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Respiratory costs and rate of protein turnover in the roots of a fast-growing (Dactylis glomerata L.) and a slow-growing (Festuca ovina L.) grass species.

Protein turnover is generally regarded as one of the most important maintenance processes in plants in terms of energy requirements. In this study, the contribution of protein turnover to the respiratory costs for maintenance in the roots of two grass species, the fast-growing D. actylis glomerata L. and the slow-growing F. estuca ovina L., is evaluated. Plants were grown under controlled-environment conditions in a nutrient solution to which NO(3)- was added at a relative addition rate of 0.2 and 0.1 mol N mol(-1) N already present in the plant d(-1) for D. glomerata and F. ovina, respectively, so as to obtain a steady exponential growth rate close to the plants' maximum relative growth rate. Pulse-chase labelling with (14)C-leucine was used to determine the rate of protein turnover in the grass roots. The rate of turnover of the total protein pool did not differ significantly between the two species. The protein degradation constant in D. glomerata and F. ovina was 0.156 and 0.116 g protein g(-1) protein d(-1), respectively, which corresponds with a total protein half-life of 4 d and 6 d. Assuming specific respiratory costs for protein turnover of 148 mmol ATP g(-1) protein, the estimated respiratory costs for protein turnover in the roots were 2.8 and 2.4 mmol ATP g(-1) root DM d(-1) in D. glomerata and F. ovina, respectively. Both the fast- and the slow-growing grass spent between 22-30% of their daily ATP production for maintenance on protein turnover, which corresponds to 11-15% of the total root ATP production per day. Note that the data presented in this abstract are based on the assumption that 50% recycling of the (14)C-labelled leucine took place in the roots of both grass species.

Adenosine Triphosphate↗

Comparison of growth performance of Lolium perenne L., Dactylis glomerata L. and Agropyron elongatum (Host.) P. Beauv. for erosion control in Turkey.

This study was carried out in plastic containers to compare growth performances of perennial ryegrass, orchardgrass and tall wheatgrass to be given priority in revegetation studies in Turkey. Three pre-germinated seeds of each grass species were planted separately into the soil in the black plastic containers. Seedlings were harvested 2, 4 and 6 months after planting pre-germinated seeds and measured for percentage of seedling emergence, rooting depth, height growth, leaf and tiller development and shoot and root weights. Germination percentage was 97.8% for perennial ryegrass, 64.1% for orchardgrass and 11.6% for tall wheatgrass and perennial ryegrass had the greatest whereas tall wheatgrass had the lowest seedling emergence. Two months old rooting depth was 25.66 cm for perennial ryegrass, 20.56 cm for orchardgrass and 30.10 cm for tall wheatgrass. At the end of the study, perennial ryegrass developed about 104 tillers per plant while they were 21.4 and 36.6 tillers per plant for orchardgrass and tall wheatgrass, respectively. Orchardgrass produced greater shoot and root biomasses than tall wheatgrass and similar to perennial ryegrass. All these meant that perennial ryegrass had a better growth performance than orchardgrass and tall wheatgrass to be used for erosion control.

Agropyron↗

Enzyme determination and RAST inhibition assays for orchard grass (Dactylis glomerata): a comparison of commericial pollen extracts.

Pollens release enzymes when they are dropped into water. Orchard Grass extracts were tested by two methods. The Api-Zym System determined 19 enzymes and colorimetric amounts of Leucine Aminopeptidases, Acid phosphatase and trypsin. These were compared with RAST inhibition assays. A good correlation was shown between the techniques. Thus the enzyme dosage may be useful for the standardization of pollen extracts.

Dose-Response Relationship, Immunologic↗

Isotypic analysis of grass pollen-specific antibodies in human plasma. 4. Biological activity of allergen-specific and autoanti-IgE antibody fractions on basophil histamine release.

