Low temperature enhances photosynthetic down-regulation in French bean (Phaseolus vulgaris L.) plants

The mechanisms of photosynthetic adaptation to different combinations of temperature and irradiance during growth, and especially the consequences of exposure to high light (2000 micro mol m(-2) s(-1) PPFD) for 5 min, simulating natural sunflecks, was studied in bean plants (Phaseolus vulgaris L.). A protocol using only short (3 min) dark pre-treatment was introduced to maximize the amount of replication possible in studies of chlorophyll fluorescence. High light at low temperature (10 degrees C) significantly down-regulated photosynthetic electron transport capacity [as measured by the efficiency of photosystem II (PSII)], with the protective acclimation allowing the simulated sunflecks to be used more effectively for photosynthesis by plants grown in low light. The greater energy dissipation by thermal processes (lower F(v)'/F(m)' ratio) at low temperature was related to increased xanthophyll de-epoxidation and to the fact that photosynthetic carbon fixation was more limiting at low than at high temperatures. A key objective was to investigate the role of photorespiration in acclimation to irradiance and temperature by comparing the effect of normal (21 kPa) and low (1.5 kPa) O(2) concentrations. Low [O(2)] decreased F(v)/F(m) and the efficiency of PSII (Phi(PSII)), related to greater PSII down-regulation in cold pre-treated plants, but minimized further inhibition by the mild 'sunfleck' treatment used. Results support the hypothesis that photorespiration provides a 'safety-valve' for excess energy.     

A major stress-inducible Mr-42000 wall glycoprotein of French bean (Phaseolus vulgaris L.)

A major wall protein of suspension-cultured cells of French bean has been isolated and characterised. It can be prepared from walls or the culture filtrate and in composition it is particularly rich in proline, valine and glutamic acid/glutamine and contains appreciable amounts of hydroxyproline. The N-terminus shows some glycosylation, while following chemical deglycosylation the first 38 residues were found to be identical to those of proline-rich proteins from soybean. However, the composition of the highly purified M_r-42000 bean protein differs considerably from the soybean proteins and must contain its own specific domains. An antibody was raised and used to demonstrate the inducibility of the M_r-42000 bean protein in response to elicitor action. The protein was found to be mainly localised in the intercellular spaces of the cortical cells of bean hypocotyls and at the wall-plasmalemma interface of xylem vessels, another potentially accessible compartment for pathogens. Following wounding, the protein was found to be generally distributed in the wall of epidermal and cortical cells of the hypocotyls. The M_r-42000 protein is cross reactive with antibodies raised to glycoproteins of the Rhizobium infection thread and the chitin-binding hydroxyproline-rich glycoprotein, potato lectin. These common epitopes together with the previously demonstrated chitin-binding properties of the bean protein indicate a role in host-microbial interactions. Furthermore, the M_r-42000 protein itself bound to the growing hyphal tips of the bean pathogen, Colletotrichum lindemuthianum.Abbreviations FITC fluorescein isothiocyanate - IgG immunoglobulin G - PAL phenylalanine ammonia-lyase We thank Dr Nick Brewin for advice on interpretation of immunolocalisations and for the gift of MCA 265. We thank Dudley Fernandino for carrying out the confocal microscopy. GPB thanks the Science and Engineering Research Council for funding.     

Light-stimulated cell expansion in bean (Phaseolus vulgaris L.) leaves

The quantity and quality of light required for light-stimulated cell expansion in leaves of Phaseolus vulgaris L. have been determined. Seedlings were grown in dim red light (RL; 4 micromoles photons m-2 s-1) until cell division in the primary leaves was completed, then excised discs were incubated in 10 mM sucrose plus 10 mM KCl in a variety of light treatments. The growth response of discs exposed to continuous white light (WL) for 16 h was saturated at 100 micromoles m-2 s-1, and did not show reciprocity. Extensive, but not continuous, illumination was needed for maximal growth. The wavelength dependence of disc expansion was determined from fluence-response curves obtained from 380 to 730 nm provided by the Okazaki Large Spectrograph. Blue (BL; 460 nm) and red light (RL; 660 nm) were most effective in promoting leaf cell growth, both in photosynthetically active and inhibited leaf discs. Far-red light (FR; 730 nm) reduced the effectiveness of RL, but not BL, indicating that phytochrome and a separate blue-light receptor mediate expansion of leaf cells.     

