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PubMed2003

Growth conditions, elemental accumulation and induced physiological changes in Chinese cabbage.

Soils contaminated with low levels of heavy metals and other trace elements are now frequently used for vegetable growing. In this situation, heavy metals and trace elements from these polluted soils may accumulate in the agricultural plants being grown in them and thereby enter the human food chain. The objectives of this study are to elucidate the effects of growth conditions, manipulated by the crop covers, on the phytoaccumulation of elements, and to investigate the conceivable influences of these conditions on the plant biochemistry. In three consecutive years of field experiments, open air (T(0)), and floating rowcover treatments (T(1): perforated polyethylene 50 micrometers; T(2): polypropylene 17 gm(-2)) were used to produce different environmental conditions for the growth of Chinese cabbage [Brassica rapa L. (Pekinensis group) cv. 'Nagaoka 50']. Five samplings (whole tops) were carried out from transplanting to harvest and measurements of B, Al, Ag, Si and Ca concentration as well as phenolics (orto-diphenols, total phenols and anthocyanins), pectic fractions, amino acids (histidine, phenylalanine and tyrosine) and polyphenol oxidase activity, were carried out in samples. The T(1) (perforated polyethylene sheet) gave greater B, Al, Ag and Si concentration and phytoextraction (in weight units) than the open-air control. These findings can help to develop new cost-effective techniques for phytoremediation as the application of plastic covers in the field. The build-up of heavy metals in those crops would make the product less suitable for human consumption.

Agriculture↗
PubMed2001

Selection and characterization of mixed starter cultures for lactic acid fermentation of carrot, cabbage, beet and onion vegetable mixtures.

An evaluation of various lactic acid bacteria (LAB) for the fermentation of cabbage, carrot and beet-based vegetable products was carried out. As part of a screening process, the growth of 15 cultures in a vegetable juice medium (VJM) was characterized by automated spectrophotometry. Acidification patterns as well as viability during storage of the LAB were also established. There were greater differences between the pure cultures than the mixed ones with respect to growth in VJM and viability during storage. Reductions in viable cell counts during storage of the fermented VJM occurred more rapidly with a Leuconostoc strain than for pediococci or lactobacilli. Inoculation of vegetables was carried out with cultures of Lactobacillus plantarum NK-312, Pediococcus acidilactici AFERM 772 and Leuconostoc mesenteroides BLAC which were rehydrated in a brine. This rehydration procedure was not detrimental to viability. During fermentation of a carrot/cabbage vegetable mix, sugar metabolism was characterized by the assimilation of both glucose and fructose, but sugars remained in the fermented vegetables when acidification stopped. The pH in the LAB-inoculated vegetables after 72 h at 20 degrees C was significantly lower (by 0.2 units) than the uninoculated control. Inoculation with LAB designed for silage fermentation resulted in the inhibition of acetic acid production, and reduced the production of ethanol during fermentation. The selection process on VJM enabled the preparation of a mixed culture that was more rapid than the silage inoculants in acidifying the medium and was more effective in reducing the production of gas during the fermentation and storage of the fermented vegetables.

Colony Count, Microbial↗
PubMed1999

Cytotoxic activity of pierisin, from the cabbage butterfly, Pieris rapae, in various human cancer cell lines.

Pierisin, a protein purified from pupae of the cabbage butterfly, Pieris rapae, exhibits cytotoxic effects against the human gastric cancer TMK-1 cell line, inducing apoptosis. The present study was performed to determine whether pierisin might exert a similar influence on nine other human cancer cell lines and human umbilical vein endothelial cells (HUVECs). Pierisin showed cytotoxic effects in all the human cells tested, with IC50 values ranging from 0.043 ng/ml to 150 ng/ml. Among the target cells, the cervical carcinoma cell line, HeLa, was the most sensitive to pierisin, showing a 1000-fold less IC50 value than that of HUVECs. While pierisin clearly induced apoptotic cell death in most cancer cell lines and HUVECs, the pathway appeared to be probably different from that involving anti-Fas, TNF-alpha and p53. Pierisin may thus be a promising new candidate for cancer therapy.

