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Chemical and Biological Technologies in Agriculture
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Characteristics of water dissolved organic matter in Zoige alpine wetlands, China

Chemical and Biological Technologies in Agriculture volume 11, Article number: 127 (2024) Cite this article

Abstract

Background

Dissolved organic matter (DOM) plays a significant role in the biogeochemical cycle of crucial elements in aquatic ecosystem. However, it is still not clear on the spectral characteristics of water DOM in different types of alpine wetlands, which have less anthropogenic influences and intensive ultraviolet radiation. Here, we collected 107 water samples from marsh, lake, and river wetlands in the Zoige plateau, China, and analyzed the chemical characteristics, compositions, and potential sources of chromophoric DOM by combining UV–vis spectroscopy and excitation–emission matrix fluorescence spectroscopy coupled with parallel factor analysis (EEMs-PARAFAC).

Results

UVC and UVA fulvic-like substances were the prevailing fluorescence components in water DOM, which accounted for 23.74–71.59% and 16.76–30.01% of the total fluorescence intensity, respectively. Compared with the lake and river wetlands, fluoresce intensities of UVC and UVA fulvic-like substances in DOM were higher in marsh wetland. Marsh wetlands possessed the highest SUVA254, E2/E3, E2/E4, and E4/E6 of DOM, suggesting higher humification degree, higher relative molecular nominal size, and higher aromaticity. And the E2/E4 ratios in most water samples were higher than 12, indicating water DOM was mainly derived from autochthonous sources in alpine wetlands.

Conclusions

Wetland types strongly affected the spectral characteristics of water DOM in Zoige plateau. These findings may be beneficial for sustainable management of alpine wetlands.

Graphical Abstract

Background

Dissolved organic matter (DOM) possesses abundant functional groups and high mobility, is widely present in lakes, rivers, and other aquatic environments. As a vital and active chemical component for aquatic environment, DOM is rich in nutrients such as C, N, and P, which participate in various physical and chemical reactions in the water environment, thereby affect the water quality and elements cycling [1]. Moreover, dissolved organic carbon in surface water have been showed an increasing trend with the combined impacts of human activities and climate change [2]. Consequently, ascertaining the characteristics of water DOM is very important to elucidate its environmental behaviors and carbon cycle.

Water DOM composition is largely depended on its sources and biogeochemical processes. It can be affected by hydrology, aquatic light intensity, plants, microbial activity and land use [3,4,5]. In particular, the concentration, composition, and source of water DOM have been showed to be mostly affected by hydrologic factors, such as hydrological connectivity [6, 7], water retention [8], flood, dry–wet periods [3]. For example, Yu et al. [7] showed that hydrological connectivity is one of key factors controlling variation of water DOM in seasonal wetlands with short hydroperiod, and the absorption coefficient at 254 nm (α254), representing chromophoric DOM (CDOM) concentration in connected wetlands is lower than that in isolated wetland in less-developed region of South Carolina, USA. Kellerman et al. [9] found that molecular composition of DOM is shaped by precipitation and water residence time, terrestrially derived DOM is selectively lost as residence time increases in Sweden lakes. Fasching et al. [4] found hydrologic variability to control DOM composition in the stream water, high-flow events increased DOM inputs from terrestrial sources (as indicated by the contributions of humic- and fulvic-like fluorescence), while summer baseflow enhanced the autochthonous imprint of DOM.

Different types of wetlands possess various hydrological connectivity, part of the wetlands have connections with the surrounding aquatic ecosystem, while the others are almost entirely geographically isolated. This might form unique environments, vegetation and microbial community structure, thereby influence the composition and sources of DOM. Of great importance are sunlight-driven photochemical processes, wherein CDOM (typically < 0.22 μm) is an important light-absorbing substance that is a mixture of humus, fulvic acid, amino acid, etc. Due to the high selectivity of CDOM, fluorescence spectroscopy has provided insights into its composition and components in detail [10]. Previous studies have showed that the combination of excitation–emission matrix (EEM) fluorescence spectroscopy and parallel factor analysis (EEMs-PARAFAC) is a powerful and economical tool to distinguish different groups of organic compounds of DOM and to determine the relative contribution of each component to the total EEM fluorescence [11]. Most previous studies about water CDOM characteristics have focused on the single type of wetlands such as lakes, rivers, and marshes, however comparative study of water CDOM of different wetland types in the same research area is relatively insufficient, especially for alpine peatland with intensive solar radiation. There are different kinds of wetlands including marsh wetland, rivers, as well as lakes, different types of wetlands with specific hydrological conditions, aquatic light intensity and vegetation type, which might result in the changes of the composition and source of DOM in Zoige alpine peatland.

