Nativa, Sinop, v. 10, n. 3, p. 319-327, 2022.

Pesquisas Agrárias e Ambientais

DOI: https://doi.org/10.31413/nativa.v10i3.13332 ISSN: 2318-7670

Soil waterlogging associated with iron excess potentiates physiological

damage to soybean leaves

Allan de Marcos LAPAZ1*, Camila Hatsu Pereira YOSHIDA2, Carlos Leonardo Pereira BOGAS3,

Liliane Santos de CAMARGOS1, Paulo Alexandre Monteiro de FIGUEIREDO3,

Jailson Vieira AGUILAR1, Ronaldo Cintra LIMA3, Rafael Simões TOMAZ

1São Paulo State University (UNESP), Ilha Solteira, SP, Brazil.

2University of Western São Paulo (UNOESTE), Presidente Prudente, SP, Brazil.

3São Paulo State University (UNESP), Dracena, SP, Brazil.

E-mail: allanlapz60@gmail.com

ORCID: (0000-0003-4798-3713; 0000-0002-8167-3324; 0000-0002-1190-6233; 0000-0002-0979-4447;

0000-0003-4505-6975; 0000-0003-3684-9180; 0000-0001-8316-7145; 0000-0002-5700-5983)

Submitted on 01/17/2022; Accepted on 07/18/2022; Published on 08/19/2022.

ABSTRACT: Many plants are exposed to soil waterlogging, including soybean plants. Soil waterlogging

exponentially increases the availability of iron (Fe) and causes O2 depletion, which may result in excessive uptake

of Fe and shortage of O2 to the roots and also nodules in leguminous plants, resulting in overproduction of

reactive oxygen species and lipid peroxidation. The present study aimed to evaluate physiological damage to

soybean leaves at the second trifoliate (V2) stage when exposed to non-waterlogged and waterlogged soils and

combined with one moderate and two toxic levels of Fe. Soybean plants were vulnerable to soil waterlogging

at all Fe levels tested, presenting the highest values of malonaldehyde, hydrogen peroxide, and Fe accumulation

in the shoot, which resulted in accentuated damage to gas exchange and chlorophyll content, consequently

leading to lower shoot dry weight. In contrast, soybean plants cultivated under optimal water availability showed

less damage caused by excess Fe, mainly at 125 mg dm-3 Fe, since the traits of net photosynthetic rate, water

use efficiency, instantaneous carboxylation efficiency, malonaldehyde, and shoot dry weight were not affected.

Keywords: chlorophylls; gas exchange; Glycine max; ferrous ion.

Encharcamento do solo associado ao excesso de ferro potencializa os danos

fisiológicos às folhas de soja

RESUMO: Muitas plantas estão expostas ao encharcamento do solo, incluindo plantas de soja. O

encharcamento do solo aumenta exponencialmente a disponibilidade de ferro (Fe) no solo e causa depleção de

O2, o que pode resultar na absorção excessiva de Fe e escassez de O2 para as raízes e também nódulos em

plantas leguminosas, resultando em superprodução de espécies reativas de oxigênio e peroxidação lipídica. O

presente estudo teve como objetivo avaliar os danos fisiológicos às folhas de soja no segundo estádio trifoliado

(V2) quando exposta a solos não encharcados e encharcados combinado com um nível moderado e dois níveis

tóxicos de Fe. As plantas de soja foram vulneráveis ao encharcamento do solo em todos os níveis de Fe testados,

apresentando os maiores valores de malonaldeído, peróxido de hidrogênio e acúmulo de Fe na parte aérea, o

que resultou em danos acentuados nas trocas gasosas e no conteúdo de clorofila, consequentemente levando a

menor peso seco de parte aérea. Em contrapartida, plantas de soja cultivadas sob disponibilidade hídrica ótima

apresentaram menos danos causados pelo excesso de Fe, principalmente a 125 mg dm-3 Fe, uma vez que as

características de taxa fotossintética líquida, eficiência do uso da água, eficiência de carboxilação instantânea,

malonaldeído e peso seca da parte aérea não foram afetados.

Palavras-chave: clorofilas; trocas gasosas; Glycine max; íon ferroso.

1. INTRODUCTION

An increase in the occurrence of extreme weather events

is expected due to variation in average rainfall and storms,

resulting in drought and waterlogging of the soil in several

regions of the world (LORETI et al., 2016). Soil waterlogging

leads to considerable losses in agricultural production

(PEDO et al., 2015; RHINE et al., 2010), especially in soils

with high water tables, compacted soils with poor drainage

(BATAGLIA; MASCARENHAS, 1981; KOKUBUN,

2013), and in lowland soils (PEDÓ et al., 2015).

Gaseous exchange between soil and atmosphere are

severely affected in waterlogged soils (GREENWAY et al.,

2006). Besides that, soil structure is destroyed, which causes

dispersion of soil aggregates; consequently, soil pores are

blocked by particles, impeding air and water movement in

soil (RODRÍGUEZ-GAMIR et al., 2011). In waterlogged

conditions, O2 has low solubility and a low diffusion rate

relative to air, which directly affects its supply to the roots

and also nodules in leguminous plants (GREENWAY et al.,

2006; VOESENEK et al., 2006).

Depending on the duration of soil waterlogging, partial

(hypoxia) or complete (anoxia) depletion of O2 may occur by

aerobic microorganisms and plants. With the absence of O2

root tissue, oxidative phosphorylation in the mitochondria is

Soil waterlogging associated with iron excess potentiates physiological damage to soybean leaves

Nativa, Sinop, v. 10, n. 3, p. 319-327, 2022.

320

negligible and ATP synthesis is restricted to substrate

phosphorylation in glycolysis via fermentative pathways

(SCHULZE et al., 2019), which consequently results in a

decline of N-fixation in leguminous plants and in ATP

synthesis (SOUZA et al., 2016).

In leaves, during waterlogging, one of the first detectable

effects is decreased CO2 availability due to reduced stomatal

opening, affecting gas exchange and water status of the plants

(ZHANG et al., 2016; YAN et al., 2018). Consequently,

blockage of the photosynthetic electron transport chain and

a limitation on CO2 assimilation by the Calvin-Benson cycle

may occur, inducing a marked production of reactive oxygen

species (ROS) production (ZHENG et al., 2017; YAN et al.,

2018). The Excess energy in the antenna complex and ion

leakage from the electron transport chain is transferred to O2,

inducing over-production of singlet oxygen (1O2) and

hydrogen peroxide (H2O2) (BARBOSA et al., 2014; ZHENG

et al., 2017). In addition, the reduced ferredoxin electron is

transferred to O2, instead of going to NADP, generating a

superoxide anion (O2•-) at the acceptor side of photosystem

I (PSI; BARBOSA et al., 2014; YAN et al., 2018).

