Environment International 175 (2023) 107939

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Increasing soil Mn abundance promotes the dissolution and oxidation of Cr

(III) and increases the accumulation of Cr in rice grains
Ming Ao a, Tenghaobo Deng b, Shengsheng Sun a, Mengyao Li a, Jingjing Li d, e, Ting Liu d, e,
Bofang Yan a, Wen-Shen Liu a, Guobao Wang a, Dedao Jing f, Yuanqing Chao a, c, Yetao Tang a, c,
Rongliang Qiu a, d, e, Shizhong Wang a, c, *
a
School of Environmental Science and Engineering, Guangdong Provincial Key Laboratory of Environmental Pollution Control and Remediation Technology, Sun Yat-sen
University, Guangzhou 510006, China
b
Institute of Quality Standard and Monitoring Technology for Agro-products of Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China
c
Guangdong Provincial Engineering Research Center for Heavy Metal Contaminated Soil Remediation, Sun Yat-sen University, Guangzhou 510006, China
d
Guangdong Provincial Key Laboratory of Agricultural & Rural Pollution Abatement and Environmental Safety, College of Natural Resources and Environment, South
China Agricultural University, Guangzhou 510642, China
e
Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou 510642, China
f
Zhenjiang Institute of Agricultural Sciences in Hilly Region of Jiangsu, Jurong 212400, China

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

Handling Editor: Adrian Covaci Hexavalent chromium (Cr(VI)) is more readily taken up by plants than trivalent chromium (Cr(III)) due to its
similar chemical structure to phosphate and sulfate. In paddy soils, Cr(VI) of natural origin are mainly produced
Keywords: from Cr(III) oxidized by O2 and Mn(III/IV) oxides, which are affected by rice radial oxygen loss (ROL) and Mn
Chromium (II)-oxidizing microorganisms (MOM). However, little is known about the effect of ROL and Mn abundance on
Rice
rice Cr uptake. Here, we investigated the effects on Cr(VI) generation and the subsequent Cr uptake and accu­
Cr(VI)
mulation with the involvement of two rice cultivars with distinct ROL capacities by increasing soil Mn abun­
Mn(III/IV) oxides
dance. Results showed that Mn(II) addition to the soil led to more Cr(III) being released into the pore water, and
the dissolved Cr(III) was oxidized to Cr(VI) by ROL and biogenic Mn(III/IV) oxides. The concentration of Cr(VI)
in soil and pore water increased linearly with the addition of Mn(II) doses. Mn(II) addition promoted the root-to-
shoot translocation and grain accumulation of Cr derived mainly from newly generated Cr(VI) in the soil. These
results emphasize that rice ROL and MOM promote the oxidative dissolution of Cr(III) at a high level of soil Mn,
resulting in more Cr accumulation in rice grains and increasing dietary Cr exposure risks.

1. Introduction Wise et al., 2022). Moreover, Cr(VI) is actively transported to accumu­

Chromium (Cr) contamination is widespread throughout the world
and can be released into the environment through human activities and
geological processes (Ao et al., 2022a; Pavesi & Moreira, 2020; Shaheen
et al., 2017; Sharma et al., 2005). Eventually, Cr-containing pollutants
accumulate in agricultural soils, increasing the risk of Cr exposure to
humans (Ao et al., 2022a; Kapoor et al., 2022; Sharma et al., 2005). In
soils, Cr toxicity and bioavailability vary with its oxidation states, with
trivalent Cr (Cr(III)) and hexavalent Cr (Cr(VI)) were the common forms
of Cr (Hai et al., 2020; Shaheen et al., 2017). Compared with Cr(III), Cr
(VI) has significantly higher toxicity, mobility and bioavailability than
Cr(III) (Aharchaou et al., 2017; Ao et al., 2022a; Shanker et al., 2005;
late in plants’ above-ground tissues due to its higher water solubility and
transmembrane efficiency (Aharchaou et al., 2017; Shanker et al., 2005)
and structure similarity to phosphate and sulfate (Ding et al., 2019;
Singh et al., 2013), ultimately endangering human health through food
chain transport (Fernando et al., 2020; Wise et al., 2022). Rice (Oryza
sativa L.), a dietary staple for approximately-three billion people glob­
ally, is more likely to accumulate Cr than other cereal crops and is Cr’s
primary food chain pathway (Ali et al., 2022; Rai et al., 2019; Xiao et al.,
2013; Xiao et al., 2015). For example, Ali et al. (2022) found that 97.4 %
of 500 rice grain samples from the leading rice-producing areas of
Pakistan exceeded the widely referred Chinese standard on the limit of
Cr concentration in food (1.0 mg/kg) (GB2762-2017), increasing Cr

* Corresponding author.
E-mail address: wshizh2@mail.sysu.edu.cn (S. Wang).

