Composition and digestibility during ageing of consecutive leaves on the main stem of Italian ryegrass plants,growing undisturbed or regrowing after cutting

For modelling of grass digestibility throughout the growing season, the impact of cutting on individual leaves needs to be quantified. Therefore, a glasshouse experiment was carried out with Italian ryegrass (Lolium multiflorum Lam) grown from seed. Half of the plants were grown undisturbed until leaf stage 8, while the other half were cut at leaf stage 4, after which they were allowed to grow until leaf stage 9. The composition and digestibility characteristics of leaves 6, 7 and 8 on the main shoot of plants from both treatments were quantified. In the uncut plants, leaf blade length and mass and specific cell wall (CW) and organic matter (OM) mass (mg cm⁻²) was higher for consecutive leaves. During leaf ageing specific CW mass remained unchanged, while specific OM mass and CW digestibility declined, resulting in a decline of OM digestibility. CW digestibility of leaves decreased to c 78%, a value also found in an earlier experiment. The newly formed leaves after cutting had a reduced leaf size and specific leaf mass (SLM, mg DM cm⁻²). Leaf 6, that was damaged by cutting, showed a lower initial CW content and a rapid decline of specific CW and OM mass and of digestibility during ageing. Its low specific mass and digestibility of CW could have been related to cessation of CW synthesis during its growth after cutting, as was evidenced by the lower CW thickness of sclerenchyma cells. The later-formed leaves, 7 and 8, had digestibility characteristics similar to those of the uncut plants. Since leaf 6 hardly contributed to the total plant dry matter mass, the cut and the uncut plants differed only slightly in digestibility of the total leaf fraction. It can be concluded that cutting sets back leaf size and growth rate but has little effect on digestibility characteristics during ageing. © 1999 Society of Chemical Industry     

Ensiling characteristics and ruminal degradation of Italian ryegrass and lucerne silages treated with cell wall-degrading enzymes

Two experiments were carried out to investigate the effect of added cell wall-degrading enzymes at ensiling on the fermentation and in situ degradation of grass and legume silages. Primary growths of Italian ryegrass (Lolium multiflorum Lam) and lucerne (Medicago sativa L) were wilted and ensiled in laboratory-scale silos with or without enzymes. Silages were opened at 2, 5, 15 and 45 days after storage; the fermentation quality and the contents of cell wall components (NDF, ADF, ADL) were determined. The 45 day silages were subjected to in situ incubation experiments, and the kinetics of DM and NDF degradation was estimated. The enzyme treatment enhanced the lactic acid production (P < 0.01) and reduced the pH value (P < 0.01) of both Italian ryegrass and lucerne silages. The contents of cell wall components, however, were not affected by enzymes, except for NDF of Italian ryegrass silage. The in situ incubation experiments revealed that added enzymes increased the rapidly degradable DM (P < 0.01) and appeared to decrease the rate of degradation of DM and NDF of Italian ryegrass silage. The rapidly degradable DM was not altered when lucerne was treated, but the rate of degradation of NDF was significantly reduced (P < 0.05). These results suggest that although added cell wall-degrading enzymes could improve the preservation of grass and legume silages, the effects on ruminal degradation may be different according to the herbage treated. Enzymatic hydrolysis during ensilage may be restricted to easily digestible cell walls, leaving relatively less digestible components that would be retained in treated silages. © 1999 Society of Chemical Industry     

Analysis of the temperature effect on the components of plant digestibility in two populations of perennial ryegrass

For the development of mechanistic models of herbage digestibility, quantitative insight into the effects of age, temperature and cultivar on digestibility characteristics of individual plant parts is needed. Towards that goal, glasshouse experiments were conducted at day/night temperatures of 13/8, 18/13 and 23/18 °C with vegetative and reproductive crops of two populations of perennial ryegrass (Lolium perenne L) selected for differences in leaf blade digestibility. Cell wall content (CWC) and true cell wall and organic matter digestibility (CWD and OMD) of vegetative and reproductive tillers were related to dimensions, mass, CWC and digestibility of separate plant parts. Compared with the vegetative tillers, the reproductive tillers had higher rates of leaf appearance, organic matter growth and CWD decline. Strikingly, for both tiller types, no direct effect of temperature on whole tiller CWD was observed, since temperature effects could be eliminated completely by relating CWD to development stage (DVS) expressed as number of leaves appearing on the main tiller. Temperature effects on CWD were restricted to its influence on tiller development rate only. The decline of CWD of individual plant parts with DVS in the reproductive tillers could be described with a negative exponential curve, which reached an asymptote that was higher for leaf blades (755 g kg^?1) than for leaf sheaths (491 g kg^?1) and stem internodes (230 g kg^?1). However, all plant parts in both tiller types had the same fractional CWD decline rate of 0.395 per leaf appearance interval, independent of plant part insertion level, population or temperature. Differences between temperature treatments in OMD were caused by the higher CWC of plant parts at higher temperature, due to a stronger decline of the specific organic matter mass than of the specific cell wall mass of plant parts at increasing temperature. Differences in whole tiller OMD between populations were observed only for vegetative tillers and were also caused by differences in CWC. It is concluded that temperature increase accelerated both the tiller development rate and the rate of decline of CWD during aging to the same extent, whereas plant parts responded similarly in the fractional CWD decline pattern as a function of DVS. These trends offer unique possibilities for modelling grass digestibility under contrasting temperature regimes. © 2003 Society of Chemical Industry     