Concomitantly, under anoxic conditions, the

microorganisms present are anaerobic and facultative

anaerobic. These microorganisms utilise alternative electron

acceptors, preferring those allowing the highest energy yields

or that are most readily available, to maintain their

metabolisms (MARANGUIT et al., 2017; LAPAZ et al.,

2022). Insoluble Fe3+ oxides are electron acceptors of

immediate risk to plants, since they are reduced into a more

soluble form (Fe2+) and released into soil pore water,

resulting in excess absorption of Fe by plants (FREI et al.,

2016; MARANGUIT et al., 2017). High levels of Fe in leaf

tissue may affect the net photosynthetic rate due to

degradation of cell membranes and disruption of

photosynthetic protein complexes such as D1 protein

(MÜLLER et al., 2017), by action of ROS via the Fenton’s

and Haber-Weiss’s reactions (BECANA et al., 1998; LAPAZ

et al., 2022).

There are no previous reports on soil waterlogging

combined with excess Fe in soybean plants in the vegetative

phenological stage. From this perspective, it was

hypothesized that the soil waterlogging combined with Fe

excess can increase the damage to the soybean crop, resulting

in the compromise of its development, since tolerance is

often a product of tolerance to anaerobiosis and to toxicities

of the excessively available elements (SINGH; SETTER,

2017). Therefore, the present study aimed to evaluate

physiological damage to leaves and biometric development

traits in soybean plants at the second trifoliate (V2) stage

(FEHR et al., 1971) when exposed to non-waterlogged and

waterlogged soils and combined with one moderate and two

toxic levels of Fe.

2. MATERIAL AND METHODS

2.1. Experimental site and cultivation conditions

The experiment was carried out under greenhouse

conditions at the College of Agricultural and Technological

Sciences, São Paulo State University (UNESP), São Paulo

State, Brazil (21° 29ʹ S, 51° 2ʹ W; 396 m above sea level). The

soybean variety ‘NS 6601 IPRO’ [Glycine max (L.) Merril] was

used.

The soil was a dystrophic Oxisol (SANTOS et al., 2018).

The soil was collected at a 0.0 – 0.2 m depth and presented

the following chemical attributes: pH (CaCl2) 4.6, organic

matter 14 g dm-3, P (resin) 4 mg dm-3, K 2.1 mmolc dm-3, Ca

7 mmolc dm-3, Mg 5 mmolc dm-3, S 5 mg dm-3, B 0.09 mg dm-

3, Cu 0.6 mg dm-3, Fe 0.25 mg dm-3, Mn 16.7 mg dm-3, Zn 1

mg dm-3, potential acidity (H + Al) 18 mmolc dm-3, Al 4

mmolc dm-3, sum of bases 14.1 mmolc dm-3, cation exchange

capacity 32.1 mmolc dm-3, and base saturation 44%.

The base saturation of soil was increased to 70%

(QUAGGIO et al., 1985) by adding CaCO3 and MgCO3,

analytical reagent (AR) grade, at a ratio of 3:1. The soil with

the carbonate salts was incubated for 30 days in pots at a

humidity of 80% of field capacity to allow it to equilibrate

(LAPAZ et al., 2020). The pots were polypropylene with a

capacity of 4 dm3, lined with a polystyrene blanket to avoid

soil loss during the experiment.

2.2. Experimental process

The following fertilisation was carried out per pot: 10 mg

dm-3 N as CO(NH2)2, 200 mg dm-3 P as Ca(H2PO4)2·H2O,

150 mg dm-3 K as K2SO4, 0.5 mg dm-3 B as H3BO3, 0.05 mg

dm-3 Co as (CoCl2·H2O), 1.0 mg dm-3 Cu as CuSO4·5H2O,

0.05 mg dm-3 Mo as H2MoO4, 0.05 mg dm-3 Ni as

NiSO4·7H2O, 5.0 mg dm-3 Mn as MnSO4, and 2.0 mg dm-3

Zn as ZnSO4 (LAPAZ et al., 2020). The K supply was split

into three equal applications, which were supplied before

sowing and at the V2 stage (FEHR et al., 1971). Fe, as

FeCl3·6H2O, was supplied at 0, 100, and 475 mg dm-3 to the

soil to give one natural level of 25 mg dm-3 Fe and two high

levels of 125 and 500 mg dm-3 Fe, respectively.

After four days, 10 soybean seeds were sown at a depth

of 3 cm, after inoculation with the N2-fixing bacterium

Bradyrhizobium japonicum (KIRCHNER, 1896) Jordan (1982)

(Bradyrhizobiaceae), strains SEMIA 587 and 5019. At the

first node with unifoliate leaf (V1; FEHR et al., 1971), the

seedlings were thinned to three representative seedlings per

pot, selecting those with greater vigour and homogeneity of

size.

At the V2 stage, the soil was waterlogged for a period of

10 days, totalling 23 days of experimental conduction from

the germination of soybean seeds. The pots undergoing a

waterlogged soil treatment were placed in larger pots with

non-draining bottoms, and the water level in these was

maintained at 2 cm above the soil surface of the inner pot.

During the experimental conduction, the replenishment

of evapotranspired water for the plots was achieved using

suspended micro-sprinklers, which were activated morning

and afternoon. For this, before sowing, field capacity of the

soil [100% water mass (g) that the soil supports] was

determined (IBAÑEZ et al., 2021). In this way, the soil

humidity was maintained at 80% of the field capacity, except

during the 10 days of stress in the pots where the soil was

waterlogged. In these days, all plants were irrigated manually,

respecting the water levels of each plot.

2.3. Measurements of gas exchange

On the 10th day of stress imposition, gas exchange traits

were determined on the first newly expanded trifoliate leaf

(counting from the apex) of two plants from each pot, using

a LCpro portable infrared gas analyser (ADC Bioscientific

Ltd., Hoddesdon, United Kingdom). Evaluations were

performed on a clear day between 10:00 and 11:30 a.m.