ADP Ribose Transferases↗
PubMed2001

Purification and characterisation of a non-plant myrosinase from the cabbage aphid Brevicoryne brassicae (L.).

Plant myrosinases and glucosinolates constitute a defence system in cruciferous plants towards pests and diseases. We have purified for the first time a non-plant myrosinase from the cabbage aphid Brevicoryne brassicae (L.) to homogeneity. The protein was N-terminally blocked and protease (trypsin and lys c) degradation gave peptides of which five were sequenced. The protein is a dimer with subunits of mass 54 kDa+/-500 Da. Western blot analysis with an anti-aphid myrosinase antibody showed a strong cross reaction with a protein extract from the Brassica specialist, B. brassicae. The anti-aphid myrosinase antibody does not cross react with plant myrosinase neither does an anti-plant myrosinase antibody cross react with aphid myrosinase.

Animals↗
PubMed2001

Structural and functional conservation and divergence among acyl-CoA desaturases of two noctuid species, the corn earworm, Helicoverpa zea, and the cabbage looper, Trichoplusia ni.

In this report, we describe the structural and functional analyses of four acyl-CoA desaturase-encoding cDNAs that we isolated from RNA expressed in the pheromone gland of the corn earworm, Helicoverpa zea. We deduced the homology relationships of the encoded proteins, designated HzPGDs1, HzPGDs2, HzPGDs3 and HzFBDs, to each other and to previously described desaturases of the cabbage looper moth, Trichoplusia ni, the fly, Drosophila melanogaster, and other more distantly related organisms. We also isolated genomic DNA fragments of the four H. zea desaturase-encoding genes, determined the locations of introns present in them, and compared them to conserved intron positions in reported desaturase genes of other species. We measured the levels of the four desaturase mRNAs in H. zea pheromone glands and larval fat bodies by RT-PCR. We established the functional identities of the deduced proteins HzPGDs1 and HzPGDs2, encoded by the two desaturase mRNAs that are differentially and abundantly expressed in pheromone glands of sexually mature adult H. zea females, by functional expression of their encoding cDNAs in a desaturase-deficient mutant, ole1, of the yeast Saccharomyces cerevisiae. We compared the unique unsaturated fatty acid profiles of HzPGDs1- and HzPGDs2-expressing transformants to those of strains expressing previously described Delta11 and Delta9 desaturases of T. ni.

Amino Acid Sequence↗
PubMed2002

A high-affinity octopamine transporter cloned from the central nervous system of cabbage looper Trichoplusia ni.

A cDNA encoding a high-affinity Na(+)/anion(-)-dependent octopamine transporter (OAT) was isolated via an RT-PCR-based approach from caterpillars of the cabbage looper, Trichoplusia ni. The deduced amino acid sequence of the OAT cDNA predicts a 670 amino acid protein bearing strong homology to previously cloned monoamine transporters. The expression pattern of OAT mRNA in the central nervous system revealed by in situ hybridization closely resembles that of OA-ergic neurons identified by the presence of mRNA for tyramine beta-hydroxylase, a marker enzyme for OA-ergic neurons in invertebrates. In vitro, insect cells infected with OAT-expressing baculovirus accumulated both (3)H-OA and (3)H-dopamine with saturation kinetics typical of carrier-mediated processes. (3)H-dopamine uptake by OAT was most inhibited by tyramine, OA, dopamine and the tricyclic antidepressants desipramine and imipramine. Substitution studies for Na(+) and Cl(-) indicate that OAT has a strong requirement for Na(+) and a less stringent requirement for Cl(-). The pharmacological profile of OAT is distinct from those of other cloned monoamine transporters and makes OAT a potential target for neuro-active pest control agents.

Adrenergic alpha-Agonists↗
PubMed2002

A protein from the cabbage looper, Trichoplusia ni, regulated by a bacterial infection is homologous to 3-dehydroecdysone 3beta-reductase.

