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PubMed1993

Partial purification of the cyanide-resistant alternative oxidase of skunk cabbage (Symplocarpus foetidus) mitochondria.

A partial purification of the cyanide-resistant, alternative oxidase from skunk cabbage (Symplocarpus foetidus L.) spadix mitochondria is described. Skunk cabbage mitochondria were solubilized in N,N-bis-(3-D-glucon-amido-propyl)deoxycholamide and the alternative oxidase was purified using a batch DEAE-cellulose treatment, followed by precipitation with Extracti-Gel and chromatography on Sephadex G-200. Following pooling and concentrating of the most active fractions from the gel filtration column, a 20- to 30-fold purification of the alternative oxidase was obtained, with no evidence of contamination by cytochrome c oxidase (complex IV) or cytochrome c reductase (complex III). Polyacrylamide gel electrophoresis of the partially purified oxidase showed major polypeptides at 36 and 29 kD, both of which react with monoclonal antibodies raised against the Sauromatum guttatum alternative oxidase. The purified oxidase fraction showed no absorbance in the visible spectral region, and addition of sodium borohydride induced no absorbance changes in the ultraviolet region. The purified alternative oxidase catalyzed the four-electron reduction of oxygen to water in the absence of citrate, but catalyzed an apparent two-electron reduction of oxygen to hydrogen peroxide in the presence of 0.7 M citrate.

Chromatography, Gel↗
PubMed2004

Emission of Plutella xylostella-induced compounds from cabbages grown at elevated CO2 and orientation behavior of the natural enemies.

Several plant species defend themselves indirectly from herbivores by producing herbivore-induced volatile compounds that attract the natural enemies of herbivores. Here we tested the effects of elevated atmospheric CO(2) (720 micromol mol(-1)) concentration on this indirect defense, physiological properties, and constitutive and induced emissions of white cabbage (Brassica oleracea ssp. capitata, cvs Lennox and Rinda). We monitored the orientation behavior of the generalist predator Podisus maculiventris (Heteroptera: Pentatomidae) and the specialist parasitoid Cotesia plutellae (Hymenoptera: Braconidae) to plants damaged by Plutella xylostella (Lepidoptera: Plutellidae) in the Y-tube olfactometer. Elevated CO(2) levels did not affect stomatal densities but reduced specific leaf area and increased leaf thickness in cv Lennox. In addition to enhanced constitutive monoterpene emission, P. xylostella-damaged cabbages emitted homoterpene (E)-4,8-dimethyl-1,3,7-nonatriene, sesquiterpene (E,E)-alpha-farnesene, and (Z)-3-hexenyl acetate. Growth at elevated CO(2) had no significant effect on the emissions expressed per leaf area, while minor reduction in the emission of homoterpene (E)-4,8-dimethyl-1,3,7-nonatriene and (E,E)-alpha-farnesene was observed at elevated CO(2) in one of two experiments. The generalist predator P. maculiventris discriminated only between the odors of intact and P. xylostella-damaged cv Rinda plants grown at ambient CO(2) concentration, preferring the odor of the damaged plants. The specialist parasitoid C. plutellae preferred the odor of damaged plants of both cultivars grown at ambient CO(2) but did not detect damaged cv Lennox plants grown at elevated CO(2). The results suggest that elevated atmospheric CO(2) concentration could weaken the plant response induced by insect herbivore feeding and thereby lead to a disturbance of signaling to the third trophic level.

Animals↗
PubMed1996

Purification and Characterization of a Cryoprotective Protein (Cryoprotectin) from the Leaves of Cold-Acclimated Cabbage.

We have purified a protein (cryoprotectin) from the leaves of cold-acclimated cabbage (Brassica oleracea L.) that protects thylakoids from nonacclimated spinach (Spinacia oleracea L.) against freeze-thaw damage. The procedure involves precipitations by heat, ammonium sulfate, and the glycosaminoglycan heparin and column chromatography on Polyamide 6 and a C18 reverse-phase matrix. After reverse-phase chromatography we obtained a single band of an apparent molecular mass of 7 kD when fractions that showed cryoprotective activity were analyzed by sodium dodecyl sulfate gel electrophoresis and silver staining. Gel-filtration experiments confirmed that the active protein is a monomer of 7 kD native molecular mass. This 7-kD protein could be purified only from cold-acclimated cabbage, but not from plants grown under nonacclimating conditions. Using peroxidase-labeled lectins, we show that cryoprotectin is a glycoprotein and that the saccharide moiety contains [alpha]1-3-linked fucose.