Photosynthetically active radiation (PAR) was standardised

to an artificial saturating light of 1000 µmol m-2 s-1, 380 μmol

Lapaz et al.

Nativa, Sinop, v. 10, n. 3, p. 319-327, 2022.

321

CO2 mol-1 air and a chamber temperature of 28°C, according

to Lapaz et al. (2020). The net photosynthetic rate (A, µmol

CO2 m-2 s-1), stomatal conductance (gs, mol H2O m-2 s-1) and

transpiration rate (E, mmol H2O m-2 s-1), water use efficiency

[WUE (A/E), μmol CO2 mmol-1 H2O], and instantaneous

carboxylation efficiency [EiC (A/Ci), mol air-1] were

obtained. Ci (µmol CO2 mol ar-1) is the internal CO2 level in

the substomatal chamber.

Subsequently, at the end of the experiment, the first

newly expanded trifoliate leaves (counting from the apex)

were collected and frozen in liquid nitrogen and the following

traits were measured:

2.4. Determination of photosynthetic pigments

Chlorophyll a (Chl a), chlorophyll b (Chl b), total

chlorophyll (Tchl), and carotenoid (CAR) contents were

quantified by spectrophotometric method described by

Lichtenthaler; Wellburn (1983). Leaves from two plants in

each pot were assayed. Fresh leaf tissue (0.5 g) was macerated

in 5 mL cold 80% acetone. The results were expressed in as

μg mL-1, and calculated as described by Aguiar et al. (2021).

2.5. Determination of hydrogen peroxide content and

lipid peroxidation amount

Hydrogen peroxide (H2O2) contents were determined

according to Alexieva et al. (2001). Leaves from two plants

of each pot were assayed. Fresh leaf tissue (0.25 g) was

macerated in 3 mL of 0.1% trichloroacetic acid (TCA) with

20% polyvinylpolypyrrolidone. Subsequently, the samples

were centrifuged at 10,000 rpm for 10 min at 4°C. For the

reaction, 0.2 mL of supernatant was added to 0.2 mL 100 mM

potassium phosphate buffer pH 7.5 and 800 μL 1 M KI

solution. The samples were kept on ice for one hour, and

then absorbance readings were taken at 390 nm. The H2O2

content was calculated based on a standard curve of H2O2,

and the results were expressed in nmol g-1 FW.

Lipid peroxidation was evaluated by determining the

amount of malondialdehyde (MDA) reactive to 2-

thiobarbituric acid (TBA), as described by Heath and Packer

(1968). The initial procedures for MDA measurement were

the same as those described above for H2O2 measurements.

Following centrifugation, 0.25 mL of supernatant was added

to 1 mL 20% TCA solution containing 0.5% TBA. The

samples were kept in a dry bath at 95°C for 30 min and then

on ice for 20 min. Subsequently, the samples were

centrifuged at 10,000 rpm for 5 min and supernatants were

read at 535 and 600 nm. The results were expressed in nmol

g-1 FW and calculated as described by Lapaz et al. (2020).

2.6. Soybean development and Fe accumulation in

shoots

The shoot was cut close to the collar with the aid of

cutting pliers. The samples were packaged in paper bags and

oven dried at 65°C for 72 h, followed by the determined of

shoot dry weight (SDW). The results were expressed as g

plant-1.

The samples were ground in a Wiley-type mill and

submitted to digestion with nitric-perchloric acid solution

(3:1) at 200°C (MALAVOLTA et al., 1997) to determine the

Fe accumulation in the shoot (FeAS). Next, Fe concentration

was determined by atomic absorption spectrophotometry.

FeAS (µg SDW-1) was calculated by multiplying Fe

concentration by the SDW (LAPAZ et al., 2020).

2.7. Experimental design and data analysis

The experimental design was completely randomised and

arranged in a 2 × 3 factorial scheme, with two water regimes

in the soil (optimum and waterlogged conditions) and three

soil Fe levels (25, 125, and 500 mg dm-3). Each pot contained

three plants, giving a final population of nine plants per

treatment.

Normality and homoscedasticity of the data were

analysed using the Shapiro-Wilk’s and Bartlett’s tests (p <

0.05). Then, the data were subjected to analysis of variance

(ANOVA) using the F test (p ≤ 0.05). The traits were

compared using the Tukey’s test (p < 0.05). Chl a was

transformed by √x. Statistical analysis was performed in the

R software (R Development Core Team, 2019).

3. RESULTS

3.1. Gas exchange and photosynthesis pigment content

A showed a significant effect of double interaction on

ANOVA (Table 1). Plants cultivated under optimal water

availability showed no difference in A. Conversely, there was

a reduction in A under waterlogged soil associated with 125

and 500 mg dm-3 Fe, which resulted in lower values when

compared to those under optimal water availability (Figure

1).

Figure 1. Net photosynthetic rate of soybean, based on the

significance of ANOVA by factorial analysis (p ≤ 0.05), comprised

of soils with two water regimes (optimum and waterlogged

conditions) and three iron levels (25, 125, and 500 mg dm-3).

Different letters indicate significant differences according to the

Tukey’s test (p < 0.05). Uppercase letters compare each water

regime between different iron levels in the soil, while lowercase

letters compare water regimes with the same iron levels. Vertical

bars represent the standard error.

Figura 1. Taxa fotossintética líquida da soja, baseada na significância

da ANOVA por análise fatorial (p ≤ 0,05), composta por solos com

dois regimes hídricos (condições ótimas e encharcadas) e três níveis

de ferro (25, 125 e 500 mg dm-3). Letras diferentes indicam

diferenças significativas de acordo com o teste de Tukey (p < 0,05).

Letras maiúsculas comparam cada regime hídrico entre os diferentes

níveis de ferro no solo, enquanto letras minúsculas comparam os

regimes hídricos com os mesmos níveis de ferro. As barras verticais

representam o erro padrão.

The traits gs, E, and WUE showed an isolated effect for

water regime and Fe level, while EiC showed an isolated

effect only for water regime on ANOVA (Table 1). Plants

subjected to waterlogged soil showed reductions in gs (36%),

E (23.9%), WUE (15.8%), and EiC (5.7%) compared to those

under optimal water availability (Table 2). In relation to Fe,

the greatest reduction in gs and WUE occurred in plants

Soil waterlogging associated with iron excess potentiates physiological damage to soybean leaves

Nativa, Sinop, v. 10, n. 3, p. 319-327, 2022.

