Agrochemical characterisation of "alperujo", a solid by-product of the two-phase centrifugation method for olive oil extraction

Introduction of the two-phase centrifugation system for olive oil extraction during the early nineties in Spain has led to the generation of approximately four million tons per year of a solid olive-mill by-product called "alperujo" (AL). Agrochemical characterisation showed that AL has a high moisture content, slightly acidic pH values and a very high content of organic matter, mainly composed by lignin, hemicellulose and cellulose. It also has a considerable proportion of fats, proteins, water-soluble carbohydrates and a small but active fraction of hydrosoluble phenolic substances. Amongst plant nutrients, AL is rich in potassium, less so in (mainly organic) nitrogen and poor in phosphorus and micronutrients. Some of those properties are not compatible with agricultural requirements, for which reason composting may be considered a suitable alternative for its disposal. AL needs to be characterised before composting, and so an attempt was made to correlate the easily determined ash content with other parameters; the regression models thus obtained are discussed.     

Effects of genotype and temperature on pollen tube growth in perennial ryegrass (Lolium perenne L.)

Summary Low yield in seed crops of perennial ryegrass is related to low fertilization efficiency and low temperature during anthesis. To study the effect of genotype and temperature on pollen performance, we conducted greenhouse experiments at controlled temperatures. Individual florets of four genotypes that are known to differ in seed production were hand pollinated at four temperatures (14°, 18°, 22°, 26° C) both in vivo and via a semiin-vitro method involving excised florets on agar. Pollen germination and tube growth were determined with UV-fluorescence microscopy and scored in six classes at 2 h after pollination in vitro and after 0.5, 2 and 5 h in vivo. In vitro, both genotype and temperature had a significant effect on the performance of self-pollen. Pollen tube growth increased with temperature. In cross-pollinations, the pistil parent had a significant effect on pollen tube growth, and there was also a significant pistil-by-temperature interaction. In vivo, genotype and temperature significantly affected pollen performance. The genotype-by-temperature interaction was only significant 5 h after pollination. One genotype with low seed yield was pseudoself-compatible and was a relatively poor mother after cross-pollination. The effects of genotype and temperature on the growth of self-pollen might be exploited in a breeding programme.A.G. Stephenson was on a sabbattical leave at SVP in 1987     

Cell growth analysis during steady and non-steady growth in leaves of perennial ryegrass (Lolium perenne L.) subject to defoliation

The effect of defoliation on leaf elongation rate (LER) and on the spatial distribution of epidermal cell lengths in the leaf growth zone was studied in vegetative main tillers of perennial ryegrass (Lolium perenne L. cv Modus) grown in a controlled environment. A new material approach was used to analyse the responses of epidermal cell expansion and production during the initial, non‐steady growth phase following defoliation. The analysis involved assigning an identity to individual expanding cells, assessing the displacement and estimating the expansion of cells with assigned identity during day 1 and day 2 after defoliation. LER decreased by 34% during the first 2 d after defoliation and did not recover to the pre‐defoliation rate within the 14 day regrowth period. Decreased LER on day 1 and day 2 after defoliation was associated with (i) a decrease in the length of the leaf growth zone; (ii) a decrease in the length at which epidermal cells stopped expanding; (iii) a reduced expansion of cells at intermediate growth stages; and (iv) a reduction in cell production (i.e. division) and an associated decrease in the number of expanding cells in the growth zone. However, defoliation had no effect on the expansion of cells located in the proximal part of the growth zone. Reduced LER at 14 d after defoliation was associated with a reduced cell production rate (27% lower than the pre‐defoliation rate) and decreased final cell size ( ? 28%).     

Effect of perennial ryegrass (Lolium perenne L.) cultivars on the milk yield of grazing dairy cows

The objective of this experiment was to investigate the effect of four perennial ryegrass cultivars: Bealey, Astonenergy, Spelga and AberMagic on the milk yield and milk composition of grazing dairy cows. Two 4 × 4 latin square experiments were completed, one during the reproductive and the other during the vegetative growth phase of the cultivars. Thirty-two Holstein–Friesian dairy cows were divided into four groups, with each group assigned 17 days on each cultivar during both experiments. Within each observation period, milk yield and milk composition, sward morphology and pasture chemical composition were measured. During the reproductive growth phase, organic matter digestibility (OMD) was greater for Bealey and Astonenergy (P < 0.001; +1.6%). AberMagic contained a higher stem proportion (P < 0.01; +0.06) and a longer sheath height (P < 0.001; +1.9 cm). Consequently, cows grazing AberMagic recorded a lower milk yield (P < 0.001; −1.5 kg/day) and a lower milk solids yield (P < 0.001; −0.13 kg/day). During the vegetative growth phase, OMD was greater (P < 0.001; +1.1%) for Bealey, whereas the differences between the cultivars in terms of sward structure were smaller and did not appear to influence animal performance. As a result, cows grazing Bealey recorded a higher milk yield (P < 0.001; +0.9 kg/day) and a higher milk solids yield (P < 0.01; +0.08 kg/day). It was concluded that grass cultivar did influence milk yield due to variations in sward structure and chemical composition.     

