Nutrient deficiency promotes male-biased apparent sex ratios at the ramet level in the dioecious plant Myrica gale var. tomentosa in oligotrophic environments in bogs.

In populations of dioecious plants, the differences in the cost of reproduction between male and female plants can promote a male-biased sex ratio. In this study, we examine the macronutrient levels in tissues of the dioecious wetland shrub Myrica gale to identify the cost of reproduction for male and female plants and to examine the effect of nutrients on the apparent sex ratio at the ramet level. We examined plants across 12 populations of M. gale inhabiting bogs and fens in Japan. For each population, we used line transects to estimate the apparent sex ratio and measured the concentrations of nitrogen (N), phosphorus (P), and potassium (K) in the leaves sampled from male and female plants and in the fruits from female plants. For five of the populations, we calculated the flowering frequency, mortality, and the recruitment rate (as the rate of clonal propagation). We found that the proportion of females was positively affected, and the male bias of sex ratios reduced, by increases in P concentration in leaves sampled from female plants. Neither mortality nor recruitment was affected by sex or by the nutrient concentration (P, K). The flowering frequency was not affected by sex or by K concentration, but decreased with decreases in the P concentration measured in leaves. This study confirmed that reproduction in M. gale is P-limited. We found no distinct differences in the flowering frequency, mortality, or recruitment rate between the male and female plants.

A Nitrogen-Saturated Plantation of and in Japan Is a Large Nonpoint Nitrogen Source.

Japanese cedar () and Japanese cypress () plantations account for approximately 30% of the total forested area in Japan. Both are arbuscular mycorrhizal trees that leach more NO in response to nitrogen (N) deposition than do forests of ectomycorrhizal trees. However, little information is available about the size of N exports from these plantations. The aim of this study was to evaluate nonpoint source N exports from a N-saturated plantation. We collected stream water samples in base-flow (25 samples) and storm-flow conditions (20 events) in a watershed (2.98 ha) where Japanese cypress and Japanese cedar were planted in 1969 (41 yr old). The annual NO export was calculated from load-discharge relationships. Atmospheric N deposition was also determined. The stream water contained high NO concentrations (160 and 165 µmol L during base flow and storm flow, respectively), indicating N saturation in the watershed. High bulk atmospheric N deposition (16.5 kg N ha yr) could explain the N saturation. There were only small variations in NO concentrations in stream water in response to discharge volume, because of the N saturation of the forest ecosystem. Consequently, there were only small errors in estimating annual NO exports from the studied watershed. The annual NO export was high (36.1 kg N ha yr), comparable to values reported for agricultural and urbanized areas. These results suggest that N-saturated forest plantations can become important nonpoint N sources. Our results also suggest that N exports from forest plantations across Japan should be quantified to evaluate nonpoint source N accurately.

Impact of soil water chemistry on the apparent sex ratio of the flowering ramets of the dioecious plant Myrica gale var. tomentosa.

We determined whether the apparent (M/Fl) sex ratio (male ramets/flowering ramets) and apparent reproductive ramet ratio (Fl/Li ratio; flowering ramets/living ramets) in 15 Myrica gale var. tomentosa populations varied with dissolved total nitrogen, dissolved total phosphorus, potassium, magnesium, calcium, or pH in the soil water. Our aim was to define the environmental factors affecting the M/Fl sex ratio and Fl/Li ratio of the populations. We also examined the habitat conditions of these populations by analyzing soil water chemistry and water dynamics. In 2007, 3 of the 15 populations had no females. The remaining 12 had significantly male-biased (M/Fl sex ratio = 0.59-0.97). Although we could not explain the absence of females by the current potassium levels alone, as potassium increased, so did the M/Fl sex ratio. As nitrogen increased and potassium decreased, Fl/Li ratio decreased. Our soil water chemistry analyses suggested that the potassium supply by soil surface erosion from flooding and the inflow of anthropogenic nitrogen were the important factors influencing the M/Fl sex ratio and Fl/Li ratio. Nitrogen management would be important in one of the endangered populations where inflow of nitrogen was the highest among 15 habitats.

