Impact of pond aquaculture effluents on seagrass performance in NE Hainan,tropical China

The impact of pond aquaculture effluents on the distribution and performance of seagrasses was examined in NE Hainan, tropical China. Samples were taken along transects in three back-reef areas with different extent of aquaculture production in their hinterland. High δ¹⁵N in seagrass leaves and epiphytes (6–9‰) similar to values in pond effluents documented aquaculture as dominant nitrogen source in the back-reefs with decreasing impact with distance from shore. Seagrass species abundance, shoot density and biomass were lower and concentrations of nutrients, chlorophyll and suspended matter were higher at nearshore sites with high and moderate pond abundance than at the control site. High epiphyte loads and low δ³⁴S in seagrass leaves suggest temporal shading and sulphide poisoning of the nearshore seagrasses. Observed gradients in environmental parameters and seagrass performance indicate that the distance from the pond outlets and size of the adjacent pond agglomeration are major determinants of seagrass degradation.     

外源氮磷负荷比增加对刺苦草（Vallisneria spinulosa）生长的影响

磷(P)是水生态系统生产力的限制因子,因此,在水体富营养化治理的过程中,许多地方采取了有效的措施控制流域P的排放;但氮(N)来源复杂,难以有效控制,导致湖泊外源的N∶P负荷比日益扩大.植物的生长需要合适的N∶P比,因此,外源N∶P负荷比的增加可能会对沉水植物的生长产生影响.采取控制实验,将刺苦草(Vallisneria spinulosa)栽种于塑料桶中,每隔3 d添加1次N、P营养盐,实验设置5个处理,P外源性负荷量固定为4 mg/(m~2·d),N外源性负荷量分别为0、40、80、120、160 mg/(m~2·d).实验周期为80 d.结果显示:随着N∶P负荷比的增加,刺苦草的叶干重、植株总干重、根干重、叶N及总N含量、叶片数、叶P及总P含量等指标基本保持不变或略有增加,单株总根长、根茎长度、块茎干重和无性系小株数目则呈现下降趋势.结果表明:随外源N∶P负荷比的增加,刺苦草个体生长指标总体呈现不变的趋势,但与种群扩张潜力相关的指标如根茎长度、块茎干重和无性系小株数目等则呈现下降趋势;刺苦草体内累积的N随外源N∶P负荷比的增加而增加,但P的累积保持不变.说明N∶P负荷比的增加对刺苦草个体生长影响不大,但对种群扩张不利;刺苦草对P的吸收也并不随N∶P比的增加而改变.     

Effects of fish farm loadings on seagrass (Posidonia oceanica) distribution, growth and photosynthesis

The spatial extent and timing of the impact of fish farms on the distribution and performance of a Posidonia oceanica meadow were examined in an embayment of the south-eastern coast of Spain (Hornillo Bay, Murcia). Changes in seagrass distribution were determined using available seagrass mapping (from 1988, i.e., before the onset of aquaculture activities and 1998) and by successive sampling in 1994 and 1998. Environmental variables (light attenuation coefficient, water-column dissolved nutrients and organic content of sediments) together with plant performance (shoot biomass, leaf growth rate, photosynthetic activity, carbohydrate reserves, the number of leaves per shoot, epiphyte loads and herbivore pressure) were measured in plants affected by organic discharges, and were compared with those found in reference healthy plants over an annual growth cycle. Since the onset of fish farm activity, 11.29 ha of P. oceanica meadow has been completely lost and 9.86 ha significantly degraded, thus resulting in a total affected area which accounts for about 53% of the former meadow, or 7-fold the fish farming area. Unequal propagation of seagrass die-off or degradation reflects the relevance of local factors such as depth and hydrodynamism on the true extent of fish farm impact. Water transparency decreases and dissolved nutrient and organic content of sediments increases in the vicinity of cages compared to distant reference stations, thus supporting the notion of environmental gradients caused by the organic release from cages, which spreads outwards. Shoot size, leaf growth rate and the number of leaves per shoot in plants close to the fish farm decreased. Moreover, low leaf growth and low rhizome carbohydrate concentration (always relative to that found in an undisturbed area) indicated carbon budget imbalances. Since light reduction in the affected area was only modest (31% of light reaching the sea surface, while at the same depth this figure was 39% at the reference site), and light availability was well above the minimum requirement estimated for this species, neither this factor nor epiphyte overgrowth (epiphyte load was lower in the affected area) seem to explain such carbon imbalances or the observed meadow regression. Alternatively, the high herbivore pressure found in the affected zone suggests that overgrazing is one of the main causes of decreasing shoot sizes and hence of carbon imbalance, reduced growth and shoot mortality. The impact of fish farms on seagrasses, therefore, seems to be highly variable and depends on complex interactions between a large number of processes.     

