Fungal endophytes in grasses from semi-arid permanent grasslands of western Spain

Abstract A survey of grass species associated with Epichloë and Neotyphodium fungal endophytes was made in permanent semi‐arid grasslands of western Spain. Endophytes were isolated from 11 of 49 grass species analysed. Infected plants of Festuca arundinacea, Festuca rubra, Dactylis glomerata and Lolium perenne were detected at most locations studied. The grasses, Agrostis castellana, Brachypodium phoenicoides and Festuca ampla, are previously unknown endophyte hosts. The results obtained show that endophyte–grass interactions are present in the grasslands where this study was conducted.     

Productivity of red clover grown alone and with companion grasses over a four-year period

Six red clover cultivars, three diploid—Essex, Sabtoron and Violetta—and three tetraploid— Teroba, Red Head and Hungaropoly—were sown alone and with each of three companion grasses—timothy (S48), tall fescue (S170) and perennial ryegrass (S24). The productivity and persistency of the red clover cultivars were compared over 4 years. Dry matter (DM) yield, DM digestibility and the crude protein (CP) concentration were assessed and botanical analyses conducted on herbage samples from each treatment at each of three harvests per annum. Annual fertilizer application consisted of 165 kg P and 312 kg K ha^-1. Comparing clover cultivars alone Essex was significantly less productive and less persistent than the other five cultivars. Yield and persistency of the five other cultivars did not differ markedly within years with the exception that the diploids were significantly less productive than the tetraploids in the fourth year. Over all 4 years mean annual total DM and clover DM yields of the five cultivars were between 12·2 and 13·2 t ha^-1 and between 9·2 (79·2% of total DM yield) and 10·2 (83·2%) t ha^-1 respectively, and differences were not significant. Up to the end of the third year there was little or no advantage gained by the inclusion of a companion grass, annual total DM yields being between 11·2 and 14·2 t ha^-1 for clover alone and between 10·2 and 14·2 t ha^-1 for clover-grass mixtures. In the fourth year there was an overall tendency for the yield of the clover alone to be lower, between 7·2 and 12·2 t ha^-1, than that of the clover-grass mixture, between 8·2 and 13·2 t ha^-1, and this was more pronounced with the diploid than with the tetraploid clover cultivars. Sown with companion grasses, Essex and Hungaropoly were lower in yield and in contribution than the other cultivars over the 4 years. The influence of the companion grass on total dry matter yield showed that the contribution of timothy was low relative to that made by tall fescue and perennial ryegrass. Perennial ryegrass made the most varied contribution from year to year. Tall fescue was the most consistent contributor with all clover cultivars and at the end of 4 years both yield and clover-grass balance had not changed materially. No pronounced differences in DM digestibility were evident between treatments. Crude protein concentration of the pure clover was similar to that of the clover-timothy treatments and both would appear to be superior to either the clover-perennial ryegrass or clover-tall fescue mixtures. It is considered that red clover dominant swards are suitable for use under a cutting regime and can provide high yields of DM at a low cost for up to 4 years. Such swards are self-sufficient in N and in addition soil N accumulation can be exploited in the production of succeeding crops.     

Changes in macro‐ and micronutrient contents of grasses and forbs following Miscanthus x giganteus feedstock,hydrochar and biochar application to temperate grassland

Biochar and hydrochar application to soil holds promise for climate change mitigation. This study provides first insights into the nutrient concentration and removal of grassland vegetation after addition of various carbon compounds together with pig slurry. Four treatments: control (no carbon application), feedstock, hydrochar and biochar from Miscanthus x giganteus were applied at a permanent grassland site near Giessen, Germany. Changes in plant functional groups, biomass production and nutrition status were monitored over 2 years. Total biomass production was not affected by the carbon amendments. However, biochar favoured growth of forbs over grasses, while legume growth was increased by all carbon amendments. The initial nutrient concentrations of the carbon compounds were enriched according to their degree of carbonization, potentially providing nutrients to plants. The plant biomass from hydro‐ and biochar amended plots, added up over 2 years, exhibited higher potassium concentrations compared to biomass from feedstock and control plots. All carbon amendments led to lower sodium concentrations in total biomass, compared to the control. Uncarbonized feedstock led to increased manganese concentrations in total biomass, while the concentrations of all other heavy metals were not influenced by any carbon amendment, compared to the control. From a plant and animal nutritional point of view, none of the carbon amendments reduced grassland yield or fodder quality. The study suggests that hydrochar and, even more so, biochar may provide a source of potassium to plants.     

