José Ignacio Querejeta

Topographic position modulates the mycorrhizal response of oak trees to interannual rainfall variability

California coast live oak (Quercus agrifolia) forms tripartite symbiotic associations with arbuscular (AMF) and ectomycorrhizal (EMF) fungi. We selected oak individuals differing in topographic position and depth to groundwater (mesic valley vs. xeric hill sites) to investigate changes of tree mycorrhizal status in response to interannual rainfall variability. EMF root colonization, as well as hyphal abundance and viability in upper rhizosphere soil (0-30 cm), were negatively affected by severe multi-year drought, although not to the same extent in each topographic location. Oak trees growing in hill sites showed EMF colonization levels <1% in upper roots during drought. By contrast, oaks in valley sites maintained much higher EMF colonization (>19%) in upper roots during drought. EMF root colonization increased sharply at both topographic positions during the ensuing wet year (78% in valley, 49% in hill), which indicates that the mycorrhizal status of roots in upper rhizosphere soil is highly responsive to interannual rainfall variability. Across sites and years, percentage EMF colonization and soil hyphal density and viability were strongly positively correlated with soil moisture potential, but percentage AMF root colonization was not. Interestingly, changes in percentage EMF root colonization and density of viable hyphae between a wet and a dry year were proportionally much greater in xeric hill sites than in mesic valley sites. The mycorrhizal status of oak trees was particularly responsive to changes in soil moisture at the hill sites, where roots in upper rhizosphere soil shifted from almost exclusively AMF during severe drought to predominantly EMF during the ensuing wet year. By contrast, the mycorrhizal status of oaks in the valley sites was less strongly coupled to current meteorological conditions, as roots in upper soil layers remained predominantly EMF during both a dry and a wet year. Canopy shading and hydraulic lift by oaks in valley sites likely contributed to maintain the integrity and viability of EMF roots and extraradical hyphae in upper rhizosphere soil during extended drought. Our results suggest that oak woodlands in water-limited ecosystems may become increasingly reliant on the AMF symbiosis under future climate change scenarios for the U.S. southwest and other world regions.

Soil water availability improved by site preparation in a Pinus halepensis afforestation under semiarid climate

This work was funded by the Spanish Comision Interministerial de Ciencia y Tecnologia (CICYT), Projects FOR-91-0352, AGF-95-0097 and AGF-99-1132.

Differential response of δ13C and water use efficiency to arbuscular mycorrhizal infection in two aridland woody plant species

During a revegetation field experiment in Southeast Spain, we measured foliar carbon isotope ratios (δ13C) and gas exchange parameters in order to evaluate the influence of arbuscular mycorrhizal (AM) infection on the water use efficiency (WUE) of two semiarid woodland species. WUE during drought was significantly enhanced by inoculation with Glomus intraradices in Olea europaea ssp sylvestris, but not in Rhamnus lycioides. While Olea is a long-lived, slow-growing evergreen tree with a conservative water use strategy, Rhamnus is a drought-deciduous shrub with a shorter lifespan; these differences may explain their dissimilar patterns of physiological response to inoculation with the same AM fungus. Differences in δ13C and WUE between Olea and Rhamnus were larger when comparing AM inoculated than non-inoculated seedlings. This result suggests that some of the interspecific variability in δ13C observed for aridland plant communities may be due to different physiological responses to mycorrhization.

Water transfer via ectomycorrhizal fungal hyphae to conifer seedlings

Little is known about water transfer via mycorrhizal hyphae to plants, despite its potential importance in seedling establishment and plant community development, especially in arid environments. Therefore, this process was investigated in the study reported in this paper in laboratory-based tripartite mesocosms containing the shrub Arctostaphylos viscida (manzanita) and young seedlings of sugar pine (Pinus lambertiana) and Douglas-fir (Pseudotsuga menziesii). The objectives were to determine whether water could be transported through mycorrhizal symbionts shared by establishing conifers and A. viscida and to compare the results obtained using two tracers: the stable isotope deuterium and the dye lucifer yellow carbohydrazide. Water containing the tracers was added to the central compartment containing single manzanita shrubs. The fungal hyphae were then collected as well as plant roots from coniferous seedlings in the other two compartments to determine whether water was transferred via fungal hyphae. In addition, the length of the hyphae and degree of mycorrhizal colonisation were determined. Internal transcribed spacer–restriction fragment length polymorphism (ITS-RFLP) analysis was used to identify the fungal species involved in dye (water) transfer. Results of the stable isotope analysis showed that water is transferred via mycorrhizal hyphae, but isotopically labelled water was only detected in Douglas-fir roots, not in sugar pine roots. In contrast, the fluorescent dye was transported via mycorrhizal hyphae to both Douglas-fir and sugar pine seedlings. Only 1 of 15 fungal morphotypes (identified as Atheliaceae) growing in the mesocosms transferred the dye. Differences were detected in the water transfer patterns indicated by the deuterium and fluorescent dye tracers, suggesting that the two labels are transported by different mechanisms in the same hyphae and/or that different fungal taxa transfer them via different routes to host plants. We conclude that both tracers can provide information on resource transfer between fungi and plants, but we cannot be sure that the dye transfer data provide accurate indications of water transfer rates and patterns. The isotopic tracer provides more direct indications of water movement and is therefore more suitable than the dye for studying water relations of plants and their associated mycorrhizal fungi.

