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Bead modeling using HYDRO and SOLPRO of the conformation of multisubunit proteins: sunflower and rape-seed 11S globulins.

Oil seed globulins from sunflower and rape seed are multi-subunit, oligomeric proteins whose native 11S form is a hexamer. In this work we try to determine the spatial structure in which the six subunits of 11S globulin are arranged. Experimental values of solution properties, including radius of gyration, sedimentation and diffusion coefficients and intrinsic viscosity, are compared with theoretical predictions for hexamers of various geometries. Bead model calculations of solution properties are carried out using the HYDRO and SOLPRO computer programs. A most compact shape, the regular octahedron, is the hexameric structure that fits best the experimental values.

Journal Article↗

Strains of the genus Serratia as beneficial rhizobacteria of oilseed rape with antifungal properties.

Isolates of Serratia have been isolated from the rhizosphere of oilseed rape. The percentage of Serratia in this microenvironment was determined as 12.4% of the total antifungal bacteria. Serratia liquefaciens, S. plymuthica and S. rubidaea were found. All of the isolates showed an antifungal activity against different phytopathogenic fungi in vitro but the efficiency of strains was different. The antifungal mechanisms of 18 selected strains were investigated. Direct antifungal effect may be based on antibiosis (production of prodigiosin and pyrrolnitrin) and production of lytic enzymes (chitinases and beta-1,3-glucanases). Potent siderophores were secreted by the strains to improve the availability of iron. No strain was able to produce cyanide. Most of the strains secrete the plant growth hormone indole-acetic-acid which can directly promote the growth of roots. The mechanisms were specific for each isolate.

Acetic Acid↗

Common themes in redox chemistry emerge from the X-ray structure of oilseed rape (Brassica napus) enoyl acyl carrier protein reductase.

BACKGROUND: Enoyl acyl carrier protein reductase (ENR) catalyzes the NAD(P)H-dependent reduction of trans-delta 2-enoyl acyl carrier protein, an essential step in de novo fatty acid biosynthesis. Plants contain both NADH-dependent and separate NADPH-dependent ENR enzymes which form part of the dissociable type II fatty acid synthetase. Highly elevated levels of the NADH-dependent enzyme are found during lipid deposition in maturing seeds of oilseed rape (Brassica napus). RESULTS: The crystal structure of an ENR-NAD binary complex has been determined at 1.9 A resolution and consists of a homotetramer in which each subunit forms a single domain comprising a seven-stranded parallel beta sheet flanked by seven alpha helices. The subunit has a topology highly reminiscent of a dinucleotide-binding fold. The active site has been located by difference Fourier analysis of data from crystals equilibrated in NADH. CONCLUSIONS: The structure of ENR shows a striking similarity with the epimerases and short-chain alcohol dehydrogenases, in particular, 3 alpha,20 beta-hydroxysteroid dehydrogenase (HSD). The similarity with HSD extends to the conservation of a catalytically important lysine that stabilizes the transition state and to the use of a tyrosine as a base--with subtle modifications arising from differing requirements of the reduction chemistry.

Amino Acid Sequence↗

Rape in Hong Kong: an overview of current knowledge.

This paper summarises all published and unpublished material available on the topic of rape in Hong Kong. Data including epidemiology, victim profile, assault characteristics, are presented. Most of the data shown are from the 15-year period between 1981 and 1995. Gaps in knowledge are highlighted so as to direct the attention of local researchers and overseas research collaborators.

Journal Article↗

False allegations of rape in adults.

The term 'false allegation', when applied to rape and assault, is a difficult concept for forensic medical examiners and the police to comprehend. Reliable evidence about how prevalent these allegations are is hard to obtain. This paper is a transcript of the oral presentation given at the 6th Cross-Channel Conference in Clinical Forensic Medicine held in Southampton, May 14-16th, 1998.

Journal Article↗

[Potential role of winter rape weeds in the extension of broomrape in Poitou-Charentes].