Recovery from the impact of light reduction on the seagrass Amphibolis griffithii, insights for dredging management

A large-scale, manipulative experiment was conducted to examine the extent and rate of recovery of meadows of the temperate Australian seagrass, Amphibolis griffithii to different light-reduction scenarios typical of dredging operations, and to identify potential indicators of recovery from light reduction stress. Shade cloth was used to mimic different intensities, durations and start times of light reduction, and then was removed to assess the recovery. The meadow could recover from 3 months of light stress (5-18% ambient) following 10 months re-exposure to ambient light, even when up to 72% of leaf biomass was lost, much faster recovery rates than has previously been observed for large seagrasses. However, when the meadow had been shaded for 6-9 months and more than 82% of leaf biomass was lost, no recovery was detected up to 23 months after the light stress had ceased, consistent with other studies. Five potential indicators of recovery were recommended.     

Physiological Responses of Five Seagrass Species to Trace Metals

Trace metal run-off associated with urban and industrial development poses potential threats to seagrasses in adjacent coastal ecosystems. Seagrass from the largest urban (Moreton Bay) and industrial (Port Curtis) coastal regions in Queensland, Australia were assessed for metal concentrations of iron (Fe), aluminium (Al), zinc (Zn), chromium (Cr) and copper (Cu). Trace metal concentrations in seagrass (Zostera capricorni) leaf and root-rhizome tissue had the following overall trend: [Fe] > [Al] > [Zn] > [Cr] > [Cu]. Rainfall events and anthropogenic disturbances appeared to influence metal concentrations in seagrasses with the exception of Al, which does not appear to bioaccumulate. In laboratory experiments, five seagrass species (Halophila ovalis, H. spinulosa, Halodule uninervis, Z. capricorni, Cymodocea serrulata) were incubated with iron (1 mg Fe l⁻¹) and copper (1 mg Cu l⁻¹) and responses assessed by changes in PSII photochemical efficiency (Fv/Fm), free amino acid content and leaf/root-rhizome metal accumulation. Iron addition experiments only affected Halophila spp, while copper additions affected other seagrass species as well. Trace metal contamination of seagrasses could have ramifications for associated trophic assemblages through metal transfer and seagrass loss. The use of photosystem II photochemical efficiency as well as amino acid concentrations and composition proved to be useful sublethal indicators of trace metal toxicity in seagrasses.     

Physiological responses of the seagrass Thalassia hemprichii (Ehrenb.) Aschers as indicators of nutrient loading

To select appropriate bioindicators for the evaluation of the influence of nutrients from human activities in a Thalassia hemprichii meadow, environmental variables and plant performance parameters were measured in Xincun Bay, Hainan Island, South China. Nutrient concentrations in the bay decreased along a gradient from west to southeast. Moreover, the nutrients decreased with an increase in the distance from the shore on the southern side of the bay. Among the candidate indicators, the P content of the tissues closely mirrored the two nutrient loading gradients. The epiphytic algae biomass and the N content in the tissues mirrored one of the two nutrient loading trends. The leaf length, however, exhibited a significant negative correlation with the nutrient gradients. We propose that changes in the P content of T. hemprichii, followed by epiphytic algae biomass and N content of the tissues, may be the useful indicators of nutrient loading to coastal ecosystems.     

