The amino acid sequence of Phaseolus aureus L. (mung-bean) cytochrome c

The amino acid sequence of Phaseolus aureus L. (mung-bean) cytochrome c has been determined. The molecule consists of a single polypeptide chain of 111 amino acid residues and is homologous with other mitochondrial cytochromes c. Comparison with the amino acid sequence of wheat-germ cytochrome c (Stevens, Glazer & Smith, 1967) shows 14 differences. On alignment with mammalian cytochromes c, mung-bean cytochrome c has an N-acetylated ;tail' of eight amino acid residues similar to that found in wheat-germ cytochrome c. Of the 22 positions in wheat-germ cytochrome c that contain amino acid residues unique to these positions, 20 were found to contain the same ones in mung-bean cytochrome c. The in-N-trimethyl-lysine residues reported for wheat-germ cytochrome c (Delange, Glazer & Smith, 1969) in positions 72 and 86 were also found in these positions in mung-bean cytochrome c. The sequence was determined from 3mumol, by using chymotryptic and tryptic peptides which were analysed by the ;dansyl'-Edman method (Gray & Hartley, 1963a), with confirmation by amino acid analysis.     

The amino acid sequence of Helianthus annuus L (sunflower) cytochrome c deduced from chymotrypic peptides

Peptides derived from digestion of 1 mumol of sunflower cytochrome c with chymotrypsin were separated by paper electrophoresis. The sequences of these peptides were determined by using the dansyl-Edman method (Gray & Hartley, 1963) and confirmed by analysis of their amino acid composition. Comparison of the set of peptides with the chymotryptic peptides of mung-bean (Thompson, Laycock, Ramshaw & Boulter, 1970) and wheat germ (Stevens, Glazer & Smith, 1967) cytochrome c shows a clear homology. The complete sequence of sunflower cytochrome c was established by alignment of the sunflower peptides with the sequences of mung bean cytochrome c and wheat germ cytochrome c.     

The amino acid sequence of rape (Brassica Napus L) cytochrome c

The amino acid sequence of rape (Brassica Napus L) cytochrome c was determined using 1 mumole of protein. Analysis of chymotryptic and tryptic peptides by the dansyl-Edman method showed that the molecule consisted of 111 residues and was homologous with other mitochondrial plant cytochromes c. The amino acid sequence was found to be identical with that previously reported for the cytochrome c from cauliflower (Brassica oleracea L).     

The switching of electron flux from the cyanide-insensitive oxidase to the cytochrome pathway in mung-bean (Phaseolus aureus L.) mitochondria.

The activities of the mung-bean (Phaseolus aureus L.) mitochondrial cyanide-insensitive oxidase and cytochrome pathways have been measured simultaneously. The results show that electrons can be diverted both from the alternative pathway to the cytochrome pathway and from the cytochrome to the alternative pathway. The competition of the two pathways for the available electron flux is discussed.     

Transesterified sesame (Sesamum indicum L.) seed oil as a biodiesel fuel

The sesame (Sesamum indicum L.) oil was extracted from the seeds of the sesame that grows in Diyarbakir, SE Anatolia of Turkey. Sesame seed oil was obtained in 58wt/wt%, by traditional solvent extraction. The methylester of sesame (Sesamum indicum L.) seed oil was prepared by transesterification of the crude oil. Transesterification shows improvement in fuel properties of sesame seed oil. This study supports the production of biodiesel from sesame seed oil as a viable alternative to the diesel fuel.     

Auxin Transport Inhibitors and the Rooting of Hypocotyl Cuttings from Etiolated Mung-bean Vigna radiata (L.) Wilczek Seedlings

The auxin transport inhibitors 2, 3, 5-triiodobenzoic acid (TIBA)and naphthylphthalamic acid (NPA) inhibited adventitious rootformation (ARF) induced by indol-3-butyric acid (IBA) on cuttingsfrom etiolated mung-bean seedlings floated on solutions of thegrowth regulators. The concentrations of TIBA and NPA requiredfor a 25 per cent reduction in ARF with 10 µM IBA wereestimated by linear interpolation to be 11.3 µm and 0.42µM respectively. NPA is a particularly potent inhibitorof IBA-induced ARF. The inhibitory effect of either compoundwas reversible by higher concentrations of IBA. NPA had no effectwhen applied after the auxin treatment. The inhibitory effects of TIBA or NPA could not be explainedby effects on the uptake or metabolism of [2-^14C]IAA. Consideringthis and other evidence, it is suggested that NPA and possiblyTIBA are acting as specific antagonists of auxin in the inductionof ARF. Vigna radiata (L.), mung-bean, root induction, hypocotyl cuttings, auxin inhibitors, indol-3-butyric acid, 2,3,5-triiodobenzoic acid, naphthylphthalamic acid, auxin uptake, auxin metabolism, adventitious roots     