Microsatellite marker diversity in common bean (Phaseolus vulgaris L.)

A diversity survey was used to estimate allelic diversity and heterozygosity of 129 microsatellite markers in a panel of 44 common bean (Phaseolus vulgaris L.) genotypes that have been used as parents of mapping populations. Two types of microsatellites were evaluated, based respectively on gene coding and genomic sequences. Genetic diversity was evaluated by estimating the polymorphism information content (PIC), as well as the distribution and range of alleles sizes. Gene-based microsatellites proved to be less polymorphic than genomic microsatellites in terms of both number of alleles (6.0 vs. 9.2) and PIC values (0.446 vs. 0.594) while greater size differences between the largest and the smallest allele were observed for the genomic microsatellites than for the gene-based microsatellites (31.4 vs. 19.1 bp). Markers that showed a high number of alleles were identified with a maximum of 28 alleles for the marker BMd1. The microsatellites were useful for distinguishing Andean and Mesoamerican genotypes, for uncovering the races within each genepool and for separating wild accessions from cultivars. Greater polymorphism and race structure was found within the Andean gene pool than within the Mesoamerican gene pool and polymorphism rate between genotypes was consistent with genepool and race identity. Comparisons between Andean genotypes had higher polymorphism (53.0%) on average than comparisons among Mesoamerican genotypes (33.4%). Within the Mesoamerican parental combinations, the intra-racial combinations between Mesoamerica and Durango or Jalisco race genotypes showed higher average rates of polymorphism (37.5%) than the within-race combinations between Mesoamerica race genotypes (31.7%). In multiple correspondance analysis we found two principal clusters of genotypes corresponding to the Mesoamerican and Andean gene pools and subgroups representing specific races especially for the Nueva Granada and Peru races of the Andean gene pool. Intra population diversity was higher within the Andean genepool than within the Mesoamerican genepool and this pattern was observed for both gene-based and genomic microsatellites. Furthermore, intra-population diversity within the Andean races (0.356 on average) was higher than within the Mesoamerican races (0.302). Within the Andean gene pool, race Peru had higher diversity compared to race Nueva Granada, while within the Mesoamerican gene pool, the races Durango, Guatemala and Jalisco had comparable levels of diversity which were below that of race Mesoamerica.     

Light-stimulated cell expansion in bean (Phaseolus vulgaris L.) leaves

Cell expansion in dicotyledonous leaves is strongly stimulated by bright white light (WL), at least in part as a result of light-induced acidification of the cell walls. It has been proposed that photosynthetic reactions are required for light-stimulated transport processes across plasma membranes of leaf cells, including proton excretion. The involvement of photosynthesis in growth and wall acidification of primary leaves of bean has been tested by inhibiting photosynthesis in two ways: by reducing chlorophyll content of intact plants with tentoxin (TX) and by treating leaf discs with 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). Exposure to bright WL stimulated growth of intact leaves of TX-treated plants. Discs excised from green as well as from TX-or DCMU-treated leaves also responded by growing faster in WL, as long as exogenous sucrose was supplied to the photosynthetically inhibited tissues. The WL caused acidification of the epidermal surface of intact TX-leaves, but acidification of the incubation medium by mesophyll cells only occurred when photosynthesis was not inhibited. It is concluded that light-stimulated cell enlargement of bean leaves, and the necessary acidification of epidermal cell walls, are mediated by a pigment other than chlorophyll. Light-induced proton excretion by mesophyll cells, on the other hand, may require both a photosynthetic product (or exogenous sugars) and a non-photosynthetic light effect.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1 -dimethylurea - OC osmotic concentration - RL red light - TX tentoxin - WL white light We thank Dr. G.E. Templeton, University of Arkansas, Fayetteville, USA, for initially supplying us with TX, and also Dr. Stephen O. Duke, Southern Weend Science Laboratory, Stoneville, Miss., USA, for suggesting this compound for our experiments. We are grateful to Professor E. Ballio for his generous gift of fusicoccin.     