Determining storm sampling requirements for improving precision of annual load estimates of nutrients from a small forested watershed.

This study sought to determine the lowest number of storm events required for adequate estimation of annual nutrient loads from a forested watershed using the regression equation between cumulative load (∑L) and cumulative stream discharge (∑Q). Hydrological surveys were conducted for 4 years, and stream water was sampled sequentially at 15-60-min intervals during 24 h in 20 events, as well as weekly in a small forested watershed. The bootstrap sampling technique was used to determine the regression (∑L-∑Q) equations of dissolved nitrogen (DN) and phosphorus (DP), particulate nitrogen (PN) and phosphorus (PP), dissolved inorganic nitrogen (DIN), and suspended solid (SS) for each dataset of ∑L and ∑Q. For dissolved nutrients (DN, DP, DIN), the coefficient of variance (CV) in 100 replicates of 4-year average annual load estimates was below 20% with datasets composed of five storm events. For particulate nutrients (PN, PP, SS), the CV exceeded 20%, even with datasets composed of more than ten storm events. The differences in the number of storm events required for precise load estimates between dissolved and particulate nutrients were attributed to the goodness of fit of the ∑L-∑Q equations. Bootstrap simulation based on flow-stratified sampling resulted in fewer storm events than the simulation based on random sampling and showed that only three storm events were required to give a CV below 20% for dissolved nutrients. These results indicate that a sampling design considering discharge levels reduces the frequency of laborious chemical analyses of water samples required throughout the year.

Effects of sex and soil water chemistry on leaf morphology and physiology of Myrica gale var. tomentosa.

Plants respond to environmental stressors, such as an oligotrophic environments, by altering the morphological and physiological functions of their leaves. Sex affects these functions because of the asymmetric cost of reproduction in dioecious plants. We compared the leaf mass per leaf area (LMA), ratio of intercellular air space in leaf mesophyll tissue (mesophyll porosity), palisade thickness, and carbon isotope ratio (δ13C) of leaves of the dioecious shrub Myrica gale based on sex and gradients of soil water chemistry across habitats in the field. The PCA showed that the first three principal components accounted for 84.5% of the variation. PC1 to PC3 were associated with the origin of soil water, nitrogen status of habitats, and sea-salt contributions, respectively. LMA varied from 5.22 to 7.13 µg/cm2, and it was positively related to PC2 and negatively related to PC3, but not to PC1 or sex, suggesting that LMA was low under poor nitrogen conditions and varied with salinity. Mesophyll porosity values were over 50% for all habitats. Mesophyll porosity was positively affected by PC3 and smaller in females than in males. This suggests that M. gale exhibits differences in mesophyll anatomy according to sex. Palisade thickness ranged from 0.466 to 0.559 mm/mm. The leaves of females had thinner palisade layers per mesophyll layer than those of males; however, the habitat did not affect the thickness of the palisade layer per mesophyll layer. The δ13C values of leaves varied from -32.14 to -30.51 ‰. We found that δ13C values were positively related to PC2 but not to PC1, PC3, and sex. Under poor nitrogen conditions, the δ13C of M. gale leaves decreased, suggesting that nutrient deficiency would decrease more under the long-term averaged ratio of photosynthesis than stomatal conductance, leading to low water use efficiency.

Long-term changes in atmospheric nitrogen deposition and stream water nitrate leaching from forested watersheds in western Japan.

Japan receives nitrogenous air pollutants via long-range transport from China. However, emissions of nitrogenous air pollutants in China have stabilized or decreased in recent years. This study examined both the long-term trends in atmospheric nitrogen (N) deposition from the 1990s to the 2010s and the response of stream water nitrate (NO3-) leaching from forested areas in western Japan. A long-term (1992-2018) temporal analysis of atmospheric N deposition in Fukuoka (western Japan) was conducted. Atmospheric bulk N deposition was collected at forested sites in a suburban forest (Swest) and a rural forest (Rwest) in western Japan during 2009-2018. Stream water samples were also collected from four locations at sites Swest and Rwest during the same period. Results showed that atmospheric N deposition in Fukuoka started to decrease from the mid-2000s at an annual rate of -2.5% yr-1. The decrease in atmospheric N deposition was attributable mainly to decreased atmospheric ammonium (NH4+) deposition, which caused greater contribution of NO3- deposition to atmospheric N deposition. Concentrations of NO3- in the stream water samples from three of the four locations decreased significantly at an annual rate of -3.7 to -0.7% yr-1. However, stream water NO3- concentrations increased in one watershed where understory vegetation has been deteriorating owing to the increased deer population. This might weaken the recovery of N leaching from forested areas.

