Ecological Indicators 155 (2023) 110952

Contents lists available at ScienceDirect

Ecological Indicators
journal homepage: www.elsevier.com/locate/ecolind

Spatial distribution of surface-sediment diatom assemblages from 45

Tibetan Plateau lakes and the development of a salinity transfer function
Siwei Yu a, b, c, Junbo Wang a, *, Kathleen M. Rühland b, Liping Zhu a, c, Jianting Ju a, Chong Liu a, d,
Qingfeng Ma a, John P. Smol b
a
State Key Laboratory of Tibetan Plateau Earth System, Environment and Resources (TPESER), Nam Co Observation and Research Station (NAMORS), Institute of
Tibetan Plateau Research, Chinese Academy of Sciences, Beijing 100101, China
b
Paleoecological Environmental Assessment and Research Laboratory (PEARL), Department of Biology, Queen’s University, Kingston, Ontario K7L3N6, Canada
c
University of Chinese Academy of Sciences, Beijing 100049, China
d
Piesat Information Technology Co., Ltd, Beijing 100195, China

A R T I C L E I N F O A B S T R A C T

Keywords: The Tibetan Plateau (TP) is often referred to as the “Asian Water Tower”, as it plays a crucial role in supplying
Diatoms water for ~1.9 billion people. Lakes in this arid mountain region have a wide salinity gradient and are sensitive
Tibetan Plateau to the complex interactions between climate and the water cycle, affecting aquatic ecosystems and species
Spatial distribution
distributions. Here, we analyzed relationships between measured environmental variables and the spatial dis­
Salinity
Transfer function
tribution of surface sediment diatom assemblages from 45 lakes located throughout the TP with the intention of
developing a diatom-inference model for salinity. Diatom assemblage composition and diversity differed among
lakes from the western (W), southern (S), northern (N), and southeastern (E) regions of the TP. The most common
taxa observed in the lake set, Pantocsekiella ocellata, Nitzschia palea, and Staurosira venter, tended to have
widespread distributions across the TP. In general, diatom diversity and richness tended to be highest in lakes
with salinity concentrations between 0.1 and 10 g/L and were lowest in high salinity lakes (>20 g/L) of the N
region. Canonical correspondence analysis (CCA) identified lake water salinity and maximum lake depth as
explaining a statistically significant portion of diatom assemblage variation. Weighted-averaging (WA) was used
to develop a diatom-based salinity inference model (R2jack = 0.71, RMSEPjack = 0.28) based on diatom optima
from freshwater to meso-hyposaline lakes of the TP. Diatom-salinity reconstructions and changes in diatom
community composition can potentially be used to track long-term patterns in climate and environmental
conditions across the highly complex mountain landscape of the TP.

1. Introduction
The Tibetan Plateau (TP), also known as “the Third Pole” (Qiu, 2008)
and “the Asian Water Tower” (Immerzeel et al., 2010), plays an
important role in the Earth’s climate system and supplies a critical water
resource for ~1.9 billion people (Immerzeel et al., 2020; Yao et al.,
2012). Because of its location, the TP is highly vulnerable to water stress,
governance, hydropolitical tensions, and to future climatic and socio­
economic changes (Immerzeel et al., 2020). This arid to semi-arid and
topographically complex mountain region is situated far inland resulting
in high evaporation rates and a wide gradient of environmental vari­
ables across lakes (Liao et al., 2020), including salinity. Salinity is
important for characterizing the limnology of TP lakes, particularly in
the numerous hydrologically closed basin lakes that are common to the
region. These characteristics play a significant role in lake ecosystem
structure, diversity, productivity (Wischnewski et al., 2011; Anslan
et al., 2020) and in aquatic community composition including diatom
species distributions (Wang et al., 2011).
Understanding the responses of aquatic organisms to multiple
stressors and identifying their relationship to key environmental vari­
ables is vital to advancing global freshwater biodiversity research in the
context of environmental change (Birk et al., 2020; Maasri et al., 2022).
In the TP and many other remote alpine regions, the complexities of
aquatic community structure and diversity, as well as biogeographic
gradients, are poorly known (Kärnä et al., 2018). Examining past
(baseline) environmental conditions will provide context to better

* Corresponding author.
E-mail address: wangjb@itpcas.ac.cn (J. Wang).

https://doi.org/10.1016/j.ecolind.2023.110952
Received 30 May 2023; Received in revised form 9 September 2023; Accepted 12 September 2023
Available online 19 September 2023
1470-160X/Crown Copyright © 2023 Published by Elsevier Ltd. This is an open access article under the CC BY license
(http://creativecommons.org/licenses/by/4.0/).
S. Yu et al.
understand contemporary ecological patterns and species distributions
(Seddon et al., 2014). For example, paleolimnological techniques can be
used to establish ecological and chemical baseline conditions, assess
how much the environment has changed relative to the baseline state,
and have the potential to qualitatively and quantitatively reconstruct
past environmental conditions (Bennion et al., 2004; Battarbee et al.,
2005; Smol, 2019). Here, we explore the modern distributions of diatom
assemblages using a calibration set of lakes in the TP to better under­
stand spatial patterns of environmental variables and diatom species
composition in this complex alpine region. We analyse the relationships
between diatom assemblages and key environmental variables to
explore the development of a diatom-based salinity transfer function for
the TP.
Quantitative reconstructions of environmental variables are based
on the relationships between the modern distributions of biotic proxies
and measured environmental variables that can then be used to estimate
past lake-water conditions using subfossil assemblages preserved in lake
sediment cores (Imbrie and Kipp, 1971; Birks et al., 2010). One of the
most commonly used and powerful indicators of environmental change
are the siliceous algae, diatoms (Bacillariophyceae). They are successful
paleo-indicators because their cell walls typically preserve well in lake
sediments and they respond quickly and sensitively to environmental
change (Smol 2009; Smol and Stoermer, 2010). Over the past few years,
several studies from the central and eastern TP have examined the
geographic distribution of diatoms in response to recent climate change
(Yu et al., 2019; Wang et al., 2020a, Wang et al., 2020b). However, there
is a lack of diatom studies in the western, southern and northern parts of
the TP. The scarcity of data is understandable given that these high-
altitude regions are remote and weather conditions are often harsh,
making fieldwork logistically challenging (Qiu et al., 2008). Examining
the spatial distributions and the environmental optima and tolerances of
diatom taxa in understudied aquatic systems from remote regions will
help to better understand and interpret environmental changes, partic­
ularly in the context of climate change (Benito et al., 2018; Rivera-
Rondón and Catalan, 2020). To date, the majority of diatom-based
paleolimnological studies of the TP have focused on qualitative in­
terpretations of environmental change during the past few centuries (e.
g., Wang et al., 2011; Wischnewski et al., 2011a) to millennia (Li et al.,
2015; Wang et al., 2018; Liao et al., 2020; Su et al., 2022).
Diatom assemblages show strong relationships to salinity, and
diatom-based models have been developed to quantitatively track past
variations in lake water salinity (reviewed in Fritz et al. 2010). Recon­
structing salinity is particularly appropriate in semi-arid to arid regions
of the world where lakes are typically subjected to relatively high
evaporation rates, making them highly susceptible to climate change
(Cumming and Smol 1993; Wilson et al. 1994; Cumming et al., 1995;
Bloom et al. 2003). However, only a few diatom-based salinity models
have been developed from lakes in the eastern Tibetan Plateau (Wang
et al. 2011), and therefore more calibration studies are required to better
represent this large and highly complex mountain region represented by
a large gradient of salinity. Here, we examined the distribution of
modern diatom assemblages from the surface sediments of 45 lakes
throughout the TP that cross gradients of altitude, lake depth, pH and
salinity. These data were then used to develop a diatom-based inference
model that can potentially reconstruct past salinity trends across the TP.
2. Study region
The TP is the highest and most extensive plateau in the world and is
characterized by a highly complex and heterogeneous landscape that is
sensitive to climate change and other environmental stressors (Liao
et al., 2020). It spans relatively large longitudinal (26.00◦ N-39.78◦ N)
and latitudinal (73.32◦ E-104.78◦ E) gradients, with the majority of the
region located at elevations above 4500 m a.s.l., covering an area of
approximately 2.5 × 106 km2 (Yao et al., 2012). The TP is characterized
by cold-dry winters and cool-humid summers (Lu et al., 2011) with
2
Ecological Indicators 155 (2023) 110952
mean annual temperature ranging from − 15 ◦ C to 5 ◦ C, relatively short
growing seasons and long periods of frost. Most areas of the TP are
characterized by an arid/semiarid climate, with annual precipitation
ranging from tens to hundreds of millimeters, mostly occurring as rain
from May to September (Gao and Liu, 2013). The southern and south­
eastern TP has comparatively high rainfall, with mean annual precipi­
tation ranging from 370 mm to over 1800 mm (Lu et al., 2011).
Lakes on the TP are relatively isolated with minimal human activ­
ities. Many TP lakes are hydrologically closed systems, a characteristic
that is known to have important implications on water quality and
aquatic biota. With the exception of the southeastern region, the vege­
tation of the TP is dominated by meadow, steppe, and shrub commu­
nities. TP vegetation is influenced by climate and altitude and is
distributed along a gradient of decreasing precipitation, changing from
forest to meadow/shrub, to steppe, then to desert with a zonal pattern
from southeast to northwest (Institute of Geography, 1990; Hou, 2001).
Vegetation in the eastern and southeastern TP is dominated by montane
coniferous and broad-leaved forests, whereas in the southern and east-
central TP, vegetation is dominated by shrubs (Institute of Geography,
1990; Ma et al., 2019). Alpine meadows are mainly distributed in the
eastern TP above 4000 m a.s.l. and dominated by sedge and grass
communities. The vegetation of the central and southern TP is domi­
nated by alpine steppe between 4400 and 5400 m a.s.l., and is mainly
comprised of Poaceae (e.g., Stipa purpurea and S. basiplumosa) and
Asteraceae (e.g., Artemisia) (Institute of Geography, 1990). Alpine
shrub, one of the dominant vegetation types on the TP, are distributed
throughout and in large tracts across the alpine forest and meadow
transition zones (Zhao et al., 2006).
3. Methods
3.1. Sample collection
The 45 lakes were sampled between 2009 and 2019 and are mainly
located in the western, southern, northern, and southeastern TP (Fig. 1).
The lakes span a latitudinal gradient of 28.96 ◦ N to 35.9 ◦ N, a longitu­
dinal gradient of 79.36 ◦ E to 98.85 ◦ E and an altitudinal gradient of
4240 m a.s.l. to 5220 m a.s.l. These lakes were strategically selected to
cover a large geographic area with the aim of acquiring a large gradient
of salinity, and to include some of the larger and deeper lakes that are
found in the western and central parts of TP. Surface sediment samples
were collected from the deepest part (center) of each lake (Fig. 1) using a
Hydro-BIOS Bottom Sampler acc. to Ekman-Birge. The top 2 cm of
sediment was carefully sub-sampled and was used to represent modern
environments. Based on the few published 210Pb-dated sediment cores
from the TP region, including 39 lakes from Wang et al. (2020) and six
lakes from Yang and Turner (2013), our surface sediment samples are
estimated to represent a few years to twenty years. At each lake, water
samples were collected with a polyethylene bottle from a depth of 1-m at
the same time and location that the surface sediments were sampled. In
the field, water samples were stored in a cooler until sent to the Institute
of Tibetan Plateau Research, Chinese Academy of Sciences for analyses.
Major cations (Na+, K+, Mg2+, Ca2+) and anions (Cl− , SO2− 4 ) were
measured using a Dionex-ICS 900 Ion Chromatography System. HCO–3
concentrations were estimated from the ion balance of major cations and
anions (Murakami et al., 2007). Water temperature, maximum lake
depth, pH, turbidity, chlorophyll a, and salinity were measured in the
field using a Yellow Springs Instrument (YSI) EXO2 multiparameter
sonde (YSI Incorporated, Yellow Springs, OH, USA).
For the purposes of this study, the TP region was divided into four
geographic categories (Fig. 1) based on atmospheric circulation and
moisture conditions (Chen et al., 2010), permafrost development (Wang
et al., 2019), and lake salinity. The western (W) TP is dominated by
westerly winds (Westerlies) (An et al., 2012), and the eastern (E) TP is
dominated by the Asian Summer Monsoon. The boundary between the
Northern (N) TP and Southern (S) TP was based on the modern position
S. Yu et al. Ecological Indicators 155 (2023) 110952

