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PubMed1987

The complete amino acid sequence of the Vigna unguiculata (L.) Walp. seed trypsin and chymotrypsin inhibitor.

The amino acid sequence of the "double-headed" trypsin and chymotrypsin inhibitor (BTCI), purified from the cowpea Vigna unguiculata (L.) Walp. cv "Seridó" was determined in our laboratory. Tryptic and chymotryptic peptides were sequenced by the Edman-Gray and Edman-Chang N-terminal manual method as well as the carboxypeptidase C-terminal method. The complete amino acid sequence of the BTCI is: Ser-Gly-His-His-Glx-Asx-Ser-Thr-Asx-Glx-Ala-Ser-Glx-Ser-Ser-Lys-Pro-Cys- Cys-Arg- Glx-Cys-Ala-Cys-Thr-Lys-Ser-Ile-Pro-Pro-Glx-Cys-Arg-Cys-Ser-Asx-Val-Arg- Leu-Asn- Ser-Cys-His-Ser-Ala-Cys-Lys-Ser-Cys-Ala-Cys-Thr-Phe-Ser-Ile-Pro-Ala-Glx- Cys-Phe- Cys-Gly-Asx-Ile-Asx-Asx-Phe-Cys-Tyr-Lys-Pro-Cys-Lys-Ser-Ser-His-Ser-Asx- Asx-Asx-Asx-Trp-Asn. BTCI presents a high degree of homology with the Bowman-Birk proteinase inhibitor family.

Amino Acid Sequence↗
PubMed1980

Inhibition of mosaic virus of Vigna sinensis Endl. with four chemicals.

Four chemicals--gibberellic acid, indoleacetic acid, 2,4-6 and thiouracil were tested for their inhibitory effect against a virus disease of Vigna sinensis (cowpea). All the four chemicals inhibited the virus to a considerable extent in in vitro studies. Pre and post-inoculation treatments were equally effective in inhibiting the virus, viz. local lesion formation on Chenopodium amaranticolor leaves. The level of inhibition subsequently decreased with lower concentration of the chemicals. Seed soaking in the chemicals controlled the virus infection.

2,4-Dichlorophenoxyacetic Acid↗
PubMed1978

Microtubules and organelle movements in the rust fungus Uromyces phaseoli var. vignae.

Direct visual observation and time lapse films of in vitro differentiating infection structures of the cowpea rust fungus Uromyces phaseoli var. vignae revealed three categories of movement: a) general movement of cytoplasm, plus organelles, into the developing portions of the fungus during which the nuclei, in particular, maintained their characteristic position with remarkable constancy, b) relatively slow movements of various organelles such that they became displaced relative to one another and to the growing fungal tip, and c) erratic, rapid, saltations of small organelles over short distances. Serial section ultrastructural analysis showed that microtubules were typically orientated parallel to the direction of cytoplasm migration. Simple statistical analyses showed that the microtubules were non-randomly associated with mitochondria but only rarely associated with lipid droplets or microbodies. All microtubules were typically short (less than 2 micrometer) and, in various parts of the cell, were often intimately associated with 3 to 6 nm diameter filaments of unidentified material. Interphase nuclei characteristically lacked microtubules emanating from their variously laterally or posteriorly located NAOs (nucleus associated organelle) but were associated with groups of laterally placed microtubules. The correlations between the observed types of movement and the ultrastructure of the cells discussed in terms of various models for organelle motility.

Basidiomycota↗
PubMedDate not supplied

[Nitrogenase activity of Rhizobium meliloti and Rhizobium vigna in a root tisse culture of leguminous and nonleguminous plants].

As was shown using the acetylene technique, the nitrogenase activity of Rhizobium meliloti and Rhizobium vigna increased when they were cultivated with the root tissue cultures of legumes (lucerne, sweetclover) and non-legumes (tobacco, glasswort, carrot), particularly in the case of the former. The maximum activity of nitrogenase was found in R. meliloti. The tissue cultures of legumes had no effect on the growth of Rhizobium whereas the tissues of non-legumes stimulated their biomass accumulation though the activity of nitrogenase in both Rhizobium cultures was low in this case. Therefore, the metabolites of legumes produced a specific action on the nitrogenase of nodule bacteria.

Culture Media↗
PubMed1995

Purification and properties of storage proteins (vicilins) from cowpea (Vigna unguiculata) seeds which are susceptible or resistant to the bruchid beetle Callosobruchus maculatus.

Vicilins (7S storage proteins) from seeds of cowpea (Vigna unguiculata) cultivars which are susceptible or resistant to the bruchid beetle C. maculatus were purified by size-exclusion and ion-exchange chromatography. The vicilins were partially characterized by polyacrylamide gel electrophoresis under both denaturing and nondenaturing conditions, by Western blotting and by amino acid analysis. The variant vicilins from C. maculatus-resistant seeds do not differ appreciably from vicilins from susceptible seeds by these criteria except that they are more strongly bound to DEAE-Sepharose, suggesting differences in charge between the various molecules.