Carbon and nitrogen deposition in expanding tissue elements of perennial ryegrass (Lolium perenne L.) leaves during non-steady growth after defoliation

The effect of defoliation on the deposition of carbon (C) and nitrogen (N) and the contribution of reserves and current assimilates to the use of C and N in expanding leaf tissue of severely defoliated perennial ryegrass (Lolium perenne L.) was assessed with a new material element approach. This included ¹³C/¹²C-and ¹⁵N/¹⁴N-steady-state labelling of all post-defoliation assimilated C and N, analysis of tissue expansion and displacement in the growth zone, and investigation of the spatial and temporal changes in substrate and label incorporation in the expanding elements prior to and after defoliation. The relationship between elemental expansion and C deposition was not altered by defoliation, but total C deposition in the growth zone was decreased due to decreased expansion of tissue at advanced developmental stages and a shortening of the growth zone. The N deposition per unit expansion was increased following defoliation, suggesting that N supply did not limit expansion. Transition from reserve- to current assimilation-derived growth was rapid (<1 d for carbohydrates and approximately 2 d for N), more rapid than suggested by label incorporation in growth zone biomass. The N deposition was highest near the leaf base, where cell division rates are greatest, whereas carbohydrate deposition was highest near the location of most active cell expansion. The contribution of reserve-derived relative to current assimilation-derived carbohydrates (or N) to deposition was very similar for elements at different stages of expansion     

Water deficit and plant competition effects on growth and water-use efficiency of white clover ( Trifolium repens,L.) and ryegrass (Lolium perenne,L.)

The combined effects of soil water deficit and above and below ground interspecific plant competition on the growth, water-use efficiency (WUE), and measured carbon isotopic composition (δ¹³C) values of white clover and ryegrass were studied. White clover and ryegrass were grown in specially designed crates 1) individually; 2) in shoot competition; or 3) in shoot + root competition and either well-watered or at a moderate or severe soil water deficit. The effects of shoot + root competition on shoot dry matter growth were substantial and benefited both white clover and ryegrass when well-watered or at a moderate soil water deficit, while severely reducing white clover shoot dry matter growth at severe soil water deficit. Plant competition did not affect the WUE of white clover or ryegrass. As soil water deficit increased, the WUE of white clover did not change whereas the WUE of ryegrass increased and was greater than that of white clover. This was attributed to the lower leaf water conductance of ryegrass which conserved water and maintained growth longer compared to white clover. A stronger correlation existed between soil water deficit and measured δ¹³C values for ryegrass at each plant competition level (P<0.001) than existed for white clover (individual: P<0.01; shoot + root: P<0.001; shoot: P<0.10). Unlike white clover, the relationship between measured δ¹³C values and shoot dry matter growth indicated that C assimilation for ryegrass was dependent on type of plant competition. That WUE remained constant for white clover while measured δ¹³C values increased as soil water deficit increased, suggests that the role below ground respiration rate played in determining δ¹³C values increased. The WUE of white clover appears to be independent of the nature of the competition between plants and the soil water deficit level at which it is grown, whereas for ryegrass, the addition of root competition to shoot competition should lead to increases in its WUE. This revised version was published online in June 2006 with corrections to the Cover Date.     

Factors affecting the efficacy of Brassica species and ryegrass (Lolium perenne L.) on root-knot nematode infestation of tomato

The effects of soil incorporation of metham-sodium and finely chopped plant residues of broccoli, cabbage, cauliflower, and grass against Meloidogyne incognito and M. javanica over increasing periods of time and a range of dose rates, with or without plastic covering were studied in growth chamber experiments. The scope of this study was to determine the direct effects of fresh organic amendments and the indirect effects of their volatiles on Meloidogyne spp. and tomato plant growth. Amendments were applied at the rate of 5 or 10 kg/m2. The most effective treatment in maintaining low numbers of nematodes was pre-treatment with volatiles of metham-sodium, which resulted in zero invasion of tomato roots. Application of fresh-chopped broccoli, grass and, in some instances, cauliflower also caused a significant reduction in the population of root-knot nematodes. Nematode reduction caused by broccoli was slightly better than by grass. On the other hand, cabbage incorporation did not suppress Meloidogyne spp. to the same extent, though a 19% reduction was observed, even at the lowest dose rate. Suppression of root-knot nematodes was greater in pots amended and covered with plastic tarp as compared to amended non-covered pots. Increased doses for all treatments were further effective in reducing the populations of root-knot nematodes. The direct effect of soil amendments was greater after 2 weeks, while the indirect effects of volatiles were lessened after 2 and 3 weeks.     

Effects of nitrogen and sulphur fertilization on the digestibility and chemical composition of perennial ryegrass (Lolium perenne L.)