Fig. 1. Map showing the location of the 45 study lakes in the Tibetan Plateau (TP) relative to the Westerlies and Indian Summer Monsoon (ISM). The study lakes are
categorized into four geographic regions within the TP (W, N, S, E) based on differences in atmospheric circulation and permafrost development (from Chen et al.,
2010; Wang et al., 2019) and are delineated by the blue dashed lines. (For interpretation of the references to colour in this figure legend, the reader is referred to the
web version of this article.)

of the Asian Summer Monsoon limit (Chen et al., 2010), the spatial
distribution of permafrost development (Wang et al., 2019), and lake
salinity concentrations (the salinity range of lakes in the S region
(average salinity: 2.4 g/L) is generally smaller than that of lakes in the N
region (average salinity: 23.5 g/L) (Fig. 2).
3.2. Diatom analysis
Diatom samples were prepared using standard preparation methods
at the Paleoecological Environmental Research Laboratory (PEARL),
Queen’s University (Canada) that closely followed Pienitz and Smol
(1993). Briefly, approximately 0.05 g of freeze-dried sediment from each
interval was placed into glass scintillation vials and 15 mL of a 50:50 M
ratio of concentrated sulfuric (H2SO4) and nitric (HNO3) acid was added
to digest the organic material. The samples were placed into a hot water
bath and heated to ~ 80 ◦ C for at least 2 h and then left to settle for
approximately 24 h. The supernatant was removed, and the slurries
were then rinsed with distilled water. This process was repeated until a
litmus test indicated that the samples were no longer acidic. The diatom
slurries were strewn onto coverslips at four different dilutions and
allowed to evaporate before being mounted onto glass microscope slides
with Naphrax (refractive index of 1.74). For each sample, at least 300
diatom valves were counted using a Leica DMR HC light microscope
using oil immersion at a magnification of 1000 X and fitted with dif­
ferential interference contrast optics. Diatom identifications were made
to the lowest taxonomic level possible (often variety) using a selection of
taxonomic sources, mainly Krammer and Lange-Bertalot (1986, 1988,
1991a, 1991b), Algaebase (Guiry and Guiry, 2023: algaebase.org) and
the Diatoms of North America online source (diatoms.org). Diatom
counts were expressed as percent relative abundances.
3.3. Numerical analyses
Prior to numerical analyses (with the exception of diatom richness
and diversity), diatom species relative abundances were square-root
transformed to downweight the influence of the dominant taxa (Leg­
endre and Gallagher, 2001). The physical, chemical and geographical
environmental variables were tested for normality using the Shapiro-
Wilk test (Shapiro and Wilk, 1965; R package ‘vegan’ v2.5–7, Oksanen
et al., 2020) (except for pH as this was already on a logarithmic scale)
and, where needed, were log10(x + 1) transformed to correct skewness.
To examine the relationships among environmental variables, a Pearson
correlation matrix was examined and significantly correlated variables
(p < 0.05, p < 0.01, and p < 0.001) were identified using the R software

3
S. Yu et al.
Fig. 2. Comparison of frequency distributions (box and whiskers plots) of key measured variables in the 45 Tibetan Plateau study lakes including: a) altitude, b)
maximum lake depth, c) lake temperature, d) pH, e) chlorophyll a concentration, and f) salinity across the four geographical regions. In each box plot, the vertical
lines represent the range in measured values and the horizontal lines depict the median values.
(R Core Team, 2018) and plotted using corrplot. Spatial variations of the
diatom diversity measurements (richness) made from the different re­
gions in our lakeset (i.e., S, N, W and E regions of the TP) were explored
using the Shannon Index and implemented using Past software (Hammer
et al., 2001).
All ordination analyses including principal component analysis
(PCA), detrended correspondence analysis (DCA), detrended canonical
correspondence analysis (DCCA), and canonical correspondence anal­
ysis (CCA) were performed with CANOCO version 5.0 (Ter Braak and
Šmilauer, 2012) and were based on diatom taxa that had a relative
abundance >1% and occurred in at least two lakes (total of 125 diatom
taxa). PCA was used to explore the relationships between environmental
variables and to identify important environmental gradients in the TP
lakes. DCA using the options of detrending by segments, nonlinear
rescaling, and no downweighting of rare taxa was used to determine
whether linear (RDA) or unimodal (CCA) direct ordination analysis
should be used. A DCA gradient length >4 SD units suggests a unimodal
model is most appropriate (Birks, 1995; Ter Braak and Šmilauer, 2012)
and because the gradient length for the TP lakes was 5.6 SD units, a CCA
was used to explore the relationships among diatom taxa and measured
environmental variables.
To identify variables that displayed multiple correlations with other
variables, CCA was used to examine high variance inflation factors
(VIFs): environmental variables with VIFs >10 were eliminated in
sequence, starting with the variable having the highest inflation factor in
4
Ecological Indicators 155 (2023) 110952
the CCA (Ter Braak and Prentice, 2004). To determine whether any of
the 45 TP lakes were outliers in terms of both environmental variables
and diatom species composition, PCA axes 1 and 2 samples scores
(environmental variables) and DCA axes 1 and 2 (species scores) were
examined. If the sample scores for both axes of a given lake were outside
one standard deviation of the mean sample scores of the lakeset in both
PCA and DCA (Fig. S1), that lake would be considered an outlier (Hall
and Smol, 1992). Based on these tests, Mang Co (located far from the
other lakes) was considered a statistical outlier, and therefore model
development was based on the remaining 44 TP lakes.
A CCA with forward selection (Ter Braak and Šmilauer, 2012) was
performed, together with a Monte Carlo permutation test, to identify
which of the remaining variables explained a statistically significant
amount of variation in the diatom data. To explore relationships among
environmental variables and diatom taxa, a final CCA was performed
with the variables that were identified as being significant run actively,
and additional variables of interest that were not deemed significant,
run passively in the ordination.
To determine the species gradient length for the forward selected
variable of interest (e.g., salinity), a detrended CCA with salinity was
used to determine its ability to maximize the dispersion of diatom scores
(Ter Braak and Šmilauer, 2012). To assess whether the forward selected
variables were statistically significant, a CCA constrained to that vari­
able (i.e., sole explanatory variable) together with a Monte Carlo per­
mutation (999 permutations, p > 0.05) was examined (Ter Braak and
S. Yu et al. Ecological Indicators 155 (2023) 110952

Šmilauer, 2012). A variety of inference models were explored for
reconstructing salinity based on modern diatom distributions across the
measured salinity gradient including Weighted Averaging (WA), Locally
Weighted Weighted Averaging (LWWA), Maximum Likelihood (ML),
Partial Least Squares (PLS) and Weighted Averaging Partial Least
Squares (WAPLS). Jackknifed cross-validation procedures were used to
estimate prediction errors and test the predictive ability of different
transfer functions (Birks, 1995). For WAPLS model options, each new
component was tested and retained only if it resulted in at least a 5%
reduction in the prediction error (in cross-validation) of the root mean
square error of prediction (RMSEP) (Birks et al., 1998). WA and WAPLS
regression were used to estimate the relationship of each taxon in the
calibration set to measured salinity providing estimates of species op­
tima. The reliability of the salinity inference model as a predictor of
diatom assemblage composition was assessed by examining the ratio
(λ1/λ2) of the constrained eigenvalue (λ1) to the first unconstrained
eigenvalue (λ2) of a DCCA with salinity included as the sole predictive
variable (Ter Braak, 1992). Estimated salinity optima were shown only
for diatom taxa that had an effective number of occurrences (Hill’s N2)
> 4 because taxa that are not well represented in the calibration set will
not have reliable optima estimates (Hill, 1973; Cumming and Smol,
1993). Optima and tolerances of diatom taxa and transfer functions were
determined using the software program C2 version 1.5 (Juggins, 2007).

Fig. 3. Piper diagram showing cation and anion concentrations of the 45 TP lakes, categorized by geographic region (colors of circles). The lower left ternary
diagram includes the concentrations of sodium, calcium, and magnesium, and the lower right ternary diagram shows cation concentrations of carbonate sulfate,
bicarbonate, and chloride. The diamond plot illustrates lake types based on the combined cation and anion concentrations.

