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[Biochemical and nutritional value of seeds of the lima bean (Phaseolus lunatus) in comparison with those of the common bean (Phaseolus vulgaris)].

Composition in nutrients and antinutritional factors, digestibility and growth in rats fed diets prepared with raw and cooked beans of Ph. vulgaris, cultivar "Tacarigua", and Ph. lunatus cultivar "Tapiramo", are compared. Grains from both cultivars are similar in appearance, taste, nutritional value, and acceptability. Protein and phosphorus contents were greater in Ph. vulgaris than in Ph. lunatus seeds. The chemical score and availability of lysine were better in Ph. lunatus. Diets prepared with raw beans from Ph. lunatus resulted non toxic for the rats during a 12-day period of feeding. All rats fed with raw beans from Ph. Vulgaris died in the same period of time. Protein efficiency was better with cooked beans of Ph. lunatus. The cultivar "Tapiramo" (Ph. lunatus) is recommended for autoconsumption by small farmers.

Animals↗

Purification, properties and comparative specificities of the enzyme prolyl-transfer ribonucleic acid synthetase from Phaseolus aureus and Polygonatum multiflorum.

1. A prolyl-s-RNA synthetase (prolyl-transfer RNA synthetase) has been purified about 250-fold from seed of Phaseolus aureus (mung bean), a species not producing azetidine-2-carboxylic acid, and more than 10-fold from rhizome apices of Polygonatum multiflorum, a liliaceous species containing azetidine-2-carboxylic acid. The latter enzyme was unstable during ammonium sulphate fractionation. 2. The enzymes exhibited different substrate specificities towards the analogue. That from Phaseolus, when assayed by the ATP-PP(i) exchange, showed azetidine-2-carboxylic acid activation at about one-third the rate with proline. Both labelled imino acids gave rise to a labelled aminoacyl-s-RNA. The enzyme from Polygonatum, however, activated only proline. 3. The enzyme from Polygonatum also formed a labelled prolyl-s-RNA with Phaseolus s-RNA but at a lower rate than when the Phaseolus enzyme was used. No reaction occurred when the Phaseolus enzyme was coupled with Polygonatum s-RNA, and only a very slight one was observed when both enzyme and s-RNA came from Polygonatum. 4. Protein preparations from seeds of Pisum sativum, another species not producing azetidine-2-carboxylic acid, also activated the analogue in addition to proline, whereas those from rhizome and seeds of Convallaria, the species from which the analogue was originally isolated, failed to activate it. However, a liliaceous species not producing the analogue, Asparagus officinalis, activated it. 5. Of the other proline analogues investigated, only 3,4-dehydro-dl-proline and l-thiazolidine-4-carboxylic acid were active with the enzyme preparation from Phaseolus. 6. pH optima of 7.9 and 8.4 were established for the enzymes from Phaseolus and Polygonatum respectively. 7. The Phaseolus enzyme was specific for ATP and PP(i). Mn(2+) partially replaced the requirement for Mg(2+) as cofactor. Preincubation with p-chloromercuribenzoate at a concentration of 0.5mm or higher produced over 99% inhibition of the Phaseolus enzyme. One-half the enzymic activity was destroyed by preheating for 5min. at 62 degrees in tris-hydrochloric acid buffer, pH7.9. 8. All experimental evidence supports the hypothesis that azetidine-2-carboxylic acid and proline are activated by the same enzyme in Phaseolus preparations, whereas the analogue was inactive in all Polygonatum preparations. The possible nature of this different substrate behaviour is discussed.

Journal Article↗

Effect of variable protein contents in diets containing Phaseolus vulgaris beans on performance, organ weights and blood variables in piglets, rats and chickens.

A comparison was made of the effects of antinutritional factors present in Phaseolus vulgaris on piglets, rats and chickens. Also the hypothesis of whether the negative effect on weight gain due to the inclusion of raw Phaseolus vulgaris in the diet can be attributed to an insufficient supply of amino acids was tested. Test diets containing 200 g raw Phaseolus beans/kg were balanced for digestible protein and amino acids; in one diet extra casein was incorporated. The main response criteria were live-weight gain and the weight of various organs including the intestine. Live-weight gain in piglets was markedly reduced during feeding 200 g raw Phaseolus vulgaris/kg in the diet, but not in rats and chickens. Addition of casein did not improve the weight gain of the piglets, indicating that a toxic factor was responsible for the reduced weight gain and not an insufficient supply of amino acids. The weights of the spleen and thymus were markedly reduced in the piglets when the diets with raw Phaseolus beans were given, but not in the rats and chickens. Additional supply of casein did not change this effect. Indications were found that when the supply of dietary protein is adequate there is no reduction in pancreas weight with raw Phaseolus beans as was observed in previous experiments. The weight of the intestine was increased in all three species due to feeding raw Phaseolus vulgaris.