水位波动的幅度与频率对刺苦草(Vallisneria spinulosa)生物量和形态特征的影响

沉水植物是维持浅水湖泊清水稳态以及湖泊生态系统结构与功能的关键生物类群.近年来,气候变化和人类活动的双重作用造成湖泊水文情势的明显改变,对沉水植物的生长、繁殖和演替有着较大的影响.本文选取2个水位波动幅度与2个波动频率(逐渐改变水位),以恒定水位作为对照,研究湿地中具有重要生态功能的沉水植物刺苦草(Vallisneria spinulosa)的生物量及地上和地下营养器官形态结构对水位波动的响应.结果表明,相对于恒定水位,波动水位(低幅度低频率、低幅度高频率、高幅度低频率、高幅度高频率)对刺苦草的总生物量和获取限制资源相关器官有显著影响.相比于对照组,水位波动能够显著降低刺苦草分株数、叶片数、根数、根长、匍匐茎数、总匍匐茎长以及叶、根、地上和地下生物量,但会显著增加其株高和比叶面积.不同波动频率(低频率和高频率)和幅度(低幅度和高幅度)对光合器官的形态特征都没有显著影响,但叶和地上部分的生物量积累均受到波动幅度的显著影响.水位波动幅度显著改变了刺苦草的根数和根长,而波动频率的影响不显著.因此,对浅水湖泊受损刺苦草群落进行重建时,应避免较大幅度的水位波动,从而有利于增强植株快速恢复的能力.     

Effects of seagrass leaf litter decomposition on sediment organic carbon composition and the key transformation processes

Seagrass leaf litters are an important source of sediment organic carbon (SOC). However, the mechanisms of seagrass leaf litter decomposition influencing SOC composition and the key transformation processes remain unknown. We performed a laboratory chamber experiment to compare the labile organic carbon (OC) composition and the enzyme activities governing SOC transformation between the seagrass group (seagrass leaf litter addition) and the control group. The results showed that the seagrass leaf litter decomposition significantly elevated the salt-extractable carbon (SEC) content and the SEC/SOC. Additionally, the levels of invertase, polyphenol oxidase, and cellulase in the seagrass leaf litters addition group were generally higher than in the control group, which could elevate recalcitrant OC decomposition. Following 24 days incubation, addition of seagrass leaf litter increased the amount of CO2 released, but decreased the SOC content. Therefore, seagrass leaf litter decomposition leached abundant dissolved OC, which enhanced the activity and transformation of SOC.     

Leaching and microbial degradation of dissolved organic matter from salt marsh plants and seagrasses

Dissolved organic matter (DOM) is outwelled from highly productive salt marshes, but its sources and fates are unclear. To examine common salt marsh plants as sources of coastal DOM, two dominant salt marsh vascular plants Spartina alterniflora and Juncus roemarianus, and two major coastal seagrasses Syringodium filiforme and Halodule wrightii, were collected from a Florida salt marsh and studied using laboratory incubation experiments. We investigated the leaching dynamics of dissolved organic carbon (DOC), total dissolved nitrogen (TDN), and chromophoric dissolved organic matter (CDOM) from these plants, in conjunction with our field investigations of the sources and outwelling of DOM from Florida salt marshes. The leaching of DOM and CDOM from the plants was a rapid process, and leaching rates were 65–288 µM/g dry weight/day for DOC and 3.8–16 µM/g dry weight/day for TDN from different plants in the bacteria-inhibited incubations. DOC was proportional to TDN in the leachates, but the quantity of C and N leached was dependent on the species and growth stage of the plants. At the end of the 25-day experiments, 5.4–23 % and 10–45 % of solid phase C and N were released into DOC and TDN pools, respectively. Bacteria played an important role during the leaching process. The majority of DOC and TDN leached from marsh plants and seagrasses was labile and highly biodegradable with 56–90 % of the leached DOC and 44–72 % of the leached TDN being decomposed at the end of the experiments. The fluorescence measurements of CDOM indicate that organic matter leached from marsh plants and seagrasses contained mainly protein-like DOM which was degraded rapidly by bacteria. Our study suggests that leaching of DOM from salt marsh plants and seagrasses provide not only major sources of DOC, TDN, and CDOM that affect many biogeochemical processes, but also as important food sources to microbial communities in the marsh and adjacent coastal waters.     

Monitoring polycyclic aromatic hydrocarbons in the Northeast Aegean Sea using Posidonia oceanica seagrass and synthetic passive samplers

The concentrations of 22 polyaromatic hydrocarbons (PAH) in Posidonia oceanica seagrass, sediments, and seawater from the Alexandroupolis Gulf in the Aegean Sea, were investigated from 2007 to 2011. Temporal trends of total PAH contents in P. oceanica and sediments were similar. PAH levels in seawater, sediments, and seagrasses generally decreased with increasing distance from Alexandroupolis Port. Leaves and sheaths of P. oceanica had higher PAH levels than roots and rhizomes. P. oceanica accumulates PAHs and has good potential as a bioindicator of spatiotemporal pollution trends. PAH concentrations were also examined using in situ passive seawater sampling and were compared to results of passive sampling in the laboratory using local sediments and seawater. Levels of high molecular weight PAHs assessed using passive samplers confirmed the decreasing gradient of pollution away from Alexandroupolis Port. Passive sampling also proved useful for investigating sources of PAHs in P. oceanica meadows.     