5
S. Yu et al.
4. Results
4.1. Trends in environmental variables
The 45 study lakes encompass a broad range in physical character­
istics including maximum lake depth (0.5 m to 109.6 m, median = 13 m)
(Fig. 2b) and lake surface area (LSA) (3.1 km2 to 2420 km2, median =
44 km2) (Table S1). There were 19 lakes (<10 m, median = 1 m), 21
lakes between 10 m and 50 m (median = 25 m), and 5 lakes that were
deeper than 50 m (median = 83.5 m) (Table S1). The relatively large
altitudinal gradient of this study (4200 m a.s.l. to 5200 m a.s.l.: Fig. 2a)
included lakes situated both above and below the timberline. Lakes in
the W region spanned the entire altitudinal range sampled in the study
with the highest median elevation (4900 m a.s.l: Fig. 2a), whereas lakes
in the S and N regions were generally located at lower and higher alti­
tudes (median elevation of 4636 m a.s.l, and 4900 m a.s.l., respectively)
(Fig. 2a). In general, the TP lakes were alkaline (median pH = 9.1, range
= 7 to 10.5) with a similar median value in the W, S and N regions
(Fig. 2d). Water temperature was highly variable ranging from 0 ◦ C to
20.1 ◦ C, with the greatest range in the N region (Fig. 2c). Measured
chlorophyll a concentrations were similar in lakes across geographic
regions with a few exceptions that were higher, ranging from 0.3 µg/L to
10.5 µg/L (median = 2 µg/L; Fig. 2e).
One of the goals of this study was to examine salinity across the TP
and therefore our sampling strategy aimed to target lakes along a
gradient of salinity concentrations. The study lakes spanned a range in
salinity from 0.1 g/L to 85.3 g/L, although most lakes were below 40
mg/L (median = 15 mg/L) (Fig. 2f). Lakes in the N region had the widest
range in salinity with most lakes having concentrations>3 g/L. In
contrast, the majority of lakes in the W and S regions had salinity con­
centrations lower than 2 g/L with notably narrower ranges than lakes in
the W region (Fig. 2f). Based on commonly applied salinity categories
(Hammer, 1986; Meybeck, 1995), the 45 lakes in this study would be
classified as follows: 21 lakes were freshwater (<1 g/L), 6 lakes were
subsaline (1–3 g/L), 13 lakes were hyposaline (3–20 g/L), and 5 lakes
were meso-hypersaline (>20 g/L). The majority of study lakes in the W
and S geographic regions, as well as the one lake in the E region, were
subsaline to hyposaline (Fig. 2f). In contrast, the lakes in the N region
(with the exception of Taiyang Lake) are closed basin systems and were
meso-hypersaline with higher concentrations of salinity (Fig. 2f) and
major ions (Fig. 3).
The concentrations of individual ions measured in the 45 TP lakes
(Table S2), indicated that sodium, chloride, and sulfate were prevalent
anions in samples from the majority of sampling sites. As shown in the
Piper diagram (Fig. 3), the 45 lakes can be divided into lakes charac­
terized by: (Type A) Ca2+-Mg2+-SO2− 2+
4 ; (Type B) Ca –HCO3 ; (Type C),
−
Na+-K+- Cl− ; and (Type D) Na+–HCO−3 . The majority of our TP sites can
be characterized as Type C (Na+-K+- Cl− ), and Type D (Na+–HCO−3 )
lakes (Fig. 3), indicating that most lakes are rich in carbonate and
chloride. The hydrochemical characteristics of the study lakes can be
categorized along a general trend from freshwater to saltwater to dry
salt-lake conditions, following the sequence from carbonate to sulfate,
magnesium, and sulfate, and finally to chloride (Wang et al., 2013).
Lakes in the N region were characterized by Na+-K+- Cl− and lakes in the
S region by Na+–HCO−3 (Fig. 3). Lakes in the W region were character­
ized by Ca2+–HCO−3 or Na+–HCO−3 . The one lake representing the E
region (Lake Mang Co) was dominated by Ca2+-Mg2+-SO2− 4 (Fig. 3).
A Pearson correlation matrix identified several significant correla­
tions among the chemical, physical, and geographical variables
measured in this study (Table 1). As expected, strong positive relation­
ships (p < 0.001) were observed between total salinity and the dominant
ions (Na+, K+, Mg2+, Cl− , and SO2− 4 , Table 1). Latitude and altitude had
a strong, positive correlation (r = 0.554, p < 0.001), altitude was
negatively associated with temperature (r = -0.513, p < 0.001) and
water depth (r = -0.328, p < 0.05) (Table 1), whereas water depth had a
strong positive relationship with lake area (r = 0.614, p < 0.001)
6
Ecological Indicators 155 (2023) 110952
(Table 1).
PCA analyses revealed that the PCA axis 1 (λ1 = 0.48) and PCA axis 2
(λ2 = 0.13) captured 61% of the variation in the environmental data
(Fig. 4). The strongest direction of variation (PCA axis 1) was a gradient
of major ions (Na+, K+, Mg2+, Cl− , and SO2− 4 ) and the second environ­
mental gradient (PCA axis 2) was characterized by positive correlations
with latitude and altitude and by negative correlations with HCO–3, pH
and maximum lake depth (Fig. 4). Lakes located in the N region (with
the exception of sites 33 and 44) tended to group together on the left side
of axis 1 indicating higher concentrations of salinity, Na+, K+, Mg2+,
Cl− , and SO2-
4 (Fig. 4), in contrast to the majority of lower salinity lakes
in the W region that grouped on the right side of PCA axis 1 (Fig. 4).
Diatom trends and relationships with environmental variables and
geographic regions
A total of 142 diatom taxa representing 48 genera were identified in
the 45 TP lakes of which 125 taxa (relative abundance of at least 1% in 2
lakes) were retained for ordination analysis and salinity transfer func­
tion development. Taxonomic trends across geographic regions were
examined for 37 of the most common diatom taxa that had relative
abundances > 15% and each of the three most common taxa had a high
optimum in different lake types (Fig. 5). Spatial patterns of the relative
abundances of the three most commonly occurring diatom taxa observed
in our lakes (Pantocsekiella.
ocellata, Staurosira venter, and Nitzschia palea) were examined across
measured gradients of altitude, maximum lake depth, water tempera­
ture, pH, and chlorophyll a (Fig. 6). The WA estimates of salinity optima
were only reported for 35 diatom taxa that had a Hill’s N2 > 4 (Table 6).
The majority of diatom taxa in the surface sediments of the 45 TP
lakes were benthic, including Navicula sensu lato (s.l.) spp., Nitzschia spp.
(e.g., N. fonticola, N. frustulum, and N. palea), Tryblionella spp.
(T. hungarica, T. angustata, and T. constricta), Staurosira spp. (Staurosira.
construens, Staurosira venter), and a variety of epiphytic taxa (e.g., Coc­
coneis, Amphora, Epithemia, Surirella, Achnanthidium (s.l.) spp., Masto­
gloia smithii and Karayevia clevei) (Fig. 5). Planktonic diatoms were
mainly represented by Pantocsekiella ocellata, Cyclotella distinguenda,
Lindavia affinis, Pantocsekiella tripartita, Lindavia bodanica and Stepha­
nodiscus medius. Lakes in the W region had diatom assemblages that were
more commonly comprised of Cocconeis lineata, Gyrosigma acuminatum,
Stephanodiscus medius and Denticula kuetzingii, often in high abundances
in lakes with salinities below 1 g/L (Fig. 5). Diatom assemblages from
lakes in the S region, where salinities were below 6 g/L, were commonly
comprised of Amphora (s.l.) spp. (e.g., A. ovalis, A. pediculus, A. libyca)
and a variety of Lindavia spp. (e.g., L. affinis, L. radiosa, and L. bodanica)
(Fig. 5). Lakes in the N region where the highest salinity concentrations
were measured (median = 12.5 g/L, maximum = 85.3 g/L) had higher
relative abundances of Anomoeoneis costata, Nitzschia palea, Tryblionella
hungarica, Cyclotella distinguenda and S. venter (Fig. 5). Cyclotella dis­
tinguenda was observed in only two lakes in the N region (NM4: salinity
= 85.3 g/L and Longwe Co: salinity = 37 g/L), whereas Cyclostephanos
dubius was only observed in Mang Co, the one lake representing the E
region (salinity = 0.2 g/L), where it dominated (>88% relative abun­
dance) the assemblage (Fig. 5).
The three most common diatom taxa observed in the 45 TP lakes,
P. ocellata, N. palea, and S. venter, often showed distinct trends across
environmental gradients and across geographic regions (Fig. 6).
P. ocellata was the most common and abundant taxon in this lake-set and
was observed in 16 of the 45 lakes (Fig. 6a). This planktonic taxon was
observed in high relative abundance in the N, S, and W geographic re­
gions across a broad gradient of altitude (Fig. 6ai), water temperature
(Fig. 6aiii) and pH (Fig. 6aiv). Although this taxon occurred in both
shallow and deep lakes, it had distinctly higher relative abundances at
maximum lake depths greater than ~ 10 m, particularly in the deeper
lakes of the S region (Fig. 6aii). In addition, P. ocellata occurred in higher
relative abundances in lakes with chlorophyll a below 3.5 µg/L
 S. Yu et al.
Table 1
Pearson’s correlation matrix of measured environmental variables in the 45 TP lakes: latitude (o), longitude (o), altitude (m), lake area (km2), salinity (g/L), water depth (m), lake temperature (oC), pH (unit), chlorophyll a
(μg/L), Na+ (mg/L), K+ (mg/L), Mg2+ (mg/L), Ca2+ (mg/L), Cl− (mg/L), SO2− 4 (mg/L) and HCO3 (mg/L). One asterisk, two asterisks, and three asterisks indicate 0.05, 0.01, and 0.001 significance levels, respectively.
–

Latitude Longitude Altitude Area Salinity Depth Temperature pH Chlorophyll a Na+ K+ Mg2+ Ca2+ Cl− SO2−
4 HCO–3

Latitude 1 − 0.45*** 0.55 − 0.24 0.05 − 0.34 * − 0.42 − 0.33 0.03 0.01 − 0.03 − 0.02 − 0.05 0.15 − 0.15 − 0.23
*** ** *
Longitude 1 − 0.25 0.34 0.15 0.31 − 0.11 − 0.26 − 0.20 − 0.05 − 0.03 0.02 − 0.02 − 0.02 0.01 − 0.02
*
Altitude 1 − 0.25 − 0.01 − 0.33 − 0.51*** − 0.13 0.02 0.08 0.08 − 0.04 0.16 0.09 0.04 − 0.06
*
Area 1 0.18 0.61 0.17 − 0.02 − 0.17 0.02 0.1 0.03 0.15 0.07 0.09 0.05
***
Salinity 1 0.04 − 0.07 0.21 0.08 0.88 0.81 0.70 − 0.01 0.82 0.71 0.55
*** *** *** *** ***
Depth 1 0.02 − 0.16 − 0.18 − 0.09 − 0.07 − 0.03 − 0.01 − 0.02 0.01 0.08
Temperature 1 0.20 − 0.06 − 0.07 − 0.03 − 0.11 − 0.08 − 0.12 0.03 − 0.08
7

pH 1 0.32 0.36* 0.38 0.27 − 0.12 0.27 0.42 0.43

* * ** **
Chlorophyll a 1 0.16 0.11 0.03 0.28 0.11 0.08 0.20
Na+ 1 0.93 0.75 0.16 0.83 0.84 0.56
*** *** *** *** ***
K+ 1 0.79 0.13 0.79 0.82 0.55
*** *** *** ***
Mg2+ 1 0.12 0.81 0.62 0.32
*** ***
2+
Ca 1 0.08 0.14 − 0.1
Cl− 1 0.71 0.28
***
SO42− 1 0.43
**
HCO–3 1

Ecological Indicators 155 (2023) 110952

S. Yu et al. Ecological Indicators 155 (2023) 110952

Fig. 4. Principal component analysis (PCA) of physical, chemical, and geographical variables from the 45 TP study lakes, categorized by geographic region (colored
circles). The corresponding lake name for the numbers beside each circle in the figure can be found in Table S2 in the Supplemental material.

(Fig. 6av) and with salinity concentrations below 5 g/L (Fig. 6avi).
Benthic S. venter occurred at highest relative abundances in S and N
region lakes located at altitudes between ~ 4600 m a.sl. to 4900 m a.s.l.
(Fig. 6bi) and in shallower N, S, and W region lakes below ~ 3 m
maximum depth (Fig. 6bii). This taxon was observed across the full
water temperature and pH gradients but was in highest abundances in
warmer and alkaline S region lakes (Fig. 6aii and 6aiv). S. venter was
most prevalent in S region lakes with chlorophyll a below ~ 3.5 µg/L
(Fig. 6bv) and salinity below 2.0 g/L (Fig. 6bvi). Nitzschia palea occurred
in high relative abundances (>10%) over a broad range of altitudes in

8
S. Yu et al.
Fig. 5. Relative abundances of 37 dominant (maximum relative abundance > 15% in at least 2 lakes, and their salinity optima were in the different lake types.)
diatom taxa in the 45 TP lake calibration set, with different colors depicting geographic categorizations. The lakes were organized by region, and lakes within each
region were ranked from low to high salinity. The relative abundances of diatoms in region W, S, N, E are shown with green, pink, orange and red bars, respectively.
(For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
the N, S, and W regions (Fig. 6ci) and over a range of lake depths from
relatively deep to shallow lakes (3.4 to 40 m) (Fig. 6cii). This taxon was
observed in highest relative abundances (>20%) in warmer waters from
both N and S region lakes (Fig. 6ciii) and in N, S, and W lakes across a
broad range of pH (Fig. 6civ). It occurred in highest relative abundances
(>10%) across a relatively wide gradient of chlorophyll a in both N and
S region lakes (Fig. 6cv) and across a wide gradient of salinity from
freshwater to 85.3 g/L in the N, S and W region (Fig. 6cvi).
Diatom richness and diversity showed distinct spatial patterns
among the lakes across the W, N, and S regions (Fig. 7). In general, lakes
with salinity concentrations between 0.1 and 10 g/L had higher diatom
diversity and richness than lakes with higher salinities (Fig. 7a,b). Lakes
with salinities >20 g/L were mainly located in the N region and were
characterized by relatively low biodiversity and richness compared with
other regions (Fig. 7). The S region had the largest range of diatom di­
versity and richness.
4.2. Ordination
Ordination analyses were performed on 125 diatom taxa (1% in 2
lakes) from the TP calibration set. Further data screening included an
examination of variance inflation factors (VIFs) through a series of CCAs
that identified Na+ and K+ as having multiple correlations (VIFs > 10)
and were therefore eliminated from further analyses. Forward selection
in CCA was performed on the remaining variables from this lake set,
which identified salinity and maximum lake depth as being significant
(Monte Carlo permutation test, p < 0.001) and explaining the highest
variation in diatom assemblage composition (4.2% and 3.7%, respec­
tively) (Table 2).
The relationships between diatom species and environmental vari­
ables were explored by running a CCA with salinity and maximum water
depth as active variables, and a suite of variables of interest that were
not found to be significant as passive variables in the ordination (i.e. pH,
chlorophyll a, longitude, altitude, lake area, HCO–3, SO2− 2+
4 , Mg , Ca ,
2+
Cl ) (Fig. 8). The first four axes of the CCA explained 17.09% of the
−
9
Ecological Indicators 155 (2023) 110952
cumulative variance of diatom species (Table 3), with axis 1 repre­
senting a gradient of salinity (and related variables) and axis 2, a
gradient of maximum lake depth (Fig. 8). The CCA ordination plots show
a clear separation of the more saline, N region lakes (except for lake 36)
that plotted in the right, lower quadrant of the ordination. Lakes in the
W and S regions were lower in salinity and clustered on the left of axis 1,
and with a gradient of lake depth along axis 2 (Fig. 8A and 8C). Altitude
(run passively) was positively related to salinity, suggesting that higher
altitude lakes were generally higher in salinity. Salinity and correlated
variables, Mg2+, Cl− and SO2− 4 (run passively) were negatively related to
maximum water depth and plotted on the opposite sides of the CCA
ordination (Fig. 8C). Lake Mang Co (45), the only lake from the E region,
displayed a strong association with longitude and Ca2+ (Fig. 8A and 8C).
Nitzschia spp. (species code: 89, 91, 96, 99, 100), Fallacia pygmaea
(49), C. distinguenda (22), and Denticula subtilis (36) plot in the right
portion of the CCA plot (Fig. 8B), showing that these taxa commonly
occur in lakes with higher concentrations of salinity and major ions
(Fig. 8B). Gomphonema spp. (54, 58), Lindavia radiosa (63), P. ocellata
(24), and Pinnularia rhombarea (104) plot in the left side of the CCA
ordination space, indicating that they tend to occur in deeper lakes
(Fig. 8B). C. dubius (26), Eolimna sp. (42), and Gomphonema spp. (59, 60)
showed positive associations with deeper TP lakes with higher Ca2+
concentrations and located at higher longitudes. The relative abundance
of Gomphonema spp., Stauroneis livingstonii (110) and Stephanodiscus
spp., which characterized diatom assemblages in the S and W regions,
increased with increasing pH and bicarbonate (both run passively)
(Fig. 8).
4.3. Diatom-based salinity transfer function
Salinity model development was based on the modern diatom dis­
tributions of 125 taxa (1% in 2 lakes) across the measured salinity
gradient of 44 lakes (excluding lake Mang Co). The results of a PCA of
the environmental variables and a DCA of the species data identified
Mang Co as an outlier (Fig. S1) and therefore was not included in model
S. Yu et al. Ecological Indicators 155 (2023) 110952

Fig. 6. Responses of selected dominant diatom taxa (Pantocsekiella ocellata, Staurosira venter and Nitzschia palea) with the highest occurrence in 45 lakes with the
different regions related to altitude, lake depth, temperature, pH, chlorophyll a and lakewater salinity. The various categories from freshwater to hypersaline are
delineated by the dashed vertical lines.