Herbage dry‐matter production and forage quality of three legumes and four non‐leguminous forbs grown in single‐species stands

Agronomic data on most broad‐leaved species of grasslands are scarce. The aim of this study was to obtain novel information on herbage DM yield and forage quality for several forb species, and on species differences and seasonal patterns across harvests and in successive years. Four non‐leguminous forbs [salad burnet (Sanguisorba minor), caraway (Carum carvi), chicory (Cichorium intybus) and ribwort plantain (Plantago lanceolata)] and three leguminous forbs [yellow sweet clover (Melilotus officinalis), lucerne (Medicago sativa) and birdsfoot trefoil (Lotus corniculatus)] and a perennial ryegrass–white clover mixture were investigated in a small‐plot cutting trial in Denmark during 2009 and 2010. Plots were harvested four times per year. On average, annual herbage yield was highest for lucerne (15·4 t DM) and grass–white clover (12·5 t DM ha^?1), and lowest for salad burnet (4·6 t DM ha^?1) and yellow sweet clover (3·9 t DM ha^?1). Ribwort plantain and lucerne had the highest concentrations of acid detergent fibre (339 and 321 g kg^?1 DM respectively) and lignin (78 and 67 g kg^?1 DM respectively); contents in other species were similar to grass–white clover (275 and 49 g kg^?1 DM respectively). No common feature was found within the functional groups of non‐leguminous forbs and leguminous forbs, other than higher crude protein contents (198–206 g kg^?1 DM) in the legumes. DM yield and fibre content were lowest in October. Digestibility declined with higher temperature and increasing fibre content. Results are discussed in terms of the potential of forbs to contribute to forage resources in farming practice.     

Influence of endosperm texture on milling,chemical composition and nutritive quality in maize

Three maize varieties namely Vijay, SO/SN composite and Shakti, which differ in their nutritional quality and endosperm texture were processed to prepare semolina (sooji) and process-flour. The nutritional quality of these products was determined and compared with whole kernel flour of the respective variety. Distinct differences in milling and chemical composition of three varieties were recorded. In quality evaluation tests lysine and tryptophan content, biological value and true digestibility were better in semolina and process-flour. Moreover, decrease in acid value in semolina and process-flour compared with whole maize flour recorded in all the varieties suggested improvement in shelf-life.     

Species interactions in a grassland mixture under low nitrogen fertilization and two cutting frequencies. II. Nutritional quality

Mixtures and pure stands of perennial ryegrass, tall fescue, white clover and red clover were grown in a three‐cut and a five‐cut system in southern Norway, at a low fertilization rate (100 kg N ha^?1 year^?1). The nutritional quality (annual weighted averages) of the dried forage from the two‐first harvesting years was analysed. There was no significant effect of species diversity on crude protein (CP) concentration. In the three‐cut system, we found a significant species diversity effect leading to 10% higher concentrations of acid detergent fibre (ADF), 20–22% lower concentrations of water‐soluble carbohydrate (WSC) and 4% lower net energy for lactation (NE_L) concentrations in mixtures compared with pure stands (averaged across the two‐first years). In the five‐cut system, similar effects were seen in the first year only. This diversity effect was associated with a reduction in WSC and NE_L concentrations and an increase in ADF, NDF and CP concentrations in the grass species, and not in red clover, when grown in mixtures. This is thought to be a combined result of better N availability and more shading in the mixtures. Species diversity reduced the intra‐annual variability in nutritional quality in both cutting systems.     