In the Poitou-Charentes district, among the 82 species of winter rape weeds identified, 22 displayed a strong affinity for this crop (Brassica napus L.). In fields, 50% of these weeds were parasitized by Orobanche ramosa, playing the role of host plants. Greenhouse co-cultures (weed/Orobanche ramosa) showed that weeds non-parasitized in fields could be attacked by broomrape, developing a more or less complete cycle. In vitro co-cultures (weed/Orobanche ramosa) revealed that root exudates of non-parasitized weeds, in fields or in greenhouse co-cultures, could induce Orobanche ramosa seed germination, but not attachment. These weeds could play the role of false hosts.

Brassica napus↗

Isolation of viable egg cells of rape (Brassica napus L.).

The isolation of viable egg cells of rape (Brassica napus L.) has been achieved from microdissected ovules. The non-gametic cells of the embryo sac, synergids and central cells have also been isolated. Their morphology corresponded to that of these cells in situ, making a discrimination from isolated sporophytic cells possible. Two hours after isolation the egg cells were still viable. Viable egg cells have been reproducibly isolated with a frequency of 25% per dissected ovule.

Brassica↗

Changes in tocopherol and plastochromanol-8 contents in seeds and oil of oilseed rape (Brassica napus L.) during storage as influenced by temperature and air oxygen.

The changes in tocopherol and plastochromanol-8 contents in seeds and oil of oilseed rape (Brassica napus L.) were studied during a storage period of 24 weeks at different incubation temperatures and exposure to air oxygen (open and closed flasks). In the extracted oil, total tocopherol content remained unaltered at 5 and 20 degrees C throughout the 24 weeks of storage. At 40 degrees C, a beginning degradation was observed already after 4 weeks in both open and closed flasks; the alpha-tocopherol content was affected most, followed by gamma-tocopherol and plastochromanol-8. After 16 weeks at 40 degrees C, the total tocopherol content in the oil was reduced by more than 90%. In intact seeds, no tocopherol degradation was observed; only the seeds incubated at 40 degrees C and in open flasks showed slightly lower tocopherol contents. However, the analysis of the tocopherol composition in the stored seeds showed a decrease in the alpha-tocopherol content and an increase in the gamma-tocopherol content, which resulted in a decreasing alpha-/gamma-tocopherol ratio. This trend was most apparent at 40 degrees C and after 24 weeks of storage. A reduction of plastochromanol-8 occurred only at 40 degrees C and was more pronounced in open flasks. At 40 degrees C and in closed flasks a gradual increase in the content of alpha-tocotrienol was observed, a compound normally not accumulated in rapeseed.

Brassica↗

Novel bacterial diversity recovered from the rhizosphere of oilseed rape (Brassica napus) determined by the analysis of 16S ribosomal DNA.

Soil was sampled to a distance of 2.5 mm beneath a root mat of oilseed rape (Brassica napus) in a model rhizosphere system. DNA was extracted and the 16S rDNA amplified, cloned and sequenced. Phylogenetic analysis of these sequences with those held on-line, revealed that 37% of the clones fell within the Holophaga /Acidobacterium phylum, 17% were within the proteobacteria, 14% of the clones were close relatives of Bacillus megaterium and 5% were related to Verrucomicrobium spinosum. An additional eleven clones (21%) could not be assigned to any known phylum and may represent novel bacterial lineages. This study highlights the diverse nature of rhizosphere soils and reinforces the role that molecular approaches play in unravelling such diversity.

Bacteria↗

Oilseed rape flour: another allergen causing occupational asthma among farmers.