10
S. Yu et al. Ecological Indicators 155 (2023) 110952

models showed the strongest predictive abilities. The performance sta­

Fig. 7. Distribution of diatom (a) taxon richness and (b) Shannon’s diversity in
the 45 TP lakes.
tistics of WA-PLS with one component (WAPLS-1) were similar to WA-
PLS with additional components (Table 5) and therefore did not show
a notable improvement over simple WA inference models. A WA model
with no tolerance down-weighting for both inverse and classical
deshrinking (R2jack = 0.71) outperformed WA models with tolerance
downweighting (R2jack = 0.49 and 0.50) (Table 5). Based on these com­
parisons of model performance, we chose a simple WA model with no
tolerance downweighting and with classical deshrinking (the mean and
maximum bias and RMSEP were slightly lower than the inverse
deshrinking model: Table 5). The salinity transfer function developed
from our TP lakes had a relatively high jackknifed coefficient of deter­
mination (R2jack = 0.71), and a relatively low RMSEPlog (0.28) (Fig. 9).
4.4. Species ecological optima and tolerances for salinity

The estimated salinity optima that form the basis of the WA-Cla
Table 2
model are included for the 35 most frequently occurring diatom taxa
Total variance explained and p-values for forward selected variables (salinity
in the TP calibration set (i.e., taxa that occurred at>1% relative
and maximum lake depth), that were actively included in the canonical corre­
spondence analysis (CCA) and that significantly and independently explained
diatom distributions in the 45-lake TP calibration set. Table 3
Eigenvalues, explained variation, species-environmental correlations, explained
Forward-selected environmental variable Variance explained (%) P-value
fitted variation for the first four canonical correspondence analysis (CCA) axes of
Salinity 4.2 0.001 the TP lakes calibration set, showing the total inertia and the proportion of the
Depth 3.7 0.001 total variation explained.
Statistic Axis 1 Axis 2 Axis 3 Axis 4 Value
development. The ratio of the eigenvalue for the first constrained DCCA Eigenvalues 0.56 0.42 0.40 0.37
axis to the eigenvalue for the second unconstrained DCCA axis for Explained variation (cumulative) 5.46 9.56 13.49 17.09
salinity (λ1/λ2 = 0.76; Table 4) suggests this variable has moderate Species-environmental 0.97 0.93 0.94 0.94
correlation
explanatory power for diatom distributions (Ter Braak, 1988) and is
Explained fitted variation 14.17 24.80 34.98 44.32
comparable to the value reported for salinity by Bloom et al. (2003) (λ1/ (cumulative)
λ2 = 0.67). Although a ratio below 1.0 suggests it is not the only envi­ Total Inertia CCA= 10.22
ronmental gradient that the diatom taxa are responding to (Juggins, Explanatory variables 38.6%
2013), it accounted for the highest variation in the diatom assemblage
composition in the TP calibration lakes (4.2 %; p < 0.05) and was
therefore explored for the development of a transfer function. Table 4
DCCA with the first axis constrained to salinity produced a gradient Results of the detrended canonical correspondence analysis
length of 5.9 SD, indicating that a unimodal inference model was (DCCA) and canonical correspondence analysis (CCA) con­
appropriate (Birks, 1995). A comparison of the performance statistics of strained to salinity for diatom taxa of the 45 TP lakes.
a variety of salinity inference models developed from our TP calibration Environmental variable Salinity
set indicated that simple weighted average (WA) models or models Gradient length (SD units) 5.9
based on WA partial least squares (WA-PLS) outperformed more com­ DCCA axis 1 λ1 0.44
plex models including locally weighted WA (LWWA), maximum likeli­ λ1/λ2 0.76
hood (ML), and PLS models (Table 5). PLS and ML models had weaker P-value 0.002
Variance explained (%) 10.22
jackknifed correlations between the observed diatom-inferred salinity
CCA axis 1 λ1 0.43
measurements and showed larger maximum bias, whereas the LWWA P-value 0.001
model had a larger average bias (Table 5). The WA and WA-PLS salinity

Fig. 8. Canonical correspondence analysis (CCA) biplots of (A) sampling sites categorized by geographic region (colored circles for W, S, N and E), (B) diatom taxa,
and (C) environmental variables in the 45 TP lakes. Table S1 provides corresponding lake names to the numbers in (A) and Table S3 provides corresponding diatom
taxonomic names to the numbers in (B). Solid lines with arrows represent forward-selected environmental variables, and dashed lines with arrows represent
environmental variables that were plotted passively in the ordination.

11
S. Yu et al. Ecological Indicators 155 (2023) 110952

Table 5
Performance statistics (R2, jackknifed R2 (R2jack) and root-mean-squared-error (RMSE) and RMSE of prediction (RMSEP) for weighted averaging (WA), locally weighted
weighted averaging (LWWA), maximum likelihood (ML), partial least squares (PLS) and weighted average partial least squares (WAPLS)-based diatom-salinity
inference models for 44 lakes on the Tibetan Plateau. The cross-validation of leave-one-out (jackknifing) was applied.
Model R2 RMSE (log10(Salinity + 1)) R2jack Average Bias Max Bias RMSEP (log10(Salinity + 1))

WA_Inv 0.90 0.16 0.71 0.03 0.33 0.29

WA_Cla 0.90 0.17 0.71 0.03 0.32 0.28
WATOL_Inv 0.87 0.19 0.49 0.1 1.05 0.39
WATOL_Cla 0.87 0.20 0.50 0.11 1.06 0.39
WAPLS-1 0.90 0.16 0.72 0.02 0.33 0.29
WAPLS-2 0.97 0.09 0.72 0.02 0.43 0.28
WAPLS-3 0.99 0.06 0.72 0.03 0.42 0.28
WAPLS-4 0.99 0.04 0.72 0.03 0.43 0.28
WAPLS-5 1 0.03 0.71 0.03 0.39 0.28
PLS-1 0.71 0.28 0.48 0.0006 0.70 0.38
PLS-2 0.89 0.17 0.61 − 0.008 0.63 0.33
PLS-3 0.94 0.13 0.64 0.002 0.64 0.31
PLS-4 0.97 0.09 0.63 0.002 0.49 0.32
PLS-5 0.99 0.06 0.6 0.006 0.42 0.33
ML 0.83 0.23 0.48 0.01 0.60 0.31
LWWA_Inv 0.92 0.15 0.7 0.27 0.63 0.30
LWWA_Cla 0.92 0.15 0.7 0.18 0.57 0.29

WA_Inv: WA (inverse deshrinking); WA_Cla: WA (classical deshrinking); WA_Inv: WA (tol. Downweighted inverse deshrinking); WATOL_Cla: WA (tol. Downweighted
classical deshrinking); LWWA_Inv: LWWA (inverse deshrinking); LWWA_Cla: LWWA (classical deshrinking).

Table 6
Tibetan Plateau diatom taxa that occurred with relative abundances>1% in at least two lakes in the 44 calibration lakes, species code, effective number of occurrences
(Hill’s N2 > 4), jackknifed WA_Cla (Weighted average with classical deshrinking) salinity optima (Jack_Opt, back-transformed salinity concentrations (g/L)), and
tolerances (Jack_Tol, back-transformed salinity concentrations (g/L)), max abundant (Max_Abund), mean abundant (Mean_Abund.), salinity category (Sa_categ) (F =
Freshwater lake; Sub = Subsaline lake; Hypo = Hyposaline lake; Meso-hype = Meso-hypersaline lake). The diatom taxa are ordered from low to high salinity optima.
NO. Diatom taxa Jack_Opt Jack_Tol Hill’s N2 Max_Abund. (%) Mean_Abund.(%) S_categ.

1 Navicula radiosa 0.44 0.73 5.89 14.49 0.77 F

2 Pinnularia viridiformis var. minor 0.50 0.81 6.85 8.24 0.6 F
3 Navicula cari 0.57 1.26 5.73 2.51 0.06 F
4 Amphora ovalis 0.68 0.71 8.41 29.34 1.61 F
5 Cymbella proxima 0.69 0.59 4.18 4.35 0.22 F
6 Staurosira venter 0.74 1.40 8.66 38.46 2.23 F
7 Cocconeis lineata 0.83 1.07 10.36 85.50 4.04 F
8 Mastogloia smithii 0.92 1.17 5.30 27.51 1.36 F
9 Epithemia sorex 0.95 0.79 5.63 4.76 0.3 F
10 Halamphora veneta 0.99 1.10 6.82 8.90 0.61 F
11 Gyrosigma acuminatum 1.00 1.46 10.12 46.01 1.95 F
12 Cymbella hantzschiana 1.07 2.44 5.11 9.49 0.47 Sub
13 Pantocsekiella ocellata 1.20 0.96 14.69 79.46 13.54 Sub
14 Encyonopsis microcephala 1.20 1.65 6.90 11.24 0.65 Sub
15 Navicula recens 1.26 1.56 4.19 11.49 0.51 Sub
16 Amphora pediculus 1.30 1.58 14.04 39.04 2.44 Sub
17 Pseudostaurosira brevistriata 1.46 0.82 4.79 2.97 0.23 Sub
18 Denticula kuetzingii 1.47 1.84 7.33 29.80 2.31 Sub
19 Encyonema silesiacum 1.48 1.21 5.92 11.96 0.48 Sub
20 Karayevia clevei 1.78 1.50 6.44 33.60 1.34 Sub
21 Caloneis bacillum 1.96 1.05 5.71 43.05 1.74 Sub
22 Staurosirella pinnata 1.97 1.69 9.80 10.14 0.82 Sub
23 Ulnaria ulna 2.00 1.78 10.35 13.90 0.98 Sub
24 Diploneis elliptica 2.00 1.38 7.84 9.24 0.65 Sub
25 Staurosira construens 2.12 1.14 9.11 8.46 0.91 Sub
26 Epithemia argus 2.51 1.48 4.38 60.26 1.65 Sub
27 Surirella crumena 2.68 0.96 7.01 21.43 1.09 Sub
28 Anomoeoneis costata 3.39 1.38 5.89 86.00 4.12 Hypo
29 Nitzschia fonticola 3.42 3.36 14.87 62.31 4.31 Hypo
30 Surirella ovalis 3.47 0.92 4.41 11.00 0.47 Hypo
31 Pantocsekiella tripartita 3.96 1.53 4.08 89.51 3.33 Hypo
32 Tryblionella hungarica 12.09 3.18 5.03 78.99 2.74 Hypo
33 Neidium ampliatum 13.61 6.44 4.52 8.88 0.43 Hypo
34 Nitzschia palea 13.69 3.19 8.48 79.47 6.28 Hypo
35 Navicula veneta 20.79 4.51 4.49 26.63 0.99 Meso-hype

abundance in at least two samples and that had a Hill’s N2 > 4) and are
presented in Table 6. The salinity optima for these common diatom taxa
ranged from 0.4 − 20.8 g/L, spanning a gradient from freshwater to
hyposaline conditions (Table 6). Of the 35 dominant diatoms, there were
eleven taxa with optima in the freshwater category (0.44 to 1.0 g/L),
sixteen with optima in the subsaline category (1.0 to 3.0 g/L), and seven
with optima in the hyposaline category (3.0 to 20.0 g/L) (Table 6). Only
one of the 35 dominant taxa, Navicula veneta, had a salinity optimum in
the meso-hypersaline range (>20.0 g/L) (Table 6). Three of the most
common and abundant taxa included Nitzschia palea (hyposaline opti­
mum of 13.7 g/L, mean relative abundance 6.3%, maximum relative
abundance 79.5%), P. ocellata (subsaline optimum of 1.2 g/L, mean

