scholarly journals VC1 catalyzes a key step in the biosynthesis of vicine from GTP in faba bean

2020 ◽  
Author(s):  
Emilie Björnsdotter ◽  
Marcin Nadzieja ◽  
Wei Chang ◽  
Leandro Escobar-Herrera ◽  
Davide Mancinotti ◽  
...  
Keyword(s):  
Faba Bean ◽  
Dietary Protein ◽  
Riboflavin Biosynthesis ◽  
Wild Type ◽  
Small Frame ◽  
Related Plant ◽  
Gtp Cyclohydrolase Ii ◽  
Protein Rich ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Major Qtl

AbstractFaba bean is a widely adapted and high-yielding legume cultivated for its protein-rich seeds1. However, the seeds accumulate the anti-nutritional pyrimidine glucosides vicine and convicine, which can cause haemolytic anaemia—favism—in the 400 million individuals genetically predisposed by a deficiency in glucose-6-phosphate dehydrogenase2. Here, we identify the first enzyme associated with vicine and convicine biosynthesis, which we name VC1. We show that VC1 co-locates with the major QTL for vicine and convicine content and that the expression of VC1 correlates highly with vicine content across tissues. We also show that low-vicine varieties express a version of VC1 carrying a small, frame-shift insertion, and that overexpression of wild-type VC1 leads to an increase in vicine levels. VC1 encodes a functional GTP cyclohydrolase II, an enzyme normally involved in riboflavin biosynthesis from the purine GTP. Through feeding studies, we demonstrate that GTP is a precursor of vicine both in faba bean and in the distantly related plant bitter gourd. Our results reveal an unexpected biosynthetic origin for vicine and convicine and pave the way for the development of faba bean cultivars that are free from these anti-nutrients, providing a safe and sustainable source of dietary protein.

A baseline study of vicine–convicine levels in faba bean (Vicia faba L.) germplasm

2013 ◽  
Vol 11 (3) ◽  
pp. 250-257 ◽  
Author(s):  
Khalil Khamassi ◽  
Fayçal Ben Jeddi ◽  
Doug Hobbs ◽  
Jose Irigoyen ◽  
Fred Stoddard ◽  
...  
Keyword(s):  
Plant Breeding ◽  
Faba Bean ◽  
Screening Method ◽  
Wild Type ◽  
Faba Beans ◽  
Collaborative Plant Breeding ◽  
Nutritional Compounds ◽  
The Uk ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Plant Breeding Programme

Vicine and convicine are anti-nutritional compounds that accumulate in the cotyledons of faba beans. When humans consume beans with high levels of these compounds, it can cause a condition called favism in individuals harbouring a deficiency in the activity of their glucose-6-phosphate dehydrogenase. When faba beans are used in animal feeds, there can be effects on performance. These concerns have resulted in increasing interest within plant breeding in developing low vicine and convicine faba bean germplasm. In order to facilitate this objective, we developed a rapid and robust screening method for vicine and convicine, capable of distinguishing between faba beans that are either high (wild type) or low in vicine and convicine. In the absence of reliable commercial reference materials, we report an adaptation of a previously published method where a biochemical assay and spectral data were used to confirm the identity of our analytes, vicine and convicine. This method could be readily adopted in other facilities and open the way to the efficient exploitation of diverse germplasm in regions where faba beans play a significant role in human nutrition. We screened a collection of germplasm of interest to a collaborative plant breeding programme developing between the National Institute for Agricultural Botany in the UK and L'Institut Nationale d'Agronomie de Tunisie in Tunisia. We report the results obtained and discuss the prospects for developing molecular markers for the low vicine and convicine trait.

scholarly journals Enhanced Abiotic Stress Tolerance of Vicia faba L. Plants Heterologously Expressing the PR10a Gene from Potato