Summary Effects of increasing rates of nitrogen (N) and sulphur (S) fertilizers on the yield, digestibility, and composition of N, S, and amino acid of perennial ryegrass (Lolium perenne L.) was studied in a pot experiment using an under-developed S-responsive (Harvey) and a developed non-S responsive (Gorge) soil. Ryegrass plants responded significantly to both N and S fertilization in Harvey soil, but only to N and not S in Gorge soil. Digestibility of the herbage was increased by N fertilization and S has no effect. Total N and S concentrations in herbage increased with increasing rates of N and S applied respectively, but decreased with each increment of S and N applied respectively. Total N: total S ratios were highest in high N low S treatments and were consistently decreased by S application, while N application shows the opposite effect. In severely S-deficientplants, unassimilated N accumulated mainly as non-protein N (NPN) (> 40 per cent total N), consisting largely of soluble N compounds (> 90 per cent NPN) such as asparagine/aspartic acid. Under S-sufficiency condition, high N fertilization resulted dominantly in glutamic acid.     

Stable transformation and long-term expression of the gusA reporter gene in callus lines of perennial ryegrass (Lolium perenne L.)

Stable transformation of perennial ryegrass (Lolium perenne L.) was achieved by biolistic bombardment of a non embryogenic cell suspension culture, using the hpt and gusA gene. The transformation yielded on the average 5 callus lines per bombardment (1.4×10⁶ cells). Stable integration of the genes into the plant genome was demonstrated by Southern analysis of DNA, isolated from hygromycin-resistant callus lines. The gusA reporter gene, which was regulated by the constitutive promoter of the rice gene GOS2, was expressed in both transient and stable transformation assays, indicating that this promoter is suitable for expression of a transferred gene in perennial ryegrass. Long-term GUS expression was observed in ca. 40% of the callus lines, whereas the other callus lines showed instability after 6 months and 1 year of culture.     

Changes in the amide and amino acid composition of xylem exudate from perennial ryegrass (Lolium perenne L.) during regrowth after defoliation

The paper presents the results of amino acid analyses in xylem sap during leaf regrowth of ryegrass plants defoliated firstly at the 8th and secondly at the 12th week of culture. The free amino acid composition of leaves, stubble and roots was also determined and some of the results are reported. Prior to defoliation, xylem sap contained a high proportion of amides, particularly glutamine. During regrowth after defoliation, the proportion of asparagine in the xylem sap increased until the third day when the highest ratios of asparagine/glutamine appeared. The results are compared with relative amounts of free amino acids in the different plant parts and discussed in relation to source-sink nitrogen transfer.     

Differential composition of bacterial communities as influenced by phenanthrene and dibenzo[a,h]anthracene in the rhizosphere of ryegrass (Lolium perenne L.)

Bioremediation technologies of Polycyclic Aromatic Hydrocarbons (PAH) are often limited by the recalcitrance to biodegradation of high molecular weight (HMW) PAH. Rhizosphere is known to increase the biodegradation of PAH but little is known about the biodegradability of these HMW compounds by␣rhizosphere bacteria. This study compared the effects of a 3 and a 5-ring PAH, phenanthrene (PHE) and␣dibenzo[a,h]anthracene (dBA) respectively, on the composition of bacterial community, the bacterial density and the biodegradation activity. Compartmentalized devices were designed to harvest three consecutive sections of the rhizosphere. Rhizosphere and non-rhizosphere compartments were filled with PHE or dBA spiked or unspiked sand and inoculated with a soil bacterial inoculum. Different bacterial communities and degradation values were found 5 weeks after spiking with PHE (41–76% biodegradation) and dBA (12–51% biodegradation). In sections closer to the root surface, bacterial populations differed as a function of the distance to roots and the PAH added, whereas in further rhizosphere sections, communities were closer to those of the non-planted treatments. Biodegradation of PHE was also a function of the distance to roots, and decreased from 76 to 42% within 9 mm from the roots. However, biodegradation of dBA was significantly higher in the middle section (3–6 mm from roots) than the others. Rhizosphere degradation of PAH varies with the nature of the PAH, and C fluxes from roots could limit the degradation of dBA.     

Lipolysis of olive oil and tallow in an emulsifier-free two-phase system by the lipase from oat seeds (Avena sativa L.)

Summary Lipase activity from whole oat seeds is dependent upon calcium ion in a two phase olive oil-water system. No other tested ion can substitute for calcium. The lipolysis of melted tallow at 46°C was successfully conducted in the presence of calcium ion; the fatty acid fraction contained predominantly oleic acid.     

Mature green waste compost enhances growth and nitrogen uptake in wheat (Triticum aestivum L.) and oilseed rape (Brassica napus L.) through the action of water-extractable factors

A series of field and pot trials were carried out to determine the effects of growing wheat and oilseed rape in soils supplemented with green waste composts and provided with additional fertilisers. It was shown consistently that the response of wheat and rape to compost and fertiliser applied together was greater than the responses to the individual additives, but only when very stable compost was used (>10 months processing). Experiments with 15N-labelled fertiliser showed that wheat was able to utilise the applied N more efficiently when cultivated in the stable compost. The enhanced growth was also demonstrated in hydroponic culture of oilseed rape with water extracts of green waste compost in the presence of compound fertiliser. However the effect was rapidly lost at higher dilutions of compost extract (>3). It was concluded that water-extractable growth promoters are present in stable green waste compost, but these only have measurable activity at high concentrations. The identity of the growth promoting factors remains to be found, but the literature suggests that water-extractable humic substances or cytokinins may be involved.