12
S. Yu et al.
Fig. 9. Plots of observed versus inferred log-transformed salinity (left) and the residuals (right) using the weighted averaging with classical deshrinking inference
model (WA-Cla).
relative abundance 13.5%, maximum relative abundance 79.5%), and
Amphora pediculus (subsaline optimum of 1.3 g/L, mean relative abun­
dance 2.5%, maximum relative abundance 39.0%).
5. Discussion
The climate of the Tibetan Plateau in central Asia is largely
controlled by Asian summer monsoons that produce strong gradients in
precipitation and moisture, resulting in lakes with a large range in
salinity (major ion) concentrations (Yang et al., 2003; Li et al., 2016).
Because diatoms respond sensitively to changes in salinity (Wilson et al.,
1996; Fritz et al., 1991), diatom-based salinity inference models have
the potential to be used to track past climate scenarios (Wilson et al.,
1996; Saunders, 2011; Wang et al., 2011). This is of particular interest in
the remote, high mountain lakes of the TP that are highly sensitive to
climate change.
5.1. General trends in environmental variables across Tibetan Plateau
lakes
Water chemistry and environmental variables examined across our
TP lakes generally reflect the large limnological diversity of lakes in this
high-altitude, heterogeneous region. The majority of our 45 TP lakes had
major ion concentrations that were substantially higher (by as much as
ten times) than what has been commonly reported in other regions of
China (Table S3). One of the clearest trends across our 45 lakes were the
large spatial differences in lake-water salinity, pH and major ion con­
centrations that generally increased with altitude (Tables S1 and S2). In
particular, the closed-basin lakes, which are most common to the N re­
gion (e.g., Tangra Yum Co, Nam Co), were chemically distinct and were
characterized by Na+-K+- Cl− (Fig. 3). The lack of inflow and outflow in
these hydrologically closed lakes make them susceptible to increased
evaporation and crystallization, greatly affecting their chemical
composition (Qiao et al., 2017; Wang et al., 2010). Therefore, closed
lakes in the N region are likely most susceptible to climate-related lake-
level changes. In contrast, the majority of lakes in the S and W regions
are hydrologically open with major ions dominated by Na+–HCO−3 and
Ca2+–HCO−3 , respectively. Water chemistry was distinct across the
various regions largely owing to differences in evaporation that would
affect ion concentrations (i.e. Na+, Cl− , Ca2+, Mg2+) and pH. For
example, the high evaporation rates of the W region resulted in high
Ca2+ concentrations precipitated from calcium carbonate. Differences in
pH among the lakes from the four different regions tended to increase
with latitude and is in agreement with previous limnological studies
13
Ecological Indicators 155 (2023) 110952
from the TP (Liu et al., 2021).
Lakes in the southern TP were generally warmer than lakes in the
northern TP, likely because they experience a shorter period of ice cover
(Pi et al., 2021). Chlorophyll a concentrations were generally similar
across the four regions, with a few exceptions within the S region where
concentrations in the two closed basin lakes (Angrenjin Co, Angzi Co)
were higher. Higher chlorophyll a concentrations in TP lakes were found
to be related to warmer conditions and shorter ice cover duration (Pi
et al., 2021). A longer and warmer growing season for algae and aquatic
plants would lead to higher primary production and chlorophyll a
concentrations. In addition to climate changes, higher chlorophyll a
concentrations in Angrenjin Co can also be explained by its proximity to
human activities in the northern part of Angren County.
5.2. Spatial distributions of diatom taxa and its environmental indication
The distribution of the main diatom taxa across the TP showed
distinct separation among the different geographic regions, particularly
between the W and N regions (Fig. 5). This clearly highlights the distinct
limnological differences among lakes in this heterogeneous mountain
landscape. The three most common and dominant diatom taxa in our
study (P. ocellata, S. venter, and N. palea) often showed clear preferences
for specific limnological conditions (Fig. 6). For example, P. ocellata
clearly dominated lakes in the S region and preferred low salinity, low
productivity, and relatively high pH conditions, but it also occurs in high
abundance across a range of elevations and water depths (Fig. 6a). This
planktonic diatom is one of the most commonly reported taxa in pale­
olimnological studies of the TP (e.g., Chen et al., 2021; Zhang et al.,
2023) and often dominated assemblages for the entire record (Wisch­
newski et al., 2014; Liao et al., 2020; Wang et al., 2020; He et al., 2022).
Although our study did not include lake-water nutrient measurements, it
is well-documented that P. ocellata prefers oligotrophic, non-acidified
conditions (Yang et al., 2003; Wang et al., 2014; Liao et al., 2020) and
based on our results, prefers to live in lakes with salinity concentrations
lower than 6.1 g/L (Fig. 6avi). Not surprisingly, benthic S. venter and
N. palea dominated the shallower lakes of the calibration set (Fig. 6bii,
6cii). Based on our results, N. palea had a clear preference for lakes with
a salinity higher than 8 g/L (Fig. 6cvi), whereas S. venter was usually
found in freshwater lakes (Fig. 6bvi).
The majority of surface sediment diatom assemblages in the 45 TP
lakes were dominated by a variety of benthic taxa, particularly in lakes
in the N and W regions, where lakes were generally shallower than lakes
in the S and E regions (Fig. 2b). In shallow lakes, light is sufficient to
allow aquatic plants and epiphytic diatoms to proliferate. Therefore,
S. Yu et al.
benthic taxa in the shallow W region lakes (i.e., Bangong Lake, Sumxi
Co, Hongshan Hu, Zongxiong Co) and N region lakes (i.e., Serling Co,
Longwe Co, Dong Co) were largely represented by epiphytic taxa such as
C. placentula var. lineata and G. acuminatum. Most of high salinity lakes
in the N region were dominated by benthic taxa, which can be explained
by high evaporation rates relative to low precipitation in this remote
inland region of the TP (Li et al., 2016).
Lakes located in the arid and semi-arid western and central TP
(Longwe Co, Dong Co and Serling Co) had distinct diatom assemblages
that appear to be mainly driven by salinity, major ions and water depth
(Fig. 8). These lakes tend to have higher salinity concentrations as they
are located far inland where the Himalayan mountains provide a barrier
from the ocean resulting in low precipitation (Liu et al., 2015). For
example, diatom assemblages in these higher salinity lakes were
comprised of a variety of brackish water taxa such as Anomoeoneis cos­
tata and Nitzschia palea. In meso-hypersaline lakes (e.g., Longwe Co,
Dong Co and Yagen Co), Nitzschia spp. and A. costata dominated the
assemblages. Our findings are in agreement with a previous study of
Serling Co that reported A. costata relative abundances were highest in
shallow lakes that are relatively high in specific conductance and
salinity (Yu et al., 2019). Therefore, specific diatom species are available
to assess the level of salinization (Stenger-Kovács et al., 2023).
Planktonic Cyclostephanos dubius was only observed in one of the 45
TP lakes (Mang Co, the only lake representing the E region) where it
dominated the assemblages. Cyclostephanos taxa are well-documented
indicators of eutrophic conditions (Bradshaw and Anderson, 2003;
Hausmann and Kienast 2006; Laperriere et al., 2008; Thienpont et al.,
2013) and, although lake-water nutrient concentrations were not
measured, the high relative abundances of C. dubius in Mang Co suggests
this lake is high in nutrients.
In general, the salinity of the southern lakes is significantly lower
than that of the northern lakes, and the southern lakes have a larger
gradient of diatom richness and diversity (Fig. 7a,b). This result suggests
that diatoms in the central TP lakes may have a latitudinal diversity
gradient (LDG). This observation is similar to that made by Hillebrand
and Azovski (2001) using 148 local and 25 regional freshwater diatom
data sets, which also showed that there was a significant diatom LDG.
5.3. Diatom-based salinity transfer function
Given the strong control that the East Asian and Indian summer
monsoons have on precipitation and moisture in the TP (An et al., 2012),
this region typically encompasses a large range in salinity from fresh­
water to hypersaline lakes (Liu et al., 2021; Wang et al., 2011). Diatom-
salinity transfer function studies typically aim to track changes in
climate over time, with the assumption that changes in precipitation and
effective moisture will be large enough to clearly alter lake water
salinity (Fritz et al., 2013) and would be registered in sedimentary
diatom records. In the arid TP region, where evaporative forcing is
strong, these conditions are easily met (Sun et al., 1990). In particular,
when evaporation is greater than precipitation, the numerous hydro­
logically closed lakes in the TP have great potential to concentrate dis­
solved salts (China salty lakes resources and environmental database,
2019).
Salinity was found to be important for explaining the distribution
and main variation in diatom assemblage composition in the TP cali­
bration lakes, a trend that has been well-documented for this high
mountain, monsoon-dominated region (Yang et al., 2003; Wang et al.,
2011; Li et al., 2021). Although salinity explained the greatest variance
in diatom assemblage composition in our TP lakes, the relative explan­
atory power of this variable to predict diatom assemblage composition
(λ1/λ2 of DCCA = 0.76) was moderate. Ter Braak (1988) suggested that
λ1/λ2 values should be at least 1.0 for reconstructions to be useful, and
following this rule, salinity was not the only variable determining
diatom distributions in our TP calibration lakes (Juggins, 2013). In
particular, diatom variation in our TP lakes was also influenced by
14
Ecological Indicators 155 (2023) 110952
differences in lake depth, the only other measured variable found to
significantly explain diatom distributions, although not correlated with
salinity (Fig. 8). In addition to the variables available for our study, it is
likely that some of the diatom variation can be explained by factors that
were not measured, such as nutrients (e.g., total phosphorus) and
climate-related changes to lake water properties. Other variables that
were not included in the ordination analysis such as hydrology (e.g.,
closed versus open-basin lakes) would also have an effect on salinity
where closed-basin systems tend to have higher salinity concentrations,
particularly when evaporation is greater than precipitation. Acknowl­
edging that diatom assemblages are likely also responding to other
variables in TP lakes, the development of a salinity transfer function is of
interest given the importance of this variable in characterizing these
lakes.
Simple weighted-averaging with classical deshrinking (WA-Cla) was
deemed to be the most robust salinity inference model for our TP lakes
(Table 5). As was found in our examination of performance statistics,