Plants ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 173
Author(s):  
Abeer F. Desouky ◽  
Ahmed H. Ahmed ◽  
Hartmut Stützel ◽  
Hans-Jörg Jacobsen ◽  
Yi-Chen Pao ◽  
...  
Keyword(s):  
Salt Stress ◽  
Abiotic Stress ◽  
Stress Tolerance ◽  
Vicia Faba ◽  
Faba Bean ◽  
Abiotic Stress Tolerance ◽  
Wild Type ◽  
Stress Injury ◽  
Bean Plants ◽  
Vicia Faba L

Pathogenesis-related (PR) proteins are known to play relevant roles in plant defense against biotic and abiotic stresses. In the present study, we characterize the response of transgenic faba bean (Vicia faba L.) plants encoding a PR10a gene from potato (Solanum tuberosum L.) to salinity and drought. The transgene was under the mannopine synthetase (pMAS) promoter. PR10a-overexpressing faba bean plants showed better growth than the wild-type plants after 14 days of drought stress and 30 days of salt stress under hydroponic growth conditions. After removing the stress, the PR10a-plants returned to a normal state, while the wild-type plants could not be restored. Most importantly, there was no phenotypic difference between transgenic and non-transgenic faba bean plants under well-watered conditions. Evaluation of physiological parameters during salt stress showed lower Na+-content in the leaves of the transgenic plants, which would reduce the toxic effect. In addition, PR10a-plants were able to maintain vegetative growth and experienced fewer photosystem changes under both stresses and a lower level of osmotic stress injury under salt stress compared to wild-type plants. Taken together, our findings suggest that the PR10a gene from potato plays an important role in abiotic stress tolerance, probably by activation of stress-related physiological processes.

scholarly journals Effects of glucose-6-phosphate dehydrogenase deficiency upon sickle cell anemia

Blood ◽  
1988 ◽  
Vol 71 (3) ◽  
pp. 748-752 ◽  
Author(s):  
MH Steinberg ◽  
MS West ◽  
D Gallagher ◽  
W Mentzer
Keyword(s):  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
G6pd Deficiency ◽  
Electrophoretic Pattern ◽  
Hemoglobin Concentration ◽  
Wild Type ◽  
Standardized Protocol ◽  
Cellulose Acetate Membranes ◽  
Male Patients ◽  
Glucose 6 Phosphate Dehydrogenase

Abstract We studied the interactions of the A- variety of glucose-6-phosphate dehydrogenase (G6PD) deficiency and sickle cell anemia (HbSS) to see if G6PD deficiency influenced laboratory and clinical features of HbSS. A total of 801 male patients over age 2 had G6PD electrophoresis on cellulose acetate membranes. Assays of both G6PD activity and hexokinase activity were then done on all samples that had an electrophoretic pattern other than the normal wild type (GdB). The collection of clinical data used a standardized protocol. Using cluster analyses we classified 10.4% males to be G6PD deficient, while 18.4% had the functionally normal GdA+ enzyme. The prevalence of G6PD deficiency did not change significantly when age was stratified by decade, suggesting little survival advantage or disadvantage of the combination of G6PD deficiency and HbSS. Compared to patients who were not G6PD deficient, there were no significant differences in the hemoglobin concentration, mean corpuscular volume, reticulocyte count, bilirubin, or SGOT level in patients with HbSS who had G6PD deficiency. The incidence of painful episodes, sepsis, or acute anemic episodes was similar in both groups. Our results are consistent with recent studies of smaller numbers of patients that have found little influence of G6PD deficiency upon HbSS. Specifically, we found no evidence that G6PD enhanced the severity of hemolysis or increased the incidence of acute anemic episodes or sepsis in HbSS.

scholarly journals The role of dietary protein in hepatic lipogenesis in the young rat

1981 ◽  
Vol 45 (3) ◽  
pp. 529-538 ◽  
Author(s):  
G. R. Herzberg ◽  
Minda Rogerson
Keyword(s):  
Fatty Acid ◽  
Fatty Acid Synthesis ◽  
Dietary Protein ◽  
Fatty Acid Synthetase ◽  
Acid Synthesis ◽  
Hepatic Lipogenesis ◽  
Hepatic Fatty Acid ◽  
Cleavage Enzyme ◽  
Glucose 6 Phosphate Dehydrogenase

