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PubMed1999

Factors enhancing diacylglycerol acyltransferase activity in microsomes from cell-suspension cultures of oilseed rape.

Several factors, including an unidentified endogenous component, were found to stimulate microsomal diacylglycerol acyltransferase (DGAT, EC 2.3.1.20) from a microspore-derived cell-suspension culture of oilseed rape (Brassica napus L. cv. Jet Neuf). At a concentration of 25 mM, MgSO4 and MgCl2 stimulated microsomal DGAT 25- and 10-fold, respectively. ATP and CoA at concentrations of 2 and 1 mM stimulated the enzyme 2.4- and 12-fold, respectively, although the effects were lessened in the presence of higher Mg2+ concentrations. Although microsomal DGAT activity was increased only slightly by the addition of exogenous sn-1,2-diacylglycerol to the reaction mixture, it was increased substantially by the addition of exogenous phosphatidate. sn-Glycerol-3-phosphate and other phospholipids tested did not have this stimulatory effect. DGAT activity did not decrease when microsomes were incubated with ATP in the presence of the cytosolic fraction. This fraction, however, contained a small organic compound(s) that stimulated microsomal DGAT activity.

Acyltransferases↗
PubMed1978

Uridine diphosphate glucose:fatty acid glucosyl transferase activity of rape (Brassica napus L.) anther tissue.

Particulate fractions from mature rape anther tissue catalysed the incorporation of glucose from UDPglucose into endogenous lipids. The UDPglucose:fatty acid glucosyl transferase was tightly bound to particulates that could be suspended in 0.1% Triton X-100. The apparent Km for UDPglucose was 3.5 . 10(-6) M. In Tris buffer the enzyme has a pH optimum of 8.0 and a temperature optimum of 30 degrees C. In the presence of NaF, the transferase activity was proportional to protein concentration, time, and other variables of the reaction mixture. ADPglucose, GDPglucose, and CDPglucose did not compete with the activity of the UDP-glucose-dependent glucosyl transferase. Gas chromatographic and high performance liquid chromatographic analyses indicated that the endogenous fatty acids glucosylated in vitro had carbon chain lengths of C16 to C20. This is the first report on the biosynthesis of non-phosphorylated glucosyl esters of fatty acids in higher plants.

Brassica↗
PubMed1995

Amino acid sequences of three acyl-binding/lipid-transfer proteins from rape seedlings.

The complete amino acid sequence of three acyl-binding/lipid-transfer proteins, AB/LTP I, AB/LTP II and AB/LTP III from germinated rape seeds were determined. AB/LTP I and AB/LTP II consist of 93 residues and the M(r) was determined as 9408 by mass spectrometry and calculated as 9406.8 from the sequence. AB/LTP III consists of 92 residues and the M(r) was determined as 9424 by mass spectrometry and calculated as 9422.8 from the sequence. The primary structures were determined by automated Edman degradations of the intact proteins and peptides obtained from digestion with trypsin and endoproteinase Asp-N and cyanogen bromide cleavage. Use of 252Cf plasma-desorption mass spectrometry facilitated the identification and verification of peptides.

Amino Acid Sequence↗
PubMed1971

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).

Amino Acid Sequence↗
PubMed1990

Biologically active ether lipids. Biotransformation of rac-1(3)-O-alkylglycerols in cell suspension cultures of rape and semisynthesis of 1-O-alkyl-2-palmitoyl-sn-glycero-3-phospho-(N-palmitoyl)ethanolamines, potent antitumor agents.