BACKGROUND: Farmers are exposed to a wide variety of sensitizers. Since occupational asthma (OA) can lead to permanent disability, exposure discontinuation is the preferred treatment. When this is not possible, the identification of the causative allergen may allow an alternative therapy. METHODS: We present three farmers diagnosed with OA as a consequence of handling fodder. We carried out skin tests with common and occupational allergens and with oilseed rape (OSR) extract. Total and specific serum IgE levels were measured. The patients underwent the OSR-bronchial provocation test (OSR-BPT). The day before and 24 h after the OSR-BPT, the methacholine (M)-BPT and induced sputum were performed. Eosinophil percentages and ECP levels were measured in the sputum samples. RESULTS: OSR sensitization (skin tests and specific serum IgE) was detected in all the patients. The OSR-BPT elicited early responses in two subjects. Methacholine sensitivity, sputum eosinophils, and sputum ECP levels increased 24 h after the OSR-BPT in all the patients. CONCLUSIONS: We have demonstrated that inhalation of OSR flour causes bronchoconstriction, induces an eosinophilic inflammatory bronchial response, and increases bronchial hyperresponsiveness in sensitized asthmatics. OSR flour contained in animal fodder should be considered another potential cause of OA among farmers.

Adult↗

Polygalacturonase (pectinase), a new oilseed rape allergen.

BACKGROUND: Type I hypersensitivity to rapeseed pollen allergens was described as the result of a cross-sensitization with various pollens that could constitute an aggravating factor in birch or grass pollen allergies. Recently, a few rapeseed pollen allergens were described. The aim of the present work was to identify new rapeseed pollen allergens by using two-dimensional gel analysis, microsequencing, and mass spectrometry. METHODS: Water extractable proteins from oilseed rape pollen or stamen were separated by two-dimensional gel electrophoresis. The proteins were then electroblotted onto a nitrocellulose (NC) sheet. The NC sheets were successively incubated with (1) individual human sera pre-selected for their immunoglobulin E (IgE) reactivity to rapeseed pollen proteins, (2) alkaline phosphatase (AP)-conjugated goat anti-human IgE and (3) AP substrate. The allergens localized by this method were then identified by microsequencing and MALDI-TOF mass spectrometry analysis. RESULTS: Of the 18 sera studied, five recognized a wide multispot zone with a molecular mass around 43 kD and pIs between 6.5 and 8.5. The results obtained with two representative sera are shown. From this zone, two isoforms of the polygalacturonase enzyme were identified by microsequencing. Confirmation was obtained through MALDI-TOF mass spectrometry analysis. CONCLUSION: The present results allow the identification of a new rapeseed allergen that can be the main allergen for some patients.

Allergens↗

Regulation of frost resistance during cold de-acclimation and re-acclimation in oilseed rape. A possible role of PSII redox state.

A possible role of photosynthetic apparatus during cold de-acclimation was studied in oilseed rape (Brassica napus var. oleifera). Plants of spring (Star) and winter (Górczañski) cultivars were cold acclimated at + 5 degrees C, and de-acclimated during 4 weeks at combinations of + 12 and + 20 degrees C operating in the light or/and dark, with a 12-h photoperiod. Evidence is presented that the photosynthetic apparatus may be involved in temperature perception during de-acclimation. De-acclimation was faster under a 20/12 degrees C (day/night) treatment than under the reverse 12/20 degrees C (day/night). De-acclimation rate was constant when the day temperature was constant, irrespective of the night temperature both under cold day temperature regimes (12/20, 12/12 degrees C (day/night) and warm-day treatments (20/12, 20/20 degrees C (day/night). The fast decrease in frost resistance observed under warm-day de-acclimation was always accompanied by an acceleration of elongation growth. In the spring cultivar, elongation growth increased starting from the second week of de-acclimation, regardless of temperature conditions. Once elongation growth had commenced during de-acclimation, it continued throughout the period necessary for re-acclimation to low temperature. Re-acclimation to the initial freezing tolerance level was only possible when plant elongation was reduced. In addition re-acclimation of the photosynthetic apparatus to low temperature was impossible in fast growing plants. A possible relationship between PSII, growth rate and frost resistance during cold acclimation and de-acclimation is discussed.

Journal Article↗

Male fitness of oilseed rape (Brassica napus), weedy B. rapa and their F(1) hybrids when pollinating B. rapa seeds.