Low nitrogen retention in a Japanese cedar plantation in a suburban area, western Japan.

This study aimed to evaluate nitrogen (N) leaching from Japanese cedar, the main plantation species in Japan, in response to elevated atmospheric N deposition. N leaching and possible factors, including soil nitrification, tree N uptake, and topographic steepness, were evaluated in mature (64-69 year) Japanese cedar trees planted on steep slopes (25°-40°) and neighboring Japanese oak plantations in suburban forests, which served as reference sites. N fertilization (50 kg N ha-1 year-1 as ammonium nitrate) was conducted to evaluate the response of N leaching to an elevated inorganic N pool in the surface soil. The soil water nitrate (NO3-) concentration below the rooting zone in the Japanese cedar forest (607 ± 59 µmol L-1) was much higher than that in the Japanese oak plantations (8.7 ± 8.1 µmol L-1) and increased immediately after fertilization, indicating high N leaching from the Japanese cedar plantations. The relatively low N uptake by Japanese cedar planted on the steep slopes could be an important contributor to the high N leaching. This study highlights the importance of vegetation composition for managing the water quality in headwater streams from forest ecosystems disturbed by atmospheric N deposition.

Comparison of spatial and temporal changes in riverine nitrate concentration from terrestrial basins to the sea between the 1980s and the 2000s in Japan: Impact of recent demographic shifts.

Nitrogen (N) is an essential nutrient but may become a pollution source in the environment when the N concentration exceeds a certain threshold for humans and nature. Nitrate is a major N species in river water with notable spatial and temporal variations under the influences of natural factors and anthropogenic N inputs. We analyzed the relationship between riverine N (focusing on nitrate) concentration and various factors (land use, climate, basin topography, atmospheric N deposition, agricultural N sources and human-derived N) in 104 rivers located throughout the Japanese Archipelago except small remote islands. We aimed to better understand processes and mechanisms to explain the spatial and temporal changes in riverine nitrate concentration. A publicly available river water quality database observed in the 1980s (1980-1989) and 2000s (2000-2009) was used. This study is the first to evaluate the long-term scale of 20 years in the latter half of Japan's economic growth period at the national level. A geographic information system (GIS) was employed to determine average values of each variable collected from multiple sources of statistical data. We then performed regression analysis and structural equation modeling (SEM) for each period. The forestland area influenced by the basin topography, climate (i.e., air temperature) and other land uses (i.e., farmland and urban area) played a major role in decreasing nitrate concentrations in both the 1980s and 2000s. Atmospheric N deposition (especially N oxides) and agricultural N sources (fertilizer and manure) were also significant variables regarding the spatial variations in riverine nitrate concentrations. The SEM results suggested that human-derived N (via food consumption) intensified by demographic shifts during the 2000s increased riverine nitrate concentrations over other variables within the context of spatial variation. These findings facilitate better decision making regarding land use, agricultural practices, pollution control and individual behaviors toward a sustainable society.

Nutrients exported from upland stream water enlarge perennial biomass crops.