WA models often outperform other more complex calibration ap­
proaches and are therefore widely used to examine the relationship
between diatom taxa and the environmental variable of interest (Birks
et al., 1998; Birks and Simpson, 2013; Cumming et al., 2015). The choice
of simple WA regression for developing a salinity model is consistent
with previous calibration studies from the Interior Plateau of British
Columbia (Wilson et al. 1994), and in Arctic (Moser et al. 2000) and
Antarctic (Verleyen et al. 2003) regions. However, our TP calibration set
tended to have more lakes in the higher end of the salinity gradient (18
lakes>3 g/L) whereas in many diatom-salinity studies from other re­
gions, fewer lakes had salinity concentrations>3 g/L. For example,
among the 57 lakes in the Sierra Nevada (California, USA), there were
only 8 lakes with salinity concentrations>3 g/L (Bloom et al., 2003),
and among the 102 lakes in British Columbia (Canada), only 36 lakes
had salinity concentrations>3 g/L (Wilson et al., 1994).
Although the inferred versus observed salinity values based on
diatom composition for the WA-Cla model exhibits a relatively strong
relationship, the lakes in the TP dataset were not evenly distributed
across the salinity gradient (Fig. 9A). The majority of lakes had salinity
values at the lower end of the gradient and lakes with moderate salinity
concentrations were not well represented in the dataset as they were
rare and, therefore, not sampled. A comparison of the observed versus
predicted salinity values indicated that our WA-Cla model tends to over-
predict salinity in freshwater lakes (salinity<1.0 g/L (log-salinity <
0.3)) and under-predict salinity in lakes with concentrations>1.0 g/L
(log-salinity < 0.3). This may be because the 44 calibration lakes were
not evenly distributed across the salinity gradient, with fewer lakes
representing the meso-hyposaline (9–30 g/L) part of the gradient
(Fig. 9). The performance results of our WA-Cla diatom salinity model
(R2 = 0.9, R2jack = 0.71, RMSEPjack = 0.28, N = 44) were similar to a
diatom-salinity transfer function developed with a WAPLS model from
lakes in the eastern TP by Wang et al. (2011) (R2jack = 0.738, RMSEPjack
= 0.38, N = 90). The range in salinity measured from lakes in our study
(0.1 to 85.3 g/L; 18 of 44 lakes (41%) with salinity > 3 g/L) were similar
to lakes from the eastern TP (0.1 to 123 g/L; 38 of 90 lakes (42%) with
salinity > 3 g/L) but the majority of our study lakes were located at
higher elevations (30 of 44 lakes (68%) above 4600 m a.s.l.) than the
eastern TP lakes (27 of 90 lakes (30%)) were above 4600 m a.s.l.). The
lower number of high elevation sites is not surprising, given the diffi­
culty in sampling these altitudes in the western and central TP. In gen­
eral, the diatom-salinity transfer function obtained in this study would
be most applicable to the semiarid and arid regions of the TP. Increasing
the number of lakes in the meso-hyposaline range would be a step to­
wards improving diatom-based salinity inferences in the TP. Given that
sampling these lakes may prove logistically difficult, future studies
might consider merging existing TP calibration sets.
Salinity optima
Paleolimnological studies provide key information that enable as­
sessments on the extent of environmental change in the context of
S. Yu et al.
natural, background conditions (Smol and Cumming, 2000). To more
accurately interpret changes in diatom assemblage composition over
time, it is necessary to define the environmental optima of taxa,
particularly in understudied regions such as the TP where diatom-based
salinity studies are scarce. The estimated salinity optima of the 35 most
common diatom taxa in the TP lakes (Table 6) form the basis of the
simple WA_Cla model developed in this study. Categorizing salinity
preferences (based on optima) for these taxa may be useful for future
researchers. Although a few diatom-based salinity models have been
developed for the TP (Yang et al., 2004; Wang et al., 2011), this is a vast,
highly heterogeneous and complex mountain region that would require
additional calibration sets from various locations to provide a more
complete distribution of lakes across the salinity gradient. Information
on diatom distributions and autecological information will undoubtedly
be refined as more lakes are examined in this understudied high
mountain region.
The estimated salinity optima of the most common diatom taxa in
this study were compared to the salinity optima estimated from other
diatom-based studies (including outside of the TP region). However,
diatom taxa from the TP lakes were often particular to the region and
therefore not all taxa in Table 6 have been reported in the available
literature on diatom salinity model construction. In general, many of our
taxa had estimated salinity optima that were comparable to optima re­
ported by other diatom-based salinity studies. For example, the most
common planktonic taxon in our calibration set (P. ocellata) had a
salinity optimum of 1.2 g/L (subsaline waters) that was slightly higher
than the estimated optimum of 0.9 g/L (freshwater) from an eastern TP
calibration set (Wang et al., 2011). Ulnaria ulna had an optimum (2 g/L)
in the subsaline category that was comparable (although slightly higher)
to this taxon’s optimum estimated from other datasets from south-west
of Australia (1.5 g/L; Taukulis and John, 2009) and in the northern
Great Plains, USA (1.6 g/L; Fritz et al., 1993). Based on the estimated
optimum from our TP calibration set, Tryblionella hungarica, Nitzschia
palea, Neidium ampliatum, and A. costata prefer hyposaline lake condi­
tions (Table 6), which is consistent with other studies that likewise
found these taxa to occur in high salinity conditions (Martínez et al.,
2003). Navicula veneta, a species considered to be a salt generalist (Veres
et al., 1995) and shown to withstand meso-hyposaline conditions
(Blanco et al., 2013), had a salinity optimum of 20.8 g/L in the western
and central TP (Table 6). This taxon is usually associated with high
conductivity waters and has also been shown to dominate mesosaline
ponds (specific conductivity: 10.9–17.3 mS/cm) in the southern Iberian
Peninsula (Fernández-Moreno et al., 2022).
6. Conclusions
The 45-lake calibration set from the Tibetan Plateau provides
important information on spatial distributions and diatom-
environmental relationships in this climatically sensitive high moun­
tain region. Diatom assemblage distributions in our TP lakes were
strongly influenced by lakewater salinity. In particular, the N region
included lakes with relatively high salinity concentrations and had
distinct diatom assemblage composition. Of the most common diatom
taxa observed in our lakes, P. ocellata and S. venter were found in
freshwater lakes, whereas N. palea had highest abundances in hyposa­
line lakes. Although salinity had a strong influence on diatom assem­
blage composition, other variables including water depth also played a
role in structuring the diatom assemblages. A simple WA model resulted
in a relatively robust salinity transfer function for TP lakes based on
estimated diatom salinity optima that cover a large gradient including
freshwater (0.1–1.0 g/L), subsaline (1.0–3.0 g/L), hyposaline (3.0–20.0
g/L), and meso-hyposaline water (>20.0 g/L) lakes. This relatively large
gradient in salinity across the TP provides an opportunity to explore past
changes in lake levels using paleo-salinity reconstructions. However, the
salinity inference model will likely be improved by including a greater
number of lakes in the meso-hyposaline range as these were under-
15
Ecological Indicators 155 (2023) 110952
represented in the lake set. The spatial distribution of diatoms in lakes
across the TP together with the diatom-salinity transfer function could
be used to track past changes in salinity and environmental conditions
that may reveal long-term patterns in climate in this complex, high
mountain region.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data availability
Data will be made available on request.
Acknowledgements
This study was jointly funded by the Strategic Priority Research
Program of the Chinese Academy of Sciences (Grant No.
XDA2007010102), the Second Tibetan Plateau Scientific Expedition and
Research (STEP) (2019QZKK0202), the National Natural Sciences
Foundation of China (Grant No. 41877168), the Science and Technology
Department of Tibet (XZ202001YD0013C) and China Scholarship
Council (Grant No. 20204910715). We thank Qiangqiang Kou and Teng
Xu, Cunlin Li for helping with the sampling in the fieldwork. We also
thank Qingmin You and Yameng Li for helping with the diatom
identification.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.ecolind.2023.110952.
References
An, Z., Colman, S.M., Zhou, W., Li, X., Brown, E.T., Jull, A.J., Cai, Y., Huang, Y., Lu, X.,
Chang, H., Song, Y., Sun, Y., Xu, H., Liu, W., Jin, Z., Liu, X., Cheng, P., Liu, Y., Ai, L.,
Li, X., Liu, X., Yan, L., Shi, Z., Wang, X., Wu, F., Qiang, X., Dong, J., Lu, F., Xu, X.,
2012. Interplay between the Westerlies and Asian monsoon recorded in Lake
Qinghai sediments since 32 ka. Sci. Rep. 2, 619. https://doi.org/10.1038/
srep00619.
Anslan, S., Azizi Rad, M., Buckel, J., Echeverria Galindo, P., Kai, J., Kang, W., Keys, L.,
Maurischat, P., Nieberding, F., Reinosch, E., Tang, H., Tran, T.V., Wang, Y.,
Schwalb, A., 2020. Reviews and syntheses: How do abiotic and biotic processes
respond to climatic variations in the Nam Co catchment (Tibetan Plateau)?
Biogeosciences 17 (5), 1261–1279.
Battarbee, R.W., Anderson, N.J., Jeppesen, E., Leavitt, P.R., 2005. Combining
palaeolimnological and limnological approaches in assessing lake ecosystem
response to nutrient reduction. Freshw. Biol. 50, 1365–2427. https://doi.org/
10.1111/j.1365-2427.2005.01427.x.
Benito, X., Fritz, S.C., Steinitz-Kannan, M., Tapia, P.M., Kelly, M.A., Lowell, T.V., Gill, J.,
2018. Geo-climatic factors drive diatom community distribution in tropical South
American freshwaters. J. Ecol. 106 (4), 1660–1672.
Bennion, H., Fluin, J., Simpson, G.L., 2004. Assessing eutrophication and reference
conditions for Scottish freshwater lochs using subfossil diatoms. J. Appl. Ecol. 41,
124–138. https://doi.org/10.1111/j.1365-2664.2004.00874.x.
Birk, S., Chapman, D., Carvalho, L., Spears, B.M., Andersen, H.E., Argillier, C., Auer, S.,
Baattrup-Pedersen, A., Banin, L., Beklioğlu, M., Bondar-Kunze, E., Borja, A.,
Branco, P., Bucak, T., Buijse, A.D., Cardoso, A.C., Couture, R.M., Cremona, F., de
Zwart, D., Feld, C.K., Ferreira, M.T., Feuchtmayr, H., Gessner, M.O., Gieswein, A.,
Globevnik, L., Graeber, D., Graf, W., Gutiérrez-Cánovas, C., Hanganu, J., Işkın, U.,
Järvinen, M., Jeppesen, E., Kotamäki, N., Kuijper, M., Lemm, J.U., Lu, S., Solheim, A.
L., Mischke, U., Moe, S.J., Nõges, P., Nõges, T., Ormerod, S.J., Panagopoulos, Y.,
Phillips, G., Posthuma, L., Pouso, S., Prudhomme, C., Rankinen, K., Rasmussen, J.J.,
Richardson, J., Sagouis, A., Santos, J.M., Schäfer, R.B., Schinegger, R., Schmutz, S.,
Schneider, S.C., Schülting, L., Segurado, P., Stefanidis, K., Sures, B., Thackeray, S.J.,
Turunen, J., Uyarra, M.C., Venohr, M., von der Ohe, P.C., Willby, N., Hering, D.,
2020. Impacts of multiple stressors on freshwater biota across spatial scales and
