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PubMed1993

Methionine-containing proteins in two Phaseolus vulgaris cultivars with different methionine bioavailabilities.

The full utilization of legumes as human food is limited by a deficiency of sulfur amino acids, low protein digestibility, low methionine bioavailability and the presence of anti-nutritional factors. A new cultivar of Phaseolus vulgaris (Carioca 80) has 56.8% available methionine, compared with 29.3% availability in the parent cultivar Carioca. The total methionine content, denaturing gel electrophoretic patterns of methionine-containing proteins, and the percentage of phaseolins (the major storage proteins in Phaseolus) relative to the total protein are similar in the two cultivars. Although the digestibility of the two cultivars is similar, the increased biological value of Carioca 80 may indicate that there are differences in overall bean composition that affect protein hydrolysis and utilization. We suggest the tentative explanation that this is due to differences in the distribution of methionine in the methionine-containing proteins of the two cultivars.

Biological Availability↗
PubMed1995

Expression pattern of uricase II gene during root nodule development in Phaseolus vulgaris.

A Phaseolus vulgaris uricase II cDNA clone has been isolated and sequenced. Comparison on the nucleotide level between this clone and the soybean uricase II clone revealed 88.8% identity. The in situ hybridization technique was used to follow the expression pattern in developing root nodules of Phaseolus vulgaris. The uricase II transcripts were localized only in the uninfected cells of the central tissue and mainly in the periphery of the cell. Uricase II mRNA is first detected in nodules 12 days after infection. A maximum level of transcripts is reached in 21-day-old nodules, followed by a considerable reduction in 28-day-old nodules.

Amino Acid Sequence↗
PubMed1999

Hybrid Weakness in Phaseolus vulgaris L. I. Disruption of Development and Hormonal Allocation.

A reduced concentration of cytokinins may cause the abnormal growth and development found in F1 hybrids between Andean and Mesoamerican races of Phaseolus vulgaris L. In this study, concentrations of the transportable cytokinin zeatin riboside (ZR) were measured by ELISA for ZR (cross reactivities dihydrozeatin, 14%, zeatin 7.6%) in roots, stems, and leaves of a Phaseolus Mesoamerican landrace (P. vulgaris L. cv. Redkloud), an Andean landrace (P. vulgaris L. cv. Batt), and their F1 hybrids. Concentrations of ZR in roots and leaves of F1 hybrids were significantly less than that found in roots and leaves of parental cultivars. Approximately 90% of the ZR found in F1 hybrids was found sequestered in the stems, whereas cytokinins of the parental cultivars were distributed throughout the plant (roots: Batt 37%, Redkloud, 44%; stems: Batt 35%, Redkloud 42%; leaves: Batt 28%, Redkloud 14%). These results suggest that abnormal growth and development of F1 hybrids may involve interruption of the regulation of cytokinin allocation, thereby disrupting the root-shoot feedback loop between root-sourced cytokinins and putative shoot-produced factors.

Journal Article↗
PubMed1985

The effect of the suspensor and gibberellic acid on Phaseolus vulgaris embryo protein content.

The role of the suspensor in the early development of the dicot embryo has not yet been defined. It has been described as merely an anchor and also as the major route of nutrients into the embryo. In order to further elucidate the role of the suspensor, early 0.2-mm and late heart stage 0.5-mm Phaseolus vulgaris (var. Taylor's Horticultural) embryos have been examined in tissue culture. It is known that Phaseolus embryos in culture at low osmotic potential will germinate precociously and that embryos in culture at high osmotic potential will either fail to grow or form callus. Optimum sucrose concentrations for continued, normal embryonic development of 0.2 mm and 0.5 mm P. vulgaris in tissue culture with Gamborg B5 medium were determined to be 12 and 6%, respectively. Protein content was examined in embryos and suspensors. Results showed that both 0.2- and 0.5-mm embryos required an attached suspensor for maximum protein content. Protein levels were substantially decreased when the embryo was cultured detached from or without the suspensor. Gibberellic acid at 10(-6) to 10(-7) M restored the protein content to that of freshly excised embryos.

Culture Techniques↗
PubMed1996

In vivo endoproteolytically cleaved phaseolin is stable and accumulates in developing Phaseolus lunatus L. seeds.