BACKGROUND: Blocking antibodies are defined as antibodies that compete with IgE for binding to allergens due to their specificity for those allergens. Thus, they may inhibit allergen-induced basophil and mast cell IgE-dependent mediator release both in vivo and in vitro. OBJECTIVE: The present study was designed to evaluate the ability of antibodies isolated from human plasma samples on a Dactylis glomerata (Cocksfoot) pollen affinity-column to inhibit the Dactylis pollen-induced histamine release from human basophils (BHR) in vitro. METHODS: Antibodies from Ig pools containing either high or low IgG4 anti-Dactylis pollen were purified on a Dactylis pollen affinity-column and then separated on an antihuman IgE column. Obtained Ig fractions were incubated for 30 min with Dactylis pollen allergens prior to incubation with basophils from Dactylis pollen-allergic donors. Cell supernatants were assessed for histamine content and the inhibition of BHR was calculated. RESULTS: Unlike control non-isolated Igs, the antibodies isolated on the Dactylis pollen column were able to inhibit efficiently and in a dose-dependent manner Dactylis pollen-induced BHR. The inhibitory activity was increased in isolated antibody samples that had high IgG4 levels. Antibodies isolated on the Dactylis pollen column, however, consisted not only of true allergen-specific (potentially blocking) antibodies but also of autoanti-IgE binding to allergen-specific IgE and mistaken for allergen-specific antibodies thus opening to question the involvement of the true allergen-specific antibodies in the BHR-inhibitory activity. Unlike the true allergen-specific antibodies, the autoanti-IgE were retained on and eluted from the anti-IgE column. Results showed that both the autoanti-IgE-depleted and the autoanti-IgE-containing fractions accounted for the inhibition observed with the related non-depleted sample that had been isolated on the Dactylis pollen column. CONCLUSION: For the first time, the true blocking activity of allergen-specific antibodies is demonstrated, that is, in the absence of the autoanti-IgE which can also inhibit BHR.

Allergens↗

Effect of smoke, charred wood, and nitrogenous compounds on seed germination of ten species from woodland in central-western Spain.

The effect of smoke, charred wood, and nitrogenous compounds on germination was tested on 10 species of the Cistaceae, Poaceae, Fabaceae, and Asteraceae, from fire-prone, shrubby woodlands in central-western Spain. Dry seeds were exposed to smoke, by watering with distilled water-charred wood suspensions, or NaNO2, KNO3, NH4Cl, and NH4NO3. Smoke enhanced germination in 9 of 10 of the species. In species of Poaceae, germination was stimulated by 20 min of smoke exposure. In Asteraceae and Fabaceae species, 10 min of smoke exposure was the most effective treatment for enhancing germination. Three species--Cistus ladanifer, Cistus crispus, and Cistus monspeliensis--had a positive response to 20 min of smoke exposure; germination of Cistus salviifolius L. was also enhanced after 10 min. The effect of charred wood was variable, with no consistent germination pattern within the families. Trifolium angustifolium and Retama sphaerocarpa showed no stimulation of germination under most of the charred wood concentrations. Similarly, germination of Senecio jacobea under the charred wood treatment did not surpass that of the control. NaNO2 promoted seed germination in Dactylis glomerata (10 mM), Cistus ladanifer (1, 10, and 25 mM), and Cistus crispus (1 and 10 mM). KNO3 enhanced germination in Dactylis glomerata (1 and 25 mM), Dittrichia viscosa (10 and 25 mM), C. ladanifer (1, 10, and 25 mM), Cistus crispus (1 and 25 mM), and C. salviifolius aud C. monspeliensis (25 mM). NH4Cl induced germination of Dactylis glomerata and Dittrichia viscosa (1 mM), and Cistus species germinated best in 25 mM of this salt. NH4NO3 induced germination only in Cistus species. Holcus lanatus had the highest level of germination regardless of treatment.

Adaptation, Physiological↗

[Investigations on the fermentation quality of silage from low-nitrate green forage].

In 3-year-experiments green forage from Dactylis glomerata and Festuca species, produced by differentiated cutting frequencies, was ensiled under laboratory conditions. The forage was almost without nitrate. Because of the higher content of water-soluble carbohydrates the fermentability coefficients of Festuca species were above of Dactylis glomerata. The fermentability coefficients of both plant species were inferior in the second and third cut, even if the first cut was delayed until July. Despite of similar fermentability coefficients of the two plant species silages contained different amounts of lactic and butyric acid. The portion of lactic acid amounted to 52 to 57% and 60 to 73% of total acids, the proportion of butyric acid 30 to 35% and 18 to 25% for Dactylis glomerata and Festuca species, respectively. According to the high fermentability coefficients silages free of butyric acid would be expected, however, by both species butyric acid always was formed, in some cases at high concentrations. In the contrary the amounts of ammonia, acetic acid and alcohol as well as the pH-value were almost extremely low. It can be concluded that the quality of silages produced of green forages with low nitrate cannot be assessed adequately by the current evaluation key.

Acetic Acid↗

Myeloperoxidase release after allergen-specific conjunctival challenge.