Nitrogen cycling in a15N-fertilized bean (Phaseolus vulgaris L.) crop

To increase our understanding of the fate of applied nitrogen inPhaseolus vulgaris crops grown under tropical conditions,¹⁵N-labelled urea was applied to bean crops and followed for three consecutive cropping periods. Each crop received 100 kg urea-N ha^?1 and 41 kg KCl?K ha^?1. At the end of each period we estimated each crop's recovery of the added nitrogen, the residual effects of nitrogen from the previous cropping period, the distribution of nitrogen in the soil profile, and leaching losses of nitrogen. In addition, to evaluate potential effects of added phosphorus on nitrogen cycling in this crop, beans were treated at planting with either 35 kg rock-phosphate-P, 35 kg superphosphate-P, or 0 kg P ha^?1. Results showed that 31.2% of the nitrogen in the first crop was derived from the applied urea, which represents a nitrogen utilization efficiency of 38.5%. 6.2% of the nitrogen in the second crop was derived from fertilizer applied to the first crop, and 1.4% of the nitrogen in the third crop. Nitrogen utilization efficiencies for these two crops, with respect to the nitrogen applied to the first crop, were 4.6 and 1.2%, respectively. In total, the three crops recovered 44.3% of the nitrogen applied to the first crop. The remainder of the nitrogen was either still in the soil profile or had been lost by leaching, volatilization or denitrification.¹⁵N enrichment of mineral-N(NO₃+NH₄) suggests that at the end of the second crop, the pulse of fertilizer applied to the first crop had probably passed the 120 cm depth.¹⁵N enrichment of organic-N suggests that root activity of beans and weeds transported nitrogen to 90–120 cm (or deeper). We could account for 109 kg fertilizer-N ha^?1 in harvested biomass, crop residue, and soil at the end of the first cropping period. This indicates an experimental error of about 10% if no nitrogen was lost by volatilization, denitrification, or leaching below 120 cm. At the end of the second and third crops, 76 and 80 kg N ha^?1, respectively, could be accounted for, suggesting that 20 to 25% of the applied-N was lost from the system over a 2-crop period. The two types of added phosphorus did not significantly differ in their effects on bean yields.     

Selection for increased nodule number in common bean (Phaseolus vulgaris L.)

Common bean (Phaseolus vulgaris L.) plants were grown for 21–28 days in plastic container-modified Leonard jar assemblies and placed in a controlled-environment room. The nodules on each plant were removed, counted; selected plants were repotted, grown and intercrossed to produce progenies for the next cycle of recurrent selection. Among the ten parent lines, Puebla 152 and WBR 22–34 produced the most nodules and Rio Tibagi and Negro Argel the fewest, when averaged over five experiments. An analysis of number of nodules on F₁ plants resulting from crosses made in a partial diallel design among the ten parents revealed highly significants variation for general combining ability (GCA) but not for specific combining ability (SCA). After three cycles of recurrent selection for nodule number per plant, the mean nodule number was 211% of the mean for the 10 parents control. Total nodule weight on selected plants also increased, but individual nodule weight decreased. Nineteen C₁ and 18 C₂ lines resulting from the individual plants selected for greater nodule number, along with the ten parents and two non-nodulating soybean lines included as non-fixing check plants were grown in a single experiment in a low-N field. C₂ lines on average accumulated significantly more N per plant than either the parents or C₁ lines, providing evidence for increased N₂ fixation measured by the N difference method. These data show that more nodules, possibly resulting from greater susceptibility to nodulation, are an important, heritable component of symbiosis and that selection for increased nodule number resulted in lines capable of fixing more atmospheric N₂.     

Auxin-induced assimilate translocation in the bean stem (Phaseolus vulgaris L.)

Decapitation of the fully-elongated fourth internode of Phaseolus vulgaris plants resulted in the disappearance from the internode of soluble acid invertase (EC 3.2.1.26). This loss was prevented by local applications to the internode of indol-3yl-acetic acid (IAA) and, at the point of IAA application, the specific activity of the enzyme increased by up to 3 times its initial value within 48 h of treatment. IAA applications stimulated the acropetal translocation to the internode of ¹⁴C-sucrose applied to the subtending (second) trifoliate leaf 30 h after decapitation and the start of the auxin treatment. Labelled assimilates accumulated in the IAA-treated region of the internode. Following decapitation the concentration of hexose sugars in the internode fell and that of sucrose rose substantially, but these trends were reversed by IAA treatment. However, small local accumulations of sucrose occurred at the point of auxin application where tissue concentrations of IAA were greatest (determined using [1-¹⁴C] IAA).Considerable quantities of starch were present in the ground parenchyma of the internodes at the start of the experiment but, in the absence of IAA, this was remobilised within 48 h of decapitation. IAA prevented starch loss at and below its point of application to the internode, but not from more distal tissues. Cambial proliferation, radial growth and lignification were stimulated in and below IAA-treated regions of the internode. These observations are discussed in relation to the hormonal regulation of assimilate translocation in the phloem.     