Therefore, the objective of this study was to identify the chemical characteristics, sources and composition of DOM in marsh wetland (MW), lake wetland (LW), and river wetland (RW) using EEMs-PARAFAC and ultraviolet–visible spectroscopy (UV–Vis).

Materials and methods

Study site

The study was carried out in the Zoige alpine peatland, which is located on the eastern edge of the Qinghai–Tibetan Plateau, China (32°20′-32°34′N, 101°30′-103°30′E, 3430–3700 m a.s.l.). The Zoige region belongs to typical humid monsoon climate in the cold-temperate plateau zone and has a mean annual temperature of 0.7–1.1 °C and a mean annual precipitation of 657 [12]. During the past four decades, the temperature and precipitation have increased by 0.04℃ yr−1 and 2.2 mm yr−1, respectively [13]. The Zoige alpine wetland is mainly consist of three types of wetlands, which are marsh, river, and lake wetlands. Among these wetlands, marsh wetlands and lake wetlands are generally hydrological-connected by rivers. The Zoige alpine wetland is the highest and largest peat marsh in China and covers an area of 6,180 km2, accounting for 31.5% of the whole Zoige plateau [14, 15].

Water sampling

Surface water samples (0–10 cm) were collected from typical marsh wetlands (MW, n = 44), lake wetlands (LW, n = 34), and river wetlands (RW, n = 29) in the Zoige region (Fig. 1). A total of 107 water samples were collected with 0.5 L plastic bottles and transported to the laboratory quickly. The water samples were passed through 0.45 μm PTFE filter membrane and were stored at 4 °C prior to analysis.

Fig. 1
figure 1

Locations of sampling sites in marsh wetlands (MW), lake wetlands (LW), and river wetlands (RW) in Zoige plateau

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Analysis of basic properties

Water pH was measured with a S20K pH meter (Mettler Toledo, Switzerland). Total organic carbon (TOC) was analyzed on a TOC/TN analyzer (Elementar, Germany). After acidification, Fe, Ca, Mg, K, and Na concentration in the water sample were detected using inductively coupled plasma optical emission spectrometer (ICP-OES, Optima 5300DV, Perkin-Elmer, USA). Total phosphorus (P) was analyzed by the ammonium molybdate spectrophotometry method.

Chromophoric DOM absorption analysis

CDOM absorption spectra were determined at 254, 365, 436, and 665 nm, respectively, using a T6 UV–Vis spectrophotometer (Persee, China). CDOM spectra for deionized water were used as a measurement blank. Specific absorbance (SUVA254) was calculated by dividing the UV absorption at wavelength of 254 nm by the TOC concentration, which was used as an index of DOM aromaticity [10, 16]. Ratios of E2/E3 (α254/α365), E2/E4 (α254/α436), and E4/E6 (α465/α665) were also calculated to indicate the relative molecular weight, source, and humification degree of the DOM [7, 17, 18].

Fluorescence EEMs-PARAFAC analysis

Three-dimensional excitation–emission matrix (EEM) spectra of CDOM were measured at room temperature (20 ± 2 °C) using a F7000 fluorescence spectrometer (Hitachi, Japan). The excitation wavelength (Ex) scanning range was 230 to 700 nm, and the emission wavelength (Em) scanning range was between 200 and 800 nm. The fluorescence spectra were scanned with a slit width of 5 nm and a scanning speed of 2400 nm/min. To ensure the comparability of fluorescence spectrum characteristics, the obtained spectra were corrected after the ultra-pure water blank was removed, thereby reducing the influence of instrument conditions and Raman scattering on fluorescence spectra. Parallel factor analysis (PARAFAC) was performed using the DOM Fluor toolbox on MATLAB R2016a (Mathworks, USA) to identify outlier and eliminate the Raman scattering effect on the EEM spectra [19]. Residual and split-half analyses were further conducted to validate fluorescence components of DOM [20].