1. The effect of varying dietary levels of casein (40–140 g/kg) on hepatic lipogenesis and the levels of hepatic fatty acid synthetase (FAS), glucose-6-phosphate dehydrogenase (EC 1.1.1.49; G6PD), malic enzyme (EC 1.1.1.40; ME), citrate cleavage enzyme (EC 4.1.3.8;CCE), acetyl CoA carboxylase (EC 6.4.1.2; AcCx), glucokinase (EC 2.7.1.2; GK), and pyruvate dehydrogenase (PDH) was examined in young, growing rats.2. The activities of AcCx, FAS, G6PD and in vivo fatty acid synthesis were generally found to increase with increased dietary protein.3. The levels of GK and PDH were not related to dietary protein.4. ME decreased with increasing dietary protein.5. The results demonstrate a dissociation between hepatic fatty acid synthesis and ME and suggest that when rats consume low-protein diets the NADPH needed for fatty acid synthesis is generated primarily by ME but that as the level of dietary protein is increased the contribution of ME is reduced while that of the phosphogluconate pathway becomes more important.

scholarly journals Deletion of the Glucose-6-Phosphate Dehydrogenase Gene KlZWF1 Affects both Fermentative and Respiratory Metabolism in Kluyveromyces lactis

2006 ◽  
Vol 6 (1) ◽  
pp. 19-27 ◽  
Author(s):  
Michele Saliola ◽  
Gina Scappucci ◽  
Ilaria De Maria ◽  
Tiziana Lodi ◽  
Patrizia Mancini ◽  
...  
Keyword(s):  
Biomass Yield ◽  
Kluyveromyces Lactis ◽  
Carbon Sources ◽  
Pentose Phosphate ◽  
Respiratory Metabolism ◽  
Wild Type ◽  
Dehydrogenase Gene ◽  
Increased Sensitivity ◽  
Dimeric Form ◽  
Glucose 6 Phosphate Dehydrogenase

ABSTRACT In Kluyveromyces lactis, the pentose phosphate pathway is an alternative route for the dissimilation of glucose. The first enzyme of the pathway is the glucose-6-phosphate dehydrogenase (G6PDH), encoded by KlZWF1. We isolated this gene and examined its role. Like ZWF1 of Saccharomyces cerevisiae, KlZWF1 was constitutively expressed, and its deletion led to increased sensitivity to hydrogen peroxide on glucose, but unlike the case for S. cerevisiae, the Klzwf1Δ strain had a reduced biomass yield on fermentative carbon sources as well as on lactate and glycerol. In addition, the reduced yield on glucose was associated with low ethanol production and decreased oxygen consumption, indicating that this gene is required for both fermentation and respiration. On ethanol, however, the mutant showed an increased biomass yield. Moreover, on this substrate, wild-type cells showed an additional band of activity that might correspond to a dimeric form of G6PDH. The partial dimerization of the G6PDH tetramer on ethanol suggested the production of an NADPH excess that was negative for biomass yield.

scholarly journals Specific Binding Sites for an Antifungal Plant Defensin from Dahlia (Dahlia merckii) on Fungal Cells Are Required for Antifungal Activity

2000 ◽  
Vol 13 (1) ◽  
pp. 54-61 ◽  
Author(s):  
Karin Thevissen ◽  
Rupert W. Osborn ◽  
David P. Acland ◽  
Willem F. Broekaert
Keyword(s):  
Antifungal Activity ◽  
Plasma Membranes ◽  
Specific Binding ◽  
Plant Defensin ◽  
Wild Type ◽  
Whole Cells ◽  
Specific Binding Sites ◽  
Plant Defensins ◽  
Related Plant ◽  
Wild Type Yeast