UV-B increases the harvest index of bean (Phaseolus vulgaris L.)

The effects of small changes in natural UV-B on the photosynthesis, pigmentation, flowering and yield of bean plants (Phaseolus vulgaris L. var. Label) were studied. To obtain a relatively natural growth environment, the plants were grown in small, half-open greenhouses of UV-transmitting Plexiglas of different thickness (3 and 5 mm), resulting in an 8% difference in the weighted UV-B reaching the plants. Although the UV-B doses used did not significantly influence photosynthesis on a leaf area basis during vegetative growth, important changes in biomass allocation were noted. A UV-B-O induced reduction in leaf area during the period of vegetative growth resulted in decreased dry weight after 57 d. During the flowering and pod-filling stages (57–79.d after planting), however, plants grown at high UV-B retained their photosynthetic capacity longer: maximal photosynthesis, chlorophyll and N content of the leaves were higher under the higher UV-B dose at a plant age of 79 d. Combined with an increased allocation under the higher UV-B dose of both N and biomass to the pods, this resulted in a small increase in yield and an important increase in harvest index with increased UV-B.     

Mapping quantitative trait loci for a common bean (Phaseolus vulgaris L.) ideotype.

Breeding a model plant that encompasses individual traits thought to enhance yield potential, known as ideotype breeding, has traditionally focused on phenotypic selection of plants with desirable morphological traits. Broadening this breeding method to the molecular level through the use of molecular markers would avoid the environmental interactions associated with phenotypic selection. A population of 110 F5 recombinant inbred lines (RILs), derived from the cross between WO3391 and 'OAC Speedvale', was used to develop a genetic linkage map consisting of 105 random amplified polymorphic DNA (RAPD), simple sequence repeat (SSR), and sequence-tagged site (STS) markers. The map has a total length of 641 cM distributed across 8 linkage groups (LGs). Five of them were aligned on the core linkage map of bean. Twenty-one quantitative trait loci (QTLs) were identified over three environments for eight agronomic and architectural traits previously defined for a bean (Phaseolus vulgaris L.) ideotype. The QTLs were mapped to seven LGs with several regions containing QTLs for multiple traits. At least one QTL was located for each trait and a maximum of four were associated with lodging. Total explained phenotypic variance ranged from 10.6% for hypocotyl diameter to 45.4% for maturity. Some of the QTLs identified will be useful for early generation selection of tall, upright, high-yielding lines in a breeding program.     

Ureidoglycolate amidohydrolase from developing French bean fruits (Phaseolus vulgaris [L.].)

Ureidoglycolate is an intermediate of allantoin catabolism in ureide-transporting legumes. This report describes the first purification of ureidoglycolate degrading activity (UGDA) from plant tissue in which the enzyme has been separated from urease. The enzyme from developing fruits of Phaseolus vulgaris has been purified 48-fold to give a preparation free of allantoinase and urease activity. UGDA was inhibited by EDTA while the Vmax was increased in the presence of Mn2+. The Km values for ureidoglycolate in the presence and the absence of Mn2+ were 2.0 and 5.4 mM, respectively. In the absence of Mn2+ UGDA was heat labile at 40 degrees C, but in the presence of Mn2+ the activity was stable up to temperatures of 60 degrees C. The Mr of UGDA was determined to be 300,000 by gel filtration chromatography and the pH optimum ranged from pH 7.0 to 8.5. Ammonia was determined to be the nitrogen-containing product of UGDA by a microdiffusion assay. This enzyme should therefore be described as ureidoglycolate amidohydrolase. The activity was shown to be associated with peroxisomes by fractionation of a crude extract on a sucrose density gradient. The products of ureidoglycolate degradation are glyoxylate, ammonia, and presumably carbon dioxide, which can be readily utilized by pathways of metabolism that are known to be present in this organelle.     