The likelihood that two species hybridise and backcross may depend strongly on environmental conditions, and possibly on competitive interactions between parents and hybrids. We studied the paternity of seeds produced by weedy Brassica rapa growing in mixtures with oilseed rape (B. napus) and their F(1) hybrids at different frequencies and densities. Paternity was determined by the presence of a transgene, morphology, and AFLP markers. In addition, observations of flower and pollen production, and published data on pollen fertilisation success, zygote survival, and seed germination, allowed us to estimate an expected paternity. The frequency and density of B. napus, B. rapa, and F(1) plants had a strong influence on flower, pollen, and seed production, and on the paternity of B. rapa seeds. Hybridisation and backcrossing mostly occurred at low densities and at high frequencies of B. napus and F(1), respectively. F(1) and backcross offspring were produced mainly by a few B. rapa mother plants. The observed hybridisation and backcrossing frequencies were much lower than expected from our compilation of fitness components. Our results show that the male fitness of B. rapa, B. napus, and F(1) hybrids is strongly influenced by their local frequencies, and that male fitness of F(1)hybrids, when pollinating B. rapa seeds, is low even when their female fitness (seed set) is high.

Brassica napus↗

A novel phototropic response to supplementary ultraviolet (UV-B and UV-A) radiation in the siliquas of oilseed rape (Brassica napus L.) grown under natural conditions.

An outdoor polychromatic response spectrum for the phototropic curvature of the seed pods (siliquas) of oilseed rape (Brassica napus L. cv. Rebel) has been measured. The plants were sown and grown at a location of 52 degrees 12' N and 0 degrees 07' E under natural daylight and ambient environmental conditions with supplementary ultraviolet (UV-B and UV-A) radiation. This radiation had central wavelengths of 313, 319, 324, 339, 348, 356 or 377 nm and was supplied daily for 10 hours at an irradiance of 0.13 W m(-2). An action maximum for phototropic curvature of the siliquas was observed at 319 and 324 nm.

Brassica napus↗

Characterization of lipases from the lipid bodies and microsomal membranes of erucic acid-free oilseed-rape (Brassica napus) cotyledons.

Lipase (triacylglycerol lipase, EC 3.1.1.3) activities have been reported previously in the lipid body and microsomal membranes of oilseed-rape (Brassica napus cv. Andor) seedlings, but conflicting data made it unclear whether there was one lipase in the lipid bodies, with the microsomal activity being attributable to fragments of lipid-body membrane, or if there were two separate lipase activities. In the present study, simultaneous characterization of the lipases under identical conditions showed they differed substantially in their pH-activity curves, kinetics and substrate specificities. (1) The kinetics of the microsomal lipase showed that the rate of lipolysis reached a plateau at concentrations above 5 mM, whereas the lipid-body lipase showed a linear increase in activity with substrate concentration up to 20 mM. (2) The pH optimum of the microsomal lipase was 7.5, whereas that of the lipid-body lipase was 9.0. The microsomal lipase was greatly inhibited at higher pH values, whereas the lipid-body lipase was much less affected. (3) Activity of the microsomal lipase was greatly diminished when substrates with longer chain length were used, and enhanced 4-fold if the substrates contained a single double bond. The lipid-body lipase was relatively unaffected by the type of fatty acid in the triacylglycerol. (4) SDS/polyacrylamide-gel electrophoresis showed little or no cross-contamination of the lipid-body and microsomal fractions. (5) The microsomal lipase activity comprised 75-80% of the total extracted.

Brassica↗

Barriers to gene flow from oilseed rape (Brassica napus) into populations of Sinapis arvensis.

One concern over growing herbicide-tolerant crops is that herbicide-tolerance genes may be transferred into the weeds they are designed to control. Brassica napus (oilseed rape) has a number of wild relatives that cause weed problems and the most widespread of these is Sinapis arvensis (charlock). Sinapis arvensis seed was collected from 102 populations across the UK, within and outside B. napus-growing areas. These populations were tested for sexual compatibility with B. napus and it was found that none of them hybridized readily in the glasshouse. In contrast to previous studies, we have found that hybrids can be formed naturally with S. arvensis as the maternal parent. Six diverse B. napus cultivars (Capricorn, Drakkar, Falcon, Galaxy, Hobson and Regent) were tested for their compatibility with S. arvensis but no cultivar hybridized readily in the glasshouse. We were unable to detect gene transfer from B. napus to S. arvensis in the field, confirming the extremely low probability of hybridization predicted from the glasshouse work.