Rawanbuki, a variety of Japanese butterbur (Petasites japonicus subsp. giganteus), grow naturally along the Rawan River, Hokkaido, northern Japan. Most plants reach 2-3 m in height and 10 cm in diameter in 2 months and are much larger than those grown along other rivers. We examined the hypothesis that nutrients exported from upland streams enhance the growth of the Rawanbuki. Nutrient concentrations, including nitrogen, phosphorus, and base cations, in the Rawan River were much higher than those in rivers of adjacent watersheds. High nutrient concentrations and moisture contents were found in soil along the Rawan River and a significant relationship was found between physicochemical soil conditions and aboveground biomass of butterburs. This indicates that extremely large Rawanbuki plants could be caused by these high nutrient concentrations and moisture contents in the soils. A manipulation experiment showed that fertilization simulated the growth environment along the Rawan River and enhanced the stem height and stem diameter of butterburs. This study concluded that the extremely large butterburs are caused by a large amount of nutrients exported from upland areas. These results are the first demonstration of the role of stream water nutrients in enlarging agricultural crops.

Organic acids and aldehydes in throughfall and dew in a Japanese pine forest.

We analyzed low molecular weight organic acids and aldehydes in throughfall under pine forest, and organic acids in dew on chemically inert surfaces and pine needle surfaces at urban- and mountain-facing sites of pine forest in western Japan. Low molecular weight organic acids and aldehydes accounted for less than 5% of the dissolved organic carbon in throughfall at both sites. Formaldehyde at both sites and formate at the mountain-facing site were found at significantly lower concentrations in throughfall than in rainfall, which may be explained by the degradation and/or retention of these components by the pine canopy as the incident precipitation passed through it. The oxalate concentration in throughfall was significantly higher than those in rainfall at both sites, suggesting that oxalate was derived from leaching from the pine foliage. At both sites, organic acid concentrations were higher in dew on the pine needles than in throughfall or dew on chemically inert surfaces. This could be due to the long contact time of dew on pine needles, during which leached substances from pine needles and dry deposits accumulated on their surfaces can dissolve into the small volume of dew. The role of enhanced concentrations of oxalate in an aqueous phase on the plant surfaces (e.g., dew) is discussed in relation to hydroxyl radical formation via the photo-Fenton reaction.

Long-term interactive effects of N addition with P and K availability on N status of Sphagnum.

Little information exists concerning the long-term interactive effect of nitrogen (N) addition with phosphorus (P) and potassium (K) on Sphagnum N status. This study was conducted as part of a long-term N manipulation on Whim bog in south Scotland to evaluate the long-term alleviation effects of phosphorus (P) and potassium (K) on N saturation of Sphagnum (S. capillifolium). On this ombrotrophic peatland, where ambient deposition was 8 kg N ha-1 yr-1, 56 kg N ha-1 yr-1 of either ammonium (NH4+, Nred) or nitrate (NO3-, Nox) with and without P and K, were added over 11 years. Nutrient concentrations of Sphagnum stem and capitulum, and pore water quality of the Sphagnum layer were assessed. The N-saturated Sphagnum caused by long-term (11 years) and high doses (56 kg N ha-1 yr-1) of reduced N was not completely ameliorated by P and K addition; N concentrations in Sphagnum capitula for Nred 56 PK were comparable with those for Nred 56, although N concentrations in Sphagnum stems for Nred 56 PK were lower than those for Nred 56. While dissolved inorganic nitrogen (DIN) concentrations in pore water for Nred 56 PK were not different from Nred 56, they were lower for Nox 56 PK than for Nox 56 whose stage of N saturation had not advanced compared to Nred 56. These results indicate that increasing P and K availability has only a limited amelioration effect on the N assimilation of Sphagnum at an advanced stage of N saturation. This study concluded that over the long-term P and K additions will not offset the N saturation of Sphagnum.

Sphagnum can 'filter' N deposition, but effects on the plant and pore water depend on the N form.

