Glomus intraradices and Gigaspora margarita arbuscular mycorrhizal associations differentially affect nitrogen and potassium nutrition of Plantago lanceolata in a low fertility dune soil

Two controlled microcosm experiments aimed at a critical re-assessment of the contributions of divergent arbuscular mycorrhizal (AM) fungi to plant mineral nutrition were established that specifically targeted Plantago lanceolata–Glomus intraradices (B.B/E) and –Gigaspora margarita (BEG 34) symbioses developed in a native, nutrient limited, coastal dune soil. Plant tissue nitrogen (N), phosphorus (P) and potassium (K) status as well as plant growth parameters and levels of mycorrhizal colonization were assessed at harvest. In addition to the general well-established mycorrhizal facilitation of P uptake, the study was able to demonstrate a G. intraradices-specific contribution to improved plant nitrogen and potassium nutrition. In the two respective experiments, G. intraradices-inoculated plants had 27.8% and 40.8% more total N and 55.8% and 23.3% more total K when compared to Gi. margarita inoculated counterparts. Dissimilar overall contribution of the two isolates to plant nutrition was identified in AM-genus specific differences in plant tissue N:P:K ratios. G. intraradices inoculated and non-mycorrhizal plants generally exhibited N:P:K ratios indicative of P limitation whereas for Gi.margarita mycorrhizal plants, corresponding ratios strongly implied either N or K limitation. The study provides further evidence highlighting AM functional biodiversity in respect to plant nutrient limitation experienced by mycorrhizal P. lanceolata in an ecologically relevant soil system.

Arbuscular mycorrhizal modulation of diazotrophic and denitrifying microbial communities in the (mycor)rhizosphere of Plantago lanceolata

Impacts of divergent arbuscular mycorrhizal (AM) fungi, Glomus intraradices and Gigaspora margarita, on denitrifying and diazotrophic bacterial communities of Plantago lanceolata in nutrient-limited dune soil were assessed. We hypothesized AM species-related modifications that were confirmed in respective bacterial nirK and nifH sequence polymorphism -based community clustering and community variance allocation. The denitrifying community appeared more responsive to AM fungi than the nitrogen-fixing community. Nevertheless, the main explanatory variable, in both cases, was plant age. We conclude that AM fungi can modify N-cycling microbial rhizosphere communities and future work should aim to verify the functional significance and mechanistic basis.

Olfactory selection of Plantago lanceolata by snails declines with seedling age

Background and Aims Despite recent recognition that (1) plant–herbivore interactions during the establishment phase, (2) ontogenetic shifts in resource allocation and (3) herbivore response to plant volatile release are each pivotal to a comprehensive understanding of plant defence, no study has examined how herbivore olfactory response varies during seedling ontogeny. Methods Using a Y-tube olfactometer we examined snail (Helix aspersa) olfactory response to pellets derived from macerated Plantago lanceolata plants harvested at 1, 2, 3, 4, 5, 6 and 8 weeks of age to test the hypothesis that olfactory selection of plants by a generalist herbivore varies with plant age. Plant volatiles were collected for 10 min using solid-phase microextraction technique on 1- and 8-week-old P. lanceolata pellets and analysed by gas chromatography coupled with a mass spectrometer. Key Results Selection of P. lanceolata was strongly negatively correlated with increasing age; pellets derived from 1-week-old seedlings were three times more likely to be selected as those from 8-week-old plants. Comparison of plant selection experiments with plant volatile profiles from GC/MS suggests that patterns of olfactory selection may be linked to ontogenetic shifts in concentrations of green leaf volatiles and ethanol (and its hydrolysis derivatives). Conclusions Although confirmatory of predictions made by contemporary plant defence theory, this is the first study to elucidate a link between seedling age and olfactory selection by herbivores. As a consequence, this study provides a new perspective on the ontogenetic expression of seedling defence, and the role of seedling herbivores, particularly terrestrial molluscs, as selective agents in temperate plant communities.

Detecting local adaptation in widespread grassland species - the importance of scale and local plant community

1 Adaptation of plant populations to local environments has been shown in many species but local adaptation is not always apparent and spatial scales of differentiation are not well known. In a reciprocal transplant experiment we tested whether: (i) three widespread grassland species are locally adapted at a European scale; (ii) detection of local adaptation depends on competition with the local plant community; and (iii) local differentiation between neighbouring populations from contrasting habitats can be stronger than differentiation at a European scale. 2 Seeds of Holcus lanatus, Lotus corniculatus and Plantago lanceolata from a Swiss, Czech and UK population were sown in a reciprocal transplant experiment at fields that exhibit environmental conditions similar to the source sites. Seedling emergence, survival, growth and reproduction were recorded for two consecutive years. 3 The effect of competition was tested by comparing individuals in weeded monocultures with plants sown together with species from the local grassland community. To compare large-scale vs. small-scale differentiation, a neighbouring population from a contrasting habitat (wet-dry contrast) was compared with the 'home' and 'foreign' populations. 4 In P. lanceolata and H. lanatus, a significant home-site advantage was detected in fitness-related traits, thus indicating local adaptation. In L. corniculatus, an overall superiority of one provenance was found. 5 The detection of local adaptation depended on competition with the local plant community. In the absence of competition the home-site advantage was underestimated in P. lanceolata and overestimated in H. lanatus. 6 A significant population differentiation between contrasting local habitats was found. In some traits, this small-scale was greater than large-scale differentiation between countries. 7 Our results indicate that local adaptation in real plant communities cannot necessarily be predicted from plants grown in weeded monocultures and that tests on the relationship between fitness and geographical distance have to account for habitat-dependent small-scale differentiation. Considering the strong small-scale differentiation, a local provenance from a different habitat may not be the best choice in ecological restoration if distant populations from a more similar habitat are available.

Potential contribution of natural enemies to patterns of local adaptation in plants

Genetic differentiation among plant populations and adaptation to local environmental conditions are well documented. However, few studies have examined the potential contribution of plant antagonists, such as insect herbivores and pathogens, to the pattern of local adaptation. Here, a reciprocal transplant experiment was set up at three sites across Europe using two common plant species, Holcus lanatus and Plantago lanceolata. The amount of damage by the main above-ground plant antagonists was measured: a rust fungus infecting Holcus and a specialist beetle feeding on Plantago, both in low-density monoculture plots and in competition with interspecific neighbours. Strong genetic differentiation among provenances in the amount of damage by antagonists in both species was found. Local provenances of Holcus had significantly higher amounts of rust infection than foreign provenances, whereas local provenances of Plantago were significantly less damaged by the specialist beetle than the foreign provenances. The presence of surrounding vegetation affected the amount of damage but had little influence on the ranking of plant provenances. The opposite pattern of population differentiation in resistance to local antagonists in the two species suggests that it will be difficult to predict the consequences of plant translocations for interactions with organisms of higher trophic levels.

The inclusion of forage mixtures in the diet of growing dairy heifers: impacts on digestion, energy utilisation, and methane emissions

Intensive farming focusing on monoculture grass species to maximise forage production has led to a reduction in the extent and diversity of species-rich grasslands. However, plant communities with higher species number (richness) are a potential strategy for more sustainable production and mitigation of greenhouse gas (GHG) emissions. Research has indicated the need to understand opportunities that forage mixtures can offer sustainable ruminant production systems. The objective of the two experiments reported here were to evaluate multiple species forage mixtures in comparison to ryegrass-dominant pasture, when conserved or grazed, on digestion, energy utilisation, N excretion, and methane emissions by growing 10–15 month old heifers. Experiment 1 was a 4 × 4 Latin square design with five week periods. Four forage treatments of: (1) ryegrass (control); permanent pasture with perennial ryegrass (Lolium perenne); (2) clover; a ryegrass:red clover (Trifolium pratense) mixture; (3) trefoil; a ryegrass:birdsfoot trefoil (Lotus corniculatus) mixture; and (4) flowers; a ryegrass:wild flower mixture of predominately sorrel (Rumex acetosa), ox-eye daisy (Leucanthemum vulgare), yarrow (Achillea millefolium), knapweed (Centaurea nigra) and ribwort plantain (Plantago lanceolata), were fed as haylages to four dairy heifers. Measurements included digestibility, N excretion, and energy utilisation (including methane emissions measured in respiration chambers). Experiment 2 used 12 different dairy heifers grazing three of the same forage treatments used to make haylage in experiment 1 (ryegrass, clover and flowers) and methane emissions were estimated using the sulphur hexafluoride (SF6) tracer technique. Distribution of ryegrass to other species (dry matter (DM) basis) was approximately 70:30 (clover), 80:20 (trefoil), and 40:60 (flowers) for experiment 1. During the first and second grazing rotations (respectively) in experiment 2, perennial ryegrass accounted for 95 and 98% of DM in ryegrass, and 84 and 52% of DM in clover, with red clover accounting for almost all of the remainder. In the flowers mixture, perennial ryegrass was 52% of the DM in the first grazing rotation and only 30% in the second, with a variety of other flower species occupying the remainder. Across both experiments, compared to the forage mixtures (clover, trefoil and flowers), ryegrass had a higher crude protein (CP) content (P < 0.001, 187 vs. 115 g kg −1 DM) and DM intake (P < 0.05, 9.0 vs. 8.1 kg day −1). Heifers in experiment 1 fed ryegrass, compared to the forage mixtures, had greater total tract digestibility (g kg −1) of DM (DMD; P < 0.008, 713 vs. 641) and CP (CPD, P < 0.001, 699 vs. 475), and used more intake energy (%) for body tissue deposition (P < 0.05, 2.6 vs. −4.9). For both experiments, heifers fed flowers differed the most compared to the ryegrass control for a number of measurements. Compared to ryegrass, flowers had 40% lower CP content (P < 0.001, 113 vs. 187 g kg −1), 18% lower DMD (P < 0.01, 585 vs. 713 g kg −1), 42% lower CPD (P < 0.001, 407 vs. 699 g kg −1), and 10% lower methane yield (P < 0.05, 22.6 vs. 25.1 g kg −1 DM intake). This study has shown inclusion of flowers in forage mixtures resulted in a lower CP concentration, digestibility and intake. These differences were due in part to sward management and maturity at harvest. Further research is needed to determine how best to exploit the potential environmental benefits of forage mixtures in sustainable ruminant production systems.