Species enrichment in an agriculturally improved grassland and its effects on botanical composition, yield and forage quality

In a field experiment, the effects of different management treatments on the botanical development and productivity of a previously species‐poor, intensively managed, lowland permanent grass sward were investigated over a 4‐year period. Fertilizer inputs were stopped and nine main treatments, based on three pre‐sowing and three post‐sowing cutting regimes, were applied. Half of each treatment plot was oversown with a mixture of forb (wildflower) species and half left unsown. The aims were: (1) to investigate the effect of cutting regimes on the establishment of introduced forb species, (2) to assess the development and dry‐matter (DM) yield potential of these introduced species, and (3) to compare the DM yield and forage quality of swards with and without introduced species. Seedling establishment was recorded until the end of the second year. From the third year onwards, a common twice‐yearly cutting regime was imposed on all treatments, and the development of the introduced species was recorded. The DM yield of the nine oversown sub‐treatments was compared with the sub‐treatments not oversown. Frequent cutting after oversowing resulted in the greatest number of established plants and three times greater harvested DM yield of introduced species, compared with infrequent cutting. Plantago lanceolata and Trifolium pratense were the best performing introduced species. By years 3 and 4, there was proportionately 0·15 and 0·23, respectively, additional herbage DM yield harvested from the oversown treatments compared with unsown treatments, and up to 0·60 more in some treatments. The introduction of forbs increased the crude protein concentration, but decreased the digestibility of the forage.     

Effect of season, soil type and fertilizer on the biomass production and chemical composition of five tropical shrub legumes with forage potential

The biomass production and the nutritive value of three tropical shrub legumes with condensed tannins (CT) ( Calliandra calothyrsus , Flemingia macrophylla , Leucaena leucocephala ) and two without CT ( Cratylia argentea , Desmodium velutinum ) as nitrogen-rich feed supplements for ruminants were assessed in two field experiments in Colombia. In one experiment, conducted on two different typical tropical soils (mollisol vs. oxisol; pH, 7·8 vs. 5·0; P content, 43 vs. 2 mg kg DM⁻¹), the effects of low and high levels of fertilizer application with P, K, Ca, Mg and S were tested in both the dry and rainy seasons. In a second experiment on the oxisol, the effect of a lower level of application of either P or S fertilizer was assessed. On the oxisol, C. calothyrsus and F. macrophylla had the highest biomass production (93 and 100 g DM plant⁻¹ in 9 weeks respectively) but the dry season caused extremely low DM yields in all species tested. Leucaena leucocephala did not grow on the oxisol but had the highest biomass production on the mollisol (454 g DM plant⁻¹ in 9 weeks). On the oxisol, the mineral concentrations of the forage were below the requirements of ruminant livestock. Fertilizer treatment had no clear effects on the nutrient composition of the forage. The non-CT shrub C. argentea had the highest concentrations of most minerals in its forage.     

1986 survey of upland grassland in England and Wales: changes in age structure and botanical composition since 1970–72 in relation to grassland management and physical features

A sample of 10000 ha of grassland in seven upland districts originally surveyed in 1970–72 was re-surveyed in 1986. Forty percent of the area was above 240 m elevation, 38% had gradients above 81/2°, 29% had poor or bad drainage and 54% was classified as having difficult or severe limitations to management. Twelve percent had been reseeded within the previous 5 years and 39% within 20 years; 6% was classified as arable grassland (in rotation with crops). Thirty-four percent was usually mown, two-thirds of it for silage. In most years 72% of grassland received fertilizer N (average application 123 kg N ha⁻¹) and 35% received organic manure; nearly 20% received neither. Older swards tended to receive lower rates of fertilizer N than reseeds, hay fields and grazed fields less than silage fields, and fields on beef/sheep farms less than those on dairy farms. The proportion of sown species in swards averaged 35%, of which Lolium perenne accounted for 23%. A good contribution of Trifolium repens was recorded in 15% of the grassland. Herb species were recorded in 30% of the grassland, mostly in situations with low fertilizer N under hay or grazing. Agrostis spp., Poa spp., Holcus lanatus and Festuca rubra were the main unsown grasses. The average contribution of L. perenne had increased since 1970–72; it was also present in two-thirds of the area from which it was absent in 1970–72, whilst the contribution of T. repens had decreased. The incidence of Cirsium arvense and Ranunculus spp. had also decreased, whilst that of Pteridium aquilinum had increased.     