Properties of Rhizopus stolonifer Polygalacturonase, an Elicitor of Casbene Synthetase Activity in Castor Bean (Ricinus communis L.) Seedlings

Some properties of the polygalacturonase-elicitor from the filtrates of Rhizopus stolonifer cultures have been examined in an attempt to understand its mode of action as an elicitor of casbene synthetase activity in castor bean seedlings. Both the polygalacturonase activity and the elicitor activity are heat-labile with similar heat-sensitivity profiles. Also, the catalytic activity of the enzyme is lost on treatment with sodium periodate, as had been shown previously for the elicitor activity. The pH optimum of the enzyme activity with polygalacturonic acid as the substrate is 4.9. Exposures of germinating castor bean seedlings to the elicitor for short-term periods of 1 to 10 minutes followed by washing and incubation in sterile, distilled water are partially effective in elicitation in comparison with the continuous exposure of the seedlings over 11 hours to the same amount of the elicitor. The initial rate of reaction catalyzed by the enzyme is about 3 times faster with polygalacturonic acid as a substrate than with partially (50%) methylated polygalacturonic acid (pectin). The K_m value of the enzyme for polygalacturonic acid is about 4.2 millimolar in terms of monomeric units and about 0.07 millimolar in terms of polymer concentration. Examination of the types of products formed by the action of the enzyme suggests that it is an endo-hydrolase. The amino acid composition of this enzyme is similar to those of other extracellular fungal proteins reported. The carbohydrate moiety of the glycoprotein polygalacturonase-elicitor is composed of 92% mannose and 8% glucosamine by gas chromatography-mass spectrometry analysis. The linkage group analysis of the carbohydrate moiety showed that mannosyl residues which are 1,2-linked comprise about 70% of the total glycosyl residues and demonstrated the presence of some 1,3,6- and 1,2,6-linked branching mannosyl residues.     

Contribution of corticular photosynthesis to bud development in African baobab (Adansonia digitata L.) and Castor bean (Ricinus communis L.) seedlings

The African baobab (Adansonia digitata L.) and castor bean (Ricinus communis L.) are drought resistant green-stemmed succulent plants which grow in the arid and semi-arid regions of Africa. Photosynthesis in the stems of green-stemmed plants is known to contribute to plant carbon gain especially during leafless periods. To study the contribution of stem photosynthesis in stem succulent plants, the height and stem diameter of baobab and castor bean plants grown in the greenhouse were measured. The plants were completely defoliated and subjected to different treatments: Watered with open stems (WO), watered and stems covered with aluminium foil (WC) to achieve 100% light exclusion, drought and open (DO) and drought and covered (DC). Stem coverage with aluminium foil resulted in a higher stem height and diameter during drought for baobab with similar trends seen in castor bean. Light exclusion resulted in a significantly lower bud DW production and enrichment in ¹³C in bud dry matter of castor bean and in stem dry matter of baobab. These show that corticular photosynthesis contributes in carbon gain in these species.     

Proteomic Analyses of Three Inflorescence Styles of Castor (<i>Ricinus communis</i> L.) at Different Developmental Stages

Castor (Ricinus communis L.) is one of ten oil crops in the world and has complex inflorescence styles. Generally, castor has three inflorescence types: single female inflorescence (SiFF), standard female inflorescence (StFF) and bisexual inflorescence (BF). StFF is realized as a restorer line and as a maintainer line, which was applied to castor hybrid breeding. However, the developmental mechanism of the three inflorescences is not clear. Therefore, we used proteomic techniques to analyze different inflorescence styles. A total of 72 diferentially abundant protein species (DAPs) were detected. These DAPs are primarily involved in carbon and energy metabolism and carbon fixation in the photosynthetic organism pathway. The results showed that DAPs are involved in photosynthesis to control the distribution of imported carbohydrates and exported photoassimilates and thus affect the inflorescence development of castor. In addition, these DAPs are also involved in cysteine and methionine metabolism. Quantitative real-time PCR (qRT-PCR) results demonstrated that the proteomics data collected in this study were reliable. Our findings indicate that the carbon cycle and amino acid metabolism influence the inflorescence development of castor.     