Amino Acids↗
PubMed1995

Consumption of diets containing raw soya beans (Glycine max), kidney beans (Phaseolus vulgaris), cowpeas (Vigna unguiculata) or lupin seeds (Lupinus angustifolius) by rats for up to 700 days: effects on body composition and organ weights.

Feeding trials have been done with rats to assess the effects of long-term (700 d) consumption of diets based on raw cowpeas (Vigna unguiculata; moderate Bowman-Birk inhibitor content, low lectin content), lupin seeds (Lupinus angustifolius; low lectin and protease inhibitor content) or soya beans (Glycine max; high Kunitz inhibitor content, moderate Bowman-Birk inhibitor content, moderate lectin content) or diets containing low levels of raw kidney bean (Phaseolus vulgaris; high lectin content, low Bowman-Birk inhibitor content) on body weight and composition and organ weights. All the legume-based diets reduced feed conversion efficiency and growth rates during the initial 250 d. However, after 250 d the weight gains by rats given legume-based diets were similar to those of controls given the same daily feed intake. Long-term consumption of diets containing low levels of kidney bean significantly altered body composition of rats. The levels of lipid in the body were significantly reduced. As a result, carcasses of these rats contained a higher proportion of muscle/protein than did controls. Small-intestine relative weight was increased by short- and long-term consumption of the kidney-bean-based diet. However, the increase in relative pancreatic weight observed at 30 d did not persist long term. None of the other legume-based diets caused any significant changes in body composition. However, long-term exposure to a soya-bean- or cowpea-based diet induced an extensive increase in the relative and absolute weights of the pancreas and caused an increase in the incidence of macroscopic pancreatic nodules and possibly pancreatic neoplasia. Long-term consumption of the cowpea-, kidney-bean-, lupin-seed- or soya-bean-based diets by rats resulted in a significant increase in the relative weight of the caecum and colon.

Animal Feed↗
PubMed1993

Cloning of ornithine delta-aminotransferase cDNA from Vigna aconitifolia by trans-complementation in Escherichia coli and regulation of proline biosynthesis.

Proline prototrophy was restored to an Escherichia coli proBA proline auxotroph by ornithine and a mothbean (Vigna aconitifolia) cDNA expression library. This novel strategy, "trans-complementation," allowed isolation of a cDNA encoding ornithine delta-aminotransferase (delta-OAT). This enzyme transaminates ornithine to glutamic-gamma-semialdehyde (GSA), thereby bypassing the block in GSA synthesis from glutamate in the proBA mutant. The identity of the mothbean enzyme was confirmed by its high sequence homology to mammalian and yeast delta-OATs as well as to a family of bacterial and fungal omega-aminotransferases and an absence of significant homology to various alpha-aminotransferases. The V. aconitifolia OAT cDNA encodes a polypeptide of 48.1 kDa. The native enzyme expressed in E. coli appears to be a monomer with Km of 2 mM for ornithine and 0.75 mM for alpha-ketoglutarate. Levels of mRNA in V. aconitifolia for delta 1-pyrroline-5-carboxylate synthetase (P5CS) and delta-OAT, the two key enzymes for proline synthesis, were monitored under different physiological conditions. Salt stress and nitrogen starvation induced P5CS mRNA levels and depressed OAT mRNA levels. Conversely, OAT mRNA level was elevated in plants supplied with excess nitrogen while the P5CS mRNA level was reduced. These data suggest that the glutamate pathway is the primary route for proline synthesis in plants during conditions of osmotic stress and nitrogen limitation whereas the ornithine pathway assumes prominence under high nitrogen input.

1-Pyrroline-5-Carboxylate Dehydrogenase↗
PubMed1996

Chitin-binding proteins from cowpea (Vigna unguiculata) seeds.

Vicilins (7S storage proteins) from cowpea (Vigna unguiculata) and other legume seeds were shown to bind to chitin, to regenerated chitin (fully acetylated chitin) and to chitosan (deacetylated chitin). Adsorbed vicilins were desorbed from these matrices by acetic and hydrochloric acids and by highly polymerized soluble chitosan. Proteins such as the lectin of common bean (PHA), soybean trypsin inhibitor (Kunitz), a beta-1,3-glucanase from cowpea seeds, bovine pancreatic alpha-chymotrypsin, chicken ovalbumin, serum albumin and rabbit gamma-globulin did not bind. The present result is the first description of vicilin binding to chitin but other proteins, such as wheat germ agglutinin (WGA), a lectin that contains the so called "chitin-binding domain", and a chitinase isolated from cowpea seeds, which are involved in the defense mechanisms of plants against insects and fungi, were also shown to bind to chitin as previously reported. The binding of vicilins to chitin is probably effected not through a "chitin-binding domain" because they do not share this sequence with the defense-related proteins cited above. We propose that this association of vicilins with chitin may be related to the effect of variant vicilins on the development of Callosobruchus maculatus (bruchid) in resistant cowpea seeds.

Chemical Fractionation↗
PubMed2000

Effect of domestic processing on flatus producing factors in ricebean (Vigna umbellata).