ecosystems. Nat. Ecol. Evol. 4, 1060–1068. https://doi.org/10.1038/s41559-020-
1216-4.
Birks, H.J.B., Simpson, G.L., 2013. ‘Diatoms and pH reconstruction’ (1990) revisited.
J. Paleolimnol. 49, 363–371. https://doi.org/10.1007/s10933-013-9697-7.
S. Yu et al.
Birks, H.J.B., Frey, D.G., Deevey, E.S., 1998. Numerical tools in palaeolimnology –
Progress, potentialities, and problems. J. Paleolimnol. 20, 307–332. https://doi.org/
10.1023/A:1008038808690.
Birks, H.J.B., Heiri, O., Seppä, H., Bjune, A.E., 2010. Strengths and weaknesses of
quantitative climate reconstructions based on Late-Quaternary. The Open Ecology
Journal 3, 68–110. https://doi.org/10.2174/1874213001003020068.
Birks, H.J.B., 1995. Quantitative palaeoenvironmental reconstructions, in: Statistical
Modelling of Quaternary Science Data, Technical Guide 5, edited by: Maddy, D., and
Brew, J. S., Quaternary Research Association, Cambridge, 271 pp.
Blanco, S., Álvarez-Blanco, I., Cejudo-Figueiras, C., De Godos, I., Bécares, E., Muñoz, R.,
Guzman, H.O., Vargas, V.A., Soto, R., 2013. New diatom taxa from high-altitude
Andean saline lakes. Diatom Res. 28 (1), 13–27.
Bloom, A.M., Moser, K.A., Porinchu, D.F., MacDonald, G.M., 2003. Diatom-inference
models for surface-water temperature and salinity developed from a 57-lake
calibration set from the Sierra Nevada, California, USA. J. Paleolimnol. 29, 235–255.
https://doi.org/10.1023/A:1023297407233.
Bradshaw, E.G., Anderson, N.J., 2003. Environmental factors that control the abundance
of Cyclostephanos dubius (Bacillariophyceae) in Danish lakes, from seasonal to
century scale. Eur. J. Phycol. 38, 265–276. https://doi.org/10.1080/
0967026031000136349.
Chen, F.H., Chen, J.H., Holmes, J., Boomer, I., Austin, P., Gates, J.B., Wang, N.L.,
Brooks, S.J., Zhang, J.W., 2010. Moisture changes over the last millennium in arid
central Asia: a review, synthesis and comparison with monsoon region. Quat. Sci.
Rev. 29, 1055–1068. https://doi.org/10.1016/j.quascirev.2010.01.005.
Chen, J., Liu, J.B., Rühland, K.M., Smol, J.P., Zhang, X.S., Zhang, Z.P., Zhou, A.F.,
Shen, Z.W., Chen, F.H., 2021. Aquatic ecosystem responses to environmental and
climatic changes in NE China since the last deglaciation (~17, 500 cal yr BP) tracked
by diatom assemblages from Lake Moon. Quat. Sci. Rev. 272, 107218 https://doi.
org/10.1016/j.quascirev.2021.107218.
China salty lakes resources and environment database, 2019. Qinghai Institute of Salty
Lakes. Chinese Academy of Sciences, Xining, China.
Cumming, B.F., Smol, J.P., 1993. Development of diatom-based salinity models for
paleoclimatic research from lakes in British Columbia (Canada). In: van Dam, H.
(Ed.), Twelfth International Diatom Symposium. Springer Netherlands, Dordrecht,
pp. 179–196.
Cumming, B.F., Wilson, S.E., Hall, R.I., Smol, J.P., 1995. Diatoms from British Columbia
(Canada) lakes and their relationship to salinity, nutrients and other limnological
variables. In: Lange-Bertalot, H. (Ed.), Bibliotheca Diatomologica, Band 31. J.
Cramer, Stuttgart, p. 207.
Fernández-Moreno, D., Sánchez-Castillo, P.M., Delgado, C., Almeida, S.F.P., 2022.
Diatom species that characterize saline ponds (Southern Spain) with the description
of a new Navicula species. Wetlands 42, 14. https://doi.org/10.1007/s13157-021-
01529-z.
Fritz, S.C., Juggins, S., Battarbee, R.W., Engstrom, D.R., 1991. Reconstruction of past
changes in salinity and climate using a diatom-based transfer function. Nature 352,
706–708. https://doi.org/10.1038/352706a0.
Fritz, S.C., Juggins, S., Battarbee, R.W., 1993. Diatom assemblages and ionic
characterization of lakes of the northern Great Plains, North America: a tool for
reconstructing past salinity and climate fluctuations. Can. J. Fish. Aquat. Sci. 50 (9),
1844–1856. https://doi.org/10.1139/f93-20.
Fritz, S.C., Cumming, B.F., Gasse, F., Laird, K., 2010. Diatoms as indicators of hydrologic
and climate change in saline lakes. In: Smol, J.P., Stoermer, E.F. (Eds.), the Diatoms:
Applications for the Environmental and Earth Sciences, 2nd ed. Cambridge
University Press, Cambridge, pp. 186–208.
Fritz, S.C., 2013. Salinity and climate reconstructions from continental lakes.
Encyclopedia of Quaternary Science, 2nd ed. (Scott A. Elias and Cary J. Mock,
editors) pp. 507-515.
Gao, Y.C., Liu, M.F., 2013. Evaluation of high-resolution satellite precipitation products
using rain gauge observations over the Tibetan Plateau. Hydrol. Earth Syst. Sci. 17,
837–849. https://doi.org/10.5194/hess-17-837-2013.
Guiry, M.D., Guiry, G.M., 2023. AlgaeBase. World-wide electronic publication, National
University of Ireland, Galway.
Hall, R.I., Smol, J.P., 1992. A weighted-averaging regression and calibration model for
inferring total phosphorus concentration from diatoms in British Columbia (Canada)
lakes. Freshw. Biol. 27, 417–434. https://doi.org/10.1111/j.1365-2427.1992.
tb00551.x.
Hammer, U.T., 1986. Saline lake ecosystems of the world, Vol. 59. Springer Science &
Business Media.
Hammer, Ø., Harper, D.A., Ryan, P.D., 2001. PAST: Paleontological statistics software
package for education and data analysis. Palaeontol. Electron. 4, 9. http://palaeo-el
ectronica.org.
Hausmann, S., Kienast, F., 2006. A diatom-inference model for nutrients screened to
reduce the influence of background variables: application to varved sediments of
Greifensee and evaluation with measured data. Palaeogeogr. Palaeoclimatol.
Palaeoecol. 233 (1–2), 96–112. https://doi.org/10.1016/j.palaeo.2005.09.019.
He, J.N., Liu, J.B., Rühland, K.M., Zhang, J.F., Chen, Z.T., Dong, H.R., Smol, J.P., 2022.
Response of lake diatoms to rapid 21st century warming on the southeastern Tibetan
Plateau. Anthropocene 39, 100345. https://doi.org/10.1016/j.ancene.2022.100345.
Hill, M.O., 1973. Diversity and evenness: a unifying notation and its consequences.
Ecology 54, 427–432. https://doi.org/10.2307/1934352.
Hillebrand, H., Azovski, A.I., 2001. Body size determines the strength of the latitudinal
diversity gradient. Ecography 24. https://doi.org/10.1034/j1600-
0587.2001.240302.x.
Hou, X. (Ed.), 2001. Vegetation Atlas of China. Science Press, Beijing in Chinese.
16
Ecological Indicators 155 (2023) 110952
Imbrie, J., Kipp, N.G., 1971. A new micropaleontological method for quantitative
paleoclimatology: Application to a Late Pleistocene Caribbean core, The Late
Cenozoic Glacial Ages K. Yale Univ. Press, New Haven, Conn. Turekian, 71–181,.
Immerzeel, W.W., Van Beek, L.P.H., Bierkens, M.F.P., 2010. Climate change will affect
the Asian water towers. Science 328, 1382–1385. https://doi.org/10.1126/
science.1183188.
Immerzeel, W.W., Lutz, A.F., Andrade, M., Bahl, A., Biemans, H., Bolch, T., Hyde, S.,
Brumby, S., Davies, B.J., Elmore, A.C., Emmer, A., Feng, M., Fernández, A.,
Haritashya, U., Kargel, J.S., Koppes, M., Kraaijenbrink, P.D.A., Kulkarni, A.V.,
Mayewski, P.A., Nepal, S., Pacheco, P., Painter, T.H., Pellicciotti, F., Rajaram, H.,
Rupper, S., Sinisalo, A., Shrestha, A.B., Viviroli, D., Wada, Y., Xiao, C., Yao, T.,
Baillie, J.E.M., 2020. Importance and vulnerability of the world’s water towers.
Nature 577, 364. https://doi.org/10.1038/s41586-019-1822-y.
Institute of Geography, 1990. Map of the Qinghai-Tibetan Plateau. Science Press, Beijing,
pp. 102–112 in Chinese.
Juggins, S., 2007. C2, Software for ecological and palaeoecological data analysis and
visualisation, User guide Version 1.5. School of Geography. Politics & Sociology,
Newcastle University, Newcastle.
Juggins, S., 2013. Quantitative reconstructions in palaeolimnology: new paradigm or
sick science? Quat. Sci. Rev. 64, 20–32. https://doi.org/10.1016/j.
quascirev.2012.12.014.
Kärnä, O.M., Heino, J., Grönroos, M., Hjort, J., 2018. The added value of geodiversity
indices in explaining variation of stream macroinvertebrate diversity. Ecol. Ind. 94,
420–429. https://doi.org/10.1016/j.ecolind.2018.06.034.
Krammer, K., Lange-Bertalot, H., 1986,1988,1991a,1991b. Bacillariophyceae.
Süßwasserflora von Mitteleuropa. Gustav Fischer Verlag, Jena.
Laperriere, L., Fallu, M.A., Hausmann, S., Pienitz, R., Muir, D., 2008. Paleolimnological
evidence of mining and demographic impacts on Lac Dauriat, Schefferville (subarctic
Québec, Canada). J. Paleolimnol. 40, 309–324. https://doi.org/10.1007/s10933-
007-9162-6.
Legendre, P., Gallagher, E.D., 2001. Ecologically meaningful transformations for
ordination of species data. Oecologia 129 (2), 271–280. https://doi.org/10.1007/
s004420100716.
Li, Z.X., Gao, Y., Wang, S.Y., Lu, Y., Sun, K., Jia, J.J., Wang, Y.F., 2021. Phytoplankton
community response to nutrients along lake salinity and altitude gradients on the
Qinghai-Tibet Plateau. Ecol. Ind. 128, 107848 https://doi.org/10.1016/j.
ecolind.2021.107848.
Li, C.D., Kang, S.C., Liu, Y.Q., Hou, J.Z., Guo, J.M., Liu, X.B., Cong, Z.Y., Zhang, Q.G.,
2016. Distribution of major ions in waters and their response to regional climatic
change in Tibetan lakes. Journal of Lake Sciences 28 (4), 743–754. In Chinese with
English abstract) DOI: 10.18307/2016.0407.
Li, Y.L., Liu, E.F., Xiao, X.Y., Zhang, E.L., Ji, M., 2015. Diatom response to Asian
monsoon variability during the Holocene in a deep lake at the southeastern margin
of the Tibetan Plateau. Boreas 44, 785–793. https://doi.org/10.1111/bor.12128.
Liao, M.N., Herzschuh, U., Wang, Y.B., Liu, X.Q., Ni, J., Li, K., 2020. Lake diatom
response to climate change and sedimentary events on the southeastern Tibetan
Plateau during the last millennium. Quat. Sci. Rev. 241, 106409 https://doi.org/
10.1016/j.quascirev.2020.106409.
Liu, X.D., Guo, Q.C., Guo, Z.T., Yin, Z.Y., Dong, B.W., Smith, R., 2015. Where were the
monsoon regions and arid zones in Asia prior to the Tibetan Plateau uplift? Natl. Sci.
Rev. 2, 403–416. https://doi.org/10.1093/nsr/nwv068.
Liu, C., Zhu, L., Wang, J., Ju, J., Ma, Q., Qiao, B., Wang, Y., Xu, T., Chen, H., Kou, Q.,
Zhang, R., Kai, J., 2021. In-situ water quality investigation of the lakes on the
Tibetan Plateau. Science Bulletin 66 (17), 1727–1730.
Lu, H.Y., Wu, N.Q., Liu, K.B., Zhu, L.P., Yang, X.D., Yao, T.D., Wang, L., Li, Q., Liu, X.Q.,
Shen, C.M., Li, X.Q., Tong, G.B., Jiang, H., 2011. Modern pollen distributions in
Qinghai-Tibetan Plateau and the development of transfer functions for
reconstructing Holocene environmental changes. Quat. Sci. Rev. 30, 947–966.
https://doi.org/10.1016/j.quascirev.2011.01.008.
Ma, Q.F., Zhu, L.P., Lü, X.M., Wang, J.B., Ju, J.T., Kasper, T., Daut, G., Haberzettl, T.,
2019. Late glacial and Holocene vegetation and climate variations at Lake Tangra
Yumco, central Tibetan Plateau. Global Planet. Change 174, 16–25. https://doi.org/
10.1016/j.gloplacha.2019.01.004.
Maasri, A., Jähnig, S.C., Adamescu, M.C., Adrian, R., Baigun, C., Baird, D.J., Batista-
Morales, A., Bonada, N., Brown, L.E., Cai, Q., Campos-Silva, J.V., Clausnitzer, V.,
Contreras-MacBeath, T., Cooke, S.J., Datry, T., Delacámara, G., De Meester, L.,
Dijkstra, K.-D., Do, V.T., Domisch, S., Dudgeon, D., Erös, T., Freitag, H., Freyhof, J.,
Friedrich, J., Friedrichs-Manthey, M., Geist, J., Gessner, M.O., Goethals, P.,
Gollock, M., Gordon, C., Grossart, H.-P., Gulemvuga, G., Gutiérrez-Fonseca, P.E.,
Haase, P., Hering, D., Hahn, H.J., Hawkins, C.P., He, F., Heino, J., Hermoso, V.,
Hogan, Z., Hölker, F., Jeschke, J.M., Jiang, M., Johnson, R.K., Kalinkat, G.,