Chemical composition, digestibility and antinutritional factors content of two wild legumes: Styphonolobium burseroides and Acacia bilimekii

The chemical composition, digestibility and toxin contents of two wild legumes: Styphnolobium burseroides and Acacia bilimekii, collected in a semi-arid zone of Mexico, were determined. Both legumes had a high fiber content. The seeds of Styphnolobium burseroides had a low protein content (14%), and the pod a high content of reducing sugars. However the seeds of Acacia bilimekii had a high protein concentration (35%). The seed proteins were low in sulphur amino acids and tryptophan in both legumes but were rich in lysine. Trypsin inhibitors and lectins were present in low concentrations; alkaloids and cyanogenic glucosides were not detected. The in vitro digestibility for monogastric animals was low but the same test with ruminal juice showed a high digestibility for both legumes. Based on their chemical composition and digestibility, these legumes could be a good alternative source in the feeding of ruminants.     

The silage yield potential of hybrid ryegrass over a 5-year period in Northern Ireland relative to other ryegrasses and timothy

The yield and persistency of grass species within a managed sward are a major consideration when determining species mixtures for either long-term or short-term sward management systems. Perennial ryegrass is lower yielding but more persistent than Italian ryegrass. Hybrid ryegrass, an interspecies cross between perennial and Italian ryegrass, may be higher yielding than perennial ryegrass and more persistent than Italian ryegrass. Therefore, the yield and persistency of hybrid ryegrass was investigated in Northern Ireland in five experiments, each harvested over five consecutive years, and compared with that of Italian ryegrass, perennial ryegrass and timothy. The performance of the five grass species were in the expected order; Italian ryegrass was higher yielding than hybrid ryegrass, which was higher than perennial ryegrass, with timothy the lowest, whereas perennial ryegrass and timothy had significantly higher sward densities than the hybrid and Italian ryegrasses. In addition, it was found that the rate of decline in yield and sward density with age was the same for all species. Consequently, this study indicated that the higher yielding Italian and hybrid ryegrasses could continue to out-perform perennial ryegrass for 5 years, opening the possibility of more extensive farming use of these species, particularly in Northern Ireland.     

Amino acid composition,relative nutritive value and in vitro protein digestibility of several Philippine indigenous legumes

This study demonstrates variability in amino acid composition among accessions of several Philippine indigenous legumes. Moreover, two accessions ofD. lablab were identified to have high level of methionine (>2%). Methionine was identified as the first limiting amino acid with leucine, and threonine as the second limiting amino acids for most legumes studies.The IVPDs of the legumes under study ranged from >70 to 79%. Raw mature seeds had relatively low RNVs of 11 to 68% which increased to 68 to 94% and 51 to 89% after boiling and roasting, respectively.     

Productivity and residual benefits of grain legumes to sorghum under semi-arid conditions in south-western Zimbabwe: Unravelling the effects of water and nitrogen using a simulation model

The APSIM model was used to assess the impact of legumes on sorghum grown in rotation in a nutrient-limited system under dry conditions in south-western Zimbabwe. An experiment was conducted at Lucydale, Matopos Research Station, between 2002 and 2005. The model was used to simulate soil and plant responses in the experiment. Sequences of cowpea (Vigna unguiculata), pigeonpea (Cajanus cajan), groundnut (Arachis hypogaea) and sorghum (Sorghum bicolor) were used in the rotations. Legumes accumulated up to 130 kg of N ha⁻¹ which was potentially available for uptake by sorghum in the following season. The APSIM model predicted total biomass, grain and N yields of the legume phase within the experimental error and performed well in predicting sorghum yield and N supplied in the rotation after cowpea and groundnut. The model generally under-predicted sorghum total biomass and grain yield after pigeonpea. Observed patterns of crop water use, evaporative losses during the dry season and re-charge of soil profile at the start of the rainy season were generally well predicted by the model. An assessment of output on sorghum N and water stresses in the rotation indicated that the legume–cereal rotation is more driven by soil nitrogen availability than water availability even under semi-arid conditions. Further legume–cereal rotation analysis using the model will assist in the understanding of other processes in the rotations in dry environments.     