The effect of various processing treatments viz., soaking, pressure cooking, open pan cooking, germination followed by pressure cooking and roasting of whole ricebean and pressure cooking, open pan cooking and fermentation and frying of fermented batter of dehulled ricebean flour was assessed for total sugars and oligosaccharide contents. Dehulling caused a significant reduction in the raffinose and stachyose contents. Pre-soaking of ricebean caused appreciable losses in the oligosaccharide content. Germination (48 h) followed by pressure cooking and fermentation (18 h) followed by frying caused the maximum losses in the raffinose and stachyose contents.

Chromatography, Paper↗
PubMed1980

Carbon and nitrogen nutrition of plant pathogenic fungi associated with basal stem rots of cowpeas, Vigna unguiculata (L) Walp in Nigeria.

Pythium aphanidermatum, Rhizoctonia bataticola (syn. = Macrophomina phaseolina), Botryodiplodia theobromae, and two strains of Sclerotium rolfsii readily utilized for growth the monosaccharides glucose, fructose and mannose, the disaccharides sucrose and maltose and the polysaccharides dextrin and starch. In addition, S. rolfsii grew appreciably well on carboxymethylcellulose (CMC). When filter paper (nature cellulose) was supplied as sole carbon source, it was decomposed by B. theobromae, S. rolfsii (SR2) and R. bataticola. In studies on nitrogen utilization, B. theobromae grew best on glutamic acid, P. aphanidermatum on glycine and the two strains of S. rolfsii on ammonium nitrate. R. bataticola utilized casein hydrolysate most effectively for growth. EDTANa2 was a poor nitrogen source for all the pathogens.

Amino Acids↗
PubMed1989

Protein and cDNA sequences of Bowman-Birk protease inhibitors from the cowpea (Vigna unguiculata Walp.).

The protein and gene sequences of the cowpea Bowman-Birk type trypsin inhibitor which confers enhanced insect resistance to transgenic tobacco plants, and of cowpea trypsin/chymotrypsin inhibitors are presented. There are regions of high conservation and high divergence within the 5' leader, mature protein and 3' non-coding regions of the Bowman-Birk inhibitors and in the genes which encode them in different members of this family within the Leguminosae. The practical implications of this finding for studies on the evolution of plants and the utilization of these genes for enhancing insect resistance is discussed.

Amino Acid Sequence↗
PubMed1990

Stored mRNA in cotyledons of Vigna unguiculata seeds: nucleotide sequence of cloned cDNA for a stored mRNA and induction of its synthesis by precocious germination.

By differential hybridization screening, we previously selected a class of cDNA clones from a lambda gt10 cDNA library that was constructed from the total poly(A)+ RNA of mature cowpea cotyledons (Plant Cell Physiol 31: 39-44, 1990). pSAS10, a clone of this class, hybridized with a cDNA probe complementary to poly(A)+ RNA from cotyledons collected 1 day after the onset of imbibition (DAI), but not with the cDNA probe from cotyledons at developmental stage II (13 to 15 days after flowering, DAF). pSAS10 mRNA was detectable only in cotyledons at developmental stage III (17 to 19 DAF) or later, and its level began to decline when seeds germinated. We have suggested that pSAS10 mRNA is likely to belong to the class of 'stored' mRNA or the mRNA that is formed at the late stage of seed maturation, is conserved in quiescent seeds and becomes functional at the early stage of germination. We determined the nucleotide sequence of pSAS10 cDNA consisting of 459 bp and an approximately 36 bp poly(A) tract, and deduced the amino acid sequence of its product, a 10-kDa cysteine-rich polypeptide. Synthesis of pSAS10 mRNA was induced just before germination began, not only in mature seeds but also in immature seeds even at stages I (9 to 11 DAF) and II (13 to 15 DAF) if they were placed under conditions suitable for germination.

Amino Acid Sequence↗
PubMed1996

Pectinmethylesterase isoforms from Vigna radiata hypocotyl cell walls: kinetic properties and molecular cloning of a cDNA encoding the most alkaline isoform.

Peptide maps and partial amino acid sequences of the 3 main pectinmethylesterases (PMEs) solubilized from mung bean hypocotyl cell walls demonstrated that these proteins were different isozymes originating from a small multigene family. A cDNA clone encoding the most alkaline PME (PE gamma) have been obtained by PCR using degenerate oligonucleotide primers. Combining the protein and nucleotide sequencing data, the complete amino acid sequence of PE gamma was determined. The nature protein is composed of 318 amino acids with a calculated Mtau of 34 677 and an estimated pI of 9.84 consistent with the values previously obtained by SDS-PAGE and IEF. It shares most of the conserved regions of previously known PMEs. Enzymatic activities of the three isoforms were differently affected by the presence of cations in the incubation medium but, in all cases, infra-optimal cation concentrations induced two opposite effects: a decrease in the Vmax and an increase in the affinity of the enzymes for their substrate. The presence of cations in the assay modulates both the number of enzyme molecules available to the demethylation reaction and the conformation of the pectin and, in turn, the affinity of the PMEs for their substrate.

Amino Acid Sequence↗