https://doi.org/10.1016/j.envint.2023.107939
Received 20 February 2023; Received in revised form 12 April 2023; Accepted 16 April 2023
Available online 20 April 2023
0160-4120/© 2023 The Author(s). Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-
nc-nd/4.0/).
M. Ao et al.
exposure risk. Previous studies have shown that Cr content in rice grains
has the highest correlation with water-soluble Cr and exchangeable Cr
(r = 0.99) (Bhattacharyya et al., 2005), among which Cr(VI) may be the
main reason for the increase of Cr accumulation in rice grains, because
Cr(VI) reduction can significantly decrease the absorption and accu­
mulation of Cr in rice grains (Xiao et al., 2021). Therefore, there is an
underlying need for process-based knowledge to reveal the effect of Cr
(III)’s dissolution and oxidation in paddy soils on rice uptake of Cr.
Under natural conditions, Cr(III) is the most stable state in the
environment with a standard potential of − 1.232 V, so considerable
energy is required to convert it to a higher oxidation state of Cr(VI)
(Shadreck & Mugadza, 2013). From a thermodynamical perspective,
natural oxidants that can oxidize Cr(III) are limited to oxygen (O2) and
manganese (Mn, III/IV) oxides, but their oxidation rates vary (Ao et al.,
2022b; Gorny et al., 2016; Namgung et al., 2014; Pan et al., 2017; Qian
et al., 2020). Studies have shown that Cr(III) oxidation rate by dissolved
O2 is prolonged at room temperature (only 3 % Cr(III) was oxidized
within two weeks), while adding MnO2 significantly increases Cr(III)
oxidation rate (Gorny et al., 2016). Studies have confirmed that the
presence of Mn(II) oxidation is also thermodynamically advantageous
for the oxidation of Cr(III) compared to the direct oxidation of Cr(III) by
O2 alone, even at low dissolved O2 concentrations (Namgung et al.,
2014), and the generated Mn(III) or Mn(IV) species provide more active
oxidants than O2 (Wang et al., 2014). Recent experiments confirmed
that active Mn(III/IV) oxides substantially promoted Cr(III) oxidation
under low O2 conditions compared with oxidation by O2 alone (Hai
et al., 2020; Namgung et al., 2014; Pan et al., 2017; Wadhawan et al.,
2015). These studies suggest that the presence of Mn(II/III/IV) is the
most efficient catalyst and oxidant leading to the oxidation of Cr(III) to
Cr(VI).
Mn(III/IV) oxides are highly reactive minerals formed by Mn(II)
oxidation and play a crucial role in the biogeochemical cycle of many
elements. In soils, Mn(III/IV) oxides are formed by the chemical and
biological oxidation of soluble Mn(II) (Tebo et al., 2004; Zhou & Fu,
2020). Although chemical Mn(II) oxidation is thermodynamically
feasible, the reaction rate is very slow at pH values below 8.0 and occurs
only in surface oxic soils (Akob et al., 2014). By contrast, many Mn(II)-
oxidizing microorganisms (MOM) can rapidly oxidize Mn(II) to Mn(III/
IV) oxides by secreting multicopper oxidase, which are 4–5 orders of
magnitude faster than the chemical processes (Toyoda & Tebo, 2016;
Zhang et al., 2015; Zhou & Fu, 2020). Especially in anoxic soils, MOM is
considered the primary driving force in the biogenic generation of Mn
(III/IV) oxides (Zhang et al., 2015; Zhou & Fu, 2020), which may affect
Cr(III) oxidation and dissolution. MOM are widely distributed in envi­
ronmental media, and Mn(II) abundance may be an important factor
dominating the generation rate and amount of Mn(III/IV) oxides (Zhang
et al., 2015; Zhou & Fu, 2020). In paddy fields, a unique wetland system,
frequent flooding and drainage promote redox reaction processes. For
example, Mn(III/IV) oxides are reduced before Fe(III) oxides during
flooding, and the generated Mn(II) provides the necessary Mn(II) ions
for its biological oxidation (Maguffin et al., 2020). During drainage, Fe
(II) is oxidized by dissolved O2 to form poorly crystalline Fe oxides
which are effective sorbents for Cr(VI) and Cr(III) in the soil (Hausladen
& Fendorf, 2017), possibly decreasing Cr(VI) release and inhibiting Cr
(III) oxidation. However, it is unclear how the generation and dissolu­
tion of Mn(III/IV)/Fe(III) oxides affect the migration and transformation
of Cr during flooding and drainage in paddy soils. In particular, the
mechanism of oxidation and competitive retention of Cr(III) between Fe
(III) and Mn(III/IV) oxides remains to be revealed.
In soil-rice systems, rice is a semi-aquatic plant. Like wetland plants,
it has an extensive root aerenchyma that can release nine times the
amount of O2 required for respiration from the air through the aeration
tissues to the rhizosphere, and this process is known as radial oxygen
loss (ROL) (Armstrong, 1971). ROL can oxidize soluble Fe(II) and Mn(II)
in pore water into Fe(III) and Mn(III/IV) oxide precipitates (Awasthi
et al., 2017; Kögel-Knabner et al., 2010; Xu et al., 2018). However, little
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Environment International 175 (2023) 107939
research investigating whether ROL driven Mn cycling leads to high
levels of Cr(VI) exposure conditions in rice rhizosphere microregions
have been undertaken. Furthermore, Fe(II/III) and Mn(II/III/IV), as
redox-active elements commonly co-occurring in soils (Li et al., 2021),
function differently for Cr. Fe oxides act mainly as sorbents and reducing
agents (Hausladen & Fendorf, 2017; Xu et al., 2018), while Mn(III/IV)
oxides have catalytic, oxidative and adsorptive functions, and is a nat­
ural strong oxidant (Ao et al., 2022a; Dai et al., 2009; Pan et al., 2017;
Qian et al., 2020; Zhou & Fu, 2020). Due to the mediation of rice ROL
and MOM, increasing the Mn abundance in soil may promote the gen­
eration of more Mn(III/IV) oxides, stimulate Cr(III) dissolution and
oxidation, and increase the ecological and environmental risks of Cr in
the soil-rice system.
This study investigates the effects and controlling factors of Cr(III)
dissolution and oxidation in two rice cultivars with different root ROL
capacities after increasing soil Mn abundance, Cr accumulation in rice’s
different tissues, and the corresponding mechanisms for this process.
Specifically, the study explored: (1) the impacts of Mn(II) addition on
pore water and soil Cr(VI) levels; and (2) the effects of Mn(II) addition
on Cr uptake and translocation in rice. The two processes were examined
in laboratory pot experiments. These results provide new insights into Cr
(VI) production and bioaccumulation processes in soil-rice systems,
facilitate the risk prediction of human dietary exposure to Cr, and pro­
vide a theoretical basis for finding effective measures to decrease Cr
uptake in rice.
2. Materials and methods
2.1. Pot experimental design
Rhizobag pot experiments were conducted in a greenhouse at
ambient temperatures (25–35 ℃) under daylight conditions (16 h of
light/8 h of darkness). Soil for pot experiments was collected from a
paddy field formed by weathering of basaltic rocks in the Leizhou
Peninsula, China. Based on our previous study (Ao et al., 2022b), the soil
from this area has slight Cr contamination and high Fe:Mn ratios,
making it favorable for Mn adjustment in the pot experiments. After
being air-dried and sieved to <0.5 cm, soil was supplemented with basal
fertilizer (N: 40 mg/kg, P: 9.6 mg/kg and K: 14 mg/kg), mixed thor­
oughly. The Cr, Mn and Fe concentrations were 256.7, 652.3 and
104,874.8 mg/kg, respectively. Before planting, the paddy soils were
adjusted with MnCl2. Specifically, 10 L of deionized water was added to
each 15 kg of sieved soil. Based on the Fe:Mn ratio of paddy soils in
different areas of the world (Table S1), the accurately calculated MnCl2
solution was added (Hernandez-Soriano et al., 2012) and fully mixed
with the soils to achieve Fe:Mn ratio at 110 (Low-Mn treatment) and 70
(High-Mn treatment), respectively, in the soils. Those soils without the
addition of MnCl2 solution were regarded as control soils (CK)
(Figure S2). The well-mixed soil was then divided equally into six
custom-made square pots (6 replicates) and aged under flooding con­
dition for two weeks. In each pot, 500 g soil was put in a rhizobag (200
mesh; 20 cm in height; 15 cm in diameter) to create a central
compartment (rhizosphere soil), leaving the peripheral compartment
(bulk soil) as an area without root effect (Figure S2). Thereafter, two rice
seedlings of Zhendao 14 (Z-14, high ROL rice cultivar) or Y-Liangyou 7