Statistical analysis

One-way analysis of variance (ANOVA) was performed to test the significant differences in water physicochemical properties and spectral characteristics of DOM under different types of wetlands. Correlation analysis (CA) and principal component analysis (PCA) between water properties and spectral indices and fluorescence components of DOM were performed using Origin Pro 2022.

Results

Basic physicochemical properties of water samples

The average pH in the marsh wetlands, lake wetlands, and river wetlands were 8.32, 8.19 and 7.98, respectively (Table 1). And the average TOC concentration in the MW, LW, and RW were 59.32, 44.28 and 42.15 mg L−1, respectively. Both water pH and TOC were the highest in the MW, whereas pH and TOC in LW were not significantly different from those in the RW (Table 1). Concentrations of P, K, Fe, Ca, and Mg in all the investigated water samples were 0.014, 11.87, 0.015, 40.88, and 16.48 mg L−1 on average. No significant difference of P, K, Fe, Ca, and Mg concentration was found among the MW, LW, and RW (Table 1).

Table 1 Basic properties of overlying water in Zoige wetland
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CDOM absorption

The values of SUVA254, the ratios of E2/E4 and E4/E6 were highest in the MW, followed by the LW and the RW (Fig. 2). The average SUVA254 in the MW, LW, and RW were 20.46, 13.65 and 9.81, respectively. The SUVA254 in MW was significantly higher than those in both LW and RW, while there was no significant difference for SUVA254 between LW and RW. The average E2/E4 in the MW, LW, and RW were 19.22, 16.85 and 14.08, respectively, and the average E4/E6 in the MW, LW, and RW were 3.95, 3.46 and 2.82, respectively, both ratios of E2/E4 and E4/E6 in the MW significantly higher than that in the RW, while E2/E4 and E4/E6 in both MW and RW were no significant differences with LW (Fig. 2).

Fig. 2
figure 2

Box plots of SUVA254 (A), E2/E3 (B), E2/E4 (C), and E4/E6 (D) for water dissolved organic matter in marsh wetlands (MW), lake wetlands (LW), and river wetlands (RW) in Zoige plateau. Different letters represent significant difference between different wetland type at P < 0.05

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EEM spectral characteristics

Typical EEMs of CDOM for water samples obtained from the Zogie alpine wetlands are shown in Fig. 3. Five fluorescence components (C1–C5) were identified by the PARAFAC of all the DOM samples (Fig. 4). C1 showed an excitation/emission (Ex/Em) maximum at 336/443 nm, representing UVC fulvic-like substances with high molecular nominal size and aromatic humic, widespread, but highest in wetlands and forested environments [21]. C2 showed Ex/Em maximum at 273(396)/485 nm, representing UVA fulvic-like substances [20]. C3 and C4 showed Ex/Em maximum at 575/732 nm and 333(470)/570 nm, respectively, which can be assigned into humic-like substances [22], and C5 (288/390 nm) fell in the region of the protein-like B peak (B-peak, algae- and microbial-derived, tyrosine-like substance) [23].

Fig. 3
figure 3

EEM spectra of dissolved organic matter in water for different wetland types: marsh wetlands (MW), river wetlands (RW), and lake wetlands (LW)

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Fig. 4
figure 4

Loading of fluorescence components (C1–C5) of dissolved organic matter in water by the EEM spectroscopy and parallel factor analysis. Excitation/emission maximum wavelengths are: C1, 336/443 nm; C2, < 273(396)/485 nm; C3, 575/732 nm; C4, 333(470)/570 nm, C5: 288/390 nm

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The fluorescence components of water DOM were mainly fulvic-like components (C1 and C2) in the overlying water of Zoige alpine wetlands. C1 accounted for 23.74–71.59% of the total fluorescence intensity, with an average value of 43.80%, and C2 accounted for 16.76–30.01%, with an average value of 23.33%. While C3 was the lowest component accounting for 3.98–16.96%, with an average value of 8.34% (Fig. 5).