BACKGROUND: Allergen-specific conjunctival challenge is a fruitful and complete tool in evaluating pathophysiological phenomena of allergic inflammation. After challenge, a significant neutrophil infiltrate occurred in allergic subjects. The primary (azurophilic) granules of neutrophils contain a variety of enzymes that might potentiate inflammation, such as myeloperoxidase (MPO). It is not known whether allergen-specific conjunctival challenge (ASCC) is able to elicit MPO release. We also investigated the possible role of immunotherapy (IT) in the release of MPO. METHOD: The groups studied included Dactylis glomerata-sensitive adult atopic patients suffering from seasonal allergic rhinoconjunctivitis, and healthy adult nonatopic volunteer controls. One group of allergic patients received no specific hyposensitization (not-IT allergic group). A second group of allergic patients had been immunotherapy-treated with Dactylis glomerata extract for the preceding three years and continued to receive a maintenance dose within the highest potency of the extract (IT-allergic group). ASCC with Dactylis glomerata was performed outside the pollen season in all subjects. Myeloperoxidase was assayed by MPO-enzyme immunoassay method. RESULTS: Thirty minutes after challenge, myeloperoxidase levels in the non-immunotherapy allergic patients were significantly higher compared than in the healthy group (p<0.001). The levels of myeloperoxidase released in the immunotherapy allergic group were significantly lower than those in the nonimmunotherapy allergic group (p<0.001) and higher than those in nonallergic subjects (p<0.001). CONCLUSION: These results indicate that after ASCC there is a release of MPO. Our study suggests that immunotherapy actively modifies the release of MPO after ASCC.

Allergens↗

An immunoblotting analysis of cross-reactivity between melon, and plantago and grass pollens.

It is known that most patients with type I allergy to pollens also suffer intolerance to fruits. Recently, an epidemiological and CAP-inhibition study has shown a new clustering of allergy between melon and Plantago and grass pollens. The aim of the present study was to confirm these results by immunoblotting analysis and inhibition of immunoblotting. Sera from 3 patients with confirmed allergy to melon, and Dactylis glomerata and Plantago lanceolata pollens were used for the in vitro studies. SDS-PAGE and immunoblotting analysis with a pool of sera revealed that several distinct protein bands were shared by the three extracts at 14, 31, and a spectrum between 40 and 70 kDa, approximately. Immunoblotting inhibition experiments, performed with extracts of melon, Plantago and Dactylis, showed that all allergens of melon blotting were almost completely inhibited by grass and Plantago pollen extracts. Inversely, the melon extract was capable of inhibiting IgE-binding to various allergens of Dactylis at high mol mass and partially to the band at 14 kDa. Moreover, the melon almost totally inhibited the IgE-binding capacity to the proteins of Plantago extract. Taken together, the results support the presence of structurally similar allergens in melon, Plantago and grass pollens, and that all allergenic epitopes of the melon are present in these pollens.

Cross Reactions↗

Alkaloids in ergot found on different Gramineae in The Netherlands.

The alkaloid content and the composition of the alkaloid complex of thirteen samples of ergot sclerotia from different gramineous host species collected in The Netherlands were investigated. Two samples collected in France were also examined. Ergot of Glyceria fluitans (L.) R.Br. did not contain alkaloids. The total alkaloid content of ergot found on the other wild grasses was more than 0.2%. The ratio between the contents of water soluble and water insoluble alkaloids of the investigated ergot sclerotia varies between 1:10 and 1:20. The composition of the alkaloid complex in the sclerotia was studied by quantitative thin-layer chromatography. The sclerotia could be grouped into three main categories: --sclerotia of Lolium perenne L., Festuca arundinacea Schreb. and Arrhenatherum elatius (L.) P.B. ex J. et C. Presl, containing predominantly ergocornine, alpha- and beta-ergokryptine and ergosine; --sclerotia of Dactylis glomerata L., Phalaris arundinacea L., Alopecurus geniculatus L. and Holcus mollis L., containing predominantly ergosine, ergocristine and ergotamine; --sclerotia of Molinia caerulea (L.) Moench, containing predominantly ergocristine, ergosine and ergometrine. A great similarity was found between the composition of the alkaloid complex in ergot of Lolium perenne L. from different locations and also in ergot of Molinia caerulea (L.) Moench from different locations. This similarity was less in ergot of Dactylis glomerata L. collected from different locations.

Chromatography, Thin Layer↗

DMISA (dissociated membrane immunosorbent assay), a new ELISA technique performed with blotted samples.

This study describes the use of electrophoretically purified antigens blotted onto nitrocellulose, as solid phase antigens for enzyme-linked immunosorbent assays. This procedure is called DMISA, for dissociated membrane immunosorbent assay. The method is illustrated using immunoblotted antigens of Dactylis glomerata grass pollen extract. The band of interest was located on a print of nitrocellulose by light staining (India ink), then the corresponding strip of nitrocellulose was cut out. Immediately after its solubilization in ethyleneglycol monomethyl ether, the antigen coated nitrocellulose was precipitated by the addition of buffer. In this way the bulk of the antigen remained bound to the membrane. The resulting suspension was carefully washed, and used as a solid phase antigen in an enzyme-linked immunosorbent assay. Two different electrophoretic methods were used to separate the Dactylis glomerata antigens. We compared the results obtained with classical immunoblot and with DMISA, for IgG4 and IgE quantification using sera from patients allergic to D. glomerata and purified blotted antigens present at the nanogram level.

