. Rats retained more iron from C (86%) than from ISP, CP or RKB test meals (73%, 75% and 67%, respectively) when the respective casein-legume habitual diets were fed before and after the test meals. With the casein habitual diet, there was no difference in retained iron from C, ISP, CP or RKB test meals (86%, 87%, 83% and 82%, respectively). Retention of iron from an RKB test meal was increased from 69.6 to 73% when about 90% of the extractable tannins were removed, but the difference was not statistically significant. Thus, feeding a casein-legume diet but not a casein diet prior to a test meal apparently predisposes the rat to lower iron absorption. However, in these studies, tannins per se did not significantly depress iron bioavailability.

Absorption

[Legume consumption among pre-school children: effect of energy density and type of preparation].

The objective of this study was to evaluate the effect of the energy density (E.D.), children's age and type of meals on the consumption of dishes based on legumes and its combination with cereals by preschool children, for the purpose of defining if they were adequate for preschool children's feeding. Thirty children aged 2 to 5 years old who lived in a foster home, were assigned to three experimental groups. Each group was formed by 10 children. Twenty-nine meals were offered to all groups, differing in their energy density: 0.85, 0.94 and 1.03 kcal/g in dishes of legumes and cereals, and 0.47, 0.62 and 0.80 kcal/g in soups. The study was conducted during 29 non-consecutive days throughout a period of 80 days. Food consumption of each child was determined by differential weighing. Food and energy consumption of the legume-cereal dishes was significantly higher than those observed in the case of soups (312 vs 223 g and 288 vs 138 kcal, respectively). The effect of age was verified only when the volume and energy intake was expressed by kg of body weight, being the group with the highest age which showed the lowest food and energy intake, either in dishes or in soups. The relationship between meal consumption and E.D. was significantly inverse. Dishes based on legume-cereal were consumed by pre-school children in sufficient amounts to satisfy their energy requirements in one meal, when these meals have an E.D. of 1.03 kcal/g (3% of added oil). The low E.D. of soups conditioned the fact that these meals were inadequate for the feeding of this age group.

Analysis of Variance

Integrating genomics, multi-omics, CRISPR and speed breeding for stress-resilient vegetable legume improvement.

Vegetable legumes are nutritionally and ecologically important crops. However, their genetic improvement has not kept pace with the increasing challenges posed by climate change due to the polygenic nature of stress tolerance, narrow genetic diversity, and the persistent gap between molecular discoveries and field-level cultivar development. Although recent reviews have examined individual genomic tools or specific stress responses, a comprehensive synthesis integrating genomics-assisted breeding, multi-omics technologies, genome editing, and speed breeding within a unified crop improvement framework has been lacking. This review addresses that gap by critically evaluating how these complementary approaches can accelerate the development of stress-resilient vegetable legumes, including pea, common bean, cowpea, faba bean, cluster bean, yard-long bean, and hyacinth bean. This review synthesizes advances in QTL mapping, genome-wide association studies, transcriptomics, metabolomics, and CRISPR-based functional genomics that have identified key regulators and pathways underlying resistance to major biotic and abiotic stresses. Rather than considering these technologies independently, the review emphasizes their convergence into a systems-level breeding framework integrating genomic discovery, functional validation, predictive breeding, and accelerated generation advancement to improve breeding efficiency. Speed breeding, enabling up to seven to eight generations annually under optimized controlled-environment experimental conditions in cowpea, is discussed as a complementary strategy with genomic selection and genome editing. The review further identifies major translational bottlenecks, including transformation recalcitrance, limited genomic resources for underutilized vegetable legumes, inadequate multi-environment validation, and fragmented omics integration, and presents an integrated systems-breeding framework to bridge the gap between gene discovery and cultivar development.

Fabaceae

Nutritional evaluation of oilseeds and legumes as protein supplements to cereals.

Several oilseed and legume protein products were fed to rats as the sole source of dietary protein, and in blends with cereals for the determination of protein efficiency ratio (PER) and biological availability of amino acids. In addition oilseed protein isolates were fed to mice for the determination of PER. Results of the mouse study revealed that the adjusted PER (casein = 100)for Target rapeseed isolate (108) was higher than those of sunflower (74), safflower (77), soybean (86) or flax (92) isolates. Results of the rat trials revealed that the adjusted PER for Twoer rapeseed meal (88) was higher than those of fababean (21), field pea (59) and soybean meal (72). Supplementation with methionine (0.2%) resulted in improved PER for fababean (84), field pea (101) and soybean meal (97). Mustard flour and rapeseed flour gave PER of 109 and 106, respectively, while the value of sunflower flour was low (56). Protein isolates of Tower rapeseed and soybean gave PER of 92 and 80, respectively. Blending of legumes and oilseeds with wheat flour (PER = 28) gave high PER values (60--85), as also occurred in rice blends (71--88). Supplementation of wheat-legume blends with lysine (0.4%), methionine (0.2%) and threonine (0.1%) brought all PER values above 100. It appeared that differences in PER of the diets paralleled the levels of the first limiting amino acid for rat growth. Results of balance trials indicated that the availability of the limiting amino acid(s) was lower than other essential amino acids for each protein source.

Amino Acids