Different morphology of Nuphar lutea in two contrasting aquatic environments and its effect on ecosystem engineering

Aquatic plants (macrophytes) can have a large effect on river hydraulics and geomorphology. Though, the extent to how plant morphological plasticity actively influences these feedbacks has received little scientific attention. The nymphaeid macrophyte species Nuphar lutea (L.) Smith is characterized by a distinct leaf duality. Floating leaves shade most of the submerged leaves thereby limiting light penetration in the water. Despite their apparent negligible photosynthetic role, submerged leaves of N. lutea remain intact during summer and contribute a significant part to the total biomass. Our results indicate that the submerged leaves are crucial in plant–flow interactions and hence in the engineering potential of the plant, i.e. the capacity to locally reduce flow velocities and to promote sedimentation, including organic matter deposition. Plant individuals growing in running river water were compared to individuals from adjacent oxbow lake water. The number and size of submerged leaves were significantly higher for river standing individuals and the accumulated sediment contained significantly more organic matter, total nitrogen and total phosphorus, and was characterized by a lower carbon/nitrogen ratio and a finer grain size. We therefore argue that the submerged N. lutea canopy in rivers has the ability to create a high‐nutrient, low hydrodynamic environment, resembling the conditions found in oxbow lakes. Copyright © 2014 John Wiley & Sons, Ltd.     

Are seagrass beds indicators of anthropogenic nutrient stress in the rocky intertidal?

It is well established that anthropogenic nutrient inputs harm estuarine seagrasses, but the influence of nutrients in rocky intertidal ecosystems is less clear. In this study, we investigated the effect of anthropogenic nutrient loading on Phyllospadix spp., a rocky intertidal seagrass, at local and regional scales. At sites along California, Washington, and Oregon, we demonstrated a significant, negative correlation of urban development and Phyllospadix bed thickness. These results were echoed locally along an urban gradient on the central California coast, where Phyllospadix shoot δ¹⁵N was negatively associated with Phyllospadix bed thickness, and experimentally, where nutrient additions in mesocosms reduced Phyllospadix shoot formation and increased epiphytic cover on Phyllospadix shoots. These findings provide evidence that coastal development can threaten rocky intertidal seagrasses through increased epiphytism. Considering that seagrasses provide vital ecosystem services, mitigating eutrophication and other factors associated with development in the rocky intertidal coastal zone should be a management priority.     

Contribution of primary producers to mercury trophic transfer in estuarine ecosystems: Possible effects of eutrophication

There is an ongoing eutrophication process in the Ria de Aveiro coastal lagoon (Portugal), with progressive replacement of rooted primary producers for macroalgae. Taking advantage of a well-defined environmental contamination gradient, we studied mercury accumulation and distribution in the aboveground and the belowground biomass of several salt marsh plants, including the seagrass species Zostera noltii and the dominant green macroalgal species Enteromorpha sp. The results of these experiments were then placed into the context of the estuarine mercury cycle and transport from the contaminated area.All salt marsh plants accumulated mercury in the root system, with Halimione portulacoides showing the highest levels, with up to 1.3 mg kg⁻¹ observed in the most contaminated area. Belowground/aboveground ratios were generally below 0.4, suggesting that salt marsh plants are efficient immobilizers and retainers of mercury agents. Moreover, due to their sediment accretion capacities, salt marsh plants seem to play an important role in the sequestration of mercury in estuarine sediments.Seagrasses, on the other hand, accumulated considerable amounts of mercury in the aboveground biomass with belowground/aboveground ratios reaching as high as 1.4. These results may be due to their different routes of uptake (roots and foliar uptake) which suggests that seagrass meadows can be an important agent in the export of mercury from contaminated areas, considering the high aboveground biomass replacement rates.Rooted macrophytes accumulate less mercury in their aboveground biomass than macroalgae. The change of primary producer dominance due to eutrophication can originate a 4- to 5-fold increase in primary producer associated mercury. This mercury would be available for export, making it bioavailable to estuarine food webs, which stresses the need to reverse the current eutrophic status of estuarine systems.     