During the screening of immune-regulated genes from the cabbage looper, Trichoplusia ni, a 3-dehydroecdysone 3beta-reductase homologue (DERH) was cloned. In the course of development, 3-dehydroecdysone 3beta-reductase mediates the conversion of 3-dehydroecdysone (3dE) secreted from the prothoracic glands to ecdysone (E), which is subsequently converted to 20-hydroxyecdysone (20E), the major insect molting hormone. The cloned gene is upregulated in fat body during development and is strongly induced after the larva is challenged with bacteria. The gene codes for a 308 amino acid residue protein which shows 42.5% identity to Spodoptera littoralis 3-dehydroecdysone 3beta-reductase. Using the baculovirus expression system, the recombinant DERH was expressed. The purified protein mediates the reduction of 3-dehydromakisterone A to makisterone A, and requires NADPH as a cofactor. Western blots using an antiserum to T. ni DERH revealed the presence of the protein in larval hemolymph and integument. The data indicate that the protein is regulated developmentally and is induced after a challenge with bacteria. Immunohistochemical studies localized the enzyme exclusively in the epidermis and the cuticle.

3-Hydroxysteroid Dehydrogenases↗
PubMed1979

Phospholipase D from savoy cabbage: purification and preliminary kinetic characterization.

Phospholipase D has been purified 680-fold from an acetone powder of savoy cabbage in an overall yield of 30%. The purification involves solubilization of the acetone powder in a Ca2+-containing buffer and subsequent ammonium sulfate fractionation. Gel filtration on Sephadex G-200 and hydrophobic affinity chromatography using a gamma-aminopropane-agarose gel complete the purification. The two chromatographic steps were conducted in buffers containing 50% ethylene glycol, which was necessary in order to maintain stability of the enzyme. Purity was established on the basis of gel electrophoresis and ultracentrifugation. A preliminary kinetic characterization of the enzyme was carried out by using lecithins with short-chain fatty acids below the critical micelle concentration. A complex series of results were obtained which demonstrated the following. (1) The enzyme is quite sensitive to ionic strength, being inhibited at high ionic strength. (2) The pH optimum depends on the concentration of Ca2+ used in the assay. At 0.5 mM Ca2+ the pH optimum is 7.25, but it is 6.0 at 50 mM Ca2+. (3) The effect of substrate concentration at a given pH and ionic strength did not show simple hyperbolic kinetics but rather regions of parabolic and hyperbolic kinetics.

Kinetics↗
PubMed2001

Partial purification of polyphenol oxidase from Chinese cabbage Brassica rapa L.

Polyphenol oxidase (PPO) was purified and characterized from Chinese cabbage by ammonium sulfate precipitation and DEAE-Toyopearl 650M column chromatography. Substrate staining of the crude protein extract showed the presence of three isozymic forms of this enzyme. The molecular weight of the purified enzyme was estimated to be approximately 65 kDa by gel filtration on Toyopearl HW-55F. On SDS-PAGE analysis, this enzyme was composed of a subunit molecular weight of 65 kDa. The optimum pH was 5.0, and this enzyme was stable at pH 6.0 but was unstable below pH 4.0 or above pH 7.0. The optimum temperature was 40 degrees C. Heat inactivation studies showed temperatures >40 degrees C resulted in loss of enzyme activity. PPO showed activity to catechol, pyrogallol, and dopamine (K(m) and V(max) values were 682.5 mM and 67.6 OD/min for catechol, 15.4 mM and 14.1 OD/min for pyrogallol, and 62.0 mM and 14.9 OD/min for dopamine, respectively). The most effective inhibitor was 2-mercaptoethanol, followed in decreasing order by ascorbic acid, glutathione, and L-cysteine. The enzyme activity of the preparation was maintained for 2 days at 4 degrees C but showed a sudden decreased after 3 days.

Brassica↗
PubMed2002

Accumulation of Zn, Cd, Cu, and Pb in Chinese cabbage as influenced by climatic conditions under protected cultivation.