The ability of Sphagnum moss to efficiently intercept atmospheric nitrogen (N) has been assumed to be vulnerable to increased N deposition. However, the proposed critical load (20kgNha(-1)yr(-1)) to exceed the capacity of the Sphagnum N filter has not been confirmed. A long-term (11years) and realistic N manipulation on Whim bog was used to study the N filter function of Sphagnum (Sphagnum capillifolium) in response to increased wet N deposition. On this ombrotrophic peatland where ambient deposition was 8kgNha(-1)yr(-1), an additional 8, 24, and 56kgNha(-1)yr(-1) of either ammonium (NH4(+)) or nitrate (NO3(-)) has been applied for 11years. Nutrient status of Sphagnum and pore water quality from the Sphagnum layer were assessed. The N filter function of Sphagnum was still active up to 32kgNha(-1)yr(-1) even after 11years. N saturation of Sphagnum and subsequent increases in dissolved inorganic N (DIN) concentration in pore water occurred only for 56kgNha(-1)yr(-1) of NH4(+) addition. These results indicate that the Sphagnum N filter is more resilient to wet N deposition than previously inferred. However, functionality will be more compromised when NH4(+) dominates wet deposition for high inputs (56kgNha(-1)yr(-1)). The N filter function in response to NO3(-) uptake increased the concentration of dissolved organic N (DON) and associated organic anions in pore water. NH4(+) uptake increased the concentration of base cations and hydrogen ions in pore water though ion exchange. The resilience of the Sphagnum N filter can explain the reported small magnitude of species change in the Whim bog ecosystem exposed to wet N deposition. However, changes in the leaching substances, arising from the assimilation of NO3(-) and NH4(+), may lead to species change.

Sources of hydroxyl radical in headwater streams from nitrogen-saturated forest.

Hydroxyl radical (HO) photoformation rate (RHO) was determined in headwater stream samples from nitrogen (N)-saturated forests, (1) to quantify the sources of HO in headwater streams and (2) to evaluate the nitrate NO3(-)-induced enhancement of HO formation in stream water caused by N saturation in forested watersheds. Stream water fulvic acid extracted from the forested watersheds was used to quantify the contribution of dissolved organic matter (DOM) to RHO. The results showed that almost all (97%; 81-109%) RHO sources in our headwater stream samples were quantitatively elucidated; the photolysis of NO3(-) (55%; 34-75%), nitrite [N(III)] (2%; 0.5-5.2%), and DOM-derived HO formation, from which photo-Fenton reactions (18%; 12-26%) and the direct photolysis of fluorescent dissolved organic matter (FDOM) (22%; 10-40%), was successfully separated. FDOM, which accounted for 53% (24-96%) of DOM in total organic carbon bases, was responsible for HO formation in our headwater streams. High NO3(-) leaching caused by N saturation in forested watersheds increased RHO in the headwaters, indicating that N-saturated forest could significantly change photoinduced and biogeochemical processes via enhanced HO formation in downstream water.

Atmospheric concentrations of nitric acid, sulfur dioxide, particulate nitrate and particulate sulfate, and estimation of their dry deposition on the urban- and mountain-facing sides of Mt. Gokurakuji, Western Japan.

Atmospheric concentrations of nitric acid (HNO3), sulfur dioxide (SO2), particulate nitrate (NO3-) and particulate sulfate (SO4(2-)) on the urban- and mountain-facing sides of Mt. Gokurakuji were measured from November 2002 to October 2003, in order to evaluate the effects of anthropogenic activity on air quality and dry deposited nitrate and sulfate on the surfaces of pine foliage. The results showed that HNO3, SO2 and NO3(-) concentrations were significantly higher (P < 0.05) on the urban-facing side (1.54, 2.48 and 0.65 microg m(-3), respectively) than the mountain-facing side (0.67, 1.19 and 0.37 microg m(-3), respectively), while SO4(2-) concentrations did not differ significantly between the two sides (urban-facing: 2.80 microg m(-3); mountain-facing: 2.05 microg m(-3)). Indirect estimates of dry deposition rates of nitrate and sulfate to the surfaces of pine foliage based on the measured concentrations approximately agreed with the measured values determined by the foliar rinsing technique in a previous study. It was found that HNO3 was the major source (approximately 80%) of dry deposited nitrate on pine foliage, while the contribution from SO4(2-) was about equal to that from SO2. In conclusion, HNO3 and SO2 appear to be dominant species reflecting higher dry deposition rates of nitrate and sulfate on the urban-facing side compared to the mountain-facing side of Mt. Gokurakuji.