An evaluation of inexpensive methods for root image acquisition when using rhizotrons.

BACKGROUND: Belowground processes play an essential role in ecosystem nutrient cycling and the global carbon budget cycle. Quantifying fine root growth is crucial to the understanding of ecosystem structure and function and in predicting how ecosystems respond to climate variability. A better understanding of root system growth is necessary, but choosing the best method of observation is complex, especially in the natural soil environment. Here, we compare five methods of root image acquisition using inexpensive technology that is currently available on the market: flatbed scanner, handheld scanner, manual tracing, a smartphone application scanner and a time-lapse camera. Using the five methods, root elongation rate (RER) was measured for three months, on roots of hybrid walnut (Juglans nigra × Juglans regia L.) in rhizotrons installed in agroforests. RESULTS: When all methods were compared together, there were no significant differences in relative cumulative root length. However, the time-lapse camera and the manual tracing method significantly overestimated the relative mean diameter of roots compared to the three scanning methods. The smartphone scanning application was found to perform best overall when considering image quality and ease of use in the field. The automatic time-lapse camera was useful for measuring RER over several months without any human intervention. CONCLUSION: Our results show that inexpensive scanning and automated methods provide correct measurements of root elongation and length (but not diameter when using the time-lapse camera). These methods are capable of detecting fine roots to a diameter of 0.1 mm and can therefore be selected by the user depending on the data required.

The Impact of Competition and Allelopathy on the Trade-Off between Plant Defense and Growth in Two Contrasting Tree Species.

In contrast to plant-animal interactions, the conceptual framework regarding the impact of secondary metabolites in mediating plant-plant interference is currently less well defined. Here, we address hypotheses about the role of chemically-mediated plant-plant interference (i.e., allelopathy) as a driver of Mediterranean forest dynamics. Growth and defense abilities of a pioneer (Pinus halepensis) and a late-successional (Quercus pubescens) Mediterranean forest species were evaluated under three different plant interference conditions: (i) allelopathy simulated by application of aqueous needle extracts of Pinus, (ii) resource competition created by the physical presence of a neighboring species (Pinus or Quercus), and (iii) a combination of both allelopathy and competition. After 24 months of experimentation in simulated field conditions, Quercus was more affected by plant interference treatments than was Pinus, and a hierarchical response to biotic interference (allelopathy < competition < allelopathy + competition) was observed in terms of relative impact on growth and plant defense. Both species modulated their respective metabolic profiles according to plant interference treatment and thus their inherent chemical defense status, resulting in a physiological trade-off between plant growth and production of defense metabolites. For Quercus, an increase in secondary metabolite production and a decrease in plant growth were observed in all treatments. In contrast, this trade-off in Pinus was only observed in competition and allelopathy + competition treatments. Although Pinus and Quercus expressed differential responses when subjected to a single interference condition, either allelopathy or competition, species responses were similar or positively correlated when strong interference conditions (allelopathy + competition) were imposed.

Experimental assessment of the water quality influence on the phosphorus uptake of an invasive aquatic plant: biological responses throughout its phenological stage.

Understanding how an invasive plant can colonize a large range of environments is still a great challenge in freshwater ecology. For the first time, we assessed the relative importance of four factors on the phosphorus uptake and growth of an invasive macrophyte Elodea nuttallii (Planch.) St. John. This study provided data on its phenotypic plasticity, which is frequently suggested as an important mechanism but remains poorly investigated. The phosphorus uptake of two Elodea nuttallii subpopulations was experimentally studied under contrasting environmental conditions. Plants were sampled in the Rhine floodplain and in the Northern Vosges mountains, and then maintained in aquaria in hard (Rhine) or soft (Vosges) water. Under these conditions, we tested the influence of two trophic states (eutrophic state, 100 µg x l(-1) P-PO4(3-) and hypertrophic state, 300 µg x l(-1) P-PO4(3-)) on the P metabolism of plant subpopulations collected at three seasons (winter, spring and summer). Elodea nuttallii was able to absorb high levels of phosphorus through its shoots and enhance its phosphorus uptake, continually, after an increase of the resource availability (hypertrophic > eutrophic). The lowest efficiency in nutrient use was observed in winter, whereas the highest was recorded in spring, what revealed thus a storage strategy which can be beneficial to new shoots. This experiment provided evidence that generally, the water trophic state is the main factor governing P uptake, and the mineral status (softwater > hardwater) of the stream water is the second main factor. The phenological stage appeared to be a confounding factor to P level in water. Nonetheless, phenology played a role in P turnover in the plant. Finally, phenotypic plasticity allows both subpopulations to adapt to a changing environment.