Effects of temperature on infection of French bean leaves (Phaseolus vulgaris L.) by lucerne mosaic virus

The effect of temperature on the number of lesions and the time of their appearance was studied by inoculating French bean leaves (Phaseolus vulgaris L. cv. Perli?ka) with lucerne mosaic virus either 24 or 48 h before or, 24 or 48 h after they were exposed to various temperatures. The temperatures tested were 23, 25, 27, 30, 33 and 36° C. Before and after such exposures the plants were kept in a constant temperature of 25° C. By increasing the temperature before inoculation the number of lesions increased in comparison with the control. The optimal temperature for the maximum number of lesions is between 27° and 30° C. There is no significant difference between those experiments when the exposure time was 24 h or 48 h before inoculation. The same temperatures applied for 24 or 48 h after inoculation have a decreasing effect upon the number of lesions formed by LMV on French bean leaves. The decrease is 30 to 75%. In this case the first necrotic local lesions appeared 42 h after inoculation when exposed to higher temperatures above 27° C for 24 h, and 60 h after inoculation when exposed to these temperatures for 48 h. The shape of lesions varied a little in both cases as the pictures show.     

Molecular cloning of the cDNA encoding a stress-inducible protein phosphatase 1 (PP1) catalytic subunit from French bean (Phaseolus vulgaris L.)

A cDNA showing high sequence similarity (>70%) to plant protein phosphatase 1 catalytic subunit variants from other species has been isolated from a cDNA library derived from mRNAs expressed in elicitor-treated suspension-cultured cells. The clone appears to be a near full-length 1431 bp with a 172 bp 5-untranslated region and a 317 bp 3-untranslated region. The open reading frame, determined by sequence similarity, codes for a protein with predicted M _r of 35552. Alternatively an ATG situated to the 5 end of the putative start site would increase the protein size by 6 amino acids.The mRNA for Pvpp1 was shown to be rapidly induced by elicitor treatment of suspension-cultured cells of French bean. The cloned cDNA represents one of the few examples of a gene product that is probably involved in dephosphorylation events arising after the initial responses to biotic stress.Abbreviations PAL phenylalanine ammonia-lyase - PP1 protein phosphatase 1 - Pvpp1 Phaseolus vulgaris protein phosphatase 1     

Development of SCAR marker linked to anthracnose resistance from Indian French bean (Phaseolus vulgaris L.) germplasm

This study was performed to identify the French bean genotypes resistant to anthracnose disease. Thirty-five RAPD primers were used for screening four resistant and nine susceptible French bean accessions. Of these, three RAPD primers, viz. OPAH16₇₀₀, OPN6₇₀₀ and OPS₉₀₀ showed polymorphic bands differentiating between resistant and susceptible genotypes. The RAPD primer OPAH16 was then selected for conversion into a SCAR marker. The polymorphic band present in the resistant line (D line) was eluted, cloned in pTZ57R/T cloning vector and was then transferred into DH5α Escherichia coli cells. The positively transformed clones were selected based on ampicillin resistance blue-white colony selection method. The plasmid DNA was isolated from transformed white colonies, sequenced and developed into SCAR marker SPAH 16. This SCAR marker SPAH 16 was then verified via PCR using the original French bean accessions.     

Inheritance of partial resistance to bean rust in the common bean (Phaseolus vulgaris L.)

The inheritance of partial resistance within eight bean cultivars to a single-pustule isolate of bean rust was studied by means of a F₁ diallel test. General combining ability (GCA) and specific combining ability (SCA) were very highly significant over two seasons and in interaction with seasons. The partitioning of the sums of squares indicated the greater importance of GCA in the inheritance of the resistance. Reciprocal effects were not significant. The estimates of narrow-sense heritability in the two seasons were 0.899 ± 0.056 and 0.603 ± 0.065.     

New gene-derived simple sequence repeat markers for common bean (Phaseolus vulgaris L.)

Common bean is an important and diverse crop legume with several wild relatives that are all part of the Phaseoleae tribe of tropical crop legumes. Sequence databases have been a good source of sequences to mine for simple sequence repeats (SSRs). The objective of this research was to evaluate 14 sequence collections from common bean for SSRs and to evaluate the diversity of the polymorphic microsatellites derived from these collections. SSRs were found in 10 of the GenBank sequence collections with an average of 11.3% of sequences containing microsatellite motifs. The most common motifs were based on tri- and dinucleotides. In a marker development programme, primers were designed for 125 microsatellites which were tested on a panel of 18 common bean genotypes. The markers were named as part of the bean microsatellite-database (BMd) series, and the average polymorphism information content was 0.404 for polymorphic markers and predicted well the genepool structure of common beans and the status of the wild and cultivated accessions that were included in the study. Therefore, the BMd series of microsatellites is useful for multiple studies of genetic relatedness and as anchor markers in future mapping of wide crosses in the species.     