Stand structure modulates the long‐term vulnerability of Pinus halepensis to climatic drought in a semiarid Mediterranean ecosystem

We investigated whether stand structure modulates the long-term physiological performance and growth of Pinus halepensis Mill. in a semiarid Mediterranean ecosystem. Tree radial growth and carbon and oxygen stable isotope composition of latewood (δ(13)C(LW) and δ(18)O(LW), respectively) from 1967 to 2007 were measured in P. halepensis trees from two sharply contrasting stand types: open woodlands with widely scattered trees versus dense afforested stands. In both stand types, tree radial growth, δ(13)C(LW) and δ(18)O(LW) were strongly correlated with annual rainfall, thus indicating that tree performance in this semiarid environment is largely determined by inter-annual changes in water availability. However, trees in dense afforested stands showed consistently higher δ(18)O(LW) and similar δ(13)C(LW) values compared with those in neighbouring open woodlands, indicating lower stomatal conductance and photosynthesis rates in the former, but little difference in water use efficiency between stand types. Trees in dense afforested stands were more water stressed and showed lower radial growth, overall suggesting greater vulnerability to drought and climate aridification compared with trees in open woodlands. In this semiarid ecosystem, the negative impacts of intense inter-tree competition for water on P. halepensis performance clearly outweigh potential benefits derived from enhanced infiltration and reduced run-off losses in dense afforested stands.

Leaf δ18O of remaining trees is affected by thinning intensity in a semiarid pine forest

Silvicultural thinning usually improves the water status of remaining trees in water-limited forests. We evaluated the usefulness of a dual stable isotope approach (δ¹³C, δ¹⁸O) for comparing the physiological performance of remaining trees between forest stands subjected to two different thinning intensities (moderate versus heavy) in a 60-year-old Pinus halepensis Mill. plantation in semiarid southeastern Spain. We measured bulk leaf δ¹³C and δ¹⁸O, foliar elemental concentrations, stem water content, stem water δ¹⁸O (δ¹⁸O(stem water)), tree ring widths and leaf gas exchange rates to assess the influence of forest stand density on tree performance. Remaining trees in low-density stands (heavily thinned) showed lower leaf δ¹⁸O, and higher stomatal conductance (g(s)), photosynthetic rate and radial growth than those in moderate-density stands (moderately thinned). By contrast, leaf δ¹³C, intrinsic water-use efficiency, foliar elemental concentrations and δ¹⁸O(stem water) were unaffected by stand density. Lower foliar δ¹⁸O in heavily thinned stands reflected higher g(s) of remaining trees due to decreased inter-tree competition for water, whereas higher photosynthetic rate was largely attributable to reduced stomatal limitation to CO₂ uptake. The dual isotope approach provided insight into the early (12 months) effects of stand density manipulation on the physiological performance of remaining trees.

Efflux of hydraulically lifted water from mycorrhizal fungal hyphae during imposed drought

Apart from improving plant and soil water status during drought, it has been suggested that hydraulic lift (HL) could enhance plant nutrient capture through the flow of mineral nutrients directly from the soil to plant roots, or by maintaining the functioning of mycorrhizal fungi. We evaluated the extent to which the diel cycle of water availability created by HL covaries with the efflux of HL water from the tips of extramatrical (external) mycorrhizal hyphae, and the possible effects on biogeochemical processes. Phenotypic mycorrhizal fungal variables, such as total and live hyphal lengths, were positively correlated with HL efflux from hyphae, soil water potential (dawn), and plant response variables (foliar 15N). The efflux of HL water from hyphae was also correlated with bacterial abundance and soil enzyme activity (P), and the moistening of soil organic matter. Such findings indicate that the efflux of HL water from the external mycorrhizal mycelia may be a complementary explanation for plant nutrient acquisition and survival during drought.