(Y-7, low ROL rice cultivar) with significant differences in root ROL
(Figure S1) were planted into the central compartment. A rhizo-sampler
(Rhizon Soil Moisture Samplers, Rhizosphere Research Products, The
Netherlands) with a nylon filter was inserted into the rhizosphere soil for
long-term in-situ collection of rhizosphere pore water (Figure S2).
According to the growth morphology and physiological character­
istics, the rice growth was divided into five stages, namely tillering stage
(Day 1–20), jointing stage (Day 21–40), flowering stage (Day 41–60),
filling stage (Day 61–75) and maturing stage (Day 76–100). Water
management was divided into two stages, i.e., continuous flooding (CF,
Day 1–60) with watering every 12 h to keep the soil continuously
M. Ao et al.
flooded, overlying water depth varied between 1 and 5 cm with time;
intermittent flooding (IF, Day 61–100) with identical watering intervals
but only allowed water to fill the soil, water level varied between 0 and
10 cm beneath the soil surface with time, thus increasing soil perme­
ability. Details of experimental design and rice cultivation could be
found in Supporting Information (SI).
2.2. Sample collection and preparation
After 10 d of rice planting, pore water samples were collected by
rhizo-sampler every 10 d (9 times in total) (Figure S2). The collected
pore water was immediately transferred to a glovebox to determine Eh
and pH values and then divided into two parts, one for Cr(VI) deter­
mination and the other for total dissolved Cr measurement after the
addition of HNO3.
The rhizosphere and bulk soil and rice plant samples were collected
at the end of each growth stage. The Eh and pH values of the rhizosphere
and bulk soils were determined. The rhizosphere/bulk soils were
divided into three parts, the first part was freeze-dried to determine soil
Cr(VI), the second part was centrifuged at 4500 rpm to obtain pore water
and to determine its Cr(VI) and total dissolved Cr concentrations, and
the third part was stored at –80 ℃ for DNA extraction. The rice plants
were cleaned and divided into parts of root, stem and leaf prior to being
dried and crushed. The grains were also crushed after hulling (brown
rice) for Cr concentration analysis.
2.3. Analytical methods
Detailed information concerning analytical methods described in the
SI include concentration determinations of total dissolved Cr and Cr(VI)
in pore water (dissolved Cr(III) = total dissolved Cr – dissolved Cr(VI)),
Cr(VI) in soil and total Cr and other elements in plant samples, Laser
Ablation ICP-MS (LA-ICP-MS) analysis of selected brown rice samples,
16S rRNA gene high-throughput sequencing of selected soil samples,
and statistical analysis of data.
2.4. Quality control
Quality control measures consisted of method blanks, duplicate
samples, and certified reference materials are described in the SI. The
recoveries of total Cr in rice tissues (GBW07603) and Cr(VI) in soil (GBW
(E)070251) were 91–98 % and 85–106 %, respectively.
2.5. Statistical analyses
SPSS (version 20.0, IBM) was used for statistical analysis. One-way
analysis of variance (ANOVA) with Fisher’s least significant difference
(LSD) test was performed to determine the statistical significance of
treatments. A significant level with P values of<0.05 (p < 0.05) was
selected. Figures were drawn through Origin software (version 9.8,
OriginLab 2021).
3. Results
3.1. Effect of Mn(II) addition on pore water and soil properties
In the pot experiments, Mn(II) addition resulted in decreased pH and
increased Eh in pore water, rhizosphere and bulk soils compared with
CK (Figures S3, S4 and S5). From tillering to maturing stages, pH and Eh
values in pore water, rhizosphere, and bulk soils showed opposing
patterns, with pH and Eh values in most treatments reaching their peak
and lowest values, respectively, at the flowering stage (Figures S3, S4
and S5). Compared with bulk soils, rhizosphere soils had higher Eh
values with the involvement of rice (Figure S5), proving that rice ROL
indeed facilitated oxidizing conditions therein. However, the rhizo­
sphere soil Eh values derived from Y-7 with lower ROL were slightly
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higher than those from Z-14 with higher ROL (Fig. S5A,C), which was
surprisingly in contrast to our expectation, i.e., higher ROL leading to
higher Eh. This phenomenon was speculated as related to the signifi­
cantly higher root biomass of Y-7 (Table S3) that potentially better
increased rhizosphere soil Eh with more O2 introduced (Li & Wang,
2013).
3.2. Increased Cr(VI) and Cr(III) levels in pore water under Mn(II)
addition
In the pot experiments, dissolved Cr(VI) concentrations were
elevated in rhizosphere pore water following Mn(II) addition (increased
averagely 7.1 (Y-7) and 5.7 (Z-14) times in High-Mn treatment, vs CK,
Fig. 1A,C), the more Mn(II) addition, the higher Cr(VI) levels. In the
High-Mn treatment, the rhizosphere pore water’s Cr(VI) concentrations
gradually increased from tillering to filling stages and decreased at
maturing stage (Fig. 1A,C). Compared with Z-14, the Cr(VI) concen­
tration in the rhizosphere pore water of the High-Mn treatment was
higher in Y-7, but reverse phenomenon was found in the Low-Mn
treatment (Fig. 1A,C).
In both Low-Mn and High-Mn treatments, the concentrations of
dissolved Cr(III) in the rhizosphere pore water significantly increased,
resulting in an increase of its proportion of total dissolved Cr (Cr(VI) and
Cr(III)) from 52 % to 70 % (vs CK, Fig. 1B,D). However, concentrations
of dissolved Cr(III) in rhizosphere pore water did not increase linearly
with increasing Mn(II) doses. During the whole growth period of rice,
they peaked at tillering stage, thereafter maintained at a lower level at
the jointing and flowering stages, and gradually increased at the filling
and maturing stages (Fig. 1B,D).
At each of the five growth stages, the addition of Mn(II) was also
found to significantly increase Cr(VI) and dissolved Cr(III) concentra­
tions in the rhizosphere and bulk pore water compared with CK
(Figure S6), with generally higher Cr(VI) concentrations in the rhizo­
sphere than bulk pore water (e.g., averagely 7.61 (Y-7) and 10.99 μg/L
(Z-14) higher in the High-Mn treatment) (Figure S6). Moreover, the
concentrations of Cr(VI) and dissolved Cr(III) in the pore water
increased with increasing Mn(II) doses, particularly the former one
(Figure S6). Interestingly, the Cr(VI) and dissolved Cr(III) concentra­
tions in the pore water in the Mn(II) addition treatment showed opposite
variation patterns throughout the rice growth stages, while the sum of
them as total dissolved Cr concentration remained relatively stable
(Figure S6). Specifically, the Cr(VI) and dissolved Cr(III) concentrations
showed inversed U-shaped and U-shaped variation patterns, all with
turning points at the jointing stage, respectively (Figure S6).
3.3. Enhanced soil Cr(VI) levels under Mn(II) addition
The results of Cr(VI) concentrations in the rhizosphere and bulk soils
at five growth stages of rice showed that Mn(II) addition resulted in
more Cr(VI) production (Fig. 2). Specifically, Cr(VI) concentrations in
the rhizosphere and bulk soils remained at low levels (0.045–0.13 mg/
kg) during the tillering and jointing stages. From the flowering to
maturing stages, Cr(VI) concentrations in the rhizosphere and bulk soils
gradually increased in the Mn(II) addition treatments, and Cr(VI) con­
centrations in bulk soils peaked at the filling stage (Y-7: 0.44 mg/kg; Z-
14: 0.55 mg/kg) (Fig. 2B,D), while Cr(VI) concentrations in rhizosphere
soils maintained an increasing trend at the maturing stage (Fig. 2A,C).
Soil Cr(VI) concentrations increased with Mn(II) doses and rice
growth. Rhizosphere soils generally had higher Cr(VI) concentrations
compared with bulk soils (Fig. 2). During the whole growth period of
rice, the Cr(VI) concentrations in rhizosphere soil of Z-14 with high ROL
were higher than Y-7 with low ROL in the same doses of Mn(II) (Fig. 2A,
C). The rhizosphere soil in CK of Z-14 also showed a slight increase in Cr
(VI) concentrations (0.016 mg/kg) compared with bulk soil (Fig. 2C,D),
while there was no significant difference between Cr(VI) in rhizosphere
soil and bulk soil in CK of Y-7 (Fig. 2A,B). Comparison of the Cr(VI)
M. Ao et al.