Spatial patterns of sub-tidal seagrasses and their tissue nutrients in the Torres Strait,northern Australia: Implications for management

The distribution and nutritional profiles of sub-tidal seagrasses from the Torres Strait were surveyed and mapped across an area of 31,000 km². Benthic sediment composition, water depth, seagrass species type and nutrients were sampled at 168 points selected in a stratified representative pattern. Eleven species of seagrass were present at 56 (33.3%) of the sample points. Halophila spinulosa, Halophila ovalis, Cymodocea serrulata and Syringodium isoetifolium were the most common species and these were nutrient profiled. Sub-tidal seagrass distribution (and associated seagrass nutrient concentrations) was generally confined to northern-central and south-western regions of the survey area (<longitude 142.60), where mean water depth was relatively shallow (approximately 13 m below MSL) and where sediments were comprised primarily muddy sand to gravelly sand. Seagrass nitrogen and starch content, the most important nutrients for marine herbivores, were significantly correlated with species and with the plant component (above or below ground). For all seagrass species, the above-ground component (shoots and leaves) possessed greater nitrogen concentrations than the below-ground component (roots and rhizomes), which possessed greater starch concentrations. S. isoetifolium had the highest total nitrogen concentrations (1.40±0.05% DW). However, it also had higher fibre concentrations (38.2±0.68% DW) relative to the other four species. H. ovalis possessed the highest starch concentrations (2.76±0.12% DW) and highest digestibility (83.24±0.66% DW) as well as the lowest fibre (27.2±0.66% DW). The high relative abundance (found at 55% of the sites that had seagrass) and nutrient quality characteristics of H. ovalis make it an important source of energy to marine herbivores that forage sub-tidally in the Torres Strait. There were two regions in Torres Strait (north-central and south-western) where sub-tidal seagrass meadows were prevalent and of relatively higher nutritional value. This spatial and nutritional information can be used by local agencies to manage and to protect the ecological, economic and cultural values of the sub-tidal seagrass ecosystems and associated fisheries of the Torres Strait.     

Nutrient enrichment does not affect diet selection by a tropical shredder species in a mesocosm experiment

The decomposition of plant litter is a fundamental ecological process in small forest streams. Litter decomposition is mostly controlled by litter characteristics and environmental conditions, with shredders playing a critical role. The aim of this study was to evaluate the effect of leaf species (Maprounea guianensis and Inga laurina, which have contrasting physical and chemical characteristics) and water nutrient enrichment (three levels) on leaf litter chemical characteristics and fungal biomass, and subsequent litter preference and consumption by Phylloicus sp. (a typical shredder in tropical streams). Maprounea guianensis leaves had lower lignin and nitrogen (N) concentrations, higher polyphenols concentration and lower lignin:N ratio than I. laurina leaves. Phosphorus concentrations were higher for both leaf species incubated at the highest water nutrient level. Fungal biomass was higher on M. guianensis than on I. laurina leaves, but it did not differ among nutrient levels. Relative consumption rates were higher when shredders fed on M. guianensis than on I. laurina leaves, due to the lower lignin:N ratio and higher fungal biomass of M. guianensis. Consumption rates on M. guianensis leaves were higher for those exposed to low water nutrient levels than for those exposed to moderate water nutrient levels. Feeding preferences by shredders were not affected by leaf species or nutrient level. The low carbon quality on I. laurina leaves makes it a less attractive substrate for microbial decomposers and a less palatable resource for shredders. Changes in litter input characteristics may be more important than short-term nutrient enrichment of stream water on shredder performance and ecosystem functioning.     

Contrasting recovery of shallow and deep water seagrass communities following climate associated losses in tropical north Queensland,Australia

Tropical seagrass decline and recovery from severe storm impacts was assessed via quarterly measurements of seagrass biomass, species composition and experimental investigations of recovery in north Queensland. Shallow and deep seagrass meadows suffered major declines. Significant recovery in the two years following loss only occurred at deeper sites. Halophila spp. in deep water areas had a high capacity for recovery through the availability of seed banks. In contrast, the shallow species did not recover quickly from experimental disturbance, had poor seed reserves and relied on asexual propagation. The potential for shallow species to recover rapidly from widespread losses was limited as seed banks were limited or non-existent. Understanding inter- and intra-specific differences in seagrass recovery and how this interacts with location is critical to predict the consequences of climate events to tropical seagrasses. This is especially important as more frequent severe storms are predicted as a consequence of climate change.     