Phaseolin is the most abundant storage protein of bean seeds. To modify its amino-acidic composition by protein engineering, for the improvement of its nutritional value, regions which could be modified without detrimental effects on structural features of the protein must be identified. Data presented here, on the characterisation of the major storage protein of lima bean (Phaseolus lunatus L.) seeds, a phaseolin-like glycoprotein, provide good indications on one of such region. Phaseolus lunatus phaseolin consists of four major oligomers containing two subunit classes. Polypeptides of one class show a molecular mass ranging from 38.5 kDa to 32 kDa, while the molecular mass of polypeptides belonging to the other class ranges from 27 kDa to 21 kDa. The subunits originate from the cleavage of precursor forms, with molecular masses of 58 kDa and 54 kDa, which are still present - in residual amounts - in the nature protein. Comparison of their N-terminal sequences with those of the subunits demonstrate that cleavage occurs in a region of the molecule that instead remains uncleaved in phaseolins of the other species. Since this region can accommodate such a drastic modification, we suggest it could be a good candidate for in vitro manipulation.

Amino Acid Sequence↗
PubMed1988

The connections between the septum-diagonal band complex and histaminergic neurons in the posterior hypothalamus of the rat. Anterograde tracing with Phaseolus vulgaris-leucoagglutinin combined with immunocytochemistry of histidine decarboxylase.

The connections between nuclei of the septum-diagonal band complex and the clusters of histaminergic neurons in the posterior hypothalamic region were studied with a dual-labeling procedure in which anterograde neuroanatomical tracing with Phaseolus vulgaris-leucoagglutinin was combined with immunohistochemistry of histidine decarboxylase. Phaseolus vulgaris-leucoagglutinin was injected in the medial and lateral septal nuclei, and in various parts of the nuclei of the diagonal band of Broca. The fibers arising from the medial and lateral septal nuclei traverse the vertical limb of the diagonal band and, in part, join the medial forebrain bundle in the preoptic area. Other fibers descend diffusely through the lateral hypothalamus to the posterior hypothalamus, or course in a bundle of fibers ensheathing the fornix. The nuclei of the diagonal band project via the medial forebrain bundle and the diffuse pathway to the posterior hypothalamic region. All the nuclei of the septum-diagonal band complex, with the exception of the medial and lateral parts of the nucleus of the horizontal limb of the diagonal band, project to clusters of histaminergic neurons. These projections exhibit the following arrangement: along the axis lateral septal nucleus-medial septal nucleus-vertical limb of the diagonal band-medial part of the horizontal limb of the diagonal band, the septohypothalamic fibers decrease in density and distribute to fewer clusters of histaminergic neurons. Varicosities on the labeled fibers are formed in close proximity to the cell bodies and dendrites of the histaminergic neurons.

Animals↗
PubMed1991

Patterns of direct projections from the hippocampus to the medial septum-diagonal band complex: anterograde tracing with Phaseolus vulgaris leucoagglutinin combined with immunohistochemistry of choline acetyltransferase.

The projections from the Ammon's horn to the cholinergic cell groups in the medial septal and diagonal band nuclei were investigated with anterograde tracing of Phaseolus vulgaris leucoagglutinin combined with immunocytochemical detection of choline acetyltransferase, in the rat. Tracer injections were placed into various fields of the septal and temporal parts of the Ammon's horn (CA1-3). These injections revealed differential distributions of Phaseolus vulgaris leucoagglutinin-labeled projections in both the lateral septal area and the medial septum-diagonal band complex. In addition to the labeling of dense axonal networks in the lateral septal area, significant numbers of arborizing fibers were labeled in the medial septal and diagonal band nuclei, in particular after tracer injections into the fields CA2-3. The distributions of the projections to the medial septum-diagonal band complex arising from the septal portion of fields CA1 and CA2-3 are similar. In contrast, the septal part and temporal portion of field CA3 project in a topographically differentiated manner to the medial septum and nuclei of the diagonal band. The septal pole of the Ammon's horn innervates the dorsal and medial parts of the medial septal nucleus and the anterior and dorsal parts of the vertical limb of the diagonal band. Axons of the temporal pole of the hippocampus reach the lateral and ventral parts of the medial septum and the intermediate, caudal and ventral parts of the vertical limb of the diagonal band. These results demonstrate direct feedback projections of the Ammon's horn to the medial septum-diagonal band complex, which show a topographic organization mainly as a function of the septotemporal level of the hippocampus. Within the medial septal and diagonal band nuclei, the labeled varicosities are formed in close proximity to the cell bodies and dendrites of the cholinergic neurons.