Accumulation of heavy metals from agricultural soils contaminated by low levels heavy metals has important implications in the understanding of heavy metal contamination in the food chain. Through field experiments (1994-1996), the influence of thermal regime under different treatments on the accumulation of zinc, cadmium, copper, and lead in Chinese cabbage [Brassica pekinensis (Lour) Rupr. cv. Nagaoka 50] grown in a Calcareous Fluvisol (Xerofluvent) in Granada (southern Spain) was examined. Two floating row covers were used: T(1) (perforated polyethylene, 50 microm thick) and T(2) (17 g m(-2) polypropylene nonwoven fleece). An uncovered cultivation (T(0)) served as control. Zn, Cd, Cu, and Pb levels in the whole tops of experimental plants were analyzed. Treatments T(1) and T(2) gave rise to differences in environmental conditions with respect to T(0). The influence of environmental factors manipulated by floating row covers (particularly under T(1)) increased total heavy metal accumulation in the above ground plant biomass with respect to the open-air crop. The total contents of Zn, Cd, Cu, and Pb were 30, 50, 90, and 40% higher in T(1), respectively, than in T(0). This technique could be used in contaminated zones for different plant species because the thermal effect favors the process of phytoextraction and thus reduces the contamination.

Agriculture↗
PubMed2004

Glucoraphanin and 4-hydroxyglucobrassicin contents in seeds of 59 cultivars of broccoli, raab, kohlrabi, radish, cauliflower, brussels sprouts, kale, and cabbage.

The importance of dietary sulforaphane in helping maintain good health continues to gain support within the health-care community and awareness among U.S. consumers. In addition to the traditional avenue for obtaining sulforaphane, namely, the consumption of appropriate cruciferous vegetables, other consumer products containing added glucoraphanin, the natural precursor to sulforaphane, are now appearing in the United States. Crucifer seeds are a likely source for obtaining glucoraphanin, owing to a higher concentration of glucoraphanin and the relative ease of processing seeds as compared to vegetative parts. Seeds of several commonly consumed crucifers were analyzed not only for glucoraphanin but also for components that might have negative health implications, such as certain indole-containing glucosinolates and erucic acid-containing lipids. Glucoraphanin, 4-hydroxyglucobrassicin, other glucosinolates, and lipid erucic acid were quantified in seeds of 33 commercially available cultivars of broccoli, 4 cultivars each of kohlrabi, radish, cauliflower, Brussels sprouts, kale, and cabbage, and 2 cultivars of raab.

Brassica↗
PubMed2005

Phenolic compounds in external leaves of tronchuda cabbage (Brassica oleracea L. var. costata DC).

Glycosylated kaempferol derivatives from the external leaves of tronchuda cabbage (Brassica oleracea L. var. costata DC) characterized by reversed-phase HPLC-DAD-MS/MS-ESI were kaempferol 3-O-sophorotrioside-7-O-glucoside, kaempferol 3-O- (methoxycaffeoyl/caffeoyl)sophoroside-7-O-glucoside, kaempferol 3-O-sophoroside-7-O-glucoside, kaempferol 3-O-sophorotrioside-7-O-sophoroside, kaempferol 3-O-sophoroside-7-O-sophoroside, kaempferol 3-O-tetraglucoside-7-O-sophoroside, kaempferol 3-O-(sinapoyl/caffeoyl)sophoroside-7-O-glucoside, kaempferol 3-O-(feruloyl/caffeoyl)sophoroside-7-O-glucoside, kaempferol 3-O-sophorotrioside, kaempferol 3-O-(sinapoyl)sophoroside, kaempferol 3-O-(feruloyl)sophorotrioside, kaempferol 3-O-(feruloyl)sophoroside, kaempferol 3-O-sophoroside, and kaempferol 3-O-glucoside. These acylated derivatives are reported for the first time in nature, with the exception of kaempferol 3-O-(sinapoyl)sophoroside. Quantification of the identified compounds was achieved by HPLC-DAD and carried out in samples cultivated under conventional or organic practices and collected at different times. In general, samples from organic production exhibited higher total phenolics content than those from conventional practices collected in the same period.