Fig. 1. Temporal changes in Cr(VI) and Cr(III) concentrations in pore water during rice growth. Day 1–20: tillering stage; Day 21–40: jointing stage; Day 41–60:
flowering stage; Day 61–75: filling stage; and Day 76–100: maturing stage. CF: continuous flooding; IF: intermittent flooding.
concentrations in pore water revealed that most of the newly generated
Cr(VI) in the Mn(II) addition treatments were retained in the soil, and
only about 15 % (Low-Mn) and 17 % (High-Mn) of Cr(VI) entered the
pore water (Fig. 2 and S6).
3.4. Cr accumulation and distribution in the plant tissues
In the pot experiment, Mn(II) addition significantly decreased the
accumulation of Cr in the rice roots compared with CK, in a pattern that
Cr accumulation decreased with increasing doses of Mn(II) (Fig. 3A,B).
Throughout the growth period of rice, the Cr concentration in the roots
of CK treatment gradually increased, while the addition of Mn(II) led to
a decrease in the Cr concentration in the roots from tillering to jointing
stages, after which the concentration of Cr in the roots changed mini­
mally (Fig. 3A,B).
In the above-ground tissues, Cr concentration in stems of Y-7 grad­
ually decreased with rice growth in the CK treatment (Fig. 3C). In the Mn
(II) addition treatment, Cr accumulation in stems (Y-7) decreased from
tillering to jointing stages, and the Cr concentration in stems gradually
increased from flowering to maturing stages, even exceeding that in the
CK treatment (Fig. 3C). Unlike Y-7, Mn(II) addition significantly
increased Cr accumulation in stems of Z-14 compared with CK (Fig. 3D).
From tillering to flowering stages, the Cr concentration in stems (Z-14)
gradually increased and peaked at the flowering stage (Fig. 3D).
Thereafter, it gradually decreased from filling to maturing stages, with
more decrease found in higher Mn(II) doses (Fig. 3D). In leaves, Cr
accumulation in Y-7 increased 2.96–8.15 times from tillering to jointing
stages in all treatments, and Cr concentration in CK gradually decreased
from flowering to maturing stages (Fig. 3E), while Cr concentration in
Mn(II) addition treatment increased significantly, even higher than that
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Environment International 175 (2023) 107939
in CK at maturing stage (Fig. 3E). In Z-14, leaf Cr concentration at each
growth stage gradually increased with rice growth (Fig. 3F). Moreover,
compared with CK treatment, Low-Mn treatment increased Cr accu­
mulation in leaves at flowering and maturing stages, while High-Mn
treatment decreased the accumulation of Cr in leaves at all growth
stages (Fig. 3F). Overall, the order of Cr accumulation in different parts
of Y-7 was leaves > roots > stems (Fig. 3A,C,E), while the Cr concen­
trations in roots, stems and leaves of Z-14 were essentially equivalent
(Fig. 3B,D,F).
3.5. Distribution of Cr and other elements in grain
In grains, Mn(II) addition resulted in a significant increase in Cr
accumulation, and increased linearly with the added Mn(II) doses and
soil Cr(VI) concentration (Fig. 4A). Among them, a strong positive cor­
relation was observed between the Cr concentration in Z-14 rice grains
and that in rhizosphere soil (r = 0.93, p < 0.01) (Fig. 4B). Given the
close-to 1:1 relationship, the increasing Cr concentration in grains was
mainly due to the soil’s increased Cr(VI) concentration following Mn(II)
addition. Although the correlation coefficient between Cr concentration
in Y-7 rice grains and Cr(VI) concentration in rhizosphere soil was lower
than that of Z-14, there was also a significantly positive correlation (r =
0.87, p < 0.01) (Fig. 4B), suggesting significant cultivar differences in
the biogeochemical processes of Cr uptake, accumulation and trans­
location between Y-7 and Z-14.
The distribution pattern of various elements in rice grain varies
considerably, as observed from both the longitudinal and transverse
sections (Fig. 5). Cr was distributed throughout the rice grains, with
higher Cr concentrations observed in the exterior portions of the endo­
sperm (Fig. 5). In contrast, Zn, Ca, and Mn were highly distributed
M. Ao et al.
Fig. 2. Temporal changes in Cr(VI) concentrations in the rhizosphere and bulk soils experiencing continuous flooding (CF) and intermittent flooding (IF) during rice
growth. Different lowercase letters indicate significant differences between different treatments (p < 0.05). Mean ± SD, n = 2.
within the aleurone layer and embryo, and Mg and K were also high
within the aleurone layer and embryo (Fig. 5). Fe and Ni were uniformly
distributed in the endosperm (Fig. 5). Phosphorous concentration was
highest in the embryo and decreased gradually from the outer part of the
endosperm toward the interior of the endosperm (Fig. 5). Comparative
analysis revealed that Cr and P distribution patterns were similar, with
high correlations between the Cr and P counts (r = 0.30, p < 0.01)
(Figure S7).
4. Discussion
To obtain a mechanistic understanding of Cr(VI) dynamics and the
risks of perturbing soil-rice systems with Mn(II), two processes were
explored that could contribute to the enhanced Cr(VI) generation in soils
(Section 4.1) and Cr fate and transformation in different tissues of rice
(Section 4.2).
4.1. Increased pore water and soil Cr(VI)
The Mn(II) addition resulted in an Mn(II)-Mn(III/IV) cycle mediated
by rice ROL, and Cr(III) oxidation by the generated Mn(III/IV) oxides
may primarily be responsible for the increased Cr(VI) levels in soils and
pore water. Compared with CK, Mn(II) addition significantly increased
pore water, rhizosphere and bulk soils Eh values, and the relatively high
Eh may originate from the gradual oxidation of Mn(II) to higher Mn(III/
IV) oxidation states. Based on the Mn (2p3/2) parameter of Mn oxides
proposed by Zhang et al. (2015) and Ilton et al. (2016), we determined
the Mn oxidation state changes and relative intensities. The XPS results
showed that the added MnCl2 was still present in the low oxidation
states of Mn(II) and Mn(III), even after 20 days of rice cultivation
(Fig. 6A–D). As the rice plants grew, Mn was mainly present as Mn(III)
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Environment International 175 (2023) 107939
and Mn(IV) in both rhizosphere and bulk soils of Y-7 rice at the flow­
ering stage, and the rhizosphere soil has higher oxidation state of Mn
(Fig. 6E,F). From tillering to maturing stages, the relative intensity of
high-valued Mn(III/IV) gradually increased, while Mn(II) gradually
decreased, e.g., Mn(II) from 33.38 % to 0 and Mn(IV) from 0 to 100 % in
the rhizosphere soil of Z-14 (Fig. 6D,H,L). XRD shows ambiguous Mn
mineral phases in soil, identified as bementite and manganite
(Figure S11), with nano-sized monocrystalline nature (Jeyaraj & Sub­
ramanian, 2022; Luo et al., 2017).
With the involvement of rice at different ROL, at the maturing stage
of all cultivars, highly oxidized states of Mn were found and they pre­
sented mainly as Mn(III) and Mn(IV) (Fig. 6I–L). Additionally, the
oxidation state of Mn was lower in Z-14 soils than in Y-7 soils at the same
stage (Fig. 6), which is consistent with the observed Eh values in soils
(Figure S5). This may be related to the difference in root biomass of the
two rice cultivars, i.e. root biomass of Y-7 with low ROL was three times
higher than that of Z-14 with high ROL (Table S3), since higher root
biomass can affect more soils (Chung & Sa, 2001; Li & Wang, 2013).
Therefore, it could be inferred that high root biomass of rice would
surpass the deficit of low ROL to favor oxidizing conditions in soil, so as
to allow more intensive Mn(II) oxidation happen therein.
Previous studies have shown that rice, as a semi-aquatic plant with
high ROL, can release nine times its respiratory requirement of O2 into
the rhizosphere soil (Kumazawa, 1984). This O2 can oxidize dissolved
Mn(II) to Mn(III/IV) and precipitate MnO2 (Shuman & Wang, 1997). As
a result, Eh values and Mn oxidation states in rhizosphere soils impacted
more by ROL were higher than those in bulk soils in this study (Fig. 5 and
S5). This phenomenon was reinforced over time, i.e., from tillering to
maturing stages, rice roots gradually increased (Table S3) and filled the
entire rhizosphere soil, ultimately promoting the oxidation of Mn(II) to
Mn(III/IV) oxides in the microregion around the roots by ROL. Thus,
M. Ao et al. Environment International 175 (2023) 107939