Animals↗
PubMed1992

The organization of the crossed geniculogeniculate pathway of the rat: a Phaseolus vulgaris-leucoagglutinin study.

The intergeniculate leaflet of the thalamus is known to give rise to neuronal projections to the suprachiasmatic nuclei and the rostral part of the pineal gland. Via these projections the intergeniculate leaflet is considered to play a role in regulation of circadian rhythms. Iontophoretic injections of the anterograde tracer Phaseolus vulgaris-leucoagglutinin were placed in various subnuclei of the lateral geniculate nucleus in order to study the topographical organization of the crossed geniculogeniculate pathway in the rat. Injections involving neurons in the intergeniculate leaflet or the medial subpart of the ventral nucleus (which presumably is part of the intergeniculate leaflet of the thalamus too) gave rise to labeled nerve fibers in the opposite lateral geniculate nucleus. The axons contained in this pathway were followed either medially via the posterior commissure, or via the optic tracts and optic chiasm, to the contralateral hemisphere. In the contralateral lateral geniculate nucleus, the intergeniculate leaflet was most densely innervated, but a substantial innervation of the ventral lateral geniculate nucleus was observed as well. Only a few labeled fibers were observed in the dorsal subnucleus. However, the dense innervation of the contralateral intergeniculate leaflet not only covered the small zone between the dorsal and ventral nuclei, but also a dorsomedial part of the ventral nucleus that merged caudally with the lateral part of the zona incerta. In the remaining part of the ventral nucleus, single Phaseolus vulgaris-leucoagglutinin-labeled fibers surrounded specific cells. The demonstration of a divergent projection between the intergeniculate leaflet and specific subparts of the contralateral geniculate nuclei indicates that the two lateral geniculate nuclei are regulating each other. The function of this pathway is suggested to be related to the regulation of circadian rhythmicity, but experimental evidence for this hypothesis is still lacking.

Animals↗
PubMed1995

Afferent projections to the rat locus coeruleus demonstrated by retrograde and anterograde tracing with cholera-toxin B subunit and Phaseolus vulgaris leucoagglutinin.

The aim of this study was to examine the afferents to the rat locus coeruleus by means of retrograde and anterograde tracing experiments using cholera-toxin B subunit and phaseolus leucoagglutinin. To obtain reliable injections of cholera-toxin B in the locus coeruleus, electrophysiological recordings were made through glass micropipettes containing the tracer and the noradrenergic neurons of the locus coeruleus were identified by their characteristic discharge properties. After iontophoretic injections of cholera-toxin B into the nuclear core of the locus coeruleus, we observed a substantial number of retrogradely labeled cells in the lateral paragigantocellular nucleus and the dorsomedial rostral medulla (ventromedial prepositus hypoglossi and dorsal paragigantocellular nuclei) as previously described. We also saw a substantial number of retrogradely labeled neurons in (1) the preoptic area dorsal to the supraoptic nucleus, (2) areas of the posterior hypothalamus, (3) the Kölliker-Fuse nucleus, (4) mesencephalic reticular formation. Fewer labeled cells were also observed in other regions including the hypothalamic paraventricular nucleus, dorsal raphe nucleus, median raphe nucleus, dorsal part of the periaqueductal gray, the area of the noradrenergic A5 group, the lateral parabrachial nucleus and the caudoventrolateral reticular nucleus. No or only occasional cells were found in the cortex, the central nucleus of the amygdala, the lateral part of the bed nucleus of the stria terminalis, the vestibular nuclei, the nucleus of the solitary tract or the spinal cord, structures which were previously reported as inputs to the locus coeruleus. Control injections of cholera-toxin B were made in areas surrounding the locus coeruleus, including (1) Barrington's nucleus, (2) the mesencephalic trigeminal nucleus, (3) a previously undefined area immediately rostral to the locus coeruleus and medial to the mesencephalic trigeminal nucleus that we named the peri-mesencephalic trigeminal nucleus, and (4) the medial vestibular nucleus lateral to the caudal tip of the locus coeruleus. These injections yielded patterns of retrograde labeling that differed from one another and also from that obtained with cholera-toxin B injection sites in the locus coeruleus. These results indicate that the area surrounding the locus coeruleus is divided into individual nuclei with distinct afferents. These results were confirmed and extended with anterograde transport of cholera-toxin B or phaseolus leucoagglutinin. Injections of these tracers in the lateral paragigantocellular nucleus, preoptic area dorsal to the supraoptic nucleus, the ventrolateral part of the periaqueductal gray, the Kölliker-Fuse nucleus yielded a substantial to large number of labeled fibers in the nuclear core of the locus coeruleus.(ABSTRACT TRUNCATED AT 400 WORDS)

