André Pornon

Complementarity in mineral nitrogen use among dominant plant species in a subalpine community

The underlying mechanisms that enable plant species to coexist are poorly understood. Complementarity in resource use is among the major mechanisms proposed that could favor species coexistence but is insufficiently documented. In alpine soil, low temperatures are a major constraint for the supply of plant nitrogen. We carried out (15)N labeling of soil mineral N to determine to what extent four major species of a subalpine community compete for N, or develop ionic (NH(4)(+) vs. NO(3)(-)) or temporal complementarity. The Poaceae took up much more (15)N per soil area unit than the ericaceous species, and all species displayed three major strategies in exploiting (15)N: (1) uptake mainly early in the growing season (Vaccinium myrtillus), (2) uptake at a slow and similar rate throughout the growing season (Rhododendron ferrugineum), and (3) uptake at high rates over the growing season (Festuca eskia and Nardus stricta). However, while F. eskia used (15)NH(4)(+) mainly early and (15)NO(3)(-) mainly late in the growing season, the reverse was observed for N. stricta. Taking into account (15)N dilution in soil NH(4)(+) and NO(3)(-) pools, we calculated that NH(4)(+) provided more than 80% of the mineral N uptake in Ericaceae and about 60% in grasses. Together, such ionic and temporal complementarity would reduce competition between species and could be a major mechanism promoting species diversity.

Using metabarcoding to reveal and quantify plant-pollinator interactions

Given the ongoing decline of both pollinators and plants, it is crucial to implement effective methods to describe complex pollination networks across time and space in a comprehensive and high-throughput way. Here we tested if metabarcoding may circumvent the limits of conventional methodologies in detecting and quantifying plant-pollinator interactions. Metabarcoding experiments on pollen DNA mixtures described a positive relationship between the amounts of DNA from focal species and the number of trnL and ITS1 sequences yielded. The study of pollen loads of insects captured in plant communities revealed that as compared to the observation of visits, metabarcoding revealed 2.5 times more plant species involved in plant-pollinator interactions. We further observed a tight positive relationship between the pollen-carrying capacities of insect taxa and the number of trnL and ITS1 sequences. The number of visits received per plant species also positively correlated to the number of their ITS1 and trnL sequences in insect pollen loads. By revealing interactions hard to observe otherwise, metabarcoding significantly enlarges the spatiotemporal observation window of pollination interactions. By providing new qualitative and quantitative information, metabarcoding holds great promise for investigating diverse facets of interactions and will provide a new perception of pollination networks as a whole.

Isolation and characterization of microsatellite loci in Rhododendron ferrugineum (Ericaceae) using pyrosequencing technology

PREMISE OF THE STUDY Microsatellite primers were developed for Rhododendron ferrugineum (Ericaceae) to evaluate genetic diversity, population genetic structure, and mating system of this self-compatible species. METHODS AND RESULTS The new-generation 454 FLX Titanium pyrosequencing technology was used to isolate 102 novel microsatellite loci. Two multiplex PCR sets were optimized to genotype nine polymorphic microsatellite loci. The level of genetic diversity was assessed in two populations from the Pyrenees (France). The mean number of alleles per locus ranged from 1.5 to 7. The mean observed and expected heterozygosities ranged from 0 to 0.76 and from 0.03 to 0.66, respectively. Cross-species amplification was successful for 13 other Rhododendron species and two additional genera of Ericaceae, with an average of seven pairs of primers amplifying per species. CONCLUSIONS These markers will facilitate further studies on the evolutionary history of the large Ericaceae family and Rhododendron in particular.

Microsatellite marker analysis reveals the complex phylogeographic history of Rhododendron ferrugineum (Ericaceae) in the Pyrenees.

Genetic variation within plant species is determined by a number of factors such as reproductive mode, breeding system, life history traits and climatic events. In alpine regions, plants experience heterogenic abiotic conditions that influence the populations genetic structure. The aim of this study was to investigate the genetic structure and phylogeographic history of the subalpine shrub Rhododendron ferrugineum across the Pyrenees and the links between the populations in the Pyrenees, the Alps and Jura Mountains. We used 27 microsatellite markers to genotype 645 samples from 29 Pyrenean populations, three from the Alps and one from the Jura Mountains. These data were used to estimate population genetics statistics such as allelic richness, observed heterozygosity, expected heterozygosity, fixation index, inbreeding coefficient and number of migrants. Genetic diversity was found to be higher in the Alps than in the Pyrenees suggesting colonization waves from the Alps to the Pyrenees. Two separate genetic lineages were found in both the Alps and Pyrenees, with a substructure of five genetic clusters in the Pyrenees where a loss of genetic diversity was noted. The strong differentiation among clusters is maintained by low gene flow across populations. Moreover, some populations showed higher genetic diversity than others and presented rare alleles that may indicate the presence of alpine refugia. Two lineages of R. ferrugineum have colonized the Pyrenees from the Alps. Then, during glaciation events R. ferrugineum survived in the Pyrenees in different refugia such as lowland refugia at the eastern part of the chain and nunataks at high elevations leading to a clustered genetic pattern.

High lifetime inbreeding depression counteracts the reproductive assurance benefit of selfing in a mass-flowering shrub

BackgroundDecreases in mate and/or pollinator availability would be expected to affect the selective pressure on plant mating systems. An increase in self-fertilization may evolve to compensate for the negative effects of pollination failure. However, the benefit of selfing in variable pollination environments depends on the relative fitnesses of selfed and outcrossed progeny. We investigated the potential for selfing to provide reproductive assurance over the lifetime of a long-lived perennial species and its variation between plant patches of various sizes. Patch size is likely to affect mate and pollinator availabilities, thereby affecting pollination success and the rate of selfing. We estimated fruit and seed set, reproductive assurance, self-compatibility, the multilocus patch selfing rate and lifetime inbreeding depression in natural patches of Rhododendron ferrugineum (Ericaceae), a mass-flowering species characterized by considerable patch size variation (as estimated by the total number of inflorescences).ResultsOpen seed set declined linearly with increasing patch size, whereas pollinator-mediated seed set (emasculated flowers) was not significantly affected. Progeny array analysis indicated that the selfing rate declined with increasing patch size, consistent with greater reproductive assurance in small sparse patches than in large, dense patches. However, fruit set and adult fitness decreased with decreasing patch size, with an estimated mean lifetime inbreeding depression of 0.9 (obtained by comparing F values in adults and progenies).ConclusionsLifetime inbreeding depression strongly counteracts the advantage of reproductive assurance due to selfing in this long-lived species. The poor fitness of selfed offspring should counteract any evolution towards selfing, despite its potential to alleviate the negative consequences of pollen limitation. This study highlights the need to estimate lifetime inbreeding depression, together with mating system and pollination parameters, if we are to understand the actual benefit of selfing and avoid the overestimation of reproductive assurance.

Subalpine Pyrenees received higher nitrogen deposition than predicted by EMEP and CHIMERE chemistry-transport models

Deposition of reactive nitrogen (N) from the atmosphere is expected to be the third greatest driver of biodiversity loss by the year 2100. Chemistry-transport models are essential tools to estimate spatially explicit N deposition but the reliability of their predictions remained to be validated in mountains. We measured N deposition and air concentration over the subalpine Pyrenees. N deposition was found to range from 797 to 1,463 mg N m−2 year−1. These values were higher than expected from model predictions, especially for nitrate, which exceeded the estimations of EMEP by a factor of 2.6 and CHIMERE by 3.6. Our observations also displayed a reversed reduced-to-oxidized ratio in N deposition compared with model predictions. The results highlight that the subalpine Pyrenees are exposed to higher levels of N deposition than expected according to standard predictions and that these levels exceed currently recognized critical loads for most high-elevation habitats. Our study reveals a need to improve the evaluation of N deposition in mountains which are home to a substantial and original part of the world’s biodiversity.

Complex interactions between a legume and two grasses in a subalpine meadow

Interactions between plants are a complex combination of positive and negative interactions, with the net outcome depending on environmental contexts. The more frequent association of Trifolium alpinum (legume) with Festuca eskia than with Nardus stricta (grasses) in many Pyrenean subalpine meadows suggests a differential ability to use nitrogen (N) derived from N(2) fixation. In the field, we investigated the interactions between the legume and grasses and, in the glasshouse, the transfer of (15)N from the legume to the grasses. In one grass-Trifolium mixture, the legume had a strong positive effect on the biomass and N content of the grass as compared to pure grass stands. When both grasses grew together with the legume, only Festuca benefited from the presence of Trifolium but, surprisingly, the benefit decreased with increasing Trifolium abundance. Leaf labeling experiments with (15)N-NH(4)(+) revealed a higher transfer of (15)N from Trifolium to Festuca than to Nardus, suggesting a more direct N pathway between the two species. This more direct pathway could prevent Nardus from benefiting from the legume N in the three-species mixtures. Thus, the positive interactions between N-fixers and nonfixers appear to be largely species-specific and to depend strongly on the species in the plant assemblage.

Changes of Humus Features along with a Successional Gradient of Rhododendron ferrugineum (L.) Populations (Subalpine Level, Northwestern Alps, France)

Abstract The modifications of humus characteristics as a result of the establishment and the increasing cover of Rhododendron ferrugineum L. (Ericaceae) in a subalpine meadow were studied in three sites in the northwestern French Alps corresponding to a west–east transect between wet and dry Alps (“Chaîne de Belledonne,” “Massif du Taillefer,” “Briançonnais”). The physical and chemical parameters of humus and biological parameters were studied during the successive studies. As opposed to other Ericaceae heathlands on siliceous soils (Vaccinium sp., Erica sp., Calluna vulgaris), with an increasing Rhododendron cover, the humus structure changed while some other parameters remain stable or increased. The development of Rhododendron populations on calcareous soils (Briançonnais) with maintenance of a calcareous humus has never been checked before. The modifications of the Rhododendron environment under its canopy led to an improvement in its growth conditions (positive feedback).