Fig. 3. Cr concentrations (mg/kg dry weight) in roots, stems and leaves of two rice cultivars (Y-7 and Z-14) under Mn(II) addition. Different lowercase letters
indicate significant differences between different treatments (p < 0.05). Mean ± SD, n = 2. CF: continuous flooding; IF: intermittent flooding.

ROL is an important factor contributing to the elevated Mn oxidation
state in the soil. Additionally, the intermittent flooding (IF) treatment
from filling to maturing stages increased O2 input and accelerated Mn
(II) oxidation in this study, which is also in line with the phenomenon
found by Xiao et al. (2015).
In contrast to ROL-mediated Mn(II) oxidation, Mn(II)-oxidizing mi­
croorganisms (MOM) may be the most important driver of Mn(II)
oxidation. In this study, four reported potential MOM strains (Bacillus,
Pseudomonas, Arthrobacter and Aeromonas) (Tebo et al., 2004; Yang
et al., 2014; Zhang et al., 2019) were detected in the rhizosphere and
bulk soils of High-Mn treatment samples by 16S rRNA gene sequences,
and they were found closely related to known functional groups (Fig. 7)
(Bohu et al., 2016). These MOM contains two Mn(II) oxidizing microbial
species (Bacillus sp. and Pseudomonas sp.) that have been identified
(Figure S9) (Tebo et al., 2004; Toyoda & Tebo, 2016), and Pseudomonas
sp. also has denitrification functions (Su et al., 2016). Studies have
confirmed that nitrate reduction in paddy soil is coupled with Mn(II)
oxidation in anoxic environment mediated by nitrate-reducing micro­
organisms (Xu et al., 2020), and Mn(II) is an important electron donor
for denitrification in paddy soil (Wang et al., 2022). In this study, the soil
was significantly acidic (pH < 6.5) (Figure S4), and the oxidation of Mn
(II) by O2 was thermodynamically unfavorable (Akob et al., 2014), and
the oxidation of Mn(II) by MOM became the main source of Mn(III/IV)
oxides in soil (Bohu et al., 2016). Previous studies have shown that MOM
is the main driver of Mn(II) oxidation under low pH conditions (Akob
et al., 2014; Bohu et al., 2016), a confirmed pathway of which is Mn(II)
being oxidized to Mn(III) (similarly found in this study as shown in
Fig. 6E,F) by synthetic multicopper oxidases of MOM, followed by Mn
(III) being oxidized to Mn(IV) (Hastings & Emerson, 1986; Tebo et al.,
2004). The produced Mn(III/IV) presented in the forms of oxides are