322

exposed to 500 mg dm-3 Fe, while E decreased in those

exposed to 125 mg dm-3 Fe (Table 2).

Chl a, Chl b, Tchl, and CAR contents showed an isolated

effect for water regime and Fe level on ANOVA (Table 1).

Plants subjected to waterlogged soil showed reductions in

Chl a (27.7%), Chl b (41.5%), Tchl (80.9%), and CAR (29.7%)

contents compared to those under optimal water availability

(Table 3). There were reductions in Chl a, Chl b, and Tchl

contents in plants cultivated at 125 and 500 mg dm-3. In

contrast, CAR content was less sensitive, showing a

reduction only at 500 mg dm-3 Fe (Table 3).

Table 1. Summary of ANOVA by factorial analysis (p ≤ 0.05) composed of soils with two water regimes (optimum and waterlogged

conditions) and three iron levels (25, 125, and 500 mg dm-3) for the soybean traits of net photosynthetic rate (A), stomatal conductance

(gs), transpiration rate (E), water use efficiency (WUE), instantaneous carboxylation efficiency (EiC), chlorophyll a (Chl a), chlorophyll b

(Chl b), total chlorophyll (Tchl), carotenoids (CAR), malondialdehyde (MDA), hydrogen peroxide (H2O2), shoot dry weight (SDW), and

iron accumulation in the shoots (FeAS).

Tabela 1. Resumo da ANOVA por análise fatorial (p ≤ 0,05) composta de solos com dois regimes hídricos (condições ótimas e encharcadas)

e três níveis de ferro (25, 125 e 500 mg dm-3) para as características de taxa fotossintética líquida (A), condutância estomática (gs), taxa de

transpiração (E), eficiência de uso de água (WUE), eficiência de carboxilação instantânea (EiC), clorofila a (Chl a), clorofila b (Chl b), clorofila

total (Tchl), carotenóides (CAR), malondialdeído (MDA), peróxido de hidrogênio (H2O2), peso seca da parte aérea (SDW) e acúmulo de

ferro na parte aérea (FeAS) da soja.

Traits Waterlogged Iron Interaction

***

WUE

***

EiC

***

Chl

***

Chl

Tchl

***

CAR

MDA

***

SDW

***

FeAS

***

Significance: ns, not significant; *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Significância: ns, não significativo; *, p < 0,05; **, p < 0,01; ***, p < 0,001.

Table 2. Stomatal conductance (gs, µmol CO2 m-2 s-1), transpiration rate (E, mmol H2O m-2 s-1), water use efficiency (WUE, μmol CO2

mmol-1 H2O), and instantaneous carboxylation efficiency (EiC, mol air-1) of soybean, based on the significance of ANOVA by factorial

analysis (p ≤ 0.05), comprised of soils with two water regimes (optimum and waterlogged conditions) and three iron levels (25, 125, and 500

mg dm-3).

Tabela 2. Condutância estomática (gs, µmol CO2 m-2 s-1), taxa de transpiração (E, mmol H2O m-2 s-1), eficiência do uso da água (WUE,

μmol CO2 mmol-1 H2O) e eficiência de carboxilação instantânea (EiC, mol air-1) da soja, baseada na significância da ANOVA por análise

fatorial (p ≤ 0,05), composta por solos com dois regimes hídricos (condições ótimas e encharcadas) e três níveis de ferro (25, 125 e 500 mg

dm-3).

Factor levels

WUE

EiC

Water regimes

Optimum

0.30A ± 0.01

5.79A ± 0.18

1.77A

± 0.31

0.035A ± 0.0009

Waterlogged

0.19B ± 0.01

4.46B ± 0.26

1.49B

± 0.13

0.024B ± 0.0015

Iron levels

0.28a ± 0.02

5.80a ± 0.26

1.65ab

± 0.36

0.033a ± 0.0016

125

0.23b ± 0.02

4.87b ± 0.37

1.78a

± 0.12

0.030a ± 0.0028

500

0.13b

± 0.02

5.16ab

± 0.30

1.46b

± 0.11

0.027a

± 0.0030

Different letters indicate significant differences according to the Tukey’s test (p < 0.05). Uppercase letters compare water regimes, while lowercase letters

compare iron levels. ± means standard error.

Letras diferentes indicam diferenças significativas de acordo com o teste de Tukey (p < 0,05). Letras maiúsculas comparam os regimes hídricos, enquanto as

letras minúsculas comparam os níveis de ferro. ± significa o erro padrão.

3.2. Toxicity and Fe accumulation in shoots

Amount of MDA and SDW showed a significant effect

of double interaction, while H2O2 content and FeAS showed

an isolated effect for water regimes and Fe levels on ANOVA

(Table 1). At 500 mg dm-3 Fe, an increase was observed in

amount of MDA in both water regimes compared to plants

at 25 mg dm-3 Fe under optimal water availability (Figure 2a).

Amount of MDA was higher in plants cultivated under

waterlogged soil than those with optimal water availability

(Figure 2a), registering an average increase of 62%.

SDW showed a decrease of 13.7% in plants cultivated

under optimal water availability only at 500 mg dm-3 Fe

compared to plants cultivated under optimal water

availability at 25 mg dm-3 Fe (Figure 2b). Excess Fe

potentiated this reduction in SDW when associated with soil

waterlogging, recording reductions of 43.1 and 78.4% at 125

and 500 mg dm-3 Fe, respectively, compared to plants at 25

mg dm-3 Fe under optimal water availability. SDW was higher

in plants cultivated under optimal water availability than in

waterlogged soil (Figure 2b).

Plants exposed to waterlogged soil showed the highest

contents of H2O2 and FeAS (12.1 and 92.8%, respectively) in

relation to those under optimal water availability (Table 4).

There was a progressive increase in H2O2 in response to

excess Fe, whereas FeAS only increased at 500 mg dm-3 Fe

(Table 4).

Lapaz et al.

Nativa, Sinop, v. 10, n. 3, p. 319-327, 2022.