Preparation and composition of mesophyll,epidermis and fibre cell walls from leaves of perennial ryegrass (lolium perenne) and italian ryegrass (lolium multiflorum)

Cells of mesophyll, epidermis and residual fibrous material were obtained from leaves of Italian and perennial ryegrass harvested at different stages of maturity by mechanical disruption of leaf tissue. Mesophyll cells were selectively removed by filtration through 0.045 mm nylon mesh and remaining non-mesophyll cells centrifuged in metrizamide solutions (56–58% wt to vol.) of known density (1.308–1.329 g cm³ at 5°C) to obtain a pure epidermis cell fraction and a residual fibre fraction. Whole mesophyll cells contributed 63–72%, epidermis 12–15% and the fibre fraction 15–24% to the total leaf dry matter. Fibre values were higher in late-cut samples. Cell walls were prepared from mesophyll and epidermis cells by disruption and washing to remove cell contents. Fibre cells were judged free of cell contents and received no further treatment. Examination of cell wall preparations by light and electron microscopy showed that both mesophyll and epidermis preparations were essentially free from contaminating material. Mesophyll cell walls were uniformly thin (200 nm) while those of epidermis ranged from 2000–3000 nm at the outer face, thinning to 300 nm or less at the inner surface. An electron-light layer (cuticle) of approximately 200 nm thickness was present covering the outer face of the epidermis. The fibre fraction largely consisted of sclerenchyma, but contained, in addition, other vascular cells, detached annular rings and heavily silicified leaf hairs. Analysis of cell walls accounted for 85–90% of dry matter. Cellulose was the major component of all cell walls examined (approximately 40% of dry matter) with xylose residues accounting for a further 11% of mesophyll, 13.5–17.5% of epidermis and 21–25% of fibre cell walls. Arabinose was low in fibre cells but was present in much higher proportions in mesophyll and epidermis walls. The ratio of arabinose to xylose was approximately 1:1.5 for mesophyll, 1:2.5 for epidermis and 1:7.0 for the fibre fraction. The molar ratio acetyl to xylose remained fairly constant at 1:4 regardless of the grass, cell type or maturity of the sample. The uronic acid content of epidermis was higher than that in other cell types and showed an increase with increasing maturity of the grass, reaching over 9% in late-cut samples. Total phenolic material represented 2–3% of mesophyll and epidermis cell walls and 6% of fibre walls. Ferulic acid alone was released from the primary cell walls by saponification and p-coumaric and ferulic acids from the secondary-thickened fibre walls. Crude protein values (NX6.25) were high in mesophyll cell wall preparations and low in epidermis and fibre cell walls. Amino acid patterns were similar for both grasses and cell types but hydroxyproline was found in greater amounts in fibre cell walls than in either epidermis or mesophyll.     

Effects of ageing on the in vitro fermentation of cell walls and cell contents of entire,fractionated and composite leaves of Italian ryegrass

Changes in fermentability of cell walls and cell contents of Italian ryegrass leaves at two stages of maturity were measured to explain the generally observed decline in herbage quality with ageing. A herbage fractionation method was developed to separate cell contents and cell walls. Cell walls were either untreated or protease‐treated. Fermentation characteristics of all cell wall and cell contents fractions, as well as whole and recombined leaves, were measured using cumulative gas and volatile fatty acid (VFA) production. The gas production profiles of all substrates contained two significant phases. After fractionation, addition of the profiles from separated cell contents and untreated cell walls resulted in the same profile as for the recombined leaf. The strongest reduction in gas and VFA production due to leaf ageing was observed for cell contents. The increased ratio between branched and straight chain VFAs suggested that there had been an increase in the protein fermentation. Treatment of cell walls may have removed some easily fermentable cell wall components, as seen in the small differences in gas and VFA production between whole and recombined leaves. It has been concluded that the phases of gas production, separable in kinetic fermentability studies of complex animal feeds, need to be interpreted with caution. The large reduction in fermentability of cell contents with plant maturity, compared with the cell walls, indicated the importance of determining the role of cell contents in herbage quality studies, as the cell contents clearly do not remain uniform. © 2000 Society of Chemical Industry