Journal Article↗
PubMed2001

Cabbage cryoprotectin is a member of the nonspecific plant lipid transfer protein gene family.

We have recently purified a protein (cryoprotectin) from the leaves of cold-acclimated cabbage (Brassica oleracea) to electrophoretic homogeneity, which protects thylakoids isolated from the leaves of nonacclimated spinach (Spinacia oleracea) from freeze-thaw damage. Sequencing of cryoprotectin showed the presence of at least three isoforms of WAX9 proteins, which belong to the class of nonspecific lipid transfer proteins. Antibodies raised against two synthetic peptides derived from the WAX9 proteins recognized a band of approximately 10 kD in western blots of crude cryoprotectin preparations. This protein and the cryoprotective activity could be precipitated from solution by the antiserum. We show further that cryoprotectin is structurally and functionally different from WAX9 isolated from the surface wax of cabbage leaves. WAX9 has lipid transfer activity for phosphatidylcholine, but no cryoprotective activity. Cryoprotectin, on the other hand, has cryoprotective, but no lipid transfer activity. The cryoprotective activity of cryoprotectin was strictly dependent on Ca(2+) and Mn(2+) and could be inhibited by chelating agents, whereas the lipid transfer activity of WAX9 was higher in the presence of ethylenediaminetetraacetate than in the presence of Ca(2+) and Mn(2+).

Amino Acid Sequence↗
PubMed1971

Isolation of spherosomes (oleosomes) from onion, cabbage, and cottonseed tissues.

Subcellular particles, identical in appearance to spherosomes observed in situ, were isolated from onions (Allium cepa L.) and cabbages (Brassica capitata L.). They were minute spherules about 1 micron in diameter, filled with an evenly stained osmiophilic matrix and delimited by a single, fineline membrane 20 to 30 A thick."Spherosomes" isolated from onions and cabbages were compared with "oil-droplets" isolated from cottonseeds. Morphologically, they were similar, even at ultrastructural levels. Their chemical compositions were also similar: both types of particles containing primarily lipid with very little phospholipid or protein. Neither type of particle possessed acid phosphatase activity.These results indicate that oil-droplets of oleaginous tissues correspond to spherosomes of nonoily tissues. Therefore, both types of particles should be referred to by the same name. Since these particles are rich in lipids, it is suggested that the name "spherosome" be abandoned in favor of "oleosome," which is also entitled to priority.

Journal Article↗
PubMed1982

Inducible cadmium binding complexes of cabbage and tobacco.

Cadmium complexes with apparent molecular weights of 10,000 were observed in aqueous extracts of Cd-treated cabbage (Brassica capitata L., cv. red danish) and tobacco (hybrid of Nicotiana glauca and N. langsdorffii) plants. The amount of complex (as Cd) recovered was found to be dependent on the concentration of the metal in the growth medium and the total time of exposure of plants to the metal. Induction of the complex at moderate levels of (112)Cd exposure was monitored after labeling the complex with (109)Cd in vitro. The constitutive nature of the ligand of the complex in cabbage and tobacco leaves was suggested when control plant extracts were exposed to (109)Cd. Such extracts contained (109)Cd, which eluted from Sephadex G-50 in the region of Cd complex. Simultaneous labeling with (112)Cd and (35)S or (32)P indicated that the complex contained sulfur but probably not phosphorus. The amount of (35)S which eluted coincident with (112)Cd complex increased during complex induction. No evidence was found for the presence of 10,000 molecular weight Cd complex in stem exudates (vascular sap) of Cd-treated plants.The results obtained are consistent with the presence in these tissues of a ligand which is both inducible and constitutive and binds Cd in mercaptide bonds. All of these properties, and others reported earlier, are characteristic of Cd-metallothionein formed in animals.

Journal Article↗
PubMed1984

Propylamine transferases in chinese cabbage leaves.

We have found spermidine synthase and spermine synthase activities in extracts of leaves of Chinese cabbage (Brassica pekinensis var. Pak Choy) and have developed an assay of the former in crude extracts. The method is based on the transfer of the propylamine moiety of decarboxylated S-adenosylmethionine to labeled putrescine, followed by ion-exchange separation of the labeled amine substrate and product, which are then converted to the 5-dimethylamino-1-napthalene sulfonyl (dansyl) derivatives and further purified and identified by thin layer chromatography. The specific radioactivity of putrescine present in the reaction mixture is determined, as is the radioactivity present in dansyl spermidine. The enzyme is also present in extracts of spinach leaves.Spermidine synthase has been purified about 160-fold from Chinese cabbage leaves. After partial purification, a rapid coupled enzymic assay has been used to study various properties of the enzyme. The plant enzyme shows maximum activity at pH 8.8 in glycine-NaOH buffer and has a molecular weight of 81,000. The K(m) values for decarboxylated S-adenosylmethionine and putrescine are 6.7 and 32 micromolar, respectively. The enzyme activity is inhibited strongly by dicyclohexylamine, cyclohexylamine, and S-adenosyl-3-thio-1, 8-diaminoctane. Of these, dicyclohexylamine is the most potent inhibitor with an I(50) at 0.24 micromolar.