Fig. 5
figure 5

Intensity (A) and percent (B) of five fluorescence components (C1–C5) of dissolved organic matter in water under different wetland types: marsh wetlands (MW), lake wetlands (LW), and river wetlands (RW)

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The intensities of each fluorescence component for different types of wetlands are shown in Fig. 5. The average fluorescence intensity of C1 in the MW, LW, and RW were 51.97, 40.77, and 29.18 R.U., respectively, there were significantly statistical differences among the MW, LW, and RW (P < 0.01). Similar to C1, the fluorescence intensity of C2 with an average value of 24.83 R.U. was highest in the MW, followed by LW with the mean of 22.02 R.U., they were both significantly higher than that in RW (P < 0.01), while there was no significant difference of C2 intensity between MW and LW. The average fluorescence intensity of C4 was 10.47, 10.27 and 9.40 R.U. in the MW, LW, and RW, respectively, C4 intensity in the MW was significantly higher than it in RW (P < 0.01). While the average fluorescence intensity of C3 was 7.00, 7.22, and 7.55 R.U. in the MW, LW, and RW, respectively, C3 intensity in MW was significantly lower than it in the RW (P < 0.05). The average fluorescence intensity of C5 was 12.21, 11.74, and 11.17 R.U. in the MW, LW, and RW, respectively. C5 intensity did not show any significant differences among three different types of wetlands (Fig. 5).

Relationships of water DOM composition and spectral indices with water properties

The PC1 and PC2 axis complained 77.08% and 20.26% of the total variance (Fig. 6A). Along the PC1, distinct separation of the water DOM compositions was found between the RW with the LW and MW, which was mainly contributed by the C2 (Fig. 6B). Water TOC did not show significant correlations with DOM compositions. C1 was positively correlated with water pH, Fe, SUVA254, E2/E4, and E4/E6 (P < 0.01), while C3, C4, and C5 negatively correlated with pH, Fe, SUVA254, E2/E4, and E4/E6 (P < 0.01, Fig. 7). There were all positive linear relationships between TOC and SUVA254, TOC and E2/E4 in three types of wetlands, whereas the linear correlation coefficient varied depending on the type of wetlands. For example, water SUVA254 sharply increased with TOC concentration increasing in marsh wetlands compared with lakes and rivers, whereas rivers showed the fast increasing rate (Fig. 8).

Fig. 6
figure 6

Scattered point (A) and loading (B) from the principal component analysis plot for the fluorescence components (C1–C5) of dissolved organic matter in water for different wetland types: marsh wetlands (MW, pink circle), river wetlands (RW, blue circle), and lake wetlands (LW, green circle)

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Fig. 7
figure 7

Heatmap showing correlation between fluorescent components (C1–C5) and basic properties, spectral indices of water dissolved organic matter in Zoige alpine wetland. * and ** represent significant correlation at P < 0.05, and P < 0.01, respectively

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Fig. 8
figure 8

The correlation between water TOC concentration and SUVA254 (A), E2/E4 (B) in marsh wetlands (MW), lake wetlands (LW), and river wetlands (RW). Grey, light pink and blue frame represent the 95% confidence band of MW, LW, RW, respectively

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Discussion

Chemical characteristics and sources of water DOM in the alpine wetland

Zoige plateau possesses a high altitude and a thin atmosphere compared to other studies. These environmental factors may have an important influence on the photochemical oxidation processes of CDOM and mineralization of DOM [10]. Similarly, among three types of wetlands, water DOM extracted from the marsh wetland possessed the highest SUVA254, E2/E4, and E4/E6, showing the highest degrees of aromaticity and humification. Index of SUVA254 was used to characterize the aromaticity of DOM. It has been reported that SUVA254 > 4 generally indicates that the DOM was mainly consisted of hydrophobic organics, while SUVA254 < 4 suggests hydrophilic organics as the major components [24]. The SUVA254 was higher than 4 in the most investigated samples, suggesting that the major components of the DOM in the surface water were hydrophobic organics [25]. In this study, we demonstrated that SUVA254 of marsh wetland, lakes and rivers were 20.46, 13.65 and 9.81, respectively, which indicated that there more aromatic and hydrophobic substances were present in marsh wetland than in lakes and rivers. The correlation results between water SUVA254 and TOC concentration in each type of wetlands showed that more chromophores associated with high molecular nominal size were reduced in marsh wetlands when compared with the LW and the RW. The high molecular nominal size substances were transferred to low molecular nominal size pools through chemical bond rupture in photolysis process, which could be accelerated by long hydraulic retention time and intensive solar radiation [10].