Acylation↗
PubMed2005

Influence of two fertilization regimens on the amounts of organic acids and phenolic compounds of tronchuda cabbage (Brassica oleracea L. Var. costata DC).

A phytochemical study was undertaken on tronchuda cabbage (Brassica oleracea L. var. costata DC) cultivated under conventional and organic practices and collected at different times. Six organic acids (aconitic, citric, ascorbic, malic, shikimic, and fumaric acids) were identified and quantified by HPLC-UV. Qualitative and quantitative differences were noted between internal and external leaves. Analysis of the phenolics of the internal leaves was achieved by HPLC-DAD, and the phenolic profile obtained was revealed to be distinct from that of the external leaves. By this means were identified and quantified 11 compounds: 3-p-coumaroylquinic acid, kaempferol 3-O-sophoroside-7-O-glucoside, kaempferol 3-O-(caffeoyl)sophoroside-7-O-glucoside, kaempferol 3-O-(sinapoyl)sophoroside-7-O-glucoside, kaempferol 3-O-(feruloyl)sophoroside-7-O-glucoside, kaempferol 3-O-sophoroside, two isomeric forms of 1,2-disinapoylgentiobiose, 1-sinapoyl-2-feruloylgentiobiose, 1,2,2'-trisinapoylgentiobiose, and 1,2'-disinapoyl-2-feruloylgentiobiose. In general, internal leaves exhibited more constant chemical profiles.

Agriculture↗
PubMed1999

High-performance liquid phase separation of glycosides. 5. Determination of individual glucosinolates in cabbage and rapeseed by laser-induced fluorescene capillary electrophoresis via the enzymatically released isothiocyanate aglycon.

A capillary electrophoresis (CE) method was developed for the profiling and determination of individual glucosinolates (GSs) via their isothiocyanate degradation products upon myrosinase digestion. The resulting isothiocyanates, the structures of which are reflective of the parent GS's, were then converted to their corresponding amines via base hydrolysis or reaction with 1, 2-benzenedithiol. Subsequently, the amines were fluorescently labeled to allow their sensitive detection by laser-induced fluorescence (LIF). The CE method involved the use of in situ charged micelles for the separation of isothiocyanates and their corresponding fluorescently labeled amines by micellar electrokinetic capillary chromatography (MECC). The term "in situ charged micelles" refers to micelles formed by complexing the polar hydroxyl groups of glycosidic surfactants with borate. The MECC method with on-column LIF detection was applied to the determination of GSs in white cabbage, rapeseed leaves, and rapeseed roots.

Brassica↗
PubMed2003

Analysis of HPRT and supF mutations caused by pierisin-1, a guanine specific ADP-ribosylating toxin derived from the cabbage butterfly.

Pierisin-1, an ADP-ribosylating toxin derived from the cabbage butterfly, Pieris rapae, induces apoptosis in various mammalian cell lines. We recently reported that the target for ADP ribosylation by pierisin-1 is the 2'-deoxyguanosine residue in DNA. To examine whether pierisin-1 would induce mutations in mammalian cell genes, we conducted a mutational analysis for the hypoxanthine-guanine phosphoribosyltransferase (HPRT) locus in pierisin-1-treated Chinese hamster lung (CHL) cells. N(2)-(ADP-ribos-1-yl)-2'-deoxyguanosine was detected by the (32)P-postlabeling method in CHL cells after treatment with pierisin-1 at doses of 2-32 ng/mL; adduct levels were 1.1-12.0 per 10(6) nucleotides. Pierisin-1 induced mutations in the HPRT gene dose-dependently, and the frequency was 38 times higher than the control, at a dose of 32 ng/mL. To confirm that mono(ADP-ribosyl)ated dG itself leads to mutations, the pierisin-1-treated DNA of plasmid pMY189 bearing the supF gene was used for mutational analysis. The mutation frequency of the supF gene treated with 2-8 micro g/mL of pierisin-1 was 17-40-fold the control value. Mutation spectrum analysis showed that single base substitutions dominated in both HPRT and supF genes. Among these, transversions were predominant, and more than 70% of the base substitutions occurred at G:C base pairs in both genes. The most frequent mutations were G:C to C:G, followed by G:C to T:A in HPRT gene, whereas G:C to T:A transversions dominated in the supF gene. Our results indicate that pierisin-1 produced N(2)-(ADP-ribos-1-yl)-2'-deoxyguanosine and this guanine-adduct could lead to mutations in the HPRT and supF genes. These findings could provide very useful information for understanding the biological significance of pierisin-1.