Afferent Pathways↗
PubMed1995

Intra-amygdaloid projections of the basolateral and basomedial nuclei in the cat: Phaseolus vulgaris-leucoagglutinin anterograde tracing at the light and electron microscopic level.

The amygdaloid complex plays an essential role in auditory fear conditioning of the Pavlovian type. The available evidence suggests that the lateral nucleus is the input station of the amygdala for auditory conditioned stimuli, whereas the central medial nucleus is the output for conditioned fear responses. However, the intrinsic pathway transmitting auditory information about the conditioned stimulus from the lateral to the central medial nuclei is unknown as there are no direct projections between these nuclei. The present study was undertaken to determine if the main intra-amygdaloid targets of the lateral nucleus, namely the basomedial and basolateral nuclei, project to the central medial nucleus. To this end, iontophoretic injections of the anterograde tracer Phaseolus vulgaris-leucoagglutinin were performed in these nuclei. To rule out the possibility that the anterograde labeling reflected passing fibers merging with the major fiber bundles that course in and around the central medial nucleus, labeled terminals and varicosities were observed in the electron microscope. It was determined that the basolateral and basomedial nuclei have partially overlapping intraamygdaloid targets. They both project to the central medial nucleus, nucleus of the lateral olfactory tract and peri-amygdaloid cortex, but have limited projections to each other. Small Phaseolus vulgaris-leucoagglutinin injections in both nuclei gave rise to prominent intranuclear projections but only the basomedial nucleus was found to project to the lateral and anterior cortical nuclei. At the electron microscopic level, all labeled axon terminals and varicosities formed asymmetric synapses (n = 245) with dendritic spines (83%) or with dendritic shafts (17%). This is the first unambiguous demonstration that the basolateral and basomedial nuclei project to the central medial nucleus. Since these nuclei constitute the main intra-amygdaloid targets of the lateral nucleus, they represent likely candidates for the transmission of auditory conditioned stimuli to the central medial nucleus in auditory fear conditioning.

Amygdala↗
PubMed1996

Layer-specific sprouting of commissural fibres to the rat fascia dentata after unilateral entorhinal cortex lesion: a Phaseolus vulgaris leucoagglutinin tracing study.

After unilateral entorhinal cortex lesion commissural fibres to the inner molecular layer of the rat fascia dentata are said to sprout into the former termination zone of entorhinal afferents. This sprouting process has not yet been demonstrated at the level of individual fibres. In the present study, Phaseolus vulgaris leucoagglutinin tracing was used to analyse the commissural projection to the inner molecular layer in rats with longstanding entorhinal cortex lesions. In comparison with controls, the commissural fibre plexus in the inner molecular layer had expanded by 20-45 microns outwards on the side of the entorhinal lesion. Unexpectedly, only a small number of axons arising from the bulk of commissural fibres in the inner molecular layer left the main fibre plexus and entered the outer molecular layer. Thus, there was still a clearly recognizable border between the Phaseolus vulgaris leucoagglutinin-labelled commissural fibre plexus in the inner molecular layer and the unstained outer molecular layer. The few commissural axons invading the outer molecular layer rarely branched but formed multiple en passant boutons, and occasionally exhibited growth cones. The data indicate that only few commissural fibres appear to be able to sprout beyond the border of their appropriate layer suggesting that the characteristic laminar specificity of hippocampal afferents is largely retained following deafferentation.

Animals↗
PubMed1973

The formation of a -(1 leads to 4)-D-galactan chain catalysed by a Phaseolus aureus enzyme.