323

Figure 2. Amount of malonaldehyde (MDA, a) and shoot dry weight (SDW, b) of soybean, based on the significance of ANOVA by factorial

analysis (p ≤ 0.05), comprised of soils with two water regimes (optimum and waterlogged conditions) and three iron levels (25, 125, and 500

mg dm-3). Different letters indicate significant differences according to the Tukey’s test (p < 0.05). Uppercase letters compare each water

regime between different iron levels in the soil, while lowercase letters compare water regimes with the same iron levels. Vertical bars

represent the standard error.

Figura 2. Quantidade de malondialdeído (MDA, a) e peso seco da parte aérea (SDW, b) da soja, baseada na significância da ANOVA por

análise fatorial (p ≤ 0,05), composta por solos com dois regimes hídricos (condições ótimas e encharcadas) e três níveis de ferro (25, 125 e

500 mg dm-3). Letras diferentes indicam diferenças significativas de acordo com o teste de Tukey (p < 0,05). Letras maiúsculas comparam

cada regime hídrico entre os diferentes níveis de ferro no solo, enquanto letras minúsculas comparam os regimes hídricos com os mesmos

níveis de ferro. As barras verticais representam o erro padrão.

Table 3. Chlorophyll a (Chl a, μg mL-1), chlorophyll b (Chl b, μg mL-1), total chlorophyll (Tchl, μg mL-1) e carotenoid (CAR, μg mL-1)

contents of soybean, based on the significance of ANOVA by factorial analysis (p ≤ 0.05), comprised of soils with two water regimes

(optimum and waterlogged conditions) and three iron levels (25, 125, and 500 mg dm-3).

Tabela 3. Conteúdo de clorofila a (Chl a, μg mL-1), clorofila b (Chl b μg mL-1), clorofila total (Tchl, μg mL-1) e carotenoides (CAR, μg mL-

1) da soja, baseada na significância da ANOVA por análise fatorial (p ≤ 0,05), composta por solos com dois regimes hídricos (condições

ótimas e encharcadas) e três níveis de ferro (25, 125 e 500 mg dm-3).

Factor levels

Chl

Tchl

CAR

Water regimes

Optimum

12.82A ± 0.72

6.22A ± 1.00

19.05A ± 1.64

8.44A ± 0.87

Waterlogged

9.27B ± 0.36

3.64B ± 0.25

12.91B ± 0.60

5.93B ± 0.37

Iron levels

12.82a ± 1.02

6.69a ± 1.47

19.52a ± 2.41

8.99a ± 1.12

125

10.22b ± 0.92

4.10b ± 0.43

14.32b ± 1.34

6.93ab ± 0.64

500

10.10b ± 0.78

3.99b ± 0.50

14.10b ± 1.27

5.64b ± 0.52

Different letters indicate significant differences according to the Tukey test (p < 0.05). Uppercase letters compare water regimes regardless of iron level, while

isolated lowercase letters compare iron levels regardless of water regime. ± means standard error.

Letras diferentes indicam diferenças significativas de acordo com o teste de Tukey (p < 0,05). Letras maiúsculas comparam cada regime hídrico entre diferentes

níveis de ferro no solo, enquanto letras minúsculas comparam os regimes hídricos com os mesmos níveis de ferro. ± significa o erro padrão.

Table 4. Hydrogen peroxide (H2O2, nmol g-1 FW) content and iron accumulation in the shoots (FeAS, µg SDW-1) of soybean, based on the

significance of ANOVA by factorial analysis (p ≤ 0.05), comprised of soils with two water regimes (optimum and waterlogged conditions)

and three iron levels (25, 125, and 500 mg dm-3).

Tabela 4. Conteúdo de peróxido de hidrogênio (H2O2, nmol g-1 FW e acúmulo de ferro na parte aérea ((FeAS, µg SDW-1) da soja, baseada

na significância da ANOVA por análise fatorial (p ≤ 0,05), composta por solos com dois regimes hídricos (condições ótimas e encharcadas)

e três níveis de ferro (25, 125 e 500 mg dm-3).

Factor levels

FeAS

Water regimes

Optimum

171.26B ± 9.15

232.65B ± 30.93

Waterlogged

192.00A ± 11.41

448.63A ± 39.57

Iron levels

145.34c ± 9.42

308.48b ± 44.42

125

181.63b ±

7.56

302.63b ± 48.10

500

209.86a ± 12.45

519.72a ± 56.15

Different letters indicate significant differences according to the Tukey’s test (p < 0.05). Uppercase letters compare water regimes, while isolated lowercase

letters compare iron levels. ± means standard error.

Letras diferentes indicam diferenças significativas de acordo com o teste de Tukey (p < 0,05). Letras maiúsculas comparam os regimes hídricos, enquanto as

letras minúsculas comparam os níveis de ferro. ± significa o erro padrão.

Soil waterlogging associated with iron excess potentiates physiological damage to soybean leaves

Nativa, Sinop, v. 10, n. 3, p. 319-327, 2022.

324

4. DISCUSSION

4.1. Impact on gas exchange

The gs and EiC showed decreases under soil

waterlogging, configuring stomatal and biochemical

limitations, which reflected in a lower A (Figure 1 and Table

2). The decrease in gs (Table 2) detected under soil

waterlogging was probably an adaptive defence measure to

prevent water loss and dehydration of tissues (YAN et al.,

2018). Although soybean plants present morphological

adaptations to tolerate waterlogging (THOMAS et al., 2005),

they suffer until they develop a sufficient aerenchymatous

network for the diffusion of O2 to the roots (SHIMAMURA

et al., 2010). With the reduction of gs, consequently the E was

reduced; however, it was not enough to optimise the WUE

(Table 2). A similar result was observed by Velasco et al.

(2019) in bean (Phaseolus vulgaris L.), they observed a reduction

in gs under waterlogging prevents water loss by E, but

culminated in a reduction in WUE in the plurality of cultivars

studied.

In relation to the increase Fe levels, gs showed a similar

reduction at 125 and 500 mg dm-3 Fe (Table 2), which may

explain the lower A when combined with waterlogged soil

(Figure 1). Biochemical limitation under Fe excess was not

verified in EiC (Table 2). Conversely, Pereira et al. (2013)

observed a decrease in A in rice (Oryza sativa L.) due to

stomatal and biochemical limitations, with biochemical

limitation more severe in the most sensitive cultivar. The

decrease in gs (Table 2) was probably linked to the increase in

FeAS (Table 4). According to Dufey et al. (2009), the

reduction in stomatal opening is a late response to increased

Fe uptake, rather than a defence mechanism in rice. As

expected, the decrease in gs was immediately reflected in a

lower E, which was crucial so as not to affect WUE at 125

mg dm-3 Fe.

