Diazotrophic nitrogen fixation through aerial roots occurs in Avicennia marina: implications for adaptation of mangrove plant growth to low-nitrogen tidal flats.

Nitrogen limitation of primary production is common in coastal ecosystems. Mangrove trees maintain high levels of nitrogen fixation around their roots. The interior aerial space of mangrove roots, in which atmospheric gas is supplied through lenticels, could be efficient sites for nitrogen fixation. We measured tidal variations of partial pressure of N2 in root aerenchyma and conducted field experiments using 15 N2 as a tracer to track N2 movement through aerial roots of Avicennia marina. We used the acetylene reduction assay to identify the root parts harboring diazotrophs. The nitrogenase activity and estimated nitrogen fixation through aerenchyma were higher in pneumatophores and absorbing roots than in cable roots. Positive correlations between root nitrogen contents and turnover rates of root nitrogen derived from N2 through aerenchyma suggested that the internal supply of N2 to diazotrophs could be the main source for nitrogen assimilation by A. marina roots. Our results confirmed that N2 is supplied to diazotrophs through aerial roots and that nitrogen fixation occurs in A. marina roots. The aerial root structures, which occur across families of mangrove plants, could be an adaptation to survival in not only low-oxygen environments but also tidal flats with little plant-available nitrogen.

Temperature dependence of O2 respiration in mangrove leaves and roots: implications for seedling dispersal phenology.

Seasonal differences in diaspore dispersal of three mangrove species, Kandelia obovata, Bruguiera gymnorrhiza and Rhizophora stylosa, suggest that respiratory energy production and demand may differ as a result of interspecific differences in temperature dependence of growth and maintenance processes during seedling establishment. We analyzed growth, temperature dependencies of respiratory O2 consumption and amounts of respiratory chain enzymes in seedlings of these species grown at various temperatures. Respiration rates measured at the low reference temperature, RREF , were highest in leaves of 15°C-grown K. obovata, whose dispersal occurs in the cold season, while root RREF of 15°C-grown R. stylosa was 60% those of the other species, possibly because of warm conditions during its establishment phase. In leaves and roots of K. obovata and leaves of R. stylosa, the overall activation energy, Eo , changed with growth temperature associated with changes in the ratios of the amount of protein in the two respiratory pathways. However, Eo of seedlings of B. gymnorrhiza, which has a long dispersal phase, were constant and independent of growth temperature. The different temperature responses of seedling respiration and growth among these three species may reflect the seasonal temperature range of seedling dispersal and establishment in each species.

Distinct responses of growth and respiration to growth temperatures in two mangrove species.

BACKGROUND AND AIMS: Mangrove plants are mostly found in tropical and sub-tropical tidal flats, and their limited distribution may be related to their responses to growth temperatures. However, the mechanisms underlying these responses have not been clarified. Here, we measured the dependencies of the growth parameters and respiration rates of leaves and roots on growth temperatures in typical mangrove species. METHODS: We grew two typical species of Indo-Pacific mangroves, Bruguiera gymnorrhiza and Rhizophora stylosa, at four different temperatures (15, 20, 25 and 30 °C) by irrigating with fresh water containing nutrients, and we measured growth parameters, chemical composition, and leaf and root O2 respiration rates. We then estimated the construction costs of leaves and roots and the respiration rates required for maintenance and growth. KEY RESULTS: The relative growth rates of both species increased with growth temperature due to changes in physiological parameters such as net assimilation rate and respiration rate rather than to changes in structural parameters such as leaf area ratio. Both species required a threshold temperature for growth (12.2 °C in B. gymnorrhiza and 18.1 °C in R. stylosa). At the low growth temperature, root nitrogen uptake rate was lower in R. stylosa than in B. gymnorrhiza, leading to a slower growth rate in R. stylosa. This indicates that R. stylosa is more sensitive than B. gymnorrhiza to low temperature. CONCLUSIONS: Our results suggest that the mangrove species require a certain warm temperature to ensure respiration rates sufficient for maintenance and growth, particularly in roots. The underground temperature probably limits their growth under the low-temperature condition. The lower sensitivity of B. gymnorrhiza to low temperature shows its potential to adapt to a wider habitat temperature range than R. stylosa. These growth and respiratory features may explain the distribution patterns of the two mangrove species.

Mangrove-diazotroph relationships at the root, tree and forest scales: diazotrophic communities create high soil nitrogenase activities in Rhizophora stylosa rhizospheres.

BACKGROUND AND AIMS: The tidal flats on which mangrove plants grow tend to have low soil nitrogen contents because nitrogen-containing litter is repeatedly washed offshore by ebb tides. Under such circumstances, it is unclear how mangrove plants acquire the nitrogen required to support their vigorous growth. In the present work, chemical and biological characteristics of diazotrophy around mangrove plant roots were surveyed under natural conditions to elucidate mangrove-diazotroph relationships. METHODS: Soil nitrogenase activity of a representative mangrove plant, Rhizophora stylosa, which has a broad geographical distribution, was measured using the acetylene reduction assay at forest, tree and prop root scales. In addition, diazotrophic community composition was compared between rhizosphere and bulk soil based on sequencing of nifH genes. KEY RESULTS: Soil nitrogenase activity was high near prop roots, and this pattern was enhanced as soil live root content increased. At the forest scale, we observed high soil nitrogenase activity (acetylene-reducing activity) inside the forest (the highest value was 90.9 µmol C2H2 min-1 cm-3, average 46.8 ± 18.2 µmol C2H2 min-1 cm-3). Rates decreased sharply from the forest to the tidal flat (range 1.2-22.2 µmol C2H2 min-1 cm-3, average 7.9 ± 4.5 µmol C2H2 min-1 cm-3). The nifH operational taxonomic unit composition differed significantly among forest and tree rhizospheres and the bulk soil (P < 0.0001). CONCLUSIONS: Our results suggest that the accumulation of diazotrophs around R. stylosa mangrove trees enhances the supply of biologically fixed nitrogen to the mangrove roots. This supply is especially important when the soil naturally contains little nitrogen. This nitrogen acquisition system may be a key process that explains the high productivity of mangrove ecosystems.