As previously mentioned, the UV–visible absorption ratio E2/E4 was commonly employed to be source indicator of DOM (i.e., autochthonous or allochthonous). Battin [26] found that E2/E4 of river water ranged from 4.37 to 11.34 in the Orinoco Basin. Such values are common for allochthonous/terrestrial DOM, which has a greater aromatic carbon content associated with the presence of humic-like substances derived from macrophyte and soil organic matter. In general, the E2/E4 ratio is relatively higher for autochthonous DOM [27]. Jaffé et al. [28] reported a median value of E2/E4 in coastal Everglades estuaries was 28.7, suggesting that most of the DOM was derived from microbial sources. In this study, E2/E4 ranged from 7.6 to 27.23 with an average of 17.12, and E2/E4 ratio for 86% of total 107 sampling sites were more than 12.0. Which might hint that water DOM in Zoige wetlands were mainly from autochthonous/ microbial sources. There was a large amount of macrophytes debris with partly decomposed by microorganisms due to long-term or periodic flooding in marsh wetlands, while water DOM in hydrologically connected wetlands, especially for rivers, could be carried to downstream areas, which might result in the significant differences of E2/E4 among three types of wetlands, MW had the highest E2/E4, followed by LW, and RW.

DOM compositions in the alpine wetlands

In this study five fluorescence components were identified by the PARAFAC of all the DOM samples, and the fluorescence components for water DOM in alpine wetlands mainly consisted of UVC and UVA fulvic-like substances representing organic substances with large molecular nominal size and complex molecular structures [29], while protein-like substances possessed lower proportion of water DOM in Zoige peatland with less human activities, which was consistent with previous studies [30]. Similarly, Yu et al. [31] showed that the DOM components in natural water bodies with less anthropogenic influence are usually dominated by humic substances. On the contrary, Li et al. [29] indicated that protein-like components were more abundant than fulvic-like substances in the northwest Taihu Lake as a eutrophic lake due to industrial and domestic wastewater discharge and microbial metabolism caused by human activities [32]. Meanwhile, soil physiochemical properties (e.g., pH) could affect the microbial activities and the decomposition of soil organic matter [33], thereafter affect soil DOM components and its fluorescence intensity [34]. In this study, fluorescence intensity of C1 was positively correlated with water pH, while C3, C4, and C5 negatively correlated with pH and Fe content. The results in this study were consistent with previous studies indicating that the fluorescence intensity of each DOM components showed different reactions to varied pH value [34, 35], and that a greater fraction of high molecular nominal size of SOM released into water body with increasing pH value [36].