When comparing the current results with those of Lapaz

et al. (2020), a study similar to this research, but at the

beginning of grain filling (R3; FEHR et al., 1971), it is notable

that the negative effect on gas exchange was more deleterious

than that observed at V2 (Figure 1 and Table 2), being more

pronounced under waterlogged soil combined with excess

Fe.

4.2. Reduction of photosynthetic pigment content

The photosystems (PSI and PSII) in plants are composed

of a core complex (Chl a and β-carotene) and a peripheral

antenna system (Chls a and b and carotenoids) (WIENTJES

et al., 2017). Under soil waterlogging and Fe excess, the

content of Chl a and Chl b behaved similarly to that of Tchl

(Table 3). Recent reports have shown a decrease in Chl a and

Chl b content, but with notably greater damage to Chl b, in

Jerusalem artichoke (Helianthus tuberosus L.; YAN et al., 2018),

mung bean (Vigna radiate L. Wilzeck; SAIRAM et al., 2009)

and sorghum (Sorghum bicolor L. Moench; ZHANG et al.,

2019). Under Fe excess, previous studies with potato

(Solanum tuberosum L.; CHATTERJEE et al., 2006) pea (Pisum

sativum L.; XU et al., 2015) and Elodea nuttallii (Planch.) H. St.

John (XING et al., 2010) also reported higher degradation of

Chl b content.

Hence, it was assumed that both stresses degraded the

chlorophylls, leading to photooxidative and oxidative damage

to photosystems (XU et al., 2015), as verified by the increase

in amount of MDA and H2O2 content (Figure 2a and Table

4). Consequently, the reduction of photosynthetic pigments

can affect the light energy utilisation and dissipation (LAPAZ

et al., 2020). Besides that, the degradation observed in

carotenoids content (Table 3) can decrease the resistance of

chloroplasts to ROS, favouring the increase of lipid

peroxidation (LAPAZ et al., 2019), since carotenoids can act

as direct quenchers of triplet chlorophyll and singlet oxygen

with simultaneous transition to the triplet state (MASLOVA

et al., 2021).

4.3. Toxicity and Fe accumulation in shoots

Under waterlogging conditions, the potential redox of the

soil solution generally decreases favouring the reduction of

Fe3+ to Fe2+ (XU et al., 2018), which is its more soluble form

and can result in absorption of excess Fe (LAPAZ et al.,

2020). To avoid Fe toxicity, plants decrease the availability of

Fe in the soil through the build-up of Fe plaque, limit the

translocation of Fe towards the shoots, and sequester Fe in

vacuoles, plastids, and cell walls as ferritin complexes

(ARAÚJO et al., 2020). The increase in H2O2 content and

amount of MDA in soybean leaves (Figure 2a; Table 4)

showed that these strategies were not sufficient to keep Fe at

a functional level in the plant, allowing it to react with O2

(LAPAZ et al., 2022). Plants subjected to waterlogged soil at

500 mg dm-3 Fe had higher FeAS values (Table 4), even with

the decrease in SDW (Table 4). In this treatment, the highest

values of H2O2 were observed (Table 4).

Soil waterlogging and Fe excess caused changes in the

dynamics of the function of the photosynthetic apparatus

(Figure 1 and Table 2), which can induce the overproduction

of ROS causing lipid peroxidation (XING et al., 2010;

ZHENG et al., 2017; YAN et al., 2018; LAPAZ et al., 2020;

LAPAZ et al., 2022). Malondialdehyde is one of membrane

lipid peroxidation products, and its amount can reflect the

stability of the cell membrane (WANG et al., 2022).

Therefore, the responses of this work indicate that the

antioxidant system was not efficient in containing the

degradation of the membranes, except at 125 mg dm-3 Fe

under optimal water availability where there was no increase

in amount of MDA (Figure 2a). Similar results under

waterlogged soil (WANG et al., 2022) and excess Fe (XING

et al., 2010) were observed in G. max (R1 – one flower at any

node; FEHR et al., 1971) and E. nuttallii, respectively.

It is important to emphasise that the effects of

waterlogging are complex, and vary depending on genotype,

environmental conditions, growth stage and the duration of

waterlogging (TIAN et al., 2019). The limitation of biological

N fixation by soybeans (Souza et al. 2016) and the

consequence of a change from oxidative phosphorylation to

glycolysis and fermentation imposed by soil waterlogging

leads to a progressive loss of biomass and performance by

plants (GREENWAY et al., 2006; SAIRAM et al., 2009;

MARTÍNEZ-ALCÁNTARA et al., 2012), corroborating

with the results verified in SDW (Figure 2b). These

aggravations were more evident in soybean plants under

waterlogged soil associated with Fe excess. According to

Müller et al. (2017), tolerance of Fe toxicity by cultivars or

species is reflected in their biomass, which is dependent in

part by an ability to mitigate excessive Fe uptake via exclusion

mechanisms and/or by storing/remobilising absorbed Fe.

5. CONCLUSIONS

In summary, soybean plants were vulnerable to soil

waterlogging at all Fe levels tested, presenting the highest

values of MDA, H2O2, and FeAS, which resulted in

Lapaz et al.

Nativa, Sinop, v. 10, n. 3, p. 319-327, 2022.

325

accentuated damage to gas exchange and chlorophyll

content, consequently leading to lower SDW. In contrast,

soybean plants cultivated under optimal water availability

showed less damage caused by excess Fe, mainly at 125 mg

dm-3 Fe, since the traits of A, WUE, EiC, MDA, and SDW

were not affected.

6. ACKNOWLEDGEMENTS

The authors are grateful to the São Paulo Research

Foundation (FAPESP; grant number #2018/17380-4,

#2018/01498-6, and #2020/12421-4), for the scholarship to

support the first author and for the support received to fund

this research.