Intraspecific variation in photosynthetic thermal acclimation in Fagus crenata seedlings from two populations growing at different elevations in northern Japan.

Plants can acclimate their photosynthesis to growth temperature, but the contribution of local adaptation to intraspecific variation in thermal acclimation of photosynthesis is not fully understood. Here, we experimentally investigated the photosynthetic thermal acclimation in Fagus crenata seedlings from two populations growing at different elevations and temperature regimes (low- and high-elevation sites) in northern Japan. We acclimated seedlings for 14-23 days at 22 °C (control) or 27 °C (warm treatment) daytime temperature and obtained photosynthetic temperature-response curves in the range of 19 to 32 °C. The optimum temperature of photosynthesis (Topt) was ca. 0.6 °C higher in seedlings acclimated at 27 °C than in those acclimated at 22 °C, and it was significantly lower in seedlings with higher stomatal sensitivity to leaf-to-air vapor pressure deficit (VPD) than in those with lower sensitivity. The effects of warm treatment, population, and treatment-population interaction on Topt were not significant in the two-way analysis of variance, but the effect of treatment became significant when stomatal sensitivity to leaf-to-air VPD was included as a covariate in the model. Structural equation modelling (SEM) indicated that seedlings with lower root biomass had lower Topt because of the high stomatal sensitivity to leaf-to-air VPD. SEM also indicated that the way of shifting the Topt differed between the two populations: seedlings from a high-elevation site depended on decreasing photosynthetic rates at low temperatures for the increase in Topt but seedlings from a low-elevation site did not. We suggest that difference in thermal acclimation of photosynthesis between the two populations may reflect adaptation to different climate regimes and that belowground traits should be considered when investigating thermal acclimation capacity, especially in seedlings.

The Impact of Phenological Gaps on Leaf Characteristics and Foliage Dynamics of an Understory Dwarf Bamboo, Sasa kurilensis.

Phenological gaps exert a significant influence on the growth of dwarf bamboos. However, how dwarf bamboos respond to and exploit these phenological gaps remain enigmatic. The light environment, soil nutrients, leaf morphology, maximum photosynthetic rate, foliage dynamics, and branching characteristics of Sasa kurilensis were examined under the canopies of Fagus crenata and Magnolia obovata. The goal was to elucidate the adaptive responses of S. kurilensis to phenological gaps in the forest understory. The findings suggest that phenological gaps under an M. obovata canopy augment the available biomass of S. kurilensis, enhancing leaf area, leaf thickness, and carbon content per unit area. However, these gaps do not appreciably influence the maximum photosynthetic rate, total leaf number, leaf lifespan, branch number, and average branch length. These findings underscore the significant impact of annually recurring phenological gaps on various aspects of S. kurilensis growth, such as its aboveground biomass, leaf morphology, and leaf biochemical characteristics. It appears that leaf morphology is a pivotal trait in the response of S. kurilensis to phenological gaps. Given the potential ubiquity of the influence of phenological gaps on dwarf bamboos across most deciduous broadleaf forests, this canopy phenomenon should not be overlooked.

Root endophytic bacterial and fungal communities in a natural hot desert are differentially regulated in dry and wet seasons by stochastic processes and functional traits.

Dryland ecosystems experience seasonal cycles of severe drought and moderate precipitation. Desert plants may develop symbiotic relationships with root endophytic microbes to survive under the repeated wet and extremely dry conditions. Although community coalescence has been found in many systems, the colonization by functional microbes and its relationship to seasonal transitions in arid regions are not well understood. Here we examined root endophytic microbial taxa, and their traits in relation to their root colonization, during the dry and wet seasons in a hot desert of the southwestern United States. We used high-throughput DNA sequencing of 16S rRNA and internal transcribed spacer gene profiling of five desert shrubs, and analyzed the seasonal change in endophytic microbial lineages. Goodness of fit to the neutral community model in relationship to microbial traits was evaluated. In summer, Actinobacteria and Bacteroidia increased, although this was not genus-specific. For fungi, Glomeraceae selectively increased in summer. In winter, Gram-negative bacterial genera, including those capable of nitrogen fixation and plant growth promotion, increased. Neutral model analysis revealed a strong stochastic influence on endophytic bacteria but a weak effect for fungi, especially in summer. The taxa with higher frequency than that predicted by neutral model shared environmental adaptability and symbiotic traits, whereas the frequency of pathogenic fungi was at or under the predicted value. These results suggest that community assembly of bacteria and fungi is regulated differently. The bacterial community was affected by stochastic and deterministic processes via bacterial response to drought (response trait), beneficial effect on plants (effect trait), and likely stable mutualistic interactions with plants suggested by the frequency of nodule bacteria. For fungi, mycorrhizal fungi were selected by plants in summer. The regulation of beneficial microbes by plants in both dry and wet seasons suggests the presence of plant-soil positive feedback in this natural desert ecosystem.