Biotransformation of rac-1(3)-O-hexadecylglycerol by photomixotrophic rape (Brassica napus) cells in suspension culture leads to 1-O-hexadecyl-2-acyl-sn-glycero-3-phosphocholines and small proportions of other ether lipids, e.g. 1-O-hexadecyl-2-acyl-sn-glycero-3-phosphoethanolamines. Reaction of the hexadecylacyl-glycerophosphocholines with ethanolamine in the presence of phospholipase D from Streptomyces chromofuscus yields additional hexadecylacylglycerophosphoethanolamines. Partial hydrolysis of the combined hexadecylacylglycerophosphoethanolamines followed by reacylation of the resulting lyso compound with palmitic anhydride gives 1-O-hexadecyl-2-palmitoyl-sn-glycero-3-phospho-(N-palmitoyl) ethanolamine, a nontoxic ether glycerophospholipid with antitumor activity. The corresponding 1-O-tetradecyl,1-O-octadecyl, and 1-O-[(Z)-9'-octadecnyl] derivatives are prepared similarly.

Antineoplastic Agents↗
PubMed1989

Inhibition and covalent modification of rape seed (Brassica napus) enoyl ACP reductase by phenylglyoxal.

The NADH-dependent enoyl-ACP reductase from oil seed rape (Brassica napus) was inactivated by treatment with phenylglyoxal, a reagent which specifically modifies arginine residues. The inhibition at various phenylglyoxal concentrations shows pseudo-first-order kinetics, with an apparent second-order rate constant of 14.2 M-1.min-1 for inactivation. The protective ability of several substrates and substrate analogues was investigated in order to ascertain if the inhibition was directed towards the active site of the enzyme. NADH and NAD+ did not protect but acyl carrier protein (ACP) and reduced coenzyme A, along with various derivatives, did protect. 9 microM ACP gave 35% protection from inactivation and 10 mM reduced coenzyme A gave 98% protection. The effectiveness of various subfragments of coenzyme A in protecting against inhibition indicates that the phosphate group is essential for preventing the binding of phenylglyoxal. The idea that phenylglyoxal is inhibiting by binding at the active site is further supported by the observation that the incorporation of 14C-labelled phenylglyoxal is directly related to the loss of activity. Extrapolation of the amount of label incorporated to give total inhibition shows that 4 mol of phenylglyoxal would be incorporated per mol of enzyme. This corresponds to the modification of two arginine side-chains with equal reactiveness towards the reagent. These results are consistent with there being two arginine residues either at the active site of the enzyme or in an environment which is protected from phenylglyoxal by a conformational change induced by coenzyme A binding.

Aldehydes↗
PubMed1991

Single and interactive effects of low levels of O3, SO2 and NO2 on the growth and yield of spring rape.

During three consecutive seasons (1987-1989), the effects of low-levels of O3, SO2 and NO2 singly and in all possible combinations (NO2 in 1988 and 1989 only) on growth and yield of potted plants of spring rape (Brassica napus L. var. napus, 'callypso') were investigated by means of factorial fumigation experiments in open-top chambers. Plants were exposed from the early vegetative stage of development until seed harvest, to charcoal-filtered air (CF; control) and CF which was supplemented for 8-h per day (8.00-16.00) with O3, for 16-h per day with NO2 (16.00-8.00) and continuously with SO2. Including the controls, the 24-h daily mean concentrations [microg m(-3)] ranged between 6-44 (O3), 9-88 (SO2) and 10-43 (NO2). The corresponding daily mean concentrations during the time of fumigation were 10-121 and 11-60 microg m(-3) for O3 and NO2, respectively. Single effects of O3 on growth and yield parameters were mostly negative and the magnitude of this effect was dependent on the season. O3 reduced plant dry weight by 11.3-18.6% and yield of seeds by 11.4-26.9%. While medium levels of SO2 stimulated the weight of pods up to 33%, higher concentrations (88 microg m(-3)) caused a decline of yield of 12.3%. From the significant interactive effects which were observed, it could be established that SO2 and NO2 alone mostly acted positively, but that their interaction with each other and especially with O3 was antagonistic, as some of the detrimental effects of O3 were mitigated by these pollutants. An important antagonistic effect between SO2 and O3 or NO2 was observed on yield. While 56 microg m(-3) SO2 increased yield by 9.9% compared to the control treatment, it aggravated the yield loss caused by O3 from -16.18% to -21.4%, and it reduced the yield stimulation caused by NO2 from +11.8% to +4.2%. Leaf area was the only parameter which was negatively affected by all pollutants, their joint action being synergistic.