Relationships between botanical composition,yield and forage quality of permanent grasslands over the first growth cycle

This study examined the relationships between botanical composition and forage parameters (yield and forage quality variables) in 153 permanent grasslands located in the Massif Central of France. Grasslands were sampled at two vegetation stages in the first growth cycle. Botanical composition, yield, ash, crude protein, neutral detergent fibre, acid detergent fibre, acid detergent lignin, organic matter digestibility (OMD) and voluntary intake (VI) were estimated for each sample. Temporal variability in species–forage parameter relationships were accounted for using innovative multivariate analyses applied mainly in ecological science. Crude protein and OMD were weakly correlated when each harvest time was analysed separately. Species–forage parameter relationships remained stable during the first growth cycle. The stability of these relationships indicates that permanent grasslands dominated by competitive species were associated with high yield and forage quality values whereas permanent grasslands composed of conservative species and/or high proportions of senescent material were associated with high structural carbohydrate values and low yield, OMD and VI values. Based on these relationships, we propose a typology of permanent grasslands along with a set of indicator species enabling non‐specialist botanists to easily classify grasslands for grassland management purposes.     

Developmental changes in composition and quality of prickly pear cactus cladodes (nopalitos)

The composition and quality of edible tender stems or cladodes of 3 Prickly Pear Cactus species (Opuntia amyclaea, O. ficus-indica, and O. inermis) were studied at different stages of development. This traditional Mexican vegetable is called nopalitos in Spanish and cactus leaves in English. Cladodes harvested when 20 cm in length have the following average composition per 100 g: 91.7 g of water, 1.1 g of protein, 0.2 g of lipid, 1.3 g of ash, 1.1 g of crude fiber, 4.6 g of complex carbohydrates and 0.82 g of simple sugars, 12.7 mg of ascorbic acid and 28.9 µg of carotenes. The cladode's juice has an average pH of 4.6, 0.45% titratable acidity and 6.9% soluble solids. The components which varied most during development of the cladodes were: carotenes, acidity and total carbohydrates which increased, and protein and crude fiber (acid-detergent) which decreased. The nutritive value of the tender cladodes in the stages of growth at which they are commonly harvested and consumed (15 to 25 cm long weighing 50 to 80 g per stem), was similar for the 3 species.     

Nutritional quality of a spun soya-bean protein: Comparison of biological and microbiological tests

A commercial spun soya-bean protein that had been subjected to mild alkali treatment in its manufacture, and the protein isolate from which it had been made, were evaluated in biological and microbiological tests. When diets were givenad lib. in tests with rats, growth was better on the parent protein; but with equal, restricted dietary intakes there was no difference between the two protein sources. The kidneys of all the rats appeared normal on histological examination and showed no signs of nephrocytomegalia, which has been found in the kidneys of rats given soya protein subjected to severe alkali treatment. Similarly in growth tests with chicks, there was no difference between the two proteins and it was concluded that the manufacturing process did not impair the nutritional quality of the spun protein. In microbiological assay tests withStreptococcus zymogenes, the available methionine of the alkali-treated material was consistently lower by about 20%. But when pronase was substituted for the papain normally used in the assay procedure there was no difference between the preparations; the methionine appeared fully available. The lower values following papain predigestion probably indicated a real though small difference in rate of proteolysis but this was clearly not significant in relation to biological availability for rats and chicks.     

The effects of rate and timing of application of fertilizer nitrogen in late summer on herbage mass and chemical composition of perennial ryegrass swards over the winter period in Northern Ireland

A small‐plot experiment was carried out in Northern Ireland on a predominantly perennial ryegrass sward over the period July 1993 to March 1994 to investigate the effect of timing and rate of fertilizer nitrogen (N) application on herbage mass and its chemical composition over the winter period. Eighty treatment combinations, involving four N fertilizer application dates (28 July, 9 and 30 August and 20 September 1993), four rates of N fertilizer (0, 30, 60 and 90 kg N ha^?1) and five harvest dates (1 October, 1 November, 1 December 1993, 1 February and 1 March 1994), were replicated three times in a randomized block design experiment. N application increased herbage mass at each of the harvest dates, but in general there was a decrease in response to N with increasing rate of N and delay in time of application. Mean responses to N applications were 13·0, 11·5 and 9·5 kg DM kg^?1 N at 30, 60 and 90 kg N ha^?1 respectively. Delaying N application, which also reduced the length of the period of growth, reduced the mean response to N fertilizer from 14·3 to 7·4 kg DM kg^?1 N for N applied on 28 July and 20 September respectively. Increasing rate of N application increased the N concentration and reduced the dry‐matter (DM) content and water‐soluble carbohydrate (WSC) concentration of the herbage but had little effect on the acid‐detergent fibre (ADF) concentration. Delaying N application increased N concentration and reduced DM content of the herbage. The effect of date of N application on WSC concentration varied between harvests. A decrease in herbage mass occurred from November onwards which was associated with a decrease in the proportion of live leaf and stem material and an increase in the proportion of dead material in the sward. It is concluded that there is considerable potential to increase the herbage mass available for autumn/early winter grazing by applying up to 60 kg N ha^?1 in early September.     