With a particulate enzyme preparation from Phaseolus aureus hypocotyls, UDP-alpha-d-[U-(14)C]galactose served as a precursor for a number of products. One of these products was characterized as a beta-(1-->4)-linked galactan. The ADP-, GDP-, TDP- and CDP- derivatives of alpha-d-galactose did not serve as biosynthetic precursors for any products insoluble in 70% ethanol, nor as substrates for a sugar nucleotide 4-epimerase which is present in the particulate enzyme preparation. The (14)C-labelled beta-(1-->4)-galactan is alkali-insoluble and was characterized by analysis of partial acetolysis products. The labelling pattern of the [(14)C]oligosaccharides derived from acetolysis indicates that (1) only slightly more than two [(14)C]galactose moieties are added to the growing polysaccharide chain on average, and (2) these additions take place at the reducing end of the polysaccharide chain. The radioactive beta-(1-->4)-linked galactan chain represented 8.5% of the radioactivity initially added, and 20% of the water- and butanol-insoluble products derived from UDP-alpha-d-[(14)C]galactose. Total hydrolysis of the alkali-insoluble fraction of Phaseolus aureus hypocotyl yielded d-glucose and d-mannose in a 5:1 ratio but no detectable quantities of d-galactose. A trace quantity of a radioactive disaccharide, identified as (1-->3)-linked galactobiose, was isolated from the partial acetolysate of the alkali-insoluble [(14)C]polysaccharide material. Also isolated from this partial acetolysate was a C-1 derivative of [(14)C]galactose, which could not be identified. An alkali-soluble galactose-containing polysaccharide was also synthesized in this enzymic reaction, and represented 20% of the water- and butanol-insoluble products derived from UDP-alpha-d-[(14)C]galactose. The spectrum of radioactive oligosaccharides produced by partial acetolysis of this alkali-soluble polysaccharide material was different from that obtained from the alkali-insoluble polysaccharide, indicating a different structure.

Carbohydrate Epimerases↗
PubMed1979

Specific interaction of human Tamm-Horsfall gylcoprotein with leucoagglutinin, a lectin from Phaseolus vulgaris (red kidney bean).

Human Tamm-Horsfall glycoprotein inhibits lymphocyte transformation induced by leucoagglutinin and haemagglutinin from Phaseolus vulgaris (red kidney bean). The glycoprotein interacts with the two lectins, giving insoluble precipitates. The interaction with leucoagglutinin is highly specific, and the shape of the precipitin curve is that of an antigen-antibody reaction; precipitation is specifically inhibited by N-acetyl-D-galactosamine. Results are discussed, and it is suggested that inhibition of lymphocyte transformation is due to competition between human Tamm-Horsfall glycoprotein and carbohydrate receptors on lymphocytes for the two lectins. The interaction between human Tamm-Horsfall glycoprotein and Phaseolus vulgaris lectins has been used to develop a one-step procedure for the separation of the two lectins by affinity chromatography on (human Tamm-Horsfall-glycoprotein)-Sepharose.

Acetylglucosamine↗
PubMed1998

Characterization and sugar-binding properties of arcelin-1, an insecticidal lectin-like protein isolated from kidney bean (Phaseolus vulgaris L. cv. RAZ-2) seeds.

Arcelin-1 is a lectin-like protein found in the seeds of wild varieties of the kidney bean (Phaseolus vulgaris). This protein displays insecticidal properties, but the mechanism of action is as yet unknown. In the present study we investigated the biochemical and biophysical properties of arcelin-1 from Phaseolus vulgaris cv. RAZ-2. Native arcelin-1 is a dimeric glycoprotein of 60 kDa, built from the non-covalent association of two identical monomers. This dimer resists dissociation by chaotropic agents and is highly resistant to proteolytic enzymes. Each subunit contains 10% (w/w) neutral sugars which belong to the high-mannose and complex-type glycans attached to three glycosylation sites. No interaction of the protein with simple sugars could be detected, but arcelin-1 displays an intrinsic specificity in binding complex glycans. Arcelin-1 therefore differs from the closely related phytohaemagglutinin lectins and alpha-amylase inhibitor in several respects: oligomerization states, sugar-binding affinities and the type and number of glycan chains. These features may be related to the toxicity of arcelin-1.