Journal Article↗
PubMed1984

Characterization of a cadmium-binding complex of cabbage leaves.

The chemical nature of a principal, inducible cadmium-binding complex which accumulates in cabbage leaves (Wagner and Trotter 1982 Plant Physiol 69: 804-809) was studied and compared with that of animal metallothionein and copper-binding proteins isolated from various organisms. The apparent molecular weight of native cabbage complex and carboxymethylated ligand of the complex under native conditions as determined by gel filtration was about 10,000 daltons. Under denaturing conditions their apparent molecular weights were about 2000 daltons. Ligand of native complex contained 37, 28, and 9 residue per cent of glutamic acid-glutamine, cysteine, and glycine, respectively, and low aromatic residue, serine and lysine content. The high acidic and low hydrophobic residue content explain the behavior of complex on electrophoresis in the presence and absence of sodium dodecyl sulfate. Its isoelectric point was below 4.0 and it bound 4 to 6 moles cadmium per mole ligand in what appear to be cadmium-mercaptide chromophores. The complex was found to be heat stable, relatively protease insensitive, and lacking in disulfide bonds. Attempts to determine the primary sequence of reduced native complex and carboxymethylated, cleaved ligand using the Edman degradation procedure were unsuccessful. An electrophoretic procedure is described for preparative isolation of purified complex and a method is described for monitoring ligand of complex as its fluorescent dibromobimane adduct.

Journal Article↗
PubMed1985

S-adenosylmethionine decarboxylase and spermidine synthase from chinese cabbage.

The enzyme, S-adenosylmethionine (SAM) decarboxylase (EC 4.1.1.50), has been demonstrated in leaves of Chinese cabbage, (Brassica pekinensis var Pak Choy). All of the enzyme can be found in extracts of the protoplasts obtained from the leaves of growing healthy or virus-infected cabbage. The protein has been purified approximately 1500-fold in several steps involving ammonium sulfate precipitation, affinity chromatography, and Sephacryl S-300 filtration. The reaction catalyzed by the purified enzyme has been shown to lead to the equimolar production of CO(2) and of decarboxylated S-adenosylmethionine (dSAM). The K(m) for SAM is 38 micromolar. The reaction is not stimulated by Mg(++) or putrescine, and is inhibited by dSAM competitively with SAM. It is also inhibited strongly by methylglyoxal bis(guanylhydrazone). The enzyme, spermidine synthase (EC 2.5.1.16), present in leaf or protoplast extracts in many fold excess over SAM decarboxylase, has been purified approximately 1900-fold in steps involving ammonium sulfate precipitation, affinity chromatography, and gel filtration on Sephacryl S-300. Standardization of the Sephacryl column by proteins of known molecular weight yielded values of 35,000 and 81,000 for the decarboxylase and synthase, respectively.

Journal Article↗
PubMed1986

Recovery of turgor by wilted, excised cabbage leaves in the absence of water uptake : a new factor in drought acclimation.

Cabbage leaves excised from a fully turgid plant wilt within 20 minutes to 2 hours (depending on plant age) with a loss of about 10% relative water content (RWC). If droughted for 2 to 4 days in a high relative humidity leaf chamber, they may acclimate, recovering their turgor without the absorption of water, in fact at a loss of 15 to 25% RWC. This turgor recovery in the absence of water uptake occurs only if (a) the rate of water loss is slow enough (about 1-5% RWC per day after the first 24 hours drought loss of about 15% RWC), (b) if the leaves are no longer growing actively. Osmotic adjustment accompanies the turgor adjustment, but cannot be the cause in the absence of water uptake. The recovery of turgor by wilted cabbage leaves in the absence of water uptake cannot be explained by (a) transfer of reserve water from apoplast to symplast either from the cell walls or from the vessel lumens by cavitation or (b) metabolic loss of dry matter and gain of water. It can be explained by a contraction of the cell walls around the partially dehydrated protoplasts, until they regain their elastic extensibility. These proposed cell wall changes during drought acclimation are therefore the opposite of those occurring during growth. This hypothesis therefore explains the long recognized inverse relation between growth and acclimation. Two predictions of this hypothesis were tested and substantiated.