Purification and characterization of the sunflower seed (Helianthus annuus L.) major aminopeptidase

The sunflower seed (Helianthus annuus L.) major peptidase was purified to molecular homogeneity. It is an 80 kDa enzyme with pI of 4.6 and optimal activity at pH 7.5–8.0 and 45–50°C. It is a thiol-dependent aminopeptidase hydrolyzing peptides in a step-by-step manner as cleaving after the N-terminal amino acid residue of the substrate. It requires substrate acyl parts with a free amino group in either α- or β-position and l-configuration of the adjacent carbon atom. The enzyme prefers amino acid residues with bulky hydrophobic side chains at P₁-position and its catalytic efficacy is affected by the structure of both P₁ and P₁′ parts of the substrate.     

Controlling the functionality of cytochrome c(1) redox potentials in the Rhodobacter capsulatus bc(1) complex through disulfide anchoring of a loop and a beta-branched amino acid near the heme-ligating methionine.

The cytochrome c(1) subunit of the ubihydroquinone:cytochrome c oxidoreductase (bc(1) complex) contains a single heme group covalently attached to the polypeptide via thioether bonds of two conserved cysteine residues. In the photosynthetic bacterium Rhodobacter (Rba.) capsulatus, cytochrome c(1) contains two additional cysteines, C144 and C167. Site-directed mutagenesis reveals a disulfide bond (rare in monoheme c-type cytochromes) anchoring C144 to C167, which is in the middle of an 18 amino acid loop that is present in some bacterial cytochromes c(1) but absent in higher organisms. Both single and double Cys to Ala substitutions drastically lower the +320 mV redox potential of the native form to below 0 mV, yielding nonfunctional cytochrome bc(1). In sharp contrast to the native protein, mutant cytochrome c(1) binds carbon monoxide (CO) in the reduced form, indicating an opening of the heme environment that is correlated with the drop in potential. In revertants, loss of the disulfide bond is remediated uniquely by insertion of a beta-branched amino acid two residues away from the heme-ligating methionine 183, identifying the pattern betaXM, naturally common in many other high-potential cytochromes c. Despite the unrepaired disulfide bond, the betaXM revertants are no longer vulnerable to CO binding and restore function by raising the redox potential to +227 mV, which is remarkably close to the value of the betaXM containing but loop-free mitochondrial cytochrome c(1). The disulfide anchored loop and betaXM motifs appear to be two independent but nonadditive strategies to control the integrity of the heme-binding pocket and raise cytochrome c midpoint potentials.     

Properties of thiamin-binding proteins from sesame seed 2S albumins

Three thiamin-binding proteins (TBPs) from sesame ( Sesamum indicum L.) seeds (STBP-I, -II and -III) were characterized. Binding of thiamin to the three STBPs was inhibited by pyrithiamin, which did not inhibit the binding of thiamin to TBPs from other plant seeds. STBP-I alone bound 2-northiamin and hydroxyethylthiamin. Isoelectric points (pIs) of STBP-I and -II both were 7.5. The pI of STBP-III was 6.5. STBPs did not have immunological homology with TBPs from rice seeds and buckwheat seeds. On the other hand, the amino acid compositions of the small and large polypeptides isolated from STBP-I, -II and -III resembled each other. Both the polypeptides contained large amounts of Glu (or Gln) and Arg. The small polypeptides contained more Ser than the large polypeptides. The N-terminal amino acid sequences (the first 29 residues) of the small polypeptides were identified. The N-terminal amino acid sequences of the three STBPs were the same. The small polypeptides had homology to castor bean 2S albumin small subunit. These results showed that STBPs were part of a plant protein superfamily and that STBPs differed from the TBPs of other plant seeds as to the binding to thiamin-related compounds and immunological properties, and further, that STBP-I, -II and -III differed in the affinity for thiamin-related compounds and pI, indicating that STBP-I, -II and -III are isomers.     

Molecular cloning and characterization of a cDNA encoding asparaginyl endopeptidase from sweet potato (Ipomoea batatas (L.) Lam) senescent leaves

Asparaginyl endopeptidase is a cysteine endopeptidase that has strict substrate specificity toward the carboxyl side of asparagine residues, and is possibly involved in the post-translational processing of proproteins. In this report one full-length cDNA, SPAE, was isolated from senescent leaves of sweet potato (Ipomoea batatas (L.) Lam). SPAE contained 1479 nucleotides (492 amino acids) in the open reading frame, and exhibited high amino acid sequence homologies (c. 61-68%) with asparaginyl endopeptidases of Vicia sativa, Phaseolus vulgaris, Canavalia ensiformis, and Vigna mungo. SPAE probably encoded a putative precursor protein. Via cleavage of the N- and C-termini, it produced a mature protein containing 325 amino acids (from the 51st to the 375th amino acid residues), the conserved catalytic residues (the 173rd His and 215th Cys amino acid residues), and the putative N-glycosylation site (the 332nd Asn amino acid residue). Semi-quantitative RT-PCR and western blot hybridization showed that SPAE gene expression was enhanced significantly in natural senescent leaves and in dark- and ethephon-induced senescent leaves, but was much less in mature green leaves, stems, and roots. Phylogenic analysis showed that SPAE displayed close association with vacuolar processing enzymes (legumains/asparaginyl endopeptidases), which function via cleavage for proprotein maturation in the protein bodies during seed maturation and germination. In conclusion, sweet potato SPAE is probably a functional, senescence-associated gene and its mRNA and protein levels were significantly enhanced in natural and induced senescent leaves. The possible role and function of SPAE associated with bulk protein degradation and mobilization during leaf senescence were also discussed.     