Karimov, B.K., Kasangaki, A., Kimirei, I.A., Kohlmann, B., Kuemmerlen, M.,
Kuiper, J.J., Kupilas, B., Langhans, S.D., Lansdown, R., Leese, F., Magbanua, F.S.,
Matsuzaki, S.-I., Monaghan, M.T., Mumladze, L., Muzon, J., Mvogo Ndongo, P.A.,
Nejstgaard, J.C., Nikitina, O., Ochs, C., Odume, O., Opperman, J.J., Patricio, H.,
Pauls, S., Raghavan, R., Ramírez, A., Rashni, B., Ross-Gillespie, V., Samways, M.J.,
Schäfer, R.B., Schmidt-Kloiber, A., Seehausen, O., Shah, D.N., Sharma, S.,
Soininen, J., Sommerwerk, N., Stockwell, J.D., Suhling, F., Tachamo Shah, R.D.,
Tharme, R.E., Thorp, J.H., Tickner, D., Tockner, K., Tonkin, J.D., Valle, M., Vitule, J.,
Volk, M., Wang, D., Wolter, C., Worischka, S., Thrall, P., 2022. A global agenda for
advancing freshwater biodiversity research. Ecol. Lett. 25 (2), 255–263.
Meybeck, M., 1995. Physics and chemistry of lakes (eds Lerman A., Imboden D.M., Gat
J.), Springer-Verlag, 36.
Moser, K.A., Korhola, A., Weckstrröm, T.B., Pienitz, R., Smol, J.P., Douglas, M.S.V.,
Hay, M.B., 2000. Paleohydrology inferred from diatoms in northern latitude regions.
J. Paleolimnol. 24, 93–107. https://doi.org/10.1023/A:1008173901591.
S. Yu et al.
Murakami, T., Terai, H., Yoshiyama, Y., Tezuka, T., Zhu, L., Matsunaka, T.,
Nishimura, M., 2007. The second investigation of Lake Puma Yum Co located in the
Southern Tibetan Plateau, China. Limnology 8, 331–335. https://doi.org/10.1007/
s10201-007-0208-2.
Oksanen, A., Savela, N., Latikka, R., Koivula, A., 2020. Trust toward robots and artificial
intelligence: an experimental approach to human–technology interactions. Front.
Psychol. 11 https://doi.org/10.3389/fpsyg.2020.568256.
Pi, X.H., Feng, L., Li, W.F., Liu, J.G., Kuang, X.X., Shi, K., Qi, W., Chen, D.L., Tang, J.,
2021. Chlorophyll-a concentrations in 82 large alpine lakes on the Tibetan Plateau
during 2003–2017: temporal-spatial variations and influencing factors. Int. J. Digital
Earth 14, 714–735. https://doi.org/10.1080/17538947.2021.1872722.
Pienitz, R., Smol, J.P., 1993. Diatom assemblages and their relationship to environmental
variables in lakes from the boreal forest-tundra ecotone near Yellowknife, Northwest
Territories, Canada. Hydrobiologia 269, 391–404. https://doi.org/10.1007/
BF00028037.
Qiao, B.J., Wang, J.B., Huang, L., Zhu, L.P., 2017. Characteristics and seasonal variations
in the hydrochemistry of the Tangra Yumco basin, central Tibetan Plateau, and
responses to the Indian summer monsoon. Environ. Earth Sci. 76, 162. https://doi.
org/10.1007/s12665-017-6479-y.
Qiu, J., 2008. The third pole. Nature 454, 393–396. https://doi.org/10.1038/454393a.
R core team,, 2018. R: A language and environment for statistical computing. R
Foundation for Statistical Computing, Vienna, Austria.
Rivera-Rondón, C.A., Catalan, J., 2020. Diatoms as indicators of the multivariate
environment of mountain lakes. Sci. Total Environ. 703, 135517 https://doi.org/
10.1016/j.scitotenv.2019.135517.
Saunders, K.M., 2011. A diatom dataset and diatom-salinity inference model for
southeast Australian estuaries and coastal lakes. J. Paleolimnol. 46 (4), 525–542.
Seddon, P.J., Griffiths, C.J., Soorae, P.S., Armstrong, D.P., 2014. Reversing defaunation:
Restoring species in a changing world. Science 345, 406–412. https://doi.org/
10.1126/science.1251818.
Smol, J.P., 2009. Pollution of lakes and rivers: a paleoenvironmental perspective. John
Wiley & Sons.
Smol, J.P., 2019. Under the radar: long-term perspectives on ecological changes in lakes.
Proc. R. Soc. B 286, 20190834. https://doi.org/10.1098/rspb.2019.0834.
Smol, J.P., Cumming, B.F., 2000. Tracking long-term changes in climate using algal
indicators in lake sediments. J. Phycol. 36, 986–1011. https://doi.org/10.1046/
j.1529-8817.2000.00049.x.
Smol, J.P., Stoermer, E.F., (editors),, 2010. The diatoms: applications for the
environmental and earth sciences. Cambridge University Press.
Stenger-Kovács, C., Béres, V.B., Buczkó, K., Tapolczai, K., Padisák, J., Selmeczy, G.B.,
Lengyel, E., 2023. Diatom community response to inland water salinization: a
review. Hydrobiologia. https://doi.org/10.1007/s10750-023-05167-w.
Su, Y.L., Li, K.Y., Zhang, Y.D., Liu, Z.W., Wang, T.J., Jeppesen, E., Middelburg, J.J.,
Smol, J.P., 2022. Effects of climate change and nutrients concentrations on carbon
sources for zooplankton in a Tibetan Plateau lake over the past millennium.
J. Paleolimnol. 68, 249–263. https://doi.org/10.1007/s10933-022-00245-.
Sun, H., Liao, K., Pan, Y., Wang, J., 1990. Altas of the Qinghai-Tibet plateau. Science
Press, Beijing, p. 17.67-69..
Taukulis, F.E., John, J., 2009. Development of a diatom-based transfer function for lakes
and streams severely impacted by secondary salinity in the south-west region of
Western Australia. Hydrobiologia 626 (1), 129–143. https://doi.org/10.1007/
s10750-009-9741-9.
Ter Braak, C.J.F., 1988. CANOCO – a FORTRAN Program for Canonical Community
Ordination by [Partial] [Detrended] [Canonical] Correspondence Analysis, Principal
Components Analysis and Redundancy Analysis (Version 2.1). Technical Report
LWA-88-02. Agricultural Mathematics Group, Wageningen.
Ter Braak, C.J.F., 1992. CANOCO a Fortran program for Canonical Community
Ordination. Microcomputer Power, New York, p. 351.
Ter Braak, C.J.F., Šmilauer, P., 2012. Canoco reference manual and user’s guide:
software for ordination, version 5.0. Microcomputer Power.
Ter Braak, C.J.F., Prentice, I.C., 2004. A theory of gradient analysis. Adv. Ecol. Res. 34,
235–282. https://doi.org/10.1016/S0065-2504(08)60183-X.
Thienpont, J.R., Ruehland, K.M., Pisaric, M.F., Kokelj, S.V., Kimpe, L.E., Blais, J.M.,
Smol, J.P., 2013. Biological responses to permafrost thaw slumping in Canadian
Arctic lakes. Freshwater Biol. 58, 337–353. https://doi.org/10.1111/fwb.12061.
Veres, A.J., Pienitz, R., Smol, J.P., 1995. Lake water salinity and periphytic diatom
succession in three subarctic lakes, Yukon Territory, Canada. Arctic 48, 63–70.
http://www.jstor.org/stable/40511618.
Verleyen, E., Hodgson, D.A., Vyverman, W., Roberts, D., McMinn, A., Vanhoutte, K.,
Sabbe, K., 2003. Modelling diatom responses to climatic induced fluctuations in the
17
Ecological Indicators 155 (2023) 110952
moisture balance in continental Antarctic lakes. J. Paleolimnol. 30, 195–215.
https://doi.org/10.1023/A1025570904093.
Wang, R., Hu, Z., Wang, Q., Xu, M., Zheng, W., Zhang, K.e., Yang, X., 2020b. Discrepancy
in the responses of diatom diversity to indirect and direct human activities in lakes of
the southeastern Tibetan Plateau, China. Anthropocene 30, 100243.
Wang, L., Jiang, W.Y., Jiang, D.B., Zou, Y.F., Liu, Y.Y., Zhang, E.L., Hao, Q.Z., Zhang, D.
G., Zhang, D.T., Peng, Z.Y., Xu, B., Yang, X.D., Lu, H.Y., 2018. Prolonged heavy
snowfall during the Younger Dryas. J. Geophys. Res. Atmos. 123 https://doi.org/
10.1029/2018JD029271.
Wang, P., Shang, Y.N., Shen, L.C., Wu, K.Y., Xiao, Q., 2013. Characteristics and evolution
of hydrochemical compositions of freshwater lake in Tibetan plateau. Huan Jing Ke
Xue 34, 874–881. In Chinese with English abstract.
Wang, M.D., Tian, Q., Li, X.M., Liang, J., He, Y., Hou, J.Z., 2020a. TEX86 as a potential
proxy of lake water pH in the Tibetan Plateau. Palaegeography, Palaeoclimatology,
Palaeoecology 538, 109381. https://doi.org/10.1016/j.palaeo.2019.109381.
Wang, T.Y., Wu, T.H., Wang, P., Li, R., Xie, C.W., Zou, D.F., 2019. Spatial distribution
and changes of permafrost on the Qinghai-Tibet Plateau revealed by statistical
models during the period of 1980 to 2010. Sci. Total Environ. 650, 661–670. https://
doi.org/10.1016/j.scitotenv. 2018.08.398.
Wang, R., Yang, X.D., Langdon, P., Zhang, E.L., 2011. Limnological responses to warming
on the Xizang Plateau, Tibet, over the past 200 years. J. Paleolimnol. 45, 257–271.
https://doi.org/10.1007/s10933-011-9496-y.
Wang, J.B., Zhu, L.P., Wang, Y., Ju, J.T., Xie, M.P., Daut, G., 2010. Comparisons between
the chemical compositions of lake water, inflowing river water, and lake sediment in
Nam Co, central Tibetan Plateau, China and their controlling mechanisms. J. Great
Lakes Res. 36, 587–595. https://doi.org/10.1016/j.jglr.2010.06.013.
Wilson, S.E., Cumming, B.F., Smol, J.P., 1994. Diatom-salinity relationships in 111 lakes
from the Interior Plateau of British Columbia, Canada: the development of diatom-
based models for paleosalinity reconstructions. J. Paleolimnol. 12 (3), 197–221.
Wilson, S.E., Cumming, B.F., Smol, J.P., 1996. Assessing the reliability of salinity
inference models from diatom assemblages: an examination of a 219-lake data set
from western North America. Journal of Fisheries and Aquatic Sciences 53 (7),
1580–1594. https://doi.org/10.1139/f96-094.
Wischnewski, J., Kramer, A., Kong, Z., Mackay, A.W., Simpson, G.L., Mischke, S.,
Herzschuh, U., 2011a. Terrestrial and aquatic responses to climate change and
human impact on the southeastern Tibetan Plateau during the past two centuries.
Glob. Chang. Biol. 17, 3376–3391. https://doi.org/10.1111/j.1365-
2486.2011.02474.x.
Wischnewski, J., Mischke, S., Wang, Y., Herzschuh, U., 2011b. Reconstructing climate
variability on the northeastern Tibetan Plateau since the last Lateglacial – a multi-
proxy, dual-site approach comparing terrestrial and aquatic signals. Quat. Sci. Rev.
30, 82–97. https://doi.org/10.1016/j.quascirev.2010.10.001.
Wischnewski, J., Herzschuh, U., Rühland, K.M., Brauning, A., Mischke, S., Smol, J.P.,
Wang, L., 2014. Recent ecological responses to climate variability and human
impacts in the Nianbaoyeze Mountains (eastern Tibetan Plateau) inferred from
pollen, diatom and tree-ring data. J. Paleolimnol. 51, 287–302. https://doi.org/
10.1007/s10933-013-9747-1.
Yang, X.D., Kamenik, C., Schmidt, R., Wang, S.M., 2003. Diatom-based conductivity and
water level inference models from eastern Tibetan (Qinghai-Xizang) Plateau lakes.
J. Paleolimnol. 30, 1–19. https://doi.org/10.1023/A:1024703012475.
Yang, H.D., Turner, S., 2013. Radiometric dating for recent lake sediments on the Tibetan
Plateau. Hydrobiologia 713, 73–86. https://doi.org/10.1007/s10750-013-1493-x.
Yao, T., Thompson, L.G., Mosbrugger, V., Zhang, F., Ma, Y., Luo, T., Xu, B., Yang, X.,
Joswiak, D.R., Wang, W., Joswiak, M.E., Devkota, L.P., Tayal, S., Jilani, R.,
Fayziev, R., 2012. Third Pole Environment (TPE). Environ. Dev. 3, 52–64. https://
doi.org/10.1016/j.envdev. 2012.04.002.
Yu, S.W., Wang, J.B., Li, Y.M., Peng, P., Kai, J.L., Kou, Q.Q., Laug, A., 2019. Spatial
distribution of diatom assemblages in the surface sediments of Selin Co, central
Tibetan Plateau, China, and the controlling factors. J. Great Lakes Res. 45,
1069–1079. https://doi.org/10.1016/j.jglr.2019.09.006.
Zhang, X.S., Liu, J.B., Rühland, K.M., Jia, X., Reed, J.M., Li, Y.L., Shen, Z.W., Zhao, J.J.,
Chen, J., Wang, H.P., Wang, X., Smol, J.P., Chen, F.H., 2023. Concurrent mid-
Holocene East Asian temperature and summer monsoon maxima forced by high- and
low-latitude interplay. Global Planet. Change 220, 104008. https://doi.org/
10.1016/j.gloplacha.2022.104008.
Zhao, L., Li, Y.N., Xu, S.X., Zhou, H.K., Gu, S., Yu, G.R., Zhao, X.Q., 2006. Diurnal,
seasonal and annual variation in net ecosystem CO2 exchange of an alpine shrubland
on Qinghai-Tibetan plateau. Glob. Chang. Biol. 12, 1940–1953. https://doi.org/
10.1111/j.1365-2486.2006.01197.x.