6
M. Ao et al. Environment International 175 (2023) 107939

Fig. 4. Cr concentration in grains of two rice cultivars (Y-7 and Z-14) under Mn(II) addition (A); the relationship between the changes in Cr concentrations in grains
and Cr(VI) in rhizosphere soil at maturing stage (B). Uppercase and lowercase letters indicate significant differences between the average values of two rice cultivars
and the same rice cultivar under different Mn(II) addition treatments, respectively (p < 0.05). Mean ± SD, n = 3.

Fig. 5. Distribution of mineral nutrients (Cr, Mn, K, Ca, Mg, Zn, P, Fe and Ni) in the longitudinal (A) and transverse (B) sections of rice grain Z-14 (High-Mn),
analyzed using LA-ICP-MS. The color scale represents different signal intensities, blue corresponds to the lowest relative intensity and red to the highest. (For
interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

effective oxidants of Cr(III) (Hausladen & Fendorf, 2017; Wu et al.,
2005). In this study, the positive correlation between MOM (Pseudo­
monas (r = 0.52, p < 0.05), Arthrobacter (r = 0.55, p < 0.05), and Aer­
omonas (r = 0.54, p < 0.05)) (Figure S8B–D) abundance and soil Cr(VI)
concentration further confirmed that MOM mediated the production of
Cr(VI) by promoting the generation of Mn(III/IV) oxides. Moreover,
Pseudomonas sp. with denitrification function (Su et al., 2016), versus
Bacillus sp. without, showed higher correlation (r = 0.39, p < 0.05,
Figure S10B) between its abundance and soil Cr(VI), implying that
denitrification reaction may indirectly stimulate Mn(II) oxidation.
Studies had also confirmed that although the abundance of MOM in soil
is low, their promotion of Mn(II) oxidation is 4–5 orders of magnitude
faster than chemical processes and are the main drivers of Mn(II)
oxidation in anoxic and acidic soils (Akob et al., 2014; Bohu et al., 2016;
Tebo et al., 2004; Wang et al., 2022; Zhou & Fu, 2020).
The widespread presence of Mn(III/IV) oxides and Cr(III) ions in pore
water provides additional reaction sites and opportunities for the
oxidation of Cr(III) to Cr(VI). Previous studies have shown that for Cr
(III) to be oxidized by Mn(III/IV) oxides in soil, Cr(III) minerals must
dissolve and migrate to the surface of Mn(III/IV) oxides, or the two
minerals must be in close contact (Hausladen & Fendorf, 2017; Pan
et al., 2017). The rate of Cr(III) oxidation is proportional to the

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M. Ao et al. Environment International 175 (2023) 107939

Fig. 6. XPS spectra of Mn(2p3/2) in rhizosphere and bulk soils of Y-7 and Z-14 in the High-Mn treatment.

Fig. 7. The abundance of Mn(II)-oxidizing microorganisms (MOM) (A: Bacillus and Pseudomonas, B: Arthrobacter and Aeromonas) in Z-14 rhizosphere and bulk soils.

concentration of dissolved Cr(III) (Oze et al., 2007). Mn(II) addition led
to more Cr(III) to be released into the pore water compared with CK
(Fig. 1), and the increased concentration of dissolved Cr(III) provided
more opportunities for its oxidation by Mn(III/IV) oxides. Previous
studies have suggested that continuous flooding (CF) would promote the
reduction of Fe(III) oxides and lead to Cr(III) dissolution and release
(Zhang et al., 2023). However, no increase in Cr(III) concentration in
pore water was observed in the CK treatment, indicating that the
contribution of Cr(III) released by reducing Fe(III) oxides is small (Fig. 1
and S6). This viewpoint was further evidenced by the results of XRD
analysis in soil samples (Figure S11) and the concentration of dissolved
Fe in pore water (Figure S12). In detail, the proportion of dissolved Fe in
pore water to the total Fe concentration in soil was ～0.08 % in the CK
treatment (Figure S12), and most Fe in soil existed as goethite during the
rice whole growth period (Figure S11). Xia et al. (2023) found that the
exchangeable and carbonate-bound Cr (available Cr fractions) increased
under anoxic phase but decreased under oxic phase, indicating the
reversibility of Cr availability, and the Fe–Mn oxide-bound Cr remained
relatively stable during anoxic–oxic alternation. Additionally, a chang­
ing trend of reciprocity was observed in Cr(VI) and dissolved Cr(III)