ADP Ribose Transferases↗
PubMed1999

Monosaccharide compositions of Danaus plexippus (monarch butterfly) and Trichoplusia ni (cabbage looper) egg glycoproteins.

Monosaccharide compositions of eggs from Danaus plexippus (monarch butterfly) and Trichoplusia ni (cabbage looper) were analyzed. Analyses were performed mainly with high performance anion exchange chromatography (HPAEC) using crude extracts of eggs or SDS-PAGE separated and PVDF-blotted protein bands. Man and GlcN were the major components in all cases, but low levels of Gal and Fuc were possibly present in some samples. Some T. ni egg glycoproteins even contained GalN. Although a peak comigrating with Neu5Ac could be detected with HPAEC-PAD or RP-HPLC (fluorometry) after derivatization with 1,2-diamino-4,5-methylenedioxy-benzene, the quantities were too small to be significant as an integral part of the analyzed glycoproteins. These data suggests that most of glycans on the glycoproteins are pauci-Man type N-glycans, but a small portion of N-glycan may be either hybrid type or complex type.

Acetylglucosamine↗
PubMed2001

Quantitative genetics of signal evolution: a comparison of the pheromonal signal in two populations of the cabbage looper, Trichoplusia ni.

Pheromones are important in reproductive isolation among populations of moths, but the genetics associated with diversification of pheromonal signals is poorly understood. To gain insight into processes that may lead to diversification we examined the genetic architecture underlying the production of the sex pheromone of the cabbage looper moth, Trichoplusia ni. We compared genetic parameters of two populations; one with a wild-type pheromone phenotype (N) and one where a single-gene mutation affecting the pheromone blend produced by females had been established (M). Using a half-sib breeding design we estimated heritabilities, coefficients of additive genetic variation, and phenotypic, genetic, and environmental correlations of the pheromone components. In both populations, narrow sense heritabilities were generally moderate and genetic correlations were mostly positive. Comparisons between the two populations showed that, while the pattern of phenotypic correlations showed significant agreement between populations, the patterns of genetic (co)variation (i.e. the shapes of the within population matrix) were dissimilar between the two populations. The presence of additive genetic variation in both populations indicates that there is the potential for further evolution of individual pheromone components. However, because of the differences between the populations in the pattern of genetic variation and covariation, the populations will evolve along different evolutionary trajectories even under identical selection pressures. These results suggest that single gene mutations, once established, can be associated with further alterations in the genetic architecture and this has implications for the evolution of pheromone communication.

Animals↗
PubMed2002

Characterization of a stamen-specific cDNA encoding a novel plant defensin in Chinese cabbage.

We isolated a stamen-specific cDNA, BSD1 (Brassica stamen specific plant defensin 1) that encodes a novel plant defensin peptide in Chinese cabbage (Brassica campestris L. ssp. pekinensis). Plant defensins are antimicrobial peptides containing eight highly conserved cysteine residues linked by disulfide bridges. In BSD1, the eight cysteine residues and a glutamate residue at position 29 are conserved whereas other amino acid residues of the plant defensins consensus sequence are substituted. BSD1 transcripts accumulate specifically in the stamen of developing flowers and its level drops as the flowers mature. The recombinant BSD1 produced in Escherichia coli showed antifungal activity against several phytopathogenic fungi. Furthermore, constitutive over-expression of the BSD1 gene under the control of the cauliflower mosaic virus (CaMV) 35S promoter conferred enhanced tolerance against the Phytophthora parasitica in the transgenic tobacco plants.

Amino Acid Sequence↗