7. REFERENCES

AGUILAR, J. V.; LAPAZ, A. DE M.; SANCHES, C. V.;

YOSHIDA, C. H. P.; CAMARGOS, L. S. D.;

FURLANI-JÚNIOR, E. Application of 2, 4-D hormetic

dose associated with the supply of nitrogen and nickel on

cotton plants. Journal of Environmental Science and

Health, Part B, v. 56, n. 9, p. 852-859, 2021.

https://doi.org/10.1080/03601234.2021.1966280

ALEXIEVA, V.; SERGIEV, I.; MAPELLI, S.; KARANOV,

E. The effect of drought and ultraviolet radiation on

growth and stress markers in pea and wheat. Plant, Cell

& Environment, v. 24, n. 12, p. 1337-1344, 2001.

https://doi.org/10.1046/j.1365-3040.2001.00778.x

ARAÚJO, T. O.; ISAURE, M. P.; CHOUBASSI, G.;

BIERLA, K.; SZPUNAR, J.; TRCERA, N.; CHAY, S.;

ALCON, C.; SILVA, L. C.; CURIE, C.; MARI, S.

Paspalum urvillei and Setaria parviflora, two grasses naturally

adapted to extreme iron-rich environments. Plant

Physiology and Biochemistry, v. 151, n. 6, p. 144-156,

2020. https://doi.org/10.1016/j.plaphy.2020.03.014

BARBOSA, M. R.; SILVA, M. M. A.; WILLADINO, L.;

ULISSES, C.; CAMARA, T. R. Geração e desintoxicação

enzimática de espécies reativas de oxigênio em plantas.

Ciência Rural, v. 44, n. 3, p. 453-460, 2014.

https://doi.org/10.1590/S0103-84782014000300011

BATAGLIA, O. C.; MASCARENHAS, H. A. A. Toxicidade

de ferro em soja. Bragantia, v. 40, n. 1, p. 199-203, 1981.

https://doi.org/10.1590/S0006-87051981000100021

BECANA, M.; MORAN, J. F.; Iturbe-Ormaetxe, I. Iron-

dependent oxygen free radical generation in plants

subjected to environmental stress: toxicity and

antioxidant protection Plant and soil, v. 201, n. 1, p. 137-

147, 1998. https://doi.org/10.1023/A:1004375732137

CHATTERJEE, C.; GOPAL, R.; DUBE, B. K. Impact of

iron stress on biomass, yield, metabolism and quality of

potato (Solanum tuberosum L.). Scientia horticulturae, v.

108, n. 1, p. 1-6, 2006.

https://doi.org/10.1016/j.scienta.2006.01.004

DUFEY, I.; HAKIZIMANA, P.; DRAYE, X.; LUTTS, S.;

BERTIN, P. QTL mapping for biomass and

physiological parameters linked to resistance mechanisms

to ferrous iron toxicity in rice. Euphytica, v. 167, n. 2, p.

143-160, 2009. https://doi.org/10.1007/s10681-008-

9870-7

FEHR, W. R.; CAVINESS, C. E.; BURMOOD, D. T.;

PENNINGTON, J. S. Stage of development

descriptions for soybeans, Glycine Max (L.) Merrill1. Crop

science, v. 11, n. 6, p. 929-931, 1971.

https://doi.org/10.2135/cropsci1971.0011183X001100

060051x

FREI, M.; TETTEH, R. N.; RAZAFINDRAZAKA, A. L.;

FUH, M. A.; WU, L. B.; BECKER, M. Responses of rice

to chronic and acute iron toxicity: genotypic differences

and biofortification aspects. Plant and Soil, v. 408, n. 1,

p. 149-161, 2016. https://doi.org/10.1007/s11104-016-

2918-x

GREENWAY, H.; ARMSTRONG, W.; COLMER, T. D.

Conditions leading to high CO2 (> 5 kPa) in

waterlogged–flooded soils and possible effects on root

growth and metabolism. Annals of Botany, v. 98, n. 1,

p. 9-32, 2006. https://doi.org/10.1093/aob/mcl076

HEATH, R. L.; PACKER, L. Photoperoxidation in isolated

chloroplasts. Archives of biochemistry and

biophysics, v. 125, n. 1, p. 189-198, 1968.

https://doi.org/10.1016/0003-9861(68)90654-1

IBAÑEZ, T. B.; SANTOS, L. F.; LAPAZ, A. M.; RIBEIRO,

I. V.; RIBEIRO, F. V.; REIS, A. R.; MOREIRA, A.;

HEINRICHS, R. Sulfur modulates yield and storage

proteins in soybean grains. Scientia Agricola, v. 78, n. 1,

p. e20190020, 2020. https://doi.org/10.1590/1678-

992X-2019-0020

KOKUBUN, M. Genetic and cultural improvement of

soybean for waterlogged conditions in Asia. Field Crops

Research, v. 152, n. 14, p. 3-7, 2013.

https://doi.org/10.1016/j.fcr.2012.09.022

LAPAZ, A. M.; CAMARGOS, L. S.; YOSHIDA, C. H. P.;

FIRMINO, A. C.; FIGUEIREDO, P. A. M.; AGUILAR,

J. V.; NICOLAI, A. B.; PAIVA, W. S.; CRUZ, V. H.;

TOMAZ, R. S. Response of soybean to soil waterlogging

associated with iron excess in the reproductive stage.

Physiology and Molecular Biology of Plants, v. 26, n.

8, p. 1635-1648, 2020. https://doi.org/10.1007/s12298-

020-00845-8

LAPAZ, A. M.; SANTOS, L. F. M.; YOSHIDA, C. H. P.;

HEINRICHS, R.; CAMPOS, M.; REIS, A. R.

Physiological and toxic effects of selenium on seed

germination of cowpea seedlings. Bragantia, v. 78, n. 4,

p. 498-508, 2019. https://doi.org/10.1590/1678-

4499.20190114

LAPAZ, A. M.; YOSHIDA, C. H. P.; GORNI, P. H.;

FREITAS-SILVA, L.; ARAÚJO, T. O.; RIBEIRO, C.

Iron toxicity: effects on the plants and detoxification

strategies. Acta bot. bras, v. 36, p. e2021abb0131, 2022.

https://doi.org/10.1590/0102-33062021abb0131

LICHTENTHALER, H. K.; WELLBURN, A. R.

Determinations of total carotenoids and chlorophylls a

and b of leaf extracts in different solvents. Biochemical

Society Transactions, v. 11, p. 591-592, 1983.

LORETI, E.; VAN VEEN, H.; PERATA, P. Plant responses

to flooding stress Current Opinion in Plant Biology, v.