Semi‐natural grassland biomass for combustion: influence of botanical composition,harvest date and site conditions on fuel composition

Bioenergy generation by combustion is an alternative utilization of biomass from semi‐natural grasslands no longer needed for forage production. Variability of fuel composition affects the suitability of grassland herbage for combustion, in terms of ash‐related problems and harmful emissions of N, S and Cl compounds. A field experiment was established at six semi‐natural grassland sites in southwest Germany to investigate the importance of botanical composition, harvest date and site effects in influencing fuel composition. The first growth at each site was harvested at five dates from June to October. Concentrations of N, S, K, Ca, Mg and ash were lower in grasses than forbs. Between June and October, K concentrations decreased and Ca concentrations increased in grasses and forbs. Ash concentration increased only in grasses, while yields remained stable. Considerable site effects were also found, particularly for K and Cl. The findings are discussed in relation to knowledge of combustion technology. Fuel composition of the investigated grassland biomass was less suitable for combustion than that of perennial energy grasses, but comparable to that of cereal straw. Most critical parameters were high N concentrations, cause of NO_x emissions, and high ash, K and Cl concentrations, responsible for particle emissions, fouling and corrosion. Although combustion of semi‐natural grassland biomass for small‐scale residential boilers cannot be recommended currently, results of this study suggest that utilization in medium‐scale combustion units may offer realistic perspectives, creating synergies between bioenergy production and biodiversity conservation.     

Establishment of legumes in grass swards: effects of pesticides on slugs,insects, legume seedling numbers and forage yield and quality*

Three legumes, red clover (Trifolium pratense L.), alfalfa (Medicago sativa L.) and birdsfoot trefoil (Lotus corniculatus L.), were planted with a minimum-tillage drill into eight grass fields, three in early spring, two in late spring and three in late summer, in Pennsylvania. The objective was to use pesticides at planting time to control slugs and insects which attack legume seedlings and ascertain whether pest control prevented seedling loss and increased yields of grass and legume dry matter (DM), in vitro DM digestibility and crude protein (CP). Methiocarb bait alone or in combination with carbofuran granules or spray applied at planting sometimes controlled the slugs Derocerus reticulatum (Müller), D. laeve (Müller) and Arion fasciatus Nilsson and improved establishment of seedling red clover and alfalfa but not birdsfoot trefoil. Increased yields of DM, digestible DM, CP and legume DM after treatment was greatest in late-spring sowings. Larvae of the clover root curculio, Sitona hispidulus (F.), attacked roots of legumes in the spring following the year of sowing. Late spring and summer sowings without pesticides sustained considerable seedling losses from pests, whereas seedlings from early spring sowings in untreated areas were injured less.     

Effects of formic acid and potassium diformate on the fermentation quality,chemical composition and aerobic stability of alfalfa silage

The objective of this study was to evaluate the efficacy of potassium diformate (KDF) as a potential additive for alfalfa silage. Fresh alfalfa was untreated or treated with formic acid (4 g/kg fresh weight, FW) or three concentrations of KDF (4, 5.5 or 7 g/kg FW). After 60 days of ensiling, the addition of formic acid and greater levels of KDF (5.5 and 7 g/kg) effectively reduced silage pH and inhibited the undesirable bacteria, indicated by lower butyric acid, ethanol, ammonia N concentrations and microbial populations (including enterobacteria, yeasts, moulds and clostridia). Additives decreased the dry‐matter loss, and more water‐soluble carbohydrates were preserved in the silages with formic acid or potassium diformate than in the control. Alfalfa silages treated with formic acid at 4 g/kg FW or potassium diformate at 5.5 or 7 g/kg FW were classified as the highest quality silage based on the higher Flieg's point (above 70) and remained stable for more than 9 days during aerobic exposure. Potassium diformate is recommended as an effective additive for alfalfa silages at a level of 5.5 or 7 g/kg FW under the humid and hot conditions of southern China.