Dm-AMP1, an antifungal plant defensin from seeds of dahlia (Dahlia merckii), was radioactively labeled with t-butoxycarbonyl-[35S]-L-methionine N-hydroxy-succinimi-dylester. This procedure yielded a 35S-labeled peptide with unaltered antifungal activity. [35S]Dm-AMP1 was used to assess binding on living cells of the filamentous fungus Neurospora crassa and the unicellular fungus Saccharomyces cerevisiae. Binding of [35S]Dm-AMP1 to fungal cells was saturable and could be competed for by preincubation with excess, unlabeled Dm-AMP1 as well as with Ah-AMP1 and Ct-AMP1, two plant defensins that are highly homologous to Dm-AMP1. In contrast, binding could not be competed for by more distantly related plant defensins or structurally unrelated antimicrobial peptides. Binding of [35S]Dm-AMP1 to either N. crassa or S. cerevisiae cells was apparently irreversible. In addition, whole cells and microsomal membrane fractions from two independently obtained S. cerevisiae mutants selected for resistance to Dm-AMP1 exhibited severely reduced binding affinity for [35S]Dm-AMP1, compared with wild-type yeast. This finding suggests that binding of Dm-AMP1 to S. cerevisiae plasma membranes is required for antifungal activity of this protein.

Effects of sugars and polyols on basidiocarp formation in a phosphoglucose isomerase mutant of Coprinus cinereus

1984 ◽  
Vol 30 (1) ◽  
pp. 45-51 ◽  
Author(s):  
Hiroshi Nyunoya ◽  
Tsuneo Takemaru ◽  
Tatsuo Ishikawa
Keyword(s):  
Culture Medium ◽  
Complete Recovery ◽  
Biochemical Properties ◽  
Phosphoglucose Isomerase ◽  
Coprinus Cinereus ◽  
Partial Recovery ◽  
Wild Type ◽  
In The Wild ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Basidiocarp Formation

Several biochemical properties of a mutant deficient in phosphoglucose isomerase (pgi) of Coprinus cinereus were examined in connection with its ability to produce basidiocarps. Mycelium of the pgi mutant accumulated glucose-6-phosphate and showed higher glucose-6-phosphate dehydrogenase activity than wild type. A conventional fruiting medium did not support basidiocarp formation by the homozygous dikaryon pgi/pgi, but the addition of a reduced amount of glucose plus a compensatory amount of fructose to the culture medium resulted in partial recovery of fruiting ability. This modification of the culture medium decreased the intracellular glucose-6-phosphate to almost the same level as in the wild type. The addition of polyols such as mannitol resulted in complete recovery of fruiting ability by the dikaryon pgi/pgi without affecting the level of glucose-6-phosphate. Mycelium of the mutant showed an elevated activity of NAD-linked polyol dehydrogenase and an elevated intracellular NAD level, irrespective of whether the mycelium was grown in the presence or absence of polyol.

scholarly journals Dietary protein content affects evolution for body size, body fat and viability in Drosophila melanogaster

2010 ◽  
Vol 7 (2) ◽  
pp. 269-272 ◽  
Author(s):  
Torsten N. Kristensen ◽  
Johannes Overgaard ◽  
Volker Loeschcke ◽  
David Mayntz
Keyword(s):  
Drosophila Melanogaster ◽  
Protein Content ◽  
Dietary Protein ◽  
Experimental Evolution ◽  
Natural Variation ◽  
Food Sources ◽  
Standard Diet ◽  
Total Body Mass ◽  
Total Body ◽  
Protein Rich

The ability to use different food sources is likely to be under strong selection if organisms are faced with natural variation in macro-nutrient (protein, carbohydrate and lipid) availabilities. Here, we use experimental evolution to study how variable dietary protein content affects adult body composition and developmental success in Drosophila melanogaster . We reared flies on either a standard diet or a protein-enriched diet for 17 generations before testing them on both diet types. Flies from lines selected on protein-rich diet produced phenotypes with higher total body mass and relative lipid content when compared with those selected on a standard diet, irrespective of which of the two diets they were tested on. However, selection on protein-rich diet incurred a cost as flies reared on this diet had markedly lower developmental success in terms of egg-to-adult viability on both medium types, suggesting a possible trade-off between the traits investigated.

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