Amino Acids↗

Molecular evolution and phylogenetic implications of internal transcribed spacer sequences of nuclear ribosomal DNA in the Phaseolus-Vigna complex.

Molecular phylogeny based on internal transcribed spacer (ITS) sequences was studied to resolve the taxonomic contradiction in Vigna and its relation to Phaseolus. The ITS region of the 18S-26S nuclear ribosomal DNA repeat was sequenced for 29 Vigna species, selected from five of the nine subgenera, and 9 species of Phaseolus. The length of ITS-1 ranged from 187 to 243 bp and 217 to 290 bp, and that of ITS-2 from 187 to 219 bp and 225 to 243 bp, within Vigna and Phaseolus species, respectively. Phylogenies derived from ITS sequences based on maximum-parsimony and neighbor-joining methods gave trees essentially of similar topology. The ITS phylogeny was generally congruent with recent classifications based largely on morphological, biochemical, cytogenetical, and palynological features, except that subgenus Plectotropis of Neotropical origin was revealed to be closely related to subgenus Vigna instead of forming a link between African (subgenus Vigna) and Asiatic (subgenus Ceratotropis) vignas, and subgenus Sigmoidotropis, featuring morphological characters of both Vigna and Phaseolus, was placed as the sister group to the Phaseolus taxa. The ITS sequences were shown to be useful for identifying wild progenitors of V. mungo, V. radiata, V. umbellata, and V. unguiculata and for clarifying taxonomy-related problems in many previously controversial cases. This study also affirms that V. umbellata and V. angularis are the diploid progenitors of the only tetraploid species (V. glabrescens) known in the genus.

Base Composition↗

A fragment of chloroplast DNA was transferred horizontally, probably from non-eudicots, to mitochondrial genome of Phaseolus.

The mitochondrial genomes of some Phaseolus species contain a fragment of chloroplast trnA gene intron, named pvs-trnA for its location within the Phaseolus vulgaris sterility sequence (pvs). The purpose of this study was to determine the type of transfer (intracellular or horizontal) that gave rise to pvs-trnA. Using a PCR approach we could not find the respective portion of the trnA gene as a part of pvs outside the Phaseolus genus. However, a BLAST search revealed longer fragments of trnA present in the mitochondrial genomes of some Citrus species, Helianthus annuus and Zea mays. Basing on the identity or near-identity between these mitochondrial sequences and their chloroplast counterparts we concluded that they had relocated from chloroplasts to mitochondria via recent, independent, intracellular DNA transfers. In contrast, pvs-trnA displayed a relatively higher sequence divergence when compared with its chloroplast counterpart from Phaseolus vulgaris. Alignment of pvs-trnA with corresponding trnA fragments from 35 plant species as well as phylogenetic analysis revealed that pvs-trnA grouped with non-eudicot sequences and was well separated from all Fabales sequences. In conclusion, we propose that pvs-trnA arose via horizontal transfer of a trnA intron fragment from chloroplast of a non-eudicot plant to Phaseolus mitochondria. This is the first example of horizontal transfer of a chloroplast sequence to the mitochondrial genome in higher plants.

Base Sequence↗

Primary structures of alpha- and beta-subunits of alpha-amylase inhibitors from seeds of three cultivars of Phaseolus beans.

The primary structures of three alpha-amylase inhibitors (TAI, DAI, and MAI-2) consisting of glycoprotein subunits alpha and beta from the respective seeds of three cultivars of Phaseolus beans, Toramame (Phaseolus vulgaris L.), Daifukumame (Phaseolus vulgaris L.), and Murasakihanamame (Phaseolus coccineus L.) were determined by sequencing the peptide fragments derived from their enzymatic digestions. Major sugar chains of the inhibitors were also assessed by analyzing glycopeptides in the enzymatic digests. The subunits, alpha and beta, were shown to be composed of 76 and 139 amino acid residues, respectively, in each inhibitor. The overall amino acid sequences of the inhibitors were slightly different from one another. Furthermore, the sequence of TAI was the same as that deduced from a cDNA clone encording alpha-amylase inhibitor-1 from the common bean (Phaseolus vulgaris L.). It was also revealed that there were two N-glycosylation sites in each alpha-subunit: PA-derivatives of the major N-glycans were estimated to be M6B at Asn(12) and M9A at Asn(65). Each beta-subunit of TAI and MAI-2 had two N-glycosylation sites, while the beta-subunit of DAI had only one site. The major N-glycans pyridylaminated were estimated to be M3X at Asn(63) in each beta-subunit and M3FX at Asn(83) in beta-subunits of TAI and MAI-2.