Isolation and characterization of complement component C3 from Atlantic cod (Gadus morhua L.) and Atlantic halibut (Hippoglossus hippoglossus L.)

Complement component C3 was isolated from the plasma of cod (Gadus morhua L.) and halibut (Hippoglossus hippoglossus L.). Fast protein liquid chromatography (FPLC) techniques, involving ion exchange and gel filtration columns, were used. The purified proteins were analysed by SDS-PAGE which showed a two-chain structure, alpha- and beta-chains, as seen in higher vertebrates. Both proteins had intra-chain thioesters located within their alpha-chains and N-terminal amino acid sequencing confirmed their identity with reference to known C3 amino acid sequences from other species. Specific antibodies were prepared against cod and halibut C3 and tested in Western blotting on sera and purified C3. The proteolytic fragmentation of C3 was tested with trypsin, pepsin, papain and the extracellular product (ECP) from the bacterium Aeromonas salmonicida ssp. achromogenes (Asa). Both trypsin and papain were successful in cleaving C3 whereas pepsin and ECP had no effect. Carbohydrate moieties were detected in the alpha- and beta-chains of cod and halibut C3 and N-linked oligosaccharides were removed from the C3 with PNGase treatment, revealing a difference in C3 glycosylation between the two species.     

The amino acid sequences of cytochrome c from four plant sources

Proposed amino acid sequences of cytochrome c from nasturtium (Tropaeolum majus L.), box-elder (Acer negundo L.), elder (Sambucus nigra L.) and parsnip (Pastinaca sativa L.) are presented. Because of the very limited amounts of cytochrome available from some plant sources, peptides derived from the cytochromes c have been sequenced by the semi-quantitative dansyl-Edman technique (Gray & Hartley, 1963) without supporting quantitative amino acid analyses. Because of the qualitative nature of the work, the sequences proposed must be regarded as tentative. Considerations of homology, although useful as a guide, have been kept to a minimum in the construction of sequences. Only the nasturtium sequence relies on considerations of homology for a complete ordering of the peptides. Where material permitted, each residue of a proposed sequence was determined at least once from both a tryptic and a chymotryptic peptide.     

Influence of four host‐plants on feeding,growth and reproduction of Diacrisia casignetum (Lepidoptera: Arctiidae)

Effects of four host‐plants, sunflower, castor, jute and sesame, on feeding, growth and reproduction of Diacrisia casignetum Kollar (Lepidoptera: Arctiidae) were studied under laboratory conditions (27 ± 0.5°C, 12 h light : 12 h dark, 65 ± 5% RH). Total larval developmental time of D. casignetum was highest on sesame than the other three host‐plants used in this study, but pupal duration was higher on sesame than sunflower but not for other dietary treatments. The longevity of females was generally longer than males. Male and female longevity was higher in sunflower than sesame (P < 0.05), but it did not differ significantly among other treatments. Fecundity was highest in sunflower followed by castor, jute and sesame. The growth and development of D. casignetum were related to nutrient and phenol contents of these four host‐plants. Total carbohydrates and amino acids were present in rich quantities in sunflower when compared to other three host leaves, while nitrogen, protein and lipid contents were comparatively higher in sunflower and castor than jute and sesame. Phenol content was greatest in sesame, and least in castor and sunflower. Higher levels of total carbohydrates, proteins, lipids, nitrogen and amino acids including water content and lower phenol content of sunflower have influenced higher growth rate and fecundity of D. casignetum.     

Ovicidal activity of Sesamum indicum (L.) oil against Spodoptera littoralis (Boisd.)