Amino Acid Sequence↗
PubMed1984

Dry beans of Phaseolus. A review. Part 2. Chemical composition: carbohydrates, fiber, minerals, vitamins, and lipids.

Beans of Phaseolus are an important food crop both economically and nutritionally, and are cultivated and consumed worldwide. With ever rising costs of meats and fresh fruits and vegetables, dry beans are expected to contribute more to the human nutrition in coming years. Traditionally, they have been referred to as "poor man's meat" and have contributed significantly to the diets of many people of several countries in Asia, Africa, Middle East, and South America. In recent years, a renewed interest in bean research in Western European countries and the U.S. is evident. In this review, certain biochemical, technological, nutritional, and toxicological aspects are discussed and the limitations and problems associated with dry beans of Phaseolus as human food are addressed.

Animals↗
PubMed1984

Dry beans of Phaseolus: a review. Part 3.

Beans of Phaseolus are important food crops both economically and nutritionally, and are cultivated and consumed world wide. With ever rising costs of meats, fresh fruits, and vegetables, dry beans are expected to contribute more to the human nutrition in coming years. Traditionally, they have been referred to as "poor man's meat" and have contributed significantly to the diets of many people of several countries in Asia, Africa, the Middle East, and South America. In recent years, a renewed interest in bean research in Western European countries and the U.S. is evident. In this review, certain biochemical, technological, nutritional, and toxicological aspects are discussed and the limitations and problems associated with dry beans of Phaseolus as human food are addressed.

Cooking↗
PubMedDate not supplied

The nodS gene of Rhizobium tropici strain CIAT899 is necessary for nodulation on Phaseolus vulgaris and on Leucaena leucocephala.

Rhizobium tropici strain CIAT899 induces nitrogen-fixing nodules on the roots of a wide range of tropical legumes, including Phaseolus vulgaris and Leucaena leucocephala. Previously, a DNA region of the CIAT899 pSym plasmid containing the common nodulation genes nodABC and one of the nodD alleles was characterized (P. van Rhijn, B. Feys, C. Verreth, and J. Vanderleyden, J. Bacteriol. 175: 438-447, 1993). As reported here, the region immediately downstream of nodC contains the nodSU genes. The nucleotide sequence of these genes is presented. CIAT899 nodS and nodU mutants were constructed. The nodS mutant was completely deficient in nodulation on the host plants P. vulgaris and L. leucocephala. The nodU mutation caused a decrease in nodulation on Leucaena but resorted no effect on Phaseolus. Introduction of the CIAT899 nodABCSU region in R. etli CE-3, a strain that only nodulates P. vulgaris, caused an extension of the host range of strain CE-3 to L. leucocephala.

Amino Acid Sequence↗
PubMed1993

Activation of bean (Phaseolus vulgaris) alpha-amylase inhibitor requires proteolytic processing of the proprotein.

Seeds of the common bean (Phaseolus vulgaris) contain a plant defense protein that inhibits the alpha-amylases of mammals and insects. This alpha-amylase inhibitor (alpha AI) is synthesized as a proprotein on the endoplasmic reticulum and is proteolytically processed after arrival in the protein storage vacuoles to polypeptides of relative molecular weight (M(r)) 15,000 to 18,000. We report two types of evidence that proteolytic processing is linked to activation of the inhibitory activity. First, by surveying seed extracts of wild accessions of P. vulgaris and other species in the genus Phaseolus, we found that antibodies to alpha AI recognize large (M(r) 30,000-35,000) polypeptides as well as typical alpha AI processing products (M(r) 15,000-18,000). Alpha AI activity was found in all extracts that had the typical alpha AI processed polypeptides, but was absent from seed extracts that lacked such polypeptides. Second, we made a mutant alpha AI in which asparagine-77 is changed to aspartic acid-77. This mutation slows down the proteolytic processing of pro-alpha AI when the gene is expressed in tobacco. When pro-alpha AI was separated from mature alpha AI by gel filtration, pro-alpha AI was found not to have alpha-amylase inhibitory activity. We interpret these results to mean that formation of the active inhibitor is causally related to proteolytic processing of the proprotein. We suggest that the polypeptide cleavage removes a conformational constraint on the precursor to produce the biochemically active molecule.

Adsorption↗