33, p. 64-71, 2016.

https://doi.org/10.1016/j.pbi.2016.06.005

MALAVOLTA, E.; VITTI, G. C.; OLIVEIRA, A. S.

Avaliação do estado nutricional de plantas:

princípios e aplicações. 2. ed. Piracicaba: Potafos, 1997.

319p

MARANGUIT, D.; GUILLAUME, T.; KUZYAKOV, Y.

Effects of flooding on phosphorus and iron mobilization

in highly weathered soils under different land-use types:

Short-term effects and mechanisms. Catena, v. 158, n.

14, p. 161-170, 2017.

https://doi.org/10.1016/j.catena.2017.06.023

MARTÍNEZ-ALCÁNTARA, B.; JOVER, S.; QUIÑONES,

A.; FORNER-GINER, M. Á.; RODRÍGUEZ-GAMIR,

Soil waterlogging associated with iron excess potentiates physiological damage to soybean leaves

Nativa, Sinop, v. 10, n. 3, p. 319-327, 2022.

326

J.; LEGAZ, F.; PRIMO-MILLO, E.; IGLESIAS, D. J.

Flooding affects uptake and distribution of carbon and

nitrogen in citrus seedlings. Journal of Plant

Physiology, v. 169, n. 12, p. 1150-1157, 2012.

https://doi.org/10.1016/j.jplph.2012.03.016

MASLOVA, T. G.; MARKOVSKAYA, E. F.; SLEMNEV,

N. N. Functions of carotenoids in leaves of higher plants.

Biology Bulletin Reviews, v. 11, n. 5, p. 476-487, 2021.

https://doi.org/10.1134/S2079086421050078

MÜLLER, C.; SILVEIRA, S. F. S.; DALOSO, D. M.;

MENDES, G. C.; MERCHANT, A.; KUKI, K. N.;

OLIVA, M. A.; LOUREIRO, M. E.; ALMEIDA, A. M.

Ecophysiological responses to excess iron in lowland and

upland rice cultivars. Chemosphere, v. 189, n. 24, p. 123-

133, 2017.

https://doi.org/10.1016/j.chemosphere.2017.09.033

PEDO, T.; KOCH, F.; MARTINAZZO, E. G.; VILLELA,

F. A.; AUMONDE, T. Z. Physiological attributes,

growth and expression of vigor in soybean seeds under

soil waterlogging. African Journal of Agricultural

Research, v. 10, n. 39, p. 3791-3797, 2015.

https://doi.org/10.5897/AJAR2015.9661

PEREIRA, E. G.; OLIVA, M. A.; ROSADO-SOUZA, L.;

MENDES, G. C.; COLARES, D. S.; STOPATO, C. H.;

ALMEIDA, A. M. Iron excess affects rice photosynthesis

through stomatal and non-stomatal limitations. Plant

Science, v. 201-202, n. 3, p. 81-92, 2013.

https://doi.org/10.1016/j.plantsci.2012.12.003

QUAGGIO, J. A.; VAN RAIJ, B.; MALAVOLTA, E.

Alternative use of the SMP‐buffer solution to determine

lime requirement of soils. Communications in Soil

Science and Plant Analysis, v. 16, n. 3, p. 245-260,

1985. https://doi.org/10.1080/00103628509367600

R Development Core Team (2019) R: a Language and

Environment for Statistical Computing. R

Foundation for Statistical Computing, Vienna,

Austria. Disponível em: https://www.R-project.org/

RHINE, M. D.; STEVENS, G.; SHANNON, G.;

WRATHER, A.; SLEPER, D. Yield and nutritional

responses to waterlogging of soybean cultivars.

Irrigation Science, v. 28, n. 2, p. 135-142, 2010.

https://doi.org/10.1007/s00271-009-0168-x

RODRÍGUEZ-GAMIR, J.; ANCILLO, G.; CARMEN

GONZÁLEZ-MAS, M.; PRIMO-MILLO, E.;

IGLESIAS, D. J.; FORNER-GINER, M. A. Root

signalling and modulation of stomatal closure in flooded

citrus seedlings. Plant Physiology and Biochemistry, v.

49, n. 6, p. 636-645, 2011.

https://doi.org/10.1016/j.plaphy.2011.03.003

SAIRAM, R. K.; DHARMAR, K.; CHINNUSAMY, V.;

MEENA, R. C. Waterlogging-induced increase in sugar

mobilization, fermentation, and related gene expression

in the roots of mung bean (Vigna radiata). Journal of

Plant Physiology, v. 166, n. 6, p. 602-616, 2009.

https://doi.org/10.1016/j.jplph.2008.09.005

SANTOS, H. G.; JACOMINE, P. K. T.; ANJOS, L. H. C.;

OLIVEIRA, V. A.; LUMBRERAS, J. F.; COELHO, M.

R.; ALMEIDA, J. A.; CUNHA, T. J. F.; OLIVEIRA, J.

B. Sistema brasileiro de classificação de solos. 3.ed.

Brasília: Embrapa, 2013. 353p.

SCHULZE E. D.; BECK E.; BUCHMANN N.; CLEMENS

S.; MÜLLER-HOHENSTEIN K.; SCHERER-

LORENZEN M. In: Plant Ecology. Springer, Berlin,

Heidelberg, 2019. p. 143-164.

https://doi.org/10.1007/978-3-662-56233-8_5

SHIMAMURA, S.; YAMAMOTO, R.; NAKAMURA, T.;

SHIMADA, S.; KOMATSU, S. Stem hypertrophic

lenticels and secondary aerenchyma enable oxygen

transport to roots of soybean in flooded soil. Annals of

botany, v. 106, n. 2, p. 277-284, 2010.

https://doi.org/10.1093/aob/mcq123

SINGH, S. P.; SETTER, T. L. Effect of waterlogging on

element concentrations, growth and yield of wheat

varieties under farmer’s sodic field conditions.

Proceedings of the National Academy of Sciences,

India Section B: Biological Sciences, v. 87, n. 2, p.

513-520, 2017. https://doi.org/10.1007/s40011-015-

0607-9

SOUZA, S. C. R.; MAZZAFERA, P.; SODEK, L. Flooding

of the root system in soybean: biochemical and molecular

aspects of N metabolism in the nodule during stress and

recovery. Amino acids, v. 48, n. 5, p. 1285-1295, 2