Journal Article↗
PubMed1994

Electron spin resonance assessment of susceptibility of roe deer (Capreolus capreolus) and red deer (Cervus elaphus) to oilseed rape (Brassica napus) poisoning.

Ex vivo studies have been carried out on roe and red deer erythrocytes using electron spin resonance spectroscopy. Free radical formation in cells challenged with the brassica-derived haemolysin, dimethyldisulphide, was measured using spin trapping techniques. Significantly greater amounts of radical were trapped in the roe deer cells which may relate to differences in the antioxidant profile of the two genotypes. Results suggest that roe deer have a greater risk of developing oilseed rape poisoning than red deer.

Animals↗
PubMed1992

Study of water and oil diffusion in rape seeds with sorption-desorption and NMR techniques.

The sorption and desorption of water in rape seeds was measured. From the sorption isotherm it follows that for water content greater than about 6% the water molecules tend to form clusters. The mutual diffusion coefficient of water into and out of the seeds was determined from the time dependence of sorption and desorption. There is a pronounced hysteresis in the sorption-desorption process, desorption proceeds faster than sorption. The self-diffusion of water (at maximum humidity of the seeds) and oil within the seeds was investigated by the pulsed field gradient NMR. The measurement of oil self-diffusion shows restricted diffusion of the oil within droplets and allows the determination of the droplet radii and their distribution width.

Brassica↗
PubMed2004

Composting of rice straw with oilseed rape cake and poultry manure and its effects on faba bean (Vicia faba L.) growth and soil properties.

Composting of rice straw with poultry manure and oilseed rape cake and its effects on growth and yield of faba bean and soil properties was studied in pot experiments at Gifu University, Japan in 2001/2002. The composts reached maturity in 90 days, were rich in organic matter and mineral nutrients, had a high level of stability, and no phytotoxicity. The addition of compost (20-200 g pot(-1)) improved selected soil chemical (increased total N, total C and CEC), physical (decreased particle density) and biological (increased soil respiration rate) properties. Application of composts at a rate of 20 g pot(-1) significantly increased growth, yield, yield components and total crude protein of faba bean plants. The benefit of this compost without chemical fertilizer demonstrated the validity and possibility of sustainable agronomic performance of faba bean using locally available recycled organic materials.

Agriculture↗
PubMed2005

[Cadmium effects on lipid metabolism of rape (Brassica napus L.)].

Treatment of rape seedlings with increasing CdCl2 concentrations in the culture medium resulted in a cadmium accumulation within plant tissues, which increased with external metal dose; such accumulation was more important in roots than in leaves. Biomass production was severely inhibited, even at low cadmium concentration. In leaves, quantities of chloroplastic lipids, monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG), sulfolipids (SL) and phosphatidylglycerol (PG) decreased sharply under metallic treatment. However, contents of extrachloroplastic lipids, mainly phosphatidylcholine (PC) and phosphatidylethanolamine (PE) increased significantly. In contrast to leaves, contents of root phospholipids decreased. Likewise, levels of tri-unsaturated fatty acids: linolenic (C18:3) and hexadecatrienoïc (C16:3) dropped in leaves of treated seedlings as compared to those of controls, suggesting that heavy metals induced an alteration in the fatty acid desaturation process or a stimulation of their peroxidation. Also, trans palmitoleic acid (C16:1-trans) level in PG decreased considerably. In roots, there was a slight decrease in C18:3 level, with a concomitant increase in the C18:2 percentage. Radioactive labelling of leaf lipids with (1-14C) acetate allowed to show that fatty acid biosynthesis was noticeably altered at the highest cadmium dose used (50 microM). Biosynthesis of tri-unsaturated fatty acids was also inhibited which may explain the decline in non-labelled lipid contents. Results showed that metallic ion seems to affect selectively chloroplastic membranes due to an inhibition of polyunsaturated fatty acid biosynthesis. Moreover, a lipid peroxidation occurred in our case because of the spectacular increase of malondialdehyde (MDA) content observed in cadmium treated leaves.