System Performance in UASB Reactors Receiving Increasing Levels of Sulfate

Glucose‐fed high‐rate UASB reactors were tested at three COD/SO₄ ratios and hydraulic retention times to promote sulfate reducing activity and observe the effects on reactor performance. Different COD/SO₄ ratios (20, 10, and 5) resulted in changes in organic matter removal, methane production, alkalinity, dissolved sulfide and biomass concentrations and profile. The COD removal dropped from 95 to 80–84 % at the lowest COD/SO₄ ratio. Sulfate was removed at 79 to 89 % at the highest ratio and dropped to 72 to 74 % with increasing sulfate loading. Alkalinity was produced at higher levels with increasing sulfate loading. Specific methane production dropped with decreasing hydraulic retention times. Sulfate‐reducing activity used a maximum of 11.7 % of organic matter at the highest sulfate loading level, producing a slight shift to sulfate‐reducing activity in the substrate competition between sulfate‐reducing bacteria and methanogens. Increased sulfate loading at COD/SO₄ ratios of 10 and 5 caused deterioration of the concentration profile of the sludge, resulting in biomass washout and decreased volatile fraction of biosolids in the reactors.     

Epiphyte loads on seagrasses and microphytobenthos abundance are not reliable indicators of nutrient availability in oligotrophic coastal ecosystems

Despite marked gradients in nutrient availability that control the abundance and species composition of seagrasses in south Florida, and the importance of nutrient availability in controlling abundance and composition of epiphytes on seagrasses in other locations, we did not find that epiphyte load on the dominant seagrass, Thalassia testudinum, or that the relative contribution of algal epiphytes to the epiphyte community, was positively correlated with nutrient availability in the water column or the sediment in oligotrophic seagrass beds. Further, the abundance of microphytobenthos, as indicated by Chlorophyll-a concentration in the sediments, was not directly correlated with concentrations of nutrients in the sediments. Our results suggest that epiphyte and microphytobenthos abundance are not unambiguous indicators of nutrient availability in relatively pristine seagrass environments, and therefore would make poor candidates for indicators of the status and trends of seagrass ecosystems in relatively low-nutrient environments like the Florida Keys.     

Future seagrass beds: Can increased productivity lead to increased carbon storage?

While carbon capture and storage (CCS) is increasingly recognised as technologically possible, recent evidence from deep-sea CCS activities suggests that leakage from reservoirs may result in highly CO₂ impacted biological communities. In contrast, shallow marine waters have higher primary productivity which may partially mitigate this leakage. We used natural CO₂ seeps in shallow marine waters to assess if increased benthic primary productivity could capture and store CO₂ leakage in areas targeted for CCS. We found that the productivity of seagrass communities (in situ, using natural CO₂ seeps) and two individual species (ex situ, Cymodocea serrulata and Halophila ovalis) increased with CO₂ concentration, but only species with dense belowground biomass increased in abundance (e.g. C. serrulata). Importantly, the ratio of below:above ground biomass of seagrass communities increased fivefold, making seagrass good candidates to partially mitigate CO₂ leakage from sub-seabed reservoirs, since they form carbon sinks that can be buried for millennia.     

Plant Species Richness Increased Belowground Plant Biomass and Substrate Nitrogen Removal in a Constructed Wetland

The effects of plant species richness on both above‐ and belowground plant biomass, plant nitrogen (N) pool size, and substrate N concentrations were studied in a full‐scale subsurface vertical‐flow constructed wetland (CW). Results showed that (i) plant species richness increased belowground plant biomass and its N pool size but had no effect on aboveground plant biomass and its N pool size; (ii) plant species richness increased substrate N removal, especially ammonium N removal; and (iii) plant species richness had no effect on plant N use efficiency, suggesting that the N pool size increased with increasing plant species richness. More N accumulation could be removed through harvesting plant biomass. We concluded that the N removal performance of the CW improved by plant species richness through increasing belowground biomass and relevant N pool size.