Effects of supplied nitrogen form on growth and water uptake of French bean (Phaseolus vulgaris L.) plants

In order to investigate the effect of N form on dry matter (DM) formation and water uptake rate, French bean (Phaseolus vulgaris L. `Sotaxa') plants were grown with a split-root system. Three treatments were compared: sole nitrate (NO^– ₃) supply (NN), sole ammonium (NH⁺ ₄) supply (AA) and spatially separated supply of NO^– ₃ and NH⁺ ₄ (NA). The pH of the nutrient solutions was kept constant at 6.3 using a pH-stat system. 9 days after onset of the treatments, NN plants had higher root (36%) and shoot dry matter (11%) than AA plants. N form drastically influenced partitioning of assimilates: in the NA treatment, the root half exposed to NO^– ₃ revealed a 170% higher DM than the root half exposed to NH⁺ ₄. N form affected stable carbon-isotope discrimination () of leaf tissue. In leaves of plants which were supplied with NH⁺ ₄ (AA; NA) was significantly more negative (–29.4, –29.6) than in NN treatment (–28.2). We explain this effect by differences in stomatal conductance. We suppose that the significantly less negative of root tissue under NH⁺ ₄ supply is most probably related to higher PEP-case activity. The water uptake rate was higher in NN than in AA grown plants. This effect was found in both, short- and long-term experiments. In case of NA plants, the water uptake in the root part being exposed to NO^– ₃ was 104% higher than in those receiving NH⁺ ₄. At least in the case of the NA treatment we can exclude shoot growth effects as being responsible for differences in water uptake. We therefore assume that differences in root hydraulic conductivity are responsible for the observed effects.     

Analysis of bean (Phaseolus vulgaris L.) proteins affected by copper stress

The effect of excess copper on the expression of soluble proteins in 10-day old Phaseolus vulgaris seedlings was studied with two-dimensional electrophoresis and mass spectrometry, to find sensitive biochemical markers of exposure. Despite major differences in root Cu contents, both 15 and 50 microM Cu treatments resulted in equal enhancements of Cu in the primary leaves. Three proteins, apparently reacting in a dose-dependent manner to Cu exposure, were identified from roots. The levels of an intracellular pathogenesis-related protein and a newly identified protein homologous to PvPR1, PvPR2, were increased with increasing Cu concentration. The level of a newly identified PR-10 protein decreased in a dose-dependent manner. No significant difference was observed in the leaf protein pattern between controls and 15 microM Cu-treated plants. However, at 50 microM Cu exposure, the appearance of PvPR1 and a homologue of Arabidopsis thaliana thylakoid lumenal 17.4kDa protein was observed. Another protein slightly enhanced by Cu treatment had sequence homology to a mitochondrial precursor of glycine cleavage system H protein of Flaveria pringlei.     

2-DE-based proteomic analysis of common bean (Phaseolus vulgaris L.) seeds

Common bean (Phaseolus vulgaris L.) is the most important grain legume for direct human consumption. Proteomic studies in legumes have increased significantly in the last years but few studies have been performed to date in P. vulgaris. We report here a proteomic analysis of bean seeds by two-dimensional electrophoresis (2-DE). Three different protein extraction methods (TCA-acetone, phenol and the commercial clean-up kit) were used taking into account that the extractome can have a determinant impact on the level of quality of downstream protein separation and identification. To demonstrate the quality of the 2-DE analysis, a selection of 50 gel spots was used in protein identification by mass spectrometry (MALDI-TOF MS and MALDI-TOF/TOF). The results showed that a considerable proportion of spots (70%) were identified in spite of incomplete genome/protein databases for bean and other legume species. Most identified proteins corresponded to storage protein, carbohydrate metabolism, defense and stress response, including proteins highly abundant in the seed of P. vulgaris such as the phaseolin, the phytohemagglutinin and the lectin-related α-amylase inhibitor.