Brassica napus↗
PubMed2026

Non-destructive prediction of lead content in oilseed rape leaves by fluorescence hyperspectral technology based on neural network.

Based on fluorescence hyperspectral imaging (FHSI), this study targeted rapid, non-destructive quantification of lead (Pb) content in oilseed rape leaves treated with varying silicon (Si) concentrations, acquiring fluorescence spectra over the 484.43-1001.61 nm wavelength range. To optimize spectral data quality, preprocessing methods (Savitzky-Golay smoothing, first derivative, detrending) were comprehensively compared. Characteristic wavelengths were then selected via interval variable iterative shrinkage, which effectively compressed data dimensionality and reduced computational load. A hybrid SE-CL1DA model, fusing a 1D convolutional neural network, a long short-term memory network and SE attention mechanism was constructed, with Bayesian optimization tuning hyperparameters to boost stability. The BO-SE-CL1DA outperformed both traditional machine learning and insufficiently optimized deep learning model (Rp2=0.9609, RMSE = 0.0377 mg/kg, RPD = 5.1736), thus enabling accurate Pb estimation, supporting Si-regulated heavy metal stress management and facilitating agricultural contamination monitoring.

Plant Leaves↗
PubMed2006

Electric and structural studies of hormone interaction with chloroplast envelope membranes isolated from vegetative and generative rape.

The electric and structural properties of envelope membranes of chloroplasts obtained from vegetative and generative plants of rape and the effect of hormone (IAA, GA(3) and zearalenone) treatment were determined by zeta potential and small-angle X-ray scattering (SAXS) methods. Chloroplasts were isolated from leaves cut off from the vegetative (before cooling) and generative apical parts of plants. The lipid composition of chloroplast envelope membranes were analyzed by chromatographic techniques. Envelopes from generative plants contained higher levels of digalactosyldiacylglycerol (DGDG) and smaller amounts of phospholipids (PLs) in comparison to those obtained from vegetative ones. Moreover, envelopes of generative plants were characterized by higher fractions of unsaturated fatty acids. The zeta potential changes caused by hormone treatment were higher for chloroplasts isolated from vegetative plants in comparison to chloroplasts isolated from generative ones. An especially strong effect was observed for chloroplasts treated with IAA. The thickness of bilayers of untreated chloroplasts from vegetative plants were larger by 0.4 nm when comparing to the thickness of layers obtained from generative ones. The effect of hormones (GA(3) and zearalenone) was detected only for vegetative chloroplasts. Both applied methods indicated differences in the properties of untreated and hormone-treated chloroplasts obtained from vegetative and generative plants.

Brassica napus↗
PubMed2005

Quantification of glucosinolates in leaves of leaf rape (Brassica napus ssp. pabularia) by near-infrared spectroscopy.

The potential of near-infrared spectroscopy (NIRS) for screening the total glucosinolate (t-GSL) content, and also, the aliphatic glucosinolates gluconapin (GNA), glucobrassicanapin (GBN), progoitrin (PRO), glucoalyssin (GAL), and the indole glucosinolate glucobrassicin (GBS) in the leaf rape (Brassica napus L. ssp. pabularia DC), was assessed. This crop is grown for edible leaves for both fodder and human consumption. In Galicia (northwestern Spain) it is highly appreciated for human nutrition and have the common name of "nabicol". A collection of 36 local populations of nabicol was analysed by NIRS for glucosinolate composition. The reference values for glucosinolates, as they were obtained by high performance liquid chromatography on the leaf samples, were regressed against different spectral transformations by modified partial least-squares (MPLS) regression. The coefficients of determination in cross-validation (r2) shown by the equations for t-GSL, GNA, GBN, PRO, GAL and GBS were, respectively, 0.88, 0.73, 0.81, 0.78, 0.37 and 0.41. The standard deviation to standard error of cross-validation ratio, were for these constituents, as follows: t-GSL, 2.96; GNA, 1.94; GBN, 2.31; PRO, 2.11; GAL, 1.27, and GBS, 1.29. These results show that the equations developed for total glucosinolates, as well as those for gluconapin, glucobrassicanapin and progoitrin, can be used for screening these compounds in the leaves of this species. In addition, the glucoalyssin and glucobrassicin equations obtained, can be used to identify those samples with low and high contents. From the study of the MPLS loadings of the first three terms of the different equations, it can be concluded that some major cell components as protein and cellulose, highly participated in modelling the equations for glucosinolates.