Amino Acid Sequence↗

Nitrate reductase activity, distribution, and response to nitrate in two contrasting Phaseolus species inoculated with Rhizobium spp.

The nitrate reductase activity distribution and response of two nodulated species of Phaseolus (Phaseolus vulgaris-common bean, and Phaseolus lunatus-lima bean) to different exogenous nitrate levels were studied during the vegetative period. These Phaseolus species showed to be very contrasting in respect to the pattern of nitrate reductase (NR) activity distribution thought the plant. The highest level of NR activity in P. vulgaris was clearly shown to occur in leaves in contrast with the lowest one detected in roots and nodules as widely seen for other tropical species of the Phaseoleae tribe. Conversely, P. lunatus had higher NR activity in the nodules, whereas its leaves exhibited a steadily decrease during the plant development. Indeed, at 32 days after emergence (pre-flowering stage), the nodulated P. vulgaris had approximately 95% of the total NR activity localized in its leaves, whereas in P. lunatus it was equally distributed in the nodules and in the leaves. Under long-term exposure to increasing exogenous level of nitrate, the leaf-NR activity of nodulated P. vulgaris presented a positive response, whereas the enzyme activity was very low and unresponsive in P. lunatus. In contrast, the nodule-NR activity showed a reverse response to the increasing NO(3)(-) level. The nodule-NR activity of P. lunatus significantly increased whereas in the P. vulgaris nodules it was very low and unresponsive. This present study suggests that P. lunatus inoculated with Rhizobium tropici presents a singular pattern of nitrate reduction distribution among leaves and nodules during the vegetative development. It is speculated that the nodulated Phaseolus lunatus may have different NR isoforms in their leaves (at least a constitutive type) and an inducible form in their nodules, responsive to long-term exposure to nitrate.

Journal Article↗

Translocation of Indole-3-acetic Acid-1'-C and Tryptophan-1-C in Seedlings of Phaseolus coccineus L. and Zea mays L.

Indole-3-acetic acid-1'-(14)C (IAA-(14)C) and tryptophan-1-(14)C injected in small amounts into cotyledons of Phaseolus coccineus L. seedlings were found to be translocated acropetally into the epicotyls and young shoots. Similarly IAA-(14)C was translocated acropetally into coleoptiles of Zea mays following injection into the endosperms. Labeled metabolites of the injected compounds were also extractable from shoot tissue. However, evidence that IAA-(14)C itself was translocated acropetally was obtained by collection in agar blocks applied to cut surfaces of coleoptiles of injected seedlings. The acropetal translocation in Phaseolus was shown not to occur in the transpiration stream but in living tissue. Cotyledons of Phaseolus coccineus and Phaseolus vulgaris contain extensive vascular tissue.Tryptophan-(14)C was not actively translocated through excised segments of Phaseolus coccineus epicotyl and Zea mays coleoptile when supplied from donor agar blocks in concentrations as high as 100 mum. The small amount of tryptophan-(14)C which did reach receiver blocks when high concentrations were used may be accounted for by passive diffusion through the fluid-filled xylem vessels. Translocation of a non-toxic dye, Light Green SF Yellowish, through xylem vessels was found to occur when supplied from donor blocks placed acropetally or basipetally. Metabolism of the supplied tryptophan-(14)C by the tissue segments was shown to occur during the 3 to 6 hour translocation experiments. IAA-(14)C was transported in a strictly basipetal manner in both tissues. Only 1 labeled compound with an R(F) value of IAA was found in receiver blocks. Composition of a simple green safelight suitable for work in plant physiology is described.

Journal Article↗

Resistance of Phaseolus species to ice crystallization at subzero temperature.