In the research for alternative tools and botanical products to control Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae). Sesamum indicum (L.) (Lamiales: Pedaliaceae) oil was assayed as an ovicide. The mortality increased with existence of fatty acids. Chemical analysis of S. indicum oil using GLC analysis showed palmitic acid as the major fatty acid (51.27%), while the major hydrocarbon and sterols were found to be heneicosane (58.63%) and β-sitosterol (2.60%), respectively. Generally, the values of LC₅₀s indicated that one-day-old egg masses are more susceptible than three-day-old eggs. Also, the leaf dip technique is more efficient than the spraying one. Results showed several features of chorionic surface deformation treated with sesame and KZ oils than control using scanning electron microscopy. Meanwhile, the tested oils caused significant reduction in both total soluble protein and transaminase enzymes as compared to control. Additionally, the oils elongated the incubation period and larval duration than control.     

Phospholipase D from Soybean (Glycine max L.) Suspension-Cultured Cells: Purification, Structural and Enzymatic Properties

Phospholipase D (phosphatidylcholine phosphatidohydrolase EC3.1.4.4 [EC] ) from soybean (Glycine max L.) suspension-cultured cellwas purified around 1,200-fold to homogeneity by acetone precipitation,Macro-Prep High Q anion exchange, and octyl-Sepharose CL-4Baffinity chromatography. The purified enzyme released 1,600µmol of choline per min per mg of protein. The enzymeis monomeric with a molecular mass of 92 kDa, as estimated bySDS-PAGE. One of the most interesting characteristics of thepurified soybean phospholipase D was the dependence of the pHoptimum on the Ca²⁺ ion concentration in the assay. With 10mM, 20 mM and 40 mM Ca²⁺ ions, the optima were at pH 7.5, 6and 5.5, respectively. The specific adsorption of phospholipaseD onto octyl-Sepharose gel suggests that the molecule becomesmore hydrophobic in the presence of Ca²⁺ ions. The amino acidsequence of the first 18 N-terminal residues of soybean phospholipaseD revealed a high degree of homology with those previously publishedfor cabbage leaf and castor bean endosperm enzymes. Westernblots of the soybean phospholipase D showed an immunoreactivitywith antibodies raised against a synthetic peptide correspondingto the 15 N-terminal aminoacid residues of phospholipase D fromcabbage leaves. (Received March 13, 1995; Accepted May 29, 1995)     

Amino acid sequence of cytochrome c from rice

The amino acid sequence of cytochrome c purified from rice, Oryza sativa L., was determined. The complete amino acid sequence of rice cytochrome c is as follows: Ac-Ala-8-Ser-Phe-Ser-Glu-Ala-Pro-Pro-Gly1-Asn-Pro-Lys-Ala-Gly-Glu-Lys-Ile-Phe10-Lys-Thr-Lys-Cys-Ala-Glx-Cys-His-Thr-Val20-Asp-Lys-Gly-Ala-Gly-His-Lys-Glx-Gly-Pro30-Asx-Leu-Asx-Gly-Leu-Phe-Gly-Arg-Glx-Ser40-Gly-Thr-Thr-Pro-Gly-Tyr-Ser-Tyr-Ser-Thr50-Ala-Asp-Lys-Asn-Met-Ala-Val-Ile-Trp-Glx60-Glx-Asx-Thr-Leu-Tyr-Asp-Tyr-Leu-Leu-Asn70-Pro-TML-Lys-Tyr-Ile-Pro-Gly-Thr-Lys-Met80-Val-Phe-Pro-Gly-Leu-TML-Lys-Pro-Glx-Glx90-Arg-Ala-Asp-Leu-Ile-Ser-Tyr-Leu-Lys-Glu100-Ala-Thr-Ser (Ac = acetyl group, TML = epsilon-N-trimethyllsine). The primary structure of rice cytochrome c was found to be homologous with those of other plant cytochromes c reported so far; it possesses general features common to plant cytochromes c, and all the invariant residues characterized in dicotyledonous cytochromes c are also conserved in the sequence of rice cytochrome c, as well as those of other monocotyledonous cytochromes c. The distinctive features of rice cytochrome c are a high content of proline residues, their unique locations in the sequence and the presence of a serine residue at position 96.     

Development of polymorphic microsatellite markers in sesame (Sesamum indicum L.)

Fifty microsatellite sequences (SSRs) were isolated from an enriched library of sesame (Sesamum indicum L.) using a modified protocol. After screening, 10 polymorphic microsatellites were used to determine their usefulness in diversity analysis among 16 sesame accessions. The number of alleles ranged from three to six alleles per locus with an average of 4.6 alleles. The fragment size varied from 150 bp to 307 bp. Expected heterozygosites (H_E) and polymorphism information contents (PICs) ranged from 0.437 to 0.858 and 0.34 to 0.80, respectively, which indicates the highly informative nature of the microsatellites reported here. These microsatellite markers will be very useful in diversity analysis among a large germplasm collection of sesame present in our Korean gene bank and also in the establishment of its core collection.