Brassica napus↗
PubMed1998

Growth stimulation of beetle larvae reared on a transgenic oilseed rape expressing a cysteine proteinase inhibitor.

The resistance of a transgenic line of oilseed rape expressing constitutively the cysteine proteinase inhibitor oryzacystatin I (OCI) was assessed against Psylliodes chrysocephala L. (Coleoptera: Chrysomelidae). The levels of OCI expression in the transformed line averaged 0.2% and 0.05% of total soluble protein in leaves and petioles respectively. In vitro analyses showed that P. chrysocephala larvae use both cysteine and serine proteinases for protein digestion, and that all the cysteine proteolytic activity is OCI-sensitive. However, bioassays showed that adults fed identically on leaf discs from control or transformed plants. When larvae were reared on transgenic plants expressing OCI, they showed an increase in weight gain compared to those reared on control plants. Furthermore, those larvae from transgenic plants exhibited a 2-fold increase in both cysteine and serine proteolytic activity as a reponse to the presence of OCI. The plasticity of insect digestive physiology and feeding behaviour are discussed, as well as the relevance of engineering a genotype expressing both types of proteinase inhibitors.

Journal Article↗
PubMed1998

Two strains of cabbage seed weevil (Coleoptera: Curculionidae) exhibit differential susceptibility to a transgenic oilseed rape expressing oryzacystatin I.

The aim of this study was to assess the potential effect of a transgenic line of oilseed rape expressing oryzacystatin I (OCI) on two strains of cabbage seed weevil. The level of OCI expression in seeds was approximately 0.05% of total soluble proteins. The insects were field-collected in two different locations, and their progeny was analyzed after a 3 week-development in pods. Both strains showed a similar pattern of proteolytic activity, and similar levels of OCI-sensitive proteinase activity in vitro. However, the larvae showed differential susceptibility to the transgenic plants. Despite inhibition of digestive proteinases in vitro by OCI in both strains, one strain showed an increased growth rate when fed the transgenic seeds, while the other strain remained unaffected. While suggesting the importance of studying individuals from different populations when assessing the effect of proteinase inhibitor-expressing plants on insect growth, our results also point out the necessity of studying the biochemical interactions taking place in vivo between the recombinant inhibitors and their target proteinases.

Journal Article↗
PubMed1998

NCC malonyltransferase catalyses the final step of chlorophyll breakdown in rape (Brassica napus).

Of the three final products of chlorophyll breakdown that in senescing cotyledons of oilseed rape are accumulated progressively, the nonfluorescent Bn-NCC-1 is the most abundant catabolite. It represents the malonylester of the minor catabolite Bn-NCC-3. The in vitro malonylation of Bn-NCC-3 into Bn-NCC-1 was investigated. Extracts from senescent as well as from presenescent cotyledons contained corresponding activities in the presence of malonyl-coenzyme A as the co-substrate. Malonyltransferase activity exhibited pH- and activation optima at 8 and 34 degrees, respectively, and it was saturable with an apparent Michaelis constant of 58 microM for Bn-NCC-3. The partially purified enzyme recognized chlorophyll catabolites as substrate specifically, provided that they had a free hydroxyl group in the ethyl side chain of pyrrole B.

Acyl-Carrier Protein S-Malonyltransferase↗