Dry bean (Phaseolus vulgaris L.) cultivars possess little or no freezing tolerance and are killed at the temperature of ice formation in their tissues. An increase in frost tolerance by 2-3 degrees C would expand dry bean production in the short growing seasons of the Canadian prairies and possibly to higher altitudes in the tropics where episodic frosts occur during the growing season. The objective of this study was to determine the differences in frost resistance of Phaseolus species in both controlled and field environments. Leaflets of dry bean cv. CDC Nighthawk, and wild relatives from the primary gene pool (P. vulgaris var. mexicanus Freytag and P. vulgaris var. aborigineus (Burkart) Baudet) and the tertiary gene pool (P. acutifolius var. tenuifolius A. Gray, P. filiformis Bentham, P. angustissimus A. Gray and P. ritensis M.E. Jones) were subjected to subzero temperatures with and without ice nucleation to determine the levels of tolerance and avoidance, respectively. The lethal temperature at which 50% of the leaflets were killed (LT(50)) was 0.5-1 degrees C lower for species of the tertiary gene pool compared to those from the primary gene pool. Leaflets of species from the tertiary gene pool were also characterized by extensive supercooling compared to leaflets of species from the primary gene pool. Resistance of Phaseolus species to spring and autumn frosts were determined on seedlings transplanted to the field. Phaseolus angustissimus, a species of the tertiary gene pool had the highest seedling survival in response to both autumn and spring frosts, when the minimum air temperatures were -5 and -7 degrees C, respectively. Frost resistance of Phaseolus angustissimus, if successfully introgressed into bean germplasm, may enable the development of frost resistant dry bean cultivars.

Journal Article↗

CENP-B autoantigen is a conserved protein from humans to higher plants: identification of the aminoterminal domain in Phaseolus vulgaris.

Centromeres are critical structures in cell division, and CENP-B is the most important protein of the centromeric complex recognized by autoantibodies from patients with scleroderma. Our major aim was to demonstrate whether CENP-B is a conserved protein along the phylogenic scale including the higher plants. Vegetal and human cell proteins were extracted from Phaseolus vulgaris and HEp-2 cells and were characterized by PAGE, Western blot, and human autoimmune sera containing anti-CENP-B autoantibodies. The aminoterminus of the gene encoding for CENP-B from HEp-2 cells and Phaseolus vulgaris was isolated by reverse transcriptase-PCR using complementary oligonucleotides to the human CENP-B gene. Also, in situ hybridization was performed on vegetal tissues and HEp-2 cells using human CENP-B box probes. Our main results were as follows: 1) Autoimmune sera were reactive to a vegetal protein of 80 kDa. 2) Affinity-purified anti-CENP-B antibodies recognized a protein from Phaseolus vulgaris with molecular mass similar to that found in human cells. Vegetal and HEp-2 cells CENP-B proteins were immunologically identical. 3) Using RT-PCR, we were able to amplify a cDNA encoding for the aminoterminus domain of CENP-B from Phaseolus vulgaris that had the same molecular behaviour as the cDNA from HEp-2 cells. 4) Complementary oligonucleotides for human CENP-B box hybridized a DNA sequence from Phaseolus vulgaris. In conclusion, CENP-B protein is a conserved protein along the phylogenic scale from humans to higher plants.

Antibody Specificity↗

Amine oxidase from Lathyrus cicera and Phaseolus vulgaris: purification and properties.

Cu-Amine oxidases (amine oxygen oxidoreductase deaminating, copper containing E.C. 1.4.3.6.) are found in all forms of life (1). They catalyze the following general reaction: R-CH2-NH2 + O2 + H2O----R-CHO + NH3 + H2O2. Cu-amine oxidases (Cu-AOs) have been extracted from different leguminosae: Pisum sativum (2-3), Lathyrus sativus (4), Lens esculenta (5), Vicia faba (6), Cicer arietinum (7), Glycine max (8) but not from Phaseolus vulgaris. Palavan and Galston (9), in a study of polyamine biosynthesis during developmental stages of Phaseolus vulgaris, did not detect diamine or polyamine oxidase activity in Phaseolus. The present paper describes the purification of Phaseolus vulgaris seedlings amine oxidase (PhSAO) and also compares the properties of this enzyme to the Lathyrus cicera enzyme (LcSAO), obtained with the same method of purification.

Amine Oxidase (Copper-Containing)↗

Influence of a natural-ingredient diet containing Phaseolus vulgaris on the colonization by segmented, filamentous bacteria of the small bowel of mice.

The appearance of segmented, filamentous bacteria (SFBs) in the small bowel of mice is influenced by the composition of the diet, but the dietary components responsible are not known. The addition of ground, whole Phaseolus vulgaris to a natural-ingredient diet at the expense of part of the skim milk powder, ground barley and wheat middlings components, caused an increase of the colonization of the mouse small bowel by SFBs. This effect was not seen when whole Phaseolus was added to a purified diet at the expense of part of the casein, corn oil, coconut fat, corn starch, dextrose and cellulose components. In an attempt to identify the fraction of Phaseolus that might contain SFB-inducing substances, the skin and kernel fraction of the bean were added to the natural-ingredient diet. The skin and kernel fraction were found to be as effective in inducing SFB appearance as was whole Phaseolus.