In this study, distribution of water DOM fluorescence components depended on wetland types. As shown in Figs. 4, 5, each wetland type had different distribution of DOM fluorescence components. The marsh wetland had the highest fluorescent intensity of UVC (C1) and UVA (C2) fulvic-like substances which have been supposed to be mainly derived from terrestrial organic matter as well as microbial reprocessing [29]. For example, Xu et al. [30] reported that the main source of fulvic acid organic matter was lignin substances, which were from the release of withered and rotting branches and leaves and soil sediment. While protein-like fluorescence (C5) was thought to be derived from autochthonous sources including algae, bacteria, picoplankton [37], and was negatively correlated with SUVA254, E2/E4, and E4/E6 in this study (Fig. 7). Even though fluoresce intensity of tyrosine-like substance (C5) had no significant differences among three types of wetlands, C5 fluoresce proportion was highest in rivers, suggesting a relatively high DOM contribution from autochthonous sources in rivers. Similar to water pH, the marsh wetland possessed the highest total fluorescence intensity, followed by lakes and rivers shown in Fig. 5, which could be explained by the molecular configuration and metal ions [35]. For example, Ghosh and Schnitzer [38] found that humic substances have linear structure at high pH and coil when pH decreases, which could explain that the fluorescence intensity of humic substances increase with increasing pH, because a spherocolloidal configuration could mask some fluorophores inside their structure at lower pH, while parts of humic substances with linear structure at higher pH were not anymore masked, following increased the fluorescence intensity. Another possible explanation could be related to the metal ions, the fluorescence intensity enhancement with increasing pH could be explained by a competition phenomena between the H+ ions and Fe ions to complex the OM in freshwater samples. This would lead some complexation–decomplexation processes directly influencing the fluorescence intensity because it is well known that Fe ions can affect the fluorescence intensity [33, 39]. In addition, each type of wetlands with unique hydrological condition and biotic community might influence the different structural diversity and processing pathways of DOM, which might be mainly driven by the oxidative dearomatization (ODA) through complementary selectivity of its photochemical, redox-initiated radical, ionic and enzymatic variants in freshwater ecosystems [40], thereby resulted in the differences of the fluoresce intensity of wetlands.

Conclusions

Here, we performed absorbance and EEM spectral analysis for the water DOM from 45 marsh wetlands, 34 lake wetlands, and 28 river wetlands in a typical alpine wetland distribution area in China to investigate the chemical characteristics, sources, and compositions of water DOM in alpine wetland. The results showed that the MW water samples possessed the highest SUVA254, E2/E3, and E4/E6 of DOM, suggesting higher humification degree, higher relative molecular nominal size, and higher aromaticity. UVC and UVA fulvic-like substances were the prevailing fluorescence components in the water DOM of alpine wetlands. Most of the investigated water sample had E2/E4 ratio higher than 12, indicating alpine water DOM was mainly derived from autochthonous sources. These findings uncover chemical nature of the DOM in alpine wetland and may deepen our knowledge to predict biochemical behavior of DOM in the alpine wetland ecosystem. The present study may also help to improve water quality management of alpine wetlands.

Availability of data and materials

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

DOM:

Dissolved organic matter

CDOM:

Chromophoric dissolved organic matter

EEM:

Excitation–emission matrix

PARAFAC:

Parallel factor analysis

MW:

Marsh wetlands

LW:

Lake wetlands

RW:

River wetlands

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Acknowledgements

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Funding

This work was supported by the financial support of the National Natural Science Foundation of China (Grant No. 41877421).

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Authors and Affiliations

  1. Institute of Wetland Research, Chinese Academy of Forestry, Beijing, 100091, China

    Jinzhi Wang, Lijuan Cui, Wei Li, Yinru Lei, Jing Li, Xiajie Zhai, Xinsheng Zhao & Rumiao Wang

  2. Beijing Key Laboratory of Wetland Services and Restoration, Chinese Academy of Forestry, Beijing, 100091, China

    Jinzhi Wang, Lijuan Cui, Wei Li, Yinru Lei, Jing Li, Xiajie Zhai, Xinsheng Zhao & Rumiao Wang

  3. Sino-Danish Centre for Education and Research, University of Chinese Academy of Sciences, Beijing, 100049, China

    Zhengyi Hu

  4. Center for Ecological and Environmental Accounting, Chinese Academy of Environmental Planning, Beijing, 100041, China

    Weishan Yang

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  1. Jinzhi Wang
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Contributions

JW, LC designed the experiment, wrote the paper and provided funding. WY, WL, YL, XZ, XZ, RW performed the experiments and analyzed data. ZH advised the study. All authors contributed to the article and approved the submitted manuscript.

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Correspondence to Lijuan Cui.

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Wang, J., Hu, Z., Cui, L. et al. Characteristics of water dissolved organic matter in Zoige alpine wetlands, China. Chem. Biol. Technol. Agric. 11, 127 (2024). https://doi.org/10.1186/s40538-024-00652-3

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