8
M. Ao et al.
concentrations in the pore water throughout the rice growth period
(Fig. 1 and S6), suggesting that increasing dissolved Cr(III) was the
critical factor in controlling Cr(VI) production. However, Cr(VI) and
dissolved Cr(III) concentrations in the pore water of the CK treatment
remained unchanged throughout the rice growth period (Figure S6),
indirectly indicating that Mn(II) addition promoted the release of Cr(III).
Collectively, the generation of natural oxidants mediated by rice ROL
and MOM accelerated the oxidation of dissolved Cr(III) (Wang et al.,
2014), ultimately leading to an increase in Cr(VI) concentrations in soil
and pore water.
Comparative analysis of Cr(VI) concentrations in pore water and soil
revealed that most of the newly generated Cr(VI) (83–85 %) was
immobilized in soil, and only about 15–17 % of Cr(VI) was released into
pore water (Fig. 2 and S6). Potentially, this is related to the weakly
acidic conditions and the high abundance of Fe(III) oxides in the test soil
(Figures S4 and S11). The surveyed literature suggests that alkaline (pH
> 8) and oxic environments are favorable for enhanced Cr(VI) mobili­
zation, as these conditions minimize competing immobilization pro­
cesses, i.e., sorption and reduction, both of which are favored under
weakly acidic and anoxic conditions (Chrysochoou et al., 2016; Rinklebe
et al., 2016; Schindler et al., 2018; Shadreck & Mugadza, 2013). In this
study, Cr(VI) generated in the weakly acid and reducing environment
(Figures S3, S4 and S5) was easily adsorbed by active minerals in the
soil. Previous studies have shown that although Cr(VI) has a strong
migration capacity, the widely distributed Fe(III) oxides in the envi­
ronmental media have a strong sorption capacity and can effectively
inhibit the release of Cr(VI) (Hai et al., 2020; Xia et al., 2023; Zhang
et al., 2023). As a result, most of the Cr(VI) was immobilized in the soil.
Although most of the Mn in the soil was present in the highly
oxidized state of Mn(III/IV) oxides during the filling and maturing stages
(Fig. 6I–L), there was no significant increase in Cr(VI) in the soil and
pore water. Instead, a decreasing pattern was observed (Fig. 1 and S6),
suggesting that the contribution of dissolved O2 oxidation of Cr(III) may
be small. During the IF stage, the concentration of dissolved Fe in pore
water gradually decreased, which indirectly indicated that the dissolved
Fe in pore water was oxidized by dissolved O2 to generate Fe(III) oxides
(Figure S12). Perhaps IF leads to the oxidation of Fe(II) to Fe(III) oxides
by dissolved O2 in the soil, and the high abundance of Fe(III) oxides in
the soil has a strong sorption capacity for Cr(III) (Hausladen & Fendorf,
2017), leading to a decrease in Cr(III) concentration in the pore water.
For example, Cr(VI) concentration is very low in dryland soils, even in
fully oxidized soils (Chung & Sa, 2001), because O2 oxidation of Cr(III) is
very slow under natural conditions (Ao et al., 2022b; Namgung et al.,
2014; Pan et al., 2017; Qian et al., 2020). Therefore, the decrease of
dissolved Cr(III) may be a key factor in inhibiting the production of Cr
(VI) in oxic environments, despite the large amount of Mn(III/IV) oxides
in soils.
4.2. Increased Cr translocation to grains
The pot experiments showed that Mn(II) addition decreased Cr
concentrations in rice roots and leaves and increased Cr concentration in
stems (Fig. 3). In contrast, Cr concentrations in rice grains increased
most significantly (2.13 (Y-7) and 4.97-fold (Z-14) higher, High-Mn vs
CK) (Fig. 4A). As shown in Table S2, Mn(II) addition significantly
increased the translocation factors (TF) of Cr from roots to stems and
leaves, linearly increasing with the Mn(II) doses. In previously reported
field surveys and pot experiments, it was generally reported that most of
the Cr absorbed by rice accumulated in the roots (70–87 %), and only
about 13–30 % of Cr was transferred to above-ground tissues (Ali et al.,
2022; Mishra et al., 1997; Yu et al., 2018b). Cr distribution in different
parts of rice is in the order of roots > stems > leaves > grains (Cervantes
et al., 2001; Sharma et al., 2005; Xie et al., 2018; Zeng et al., 2010).
Similarly, in this study, Cr in the roots, stems, and leaves of two rice
cultivars followed the same distribution order at the tillering stage.
However, Cr concentration in above-ground tissues (stems and leaves)
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Environment International 175 (2023) 107939
increased obviously in Mn(II) addition treatments from the jointing
stage, but the Cr concentration in the stems was significantly lower than
that in the leaves (Fig. 3). Thus, Mn(II) addition seems to boost Cr
translocation within rice plant tissues (Table S2).
The increased Cr distribution in above-ground tissues (especially in
grains) after Mn(II) addition may be related to the Cr(VI) concentration
in soil and pore water, as well as the high affinity and transmembrane
efficiency of Cr(VI) (Aharchaou et al., 2017; Shanker et al., 2005). Plants
can take up both Cr(III) and Cr(VI) (Ao et al., 2022a). However, there are
significant differences in the pathways and efficiency of their entry into
epidermal root cells (Aharchaou et al., 2017; Ao et al., 2022a). As the
uptake of Cr(III) by plants is a passive process with no energy con­
sumption, Cr(III) is mainly taken up by the root system through ion
channels of essential elements (e.g. Fe, Ca, Mg and K) or by the simple
diffusion of cation exchange sites in the cell wall, which are slow uptake
of Cr(III) (Singh et al., 2013). As an anionic Cr species, Cr(VI) uptake by
plants is an active process and relies on sulphate or phosphate carriers
due to the structural similarity to sulphate and phosphate, and has
higher water solubility and higher transmembrane efficiency (Aharch­
aou et al., 2017; Ding et al., 2019; Shanker et al., 2005; Singh et al.,
2013). The similar distribution pattern (Fig. 5) and the significant pos­
itive correlation (r = 0.30, p < 0.01, Figure S8) of Cr and P in grains also
suggested that Cr(VI) uptake by rice was related to phosphate carriers.
After being absorbed by root tip cells, Cr(III/VI) is transported to aerial
parts through the xylem, where Cr(VI) has fast mobility (Cervantes et al.,
2001), however, Cr(III) is easily absorbed and immobilized by pectins,
proteins, and polysaccharides (rich in carboxyl groups, aldehydes and
hydroxyl groups) in the cell wall and even forms a binuclear insoluble
complex (Chanda & Parmar, 2003; Mishra et al., 1997; Sinha et al.,
2018). Subsequently, much of the Cr(III) accumulates in the cell wall
and becomes an important barrier to decreasing its translocation (Zeng
et al., 2011).
Additionally, previous studies found that the KM values (Michaelis
constant, reflecting affinity between substrate and carrier) of rice for Cr
(VI) and Cr(III) were 0.88 and 1.58 mg/g, respectively, revealing a much
greater affinity of rice seedlings for Cr(VI) than Cr(III) (Yu et al., 2018b).
They found that more Cr was detected in shoots of rice seedlings exposed
to Cr(VI), while Cr(III) treatment limited the root-to-shoot Cr trans­