Animal Feed↗

Local (gut) and systemic metabolism of rats is altered by consumption of raw bean (Phaseolus vulgaris L var athropurpurea).

The composition of the raw legume Phaseolus vulgaris L. var. athropurpurea (PhVa) and its effects on the metabolism of young growing rats have been evaluated. The levels of protein, unsaturated fatty acids, carbohydrate, fibre and bioactive factors present in PhVa were comparable with those in other Phaseolus vulgaris varieties. However, the lectins of PhVa were predominantly of the leucoagglutinating type, and concentrated in the albumin protein fraction. Rats fed a diet (110 g total protein, 16.0 MJ/kg) in which PhVa meal provided about half of the protein excreted high levels of N in faeces and urine, and grew more slowly, than rats fed a high-quality control diet (ad libitum or pair-fed). Small intestine, large intestine and pancreas weights were increased (by almost 100 %, P<0.05), whilst skeletal muscle, thymus and spleen weights were reduced. Blood insulin (16.20 v. 0.50 mU/l, P<0.05, thyroxine, glucose, protein (60.5 v. 48.3 g/l, P<0.05) and LDL-cholesterol were lowered, whilst glucagon (155.3 v. 185.4 ng/l, P<0.05), triiodothyronine and urea were elevated, as were urinary urea, creatinine and glucose. These changes in the local (gut) and systemic metabolism of rats were probably mediated primarily by lectins in PhVa, which were concentrated in the albumin protein fraction, whereas in many other Phaseolus vulgaris lines they are distributed across the globulin and albumin fractions.

Animal Nutritional Physiological Phenomena↗

Agrobacterium rhizogenes transformation of the Phaseolus spp.: a tool for functional genomics.

A fast, reproducible, and efficient transformation procedure employing Agrobacterium rhizogenes was developed for Phaseolus vulgaris L. wild accessions, landraces, and cultivars and for three other species belonging to the genus Phaseolus: P. coccineus, P. lunatus, and P. acutifolius. Induced hairy roots are robust and grow quickly. The transformation frequency is between 75 and 90% based on the 35-S promoter-driven green fluorescent protein and beta-glucuronidase expression reporter constructs. When inoculated with Rhizobium tropici, transgenic roots induce normal determinate nodules that fix nitrogen as efficiently as inoculated standard roots. The A. rhizogenes-induced hairy root transformation in the genus Phaseolus sets the foundation for functional genomics programs focused on root physiology, root metabolism, and root-microbe interactions.

Blotting, Southern↗

Diversity in the rhizobia associated with Phaseolus vulgaris L. in Ecuador, and comparisons with Mexican bean rhizobia.

Common beans (Phaseolus vulgaris L.) have centers of origin in both Mesoamerica and Andean South America, and have been domesticated in each region for perhaps 5000 years. A third major gene pool may exist in Ecuador and Northern Peru. The diversity of the rhizobia associated with beans has also been studied, but to date with an emphasis on the Mesoamerican center of origin. In this study we compared bean rhizobia from Mexico and Andean South America using both phenotypic and phylogenetic approaches. When differences between the rhizobia of these two regions were shown, we then examined the influence of bean cultivar on the most probable number (MPN) count and biodiversity of rhizobia recovered from different soils. Three clusters of bean rhizobia were distinguished using phenotypic analysis and principal-component analysis of Box AIR-PCR banding patterns. They corresponded principally to isolates from Mexico, and the northern and southern Andean regions, with isolates from southern Ecuador exhibiting significant genetic diversity. Rhizobia from Dalea spp., which are infective and effective on beans, may have contributed to the apparent diversity of rhizobia recovered from the Mesoamerican region, while the rhizobia of wild Phaseolus aborigineus from Argentina showed only limited similarity to the other bean rhizobia tested. Use of P. vulgaris cultivars from the Mesoamerican and Andean Phaseolus gene pools as trap hosts did not significantly affect MPN counts of bean rhizobia from the soils of each region, but did influence the diversity of the rhizobia recovered. Such differences in compatibility of host and Rhizobium could be a factor in the poor reputation for nodulation and N2 fixation in this crop.

Colony Count, Microbial↗