Blotting, Southern↗

Characterization of low-molecular-mass trypsin isoinhibitors from oil-rape (Brassica napus var. oleifera) seed.

A new low-molecular-mass (6767.8 Da) serine proteinase isoinhibitor has been isolated from oil-rape (Brassica napus var. oleifera) seed, designated 5-oxoPro1-Gly62-RTI-III. The 5-oxoPro1-Gly62-RTI-III isoinhibitor is longer than the Asp2-Pro61-RTI-III and the Ser3-Pro61-RTI-III forms, all the other amino acid residues being identical. In RTI-III isoinhibitors, the P1-P1' reactive site bond (where residues forming the reactive site have been identified as PnellipsisP1 and P1'ellipsisPn', where P1-P1' is the inhibitor scissile bond) has been identified at position Arg21-Ile22. The inhibitor disulphide bridges pattern has been determined as Cys5-Cys27, Cys18-Cys31, Cys42-Cys52 and Cys54-Cys57. The disulphide bridge arrangement observed in the RTI-III isoinhibitors is reminiscent of that found in a number of toxins (e.g. erabutoxin b). Moreover, the organization of the three disulphide bridges subset Cys5-Cys27, Cys18-Cys31 and Cys42-Cys52 is reminiscent of that found in epidermal growth factor domains. Preliminary 1H-NMR data indicates the presence of alphaalphaNOEs and 3JalphaNH coupling constants, typical of the beta-structure(s). These data suggest that the three-dimensional structure of the RTI-III isoinhibitors may be reminiscent of that of toxins and epidermal growth factor domains, consisting of three-finger shaped loops extending from the crossover region. Values of the apparent association equilibrium constant for RTI-III isoinhibitors binding to bovine beta-trypsin and bovine alpha-chymotrypsin are 3.3 x 109 m-1 and 2.4 x 106 m-1, respectively, at pH 8.0 and 21.0 degrees C. The serine proteinase : inhibitor complex formation is a pH-dependent entropy-driven process. RTI-III isoinhibitors do not show any similarity to other serine proteinase inhibitors except the low molecular mass white mustard trypsin isoinhibitor, isolated from Sinapis alba L. seed (MTI-2). Therefore, RTI-III and MTI-2 isoinhibitors could be members of a new class of plant serine proteinase inhibitors.

Amino Acid Sequence↗

Lack of stable inheritance of introgressed transgene from oilseed rape in wild radish.

Hybridization of Brassica napus L. (oilseed rape) and Raphanus raphanistrum L. (wild radish) has been demonstrated, and may be the first step towards introgression of transgenes in this wild relative. If wild radish were to display a new adaptive advantage by expressing the transgene, this could modify the ecological balance of species within the agro-ecosystem. To determine if transgenes remained stable in the hybrid, the frequency of herbicide resistance was studied over four advanced generations of hybrid progeny (G8 to G11) that were subjected to herbicide selection pressure. It is expected that hemizygous resistant plants containing an herbicide resistance transgene back-crossed to wild radish would have 50% resistant progeny. In each of the G8 to G11 generations, only 18% of the progeny from resistant plants were resistant. The chromosome complement of herbicide-susceptible progenies, analyzed at G9, was not different from that of wild populations of wild radish. Herbicide-resistant G9 progeny showed higher chromosome instability, and one third of the progeny contained a supernumerary chromosome. These results suggest that in the presence of herbicide selection pressure, the transgene for herbicide resistance would be maintained despite a lack of stabilized introgression. In the absence of selection, the frequency of resistance in the population is expected to decline.

Agriculture↗