Allergens↗

Characterization with monoclonal and polyclonal antibodies of a new major allergen from grass pollen in the group I molecular weight range.

We have purified a 24/25 kd allergen from orchard grass pollen (Dactylis glomerata) that has an allergenic potency similar to that of the major group I allergen. We provisionally named this allergen grass 4B1 after the monoclonal antibody used for its identification and purification. This monoclonal antibody was obtained by immunizing mice with whole Lolium perenne-pollen extract and by screening the antibody producing hybrids for reactivity with Dactylis glomerata-pollen extract. Grass 4B1 is physicochemically separable from the group I allergen. Polyclonal rabbit antibodies to grass 4B1 do not react with group I allergen or vice versa. Ninety-five sera with IgE antibodies to grass pollen were tested for IgE antibodies to grass 4B1, and greater than 90% was positive in this test. The median response to grass 4B1 was 70% of that to Lol p I.

Allergens↗

Environmental priming influences allergen-specific nasal reactivity.

BACKGROUND: Preceding mucosal response to one allergen leads to the priming of the nasal mucosal response to another allergen. This study aimed to determine whether environmental allergens, especially ubiquitous animal dander, can induce nasal priming. METHODS: We investigated 26 grass-pollen-allergic subjects with additional sensitization to other aeroallergens. We performed continuous allergen challenge for 2 h with 1500 Dactylis glomerata pollen/m3 in the Vienna challenge chamber. The nasal flow at 150 Pa was examined, and subjective scores were obtained every 15 min. Statistical analysis was calculated from the area under curve of nasal flow reduction by Student's t-test and the Mann-Whitney U-test. Alpha was 0.05. RESULTS: In subjects with positive cat-dander RAST (class of > or = 3), besides grass-pollen allergy, the specific nasal allergic reaction to Dactylis challenge was significantly pronounced (P < 0.01), and an earlier onset of reaction was evident. The same results were obtained with additional sensitization to dog dander (P < 0.05). Concomitant sensitization to mugwort also led to escalating symptoms (P < 0.05). CONCLUSIONS: These results indicate that a specific nasal allergic reaction is augmented by environmental priming caused by ubiquitous animal dander and possibly is influenced by the daily use of spices.

Adult↗

Induction of a passive cutaneous anaphylaxis inhibitory factor in mice.

The administration of a purified protein from Dactylis glomerata pollen, named Dactylis inhibitory protein (DIP), to Balb/C mice results in the production of a heat-resistant seric factor that blocks the passive cutaneous anaphylaxis (PCA) reaction in rat skin. This PCA inhibitory factor (PCA-IF) was purified following different purification procedures and analyzed in sodium dodecyl sulfate polyacrylamide gel. Amongst the array of bands detected it was possible to ascribe the inhibitory activity to a protein band of 76,000 daltons but only when it was associated either with another protein band of 69,000 daltons (obtained after a one-step purification procedure) or two other protein bands of 55,000 and 26,000 daltons (obtained after a three-step purification procedure). The inhibitory activity of the PCA-IF is present at a low level in normal Balb/C mice sera and is enhanced after DIP treatment of the mice.

Animals↗

Isotypic analysis of grass pollen-specific immunoglobulins in human plasma. 1. Specialization of certain classes and subclasses in the immune response.

The specificity and isotypic profile of humoral immune responses to Dactylis glomerata (Cocksfoot) pollen was studied by isoelectric focusing (IEF)-immunoprint analysis using 26 human plasma samples with high levels of Dactylis pollen-specific IgG4 (IgG4+ plasma) and 25 human plasma samples with low levels of specific IgG4 (normal plasma). Over 60 individual protein components in an aqueous pollen extract were separated by IEF and immunoprinted onto nitrocellose (NC). Following plasma incubation, bound IgE, IgG1-4, IgA1, IgA2 and IgM antibodies were detected on separate immunoprints with isotype-specific antibodies. Binding patterns of IgG4 and the majority of IgG1 and IgA2 antibodies in the IgG4+ plasma group very closely paralleled the binding patterns produced by the IgE antibodies from the same plasma and are described as the 'allergen repertoire'. In contrast, IgE, IgG4, IgG1 and IgA2 antibody reactivities to the 'allergen repertoire' were insignificant in the normal plasma group. These results suggest a qualitative, as well as a quantitative relationship between the immune responses which involve these 4 isotypes. Characteristic IgG2 and IgM antibody binding patterns, predominantly to non-allergenic antigens, were shared by the plasma from both groups, while IgG3 and IgA1 antibody binding patterns were highly variable from one plasma to another in both groups. One possible origin of the allergic diseases at the immunoglobulin heavy chain gene level is discussed.

Antibody Formation↗