Bulk leaf δ18O and δ13C reflect the intensity of intraspecific competition for water in a semi-arid tussock grassland

We investigated the extent to which plant water and nutrient status are affected by intraspecific competition intensity and microsite quality in a monodominant tussock grassland. Leaf gas exchange and stable isotope measurements were used to assess the water relations of Stipa tenacissima tussocks growing along a gradient of plant cover and soil depth in a semi-arid catchment of Southeast Spain. Stomatal conductance and photosynthetic rate decreased with increasing intensity of competition during the wet growing season, leading to foliar delta(18)O and delta(13)C enrichment. A high potential for runoff interception by upslope neighbours exerted strong detrimental effects on the water and phosphorus status of downslope S. tenacissima tussocks. Foliar delta(15)N values became more enriched with increasing soil depth. Multiple stepwise regression showed that competition potential and/or rhizosphere soil depth accounted for large proportions of variance in foliar delta(13)C, delta(18)O and delta(15)N among target tussocks (57, 37 and 64%, respectively). The results presented here highlight the key role that spatial redistribution of resources (water and nutrients) by runoff plays in semi-arid ecosystems. It is concluded that combined measurement of delta(13)C, delta(18)O and nutrient concentrations in bulk leaf tissue can provide insight into the intensity of competitive interactions occurring in natural plant communities.

Plant isotopic composition provides insight into mechanisms underlying growth stimulation by AM fungi in a semiarid environment

We hypothesised that improved plant water status and enhanced transpiration are key mechanisms involved in plant growth stimulation by native arbuscular mycorrhizal fungi (AMF) in semiarid calcareous soils. Seedlings of the dryland shrubs Pistacia lentiscus L. and Retama sphaerocarpa L. were pre-inoculated with a mixture of eight native Glomus spp. fungi, or left un-inoculated, before transplanting into a degraded site in south-eastern Spain. Pre-inoculated Pistacia and Retama shrubs grew faster after transplanting, despite spontaneous colonisation of control plants by local AMF. Pre-inoculation enhanced shoot water content and shoot δ15N in both shrub species. Increased potassium uptake and improved water relations were key mechanisms behind growth stimulation by native AMF in Pistacia. Shoot δ18O (a proxy measure of stomatal conductance) was significantly less negative in AMF-inoculated than in control Pistacia seedlings, indicating enhanced cumulative transpiration in the former. In contrast, shoot δ18O was unaffected by AMF inoculation in Retama, a leafless leguminous shrub with photosynthetic stems. Growth stimulation by native AMF in Retama was attributed to increased phosphorus uptake, enhanced atmospheric nitrogen fixation and a largely nutrient-mediated improvement of plant water status. Shoot δ13C was not significantly influenced by AMF inoculation in either shrub species, thus suggesting roughly parallel upshifts in photosynthetic and transpiration rates which did not affect plant water use efficiency.

Changes in soil hyphal abundance and viability can alter the patterns of hydraulic redistribution by plant roots

Background and aimsWe conducted a mesocosm study to investigate the extent to which the process of hydraulic redistribution of soil water by plant roots is affected by mycorrhizosphere disturbance.MethodsWe used deuterium-labeled water to track the transfer of hydraulically lifted water (HLW) from well-hydrated donor oaks (Quercus agrifolia Nee.) to drought-stressed receiver seedlings growing together in mycorrhizal or fungicide-treated mesocosms. We hypothesized that the transfer of HLW from donor to receiver plants would be enhanced in undisturbed (non-fungicide-treated) mesocosms where an intact mycorrhizal hyphal network was present.ResultsContrary to expectations, both upper soil and receiver seedlings contained significantly greater proportions of HLW in mesocosms where the abundance of mycorrhizal hyphal links between donor and receiver roots had been sharply reduced by fungicide application. Reduced soil hyphal density and viability likely hampered soil moisture retention properties in fungicide-treated mesocosms, thus leading to faster soil water depletion in upper compartments. The resulting steeper soil water potential gradient between taproot and upper compartments enhanced hydraulic redistribution in fungicide-treated mesocosms.ConclusionsBelowground disturbances that reduce soil hyphal density and viability in the mycorrhizosphere can alter the patterns of hydraulic redistribution by roots through effects on soil hydraulic properties.