location (Yu et al., 2018b). These results suggest that Cr(VI) has a higher
translocation efficiency in rice plants. In this study, although Mn(II)
addition significantly increased the Cr(III) concentration in pore water
at tillering stage, the Cr concentration in rice roots did not increase
significantly (Fig. 3A,B), which may be related to the Cr(III) uptake and
translocation efficiency by rice or the oxidation of Cr(III) to Cr(VI) in
pore water. Meanwhile, Cr(III) in rhizosphere pore water was oxidized
to Cr(VI) after the tillering stage, resulting in the decrease of Cr(III)
uptake by rice (Fig. S6A,B,E,F). Furthermore, given the close 1:1 rela­
tionship (Fig. 4B) between soil Cr(VI) levels and Cr in grains (Z-14)
induced by Mn(II) addition, it could be inferred that high levels of Cr in
grains might originate from Cr(VI) in soils, while the contribution of Cr
(III) in pore water may be minor. Consistent with previous studies, they
found that CF significantly decreased Cr(VI) in soil and Cr accumulation
in grains, while alternating wetting and drying treatments contributed
to higher Cr(VI) levels in soil and ultimately more Cr accumulation in
grains (Xiao et al., 2015). These results indicate that Cr(VI) generation
can increase the Cr uptake and accumulation in rice, and decreased by
reduction (Xiao et al., 2021).
Besides root uptake, Cr accumulated in grain may also come from the
remigration of Cr accumulated in the stem. Yu et al. (2018) found that
the cell wall was the primary storage compartment for Cr in rice seed­
lings treated with Cr(III), whereas the cytoplasm was the largest Cr
storage compartment in Cr(VI) treatment. Moreover, the Cr stored in
cytoplasm had higher translocation activity (Bauenova et al., 2021), and
the cell wall of stems have a lower binding capacity for Cr than roots and
leaves (Sinha et al., 2018; Zeng et al., 2011), which collectively resulted
in more Cr transferring to the grain. The filling stage is the most
M. Ao et al.
important stage for Cr to transport from different tissues to grains of rice.
In Z-14, a gradual decrease in Cr concentration in stems was observed
from filling to maturing stages, especially when Mn(II) addition
decreased by 2.25 (Low-Mn) and 4.48 (High-Mn) mg/kg, respectively
(Fig. 3D). When the binding sites of Cr in the stem cell wall were satu­
rated during the filling and maturing stages, excess Cr was subsequently
transferred to the grains, leaves and other cellular organs (Sinha et al.,
2018). However, this phenomenon was not observed in Y-7, which may
suggest that there are differences in the remigration ability and redis­
tribution stage of Cr in different rice varieties and tissues (Yu et al.,
2018a; Zeng et al., 2014).
4.3. Environmental implications
Collectively, current data suggest that increased Mn abundance in
soil may disrupt the redox balance between Cr(III) and Cr(VI), resulting
in a significantly higher net yield of Cr(VI) in soil and pore water. These
Cr(VI) are mainly generated by the oxidation of Cr(III) by Mn(III/IV)
oxides mediated by MOM. The newly generated Cr(VI) is readily
absorbed by rice and transferred to above-ground tissues, eventually
accumulating in the grains, thereby increasing the risk of Cr exposure.
Elevated Cr levels in rice grains are a health concern especially in Asia,
the most important rice producing and consuming region in the world.
In addition, it should be noted that the widespread MOM can mediate
Mn(II) oxidation even without the involvement of rice. Therefore,
increasing soil Mn abundance can greatly increase the production of Cr
(VI) in soil and accelerate the release of Cr(III) as a “source” of Cr(VI).
These results can provide important insights into the interaction of Cr
(III/VI) and Mn(III/IV) in soils and sediments of similar systems under
periodic redox fluctuations, such as river floodplains, wetlands and
peatlands.
However, many questions regarding Mn-promoted Cr(III) dissolution
and oxidation in soil-rice systems remain unanswered and should be the
focus of future research. These include the following: (1) the mechanism
of Mn(II) oxidation in anoxic soils and sediments; (2) the mechanism of
dissolution and oxidation of Cr(III) by the MOM mediated Mn(II)
oxidation in anoxic environments; (3) the key factors affecting the dy­
namic redox balance of Cr(III) and Cr(VI) in anoxic environments; (4)
the oxidation and competition retention of Cr(III) between Fe(III) and
Mn(III/IV) oxides; (5) the contribution of dissolved Cr(III) and Cr(VI) in
soil to their levels within different rice tissues.
5. Conclusions
In the soil-rice system, Mn(II) addition promoted the dissolution of
Cr(III) and provided a prerequisite for the oxidation of Cr(III), resulting
in a significantly higher net yield of Cr(VI) in soil and pore water. The
concentration of Cr(VI) in soil and pore water increased linearly with the
addition of Mn(II) doses. With the involvement of rice, soil Cr(VI) con­
centrations in rhizosphere were slightly higher than that in bulk.
Furthermore, even in the absence of rice, increased Mn abundance in the
soil leads to more Cr(VI) production, possibly by the oxidation of Cr(III)
by Mn(II)-oxidizing microorganisms (Pseudomonas sp. and Bacillus sp.)
mediated generation of Mn(III/IV) oxides. Comparison with pore water
revealed that most (83–85 %) of the newly generated Cr(VI) remained in
the soil, which may be related to the high abundance of Fe(III) oxide
adsorbed in the soil. In rice plants, Mn(II) addition causes more Cr to be
transferred to above-ground tissues, and high ROL rice cultivar had
stronger Cr accumulation capacity. The Cr accumulated in grains may
mainly originate from the newly generated Cr(VI) in the soil, which may
include the uptake of Cr(VI) by the roots at the filling and maturing
stages, and the re-migration of accumulated Cr from the stems.
CRediT authorship contribution statement
Ming Ao: Conceptualization, Validation, Formal analysis,
10
Environment International 175 (2023) 107939
Investigation, Writing – review & editing. Tenghaobo Deng: Investi­
gation, Writing – review & editing. Shengsheng Sun: Methodology,
Formal analysis, Visualization. Mengyao Li: Formal analysis, Investi­
gation. Jingjing Li: Formal analysis, Investigation. Ting Liu: Formal
analysis, Investigation. Bofang Yan: Investigation, Writing – review &
editing. Wen-Shen Liu: Investigation, Writing – review & editing.
Guobao Wang: Investigation, Writing – review & editing. Dedao Jing: .
Yuanqing Chao: . Yetao Tang: Writing – review & editing. Rongliang
Qiu: Funding acquisition, Supervision, Writing – review & editing.
Shizhong Wang: Conceptualization, Validation, Supervision, Funding
acquisition, Project administration, Writing – review & editing.
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.
Acknowledgments
This work is partially supported by the National Key R&D Program of
China (2019YFC1805300), and the National Natural Science Founda­
tion of China (41671313 and 41977118).
Appendix A. Supplementary material
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.envint.2023.107939.
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