Journal Article↗
PubMed1989

Comparison of Photomorphogenic Responses to UV Light in Red and White Cabbage (Brassica oleracea L.).

Photoinhibition of hypocotyl growth in white cabbage (Brassica oleracea L., cv "Bianco Brunswick") is controlled by UV absorbing receptor(s) and the phytochrome system, while in red cabbage (cv "Rosso Olandese tardivo invernale") phytochrome can act without any requirement for the action of a specific UV receptor. Similar results have been obtained for the photoregulation of anthocyanin production. Twenty-four hour preirradiations with UV light or 692 nanometers light lead to the same increase in responsiveness of the system toward Pfr in a following dark period, suggesting a phytochrome promotion of subsequent light induction for both.

Journal Article↗
PubMed2006

Detection of Bacillus thuringiensis kurstaki HD1 on cabbage for human consumption.

The objectives of the study were to develop a specific procedure for quantification and identification of Bacillus thuringiensis kurstaki HD1, which is used as a biopesticide, and to quantify its presence in different kinds of cabbage for human consumption. We found that B. thuringiensis kurstaki HD1 can be distinguished from other B. thuringiensis strains by its unique random amplification of polymophic DNA-PCR pattern with the OPA9 primer and the presence of the flagellin genes, as detected by the primers FLAB1 and FLAB2. We detected from one to 100 Bacillus cereus-like bacteria in 10 batches of five different cabbage products for consumption. As many as 73 out of 134 isolates (53.7%) were identical with B. thuringiensis kurstaki HD1. The results show that B. thuringiensis kurstaki HD1 from biopesticides can be found in vegetables for human consumption.

Bacillus cereus↗
PubMed1986

Effect of temperature, sodium chloride, and pH on growth of Listeria monocytogenes in cabbage juice.

Human illness and death have resulted from the consumption of milk, cheese, and cole slaw contaminated with Listeria monocytogenes. Since the effects of temperature, NaCl, and pH on the growth of the organism in cabbage were unknown, a series of experiments was designed to investigate these factors. Two strains (LCDC 81-861 and Scott A, both serotype 4b) were examined. At 30 degrees C, the viable population of the LCDC 81-861 strain increased in sterile unclarified cabbage juice (CJ) containing 0 to 1.5% NaCl; a decrease in the population of both strains occurred in juice containing greater than or equal to 2% NaCl. At 5 degrees C, the population of the Scott A strain in CJ containing up to 5% NaCl was reduced by about 90% over a 70-day period; the LCDC 81-861 strain was more sensitive to refrigeration but remained viable in CJ containing less than or equal to 3.5% NaCl for 70 days. Growth in CJ at 30 degrees C resulted in a decrease in pH from 5.6 to 4.1 within 8 days. Death of L. monocytogenes occurred at 30 degrees C when the organism was inoculated into sterile CJ adjusted to pH less than or equal to 4.6 with lactic acid. No viable cells were detected after 3 days at pH less than or equal to 4.2. At 5 degrees C, the rate of death at pH less than or equal to 4.8 was slower than at 30 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)

Brassica↗
PubMed1993

Isolation and Characterization of Nisin-Resistant Leuconostoc mesenteroides for Use in Cabbage Fermentations.

Leuconostoc mesenteroides strains that are resistant to high levels of nisin (up to 25,000 IU/ml in broth) were isolated. These nisin-resistant mutants were evaluated to determine their potential use as starter culture strains for cabbage fermentations. We found that some L. mesenteroides strains could be adapted to high levels of nisin resistance, while others could not. The nisin resistance trait was found to be stable for at least 35 generations, in the absence of nisin selection, for all mutants tested. The effects of nisin and salt, separately and in combination, on growth kinetics of the nisin-resistant strains were determined. Salt was the most influential factor on the specific growth rates of the mutants, and no synergistic effect between nisin and salt on specific growth rates was observed. The nisin-resistant strains were unimpaired in their ability to rapidly produce normal heterolactic fermentation end products. The use of these L. mesenteroides mutants as starter cultures in combination with nisin may extend the heterolactic phase of cabbage fermentations.

Journal Article↗
PubMed2004

Interaction of the movement protein NSP and the Arabidopsis acetyltransferase AtNSI is necessary for Cabbage leaf curl geminivirus infection and pathogenicity.

DNA viruses can modulate the activity of cellular acetyltransferases to regulate virus gene expression and to affect cell cycle progression in order to support virus replication. A role for protein acetylation in regulating the nuclear export of the bipartite geminivirus DNA genome was recently suggested by the findings that the viral movement protein NSP, which shuttles the viral genome between the nucleus and the cytoplasm, interacts with a novel Arabidopsis acetyltransferase, AtNSI, and the increased expression of AtNSI enhances susceptibility to Cabbage leaf curl virus infection. To further investigate the interaction of NSP and AtNSI and to establish the importance of this interaction in virus infections, we used a reverse yeast two-hybrid selection and deletion analysis to identify NSP mutants that were impaired in their ability to bind AtNSI. These mutants identified a 38-amino-acid region of NSP, to which no function had so far been assigned, as being necessary for NSP-AtNSI interaction. Three NSP missense mutants were analyzed in detail and were found to be comparable to wild-type NSP in their levels of accumulation, nucleocytoplasmic shuttling, DNA binding, and cooperative interaction with the viral cell-to-cell movement protein MP. Despite this, Cabbage leaf curl virus that expressed each mutated NSP was defective in its ability to infect Arabidopsis, exhibiting lower levels of infectivity than the wild-type virus, and delayed systemic spread of the virus and attenuated disease symptoms. Our data demonstrate the importance of the interaction of NSP with AtNSI for virus infection and pathogenicity.

Amino Acid Sequence↗
PubMed1998

Effects of cabbage leaf extract on breast engorgement.

The effectiveness of cabbage leaf extract was compared with that of a placebo in treating breast engorgement in lactating women. In a double-blind experiment with a pretest/posttest design, 21 participants received a cream containing cabbage leaf extract, while 18 received placebo cream. The placebo group received equal relief to the treated group, with the two groups showing no difference on all outcome measures. However, mothers perceived both creams to be effective in relieving discomfort. Feeding had a greater effect than the application of cream on relieving discomfort and decreasing tissue hardness. It is therefore recommended that lactation consultants encourage mothers to breastfeed if possible to relieve the discomfort of breast engorgement.

Administration, Topical↗
PubMed2000

Preventive effects of dietary cabbage acylated anthocyanins on paraquat-induced oxidative stress in rats.

The preventive effects of acylated anthocyanins from red cabbage on paraquat-induced oxidative stress were determined in rats. Decreased food intake and body weight gain, and increased lung weight and atherogenic index by feeding the rats on a diet containing paraquat were clearly suppressed by supplementing acylated anthocynins to the paraquat diet. Paraquat feeding increased the concentration of thiobarbituric acid-reactive substances (TBARS) in liver lipids, and decreased the liver triacylglycerol level. These effects tended to be suppressed by supplementing acylated anthocynins to the paraquat diet. In addition, the catalase activity in the liver mitochondrial fraction was markedly decreased by feeding on the paraquat diet, this decrease being partially suppressed by supplementing the paraquat diet with acylated anthocyanins. An increase in the NADPH-cytochrome-P450-reductase activity in the liver microsome fraction by paraquat was suppressed by supplementing the paraquat diet with acylated anthocyanins. These results suggest that acylated anthocyanins from red cabbage acted preventively against the oxidative stress in vivo that may have been due to active oxygen species formed through the action of paraquat.

Acylation↗
PubMed2000

Host selection, development, survival, and reproduction of turnip aphid (Homoptera: Aphididae) on green and red cabbage varieties.

Host selection, development, survival, and reproduction of the turnip aphid, Lipaphis erysimi (Kaltenbach), on selected green and red cabbage varieties, Brassicae oleracea L. variety capitata, were studied in the laboratory. Although numbers of alate L. erysimi on different varieties varied generally, more alate adults and nymphs were observed on green than on red varieties on day 3; whereas, these differences were not so significant on day 15. There were no significant differences in alate adults on all six varieties on day 15. Numbers of nymphs and apterous adults of L. erysimi varied among the varieties. When mean numbers of those aphids on green varieties were compared with those on red varieties, the green varieties had more nymphs and apterous adults than those on the red varieties. Developmental durations of L. erysimi nymphs also varied among all varieties, and the nymphs fed on green varieties developed longer than those fed on red varieties. Adult longevities on these cabbage varieties were not significantly different; whereas, the adult fecundities were. Survival rates of L. erysimi varied greatly among all varieties, but there are no significant differences between the green and red varieties. Factors that might affect aphid host selection, growth, longevity, fecundity, and survivorship were discussed.

Animals↗