Selection of wild macrophytes for use in constructed wetlands for phytoremediation of contaminant mixtures.

Constructed wetlands (CWs) offer an alternative to traditional industrial wastewater treatment systems that has been proved to be efficient, cost-effective and environmentally friendly. Most of the time, CWs are planted with proliferative species such as Phragmites australis or with plants originating from nurseries, both representing a risk for the natural biodiversity conservation of aquatic ecosystems located downstream of the CWs. For the removal of metals and organic pollutant mixtures present in industrial effluents, it is necessary to select tolerant plant species that are able to produce a high aboveground biomass and to develop a healthy belowground system. Wild plant species growing in aquatic bodies at industrial outfalls could constitute suitable tolerant species to use in CWs for industrial effluent treatment. To test this hypothesis, we assessed, under laboratory conditions (using an experimental design), the tolerance to mixtures of metals (Al, As, Cd, Cu, Cr, Fe, Mn, Ni, Pb, Sn, Zn) or/and organic pollutants (THC, PHE, PYR, LAS) of five European sub-cosmopolitan native macrophytes (Alisma lanceolatum, Carex cuprina, Epilobium hirsutum, Iris pseudacorus and Juncus inflexus) that had been collected in a polluted Mediterranean wetland, after a field study (crossing ecological relevés and analyses of contaminant concentrations in water and sediments). Our results demonstrated that research on phytoremediation of industrial effluents should focus much more on the use of native macrophytes growing at short distances from industrial discharges (such as C. cuprina in this study), and that root/shoot ratio, aerial height and proportion of green leaves are good and cost-effective indicators of plant tolerance to metals and organic pollutant mixtures in laboratory studies.

Allelochemicals of Pinus halepensis as drivers of biodiversity in Mediterranean open mosaic habitats during the colonization stage of secondary succession.

The Mediterranean region is recognized as a global biodiversity hotspot. However, over the last 50 years or so, the cessation of traditional farming has given way to strong afforestation at the expense of open habitats. Pinus halepensis Miller, known to synthesize a wide range of secondary metabolites, is a pioneer expansionist species colonizing abandoned agricultural land that present high species richness. Here, laboratory bioassays were used to study the potential impact of P. halepensis on plant diversity through allelopathy, and the role of microorganisms in these interactions. Germination and growth of 12 target species naturally present in fallow farmlands were tested according to concentration of aqueous extracts obtained from shoots of young pines (aged about 5 years), with or without the presence of soil microorganisms (autoclaved or natural soil). Under the highest concentrations and autoclaved soil, more than 80 % of target species were germination and/or growth-inhibited, and only two species were non-sensitive. Under more natural conditions (lower extracts concentrations and natural soil with microorganisms), only 50 % of species were still inhibited, one was non-sensitive, and five were stimulated. Thus, microorganisms alter the expression of allelochemicals released into the ecosystem, which highlights their key role in chemical plant-plant interactions. The results of allelopathic experiments conducted in the lab are consistent with the community patterns observed in the field. These findings suggest that allelopathy is likely to shape vegetation composition and participate to the control of biodiversity in Mediterranean open mosaic habitats.

Variations in allelochemical composition of leachates of different organs and maturity stages of Pinus halepensis.

We investigated changes in the occurrence of allelochemicals from leachates of different Pinus halepensis organs taking into account the stages of pine stand age (i.e., young < 15-years-old, middle age +/- 30-years, and old > 60-years-old). GC-MS analysis of aqueous extracts revealed approx. 59 components from needles and roots. The major constituents were divided into different phytochemical groups-phenolics (50%), fatty acids (44%), and terpenoids. Further analyses were carried out to characterize the distribution of allelochemicals in different organs and P. halepensis successional stages. Roots and needles had two distinct chemical profiles, while needle leachates were composed mainly of oxygenated terpenoids (e.g., alpha-eudesmol, alpha-cadinol, and alpha-terpineol). Roots mainly contained fatty acids. Needles from young pine stands had the highest content of monoterpenes, suggesting their role as potential allelochemicals that could help young pine stands to establish. Pooling the different functional chemical groups showed that needles and, to a lesser extent, old roots, had higher chemical diversity than the roots of young and medium-aged pines. The highest diversity in phenolic constituents and fatty acids was in young needles (D(chem) = 2.38). Finally, caffeic acid, a compound that has allelopathic properties was found in aqueous extracts at high concentrations in both young needles and old roots. The role of this compound in mediation of biological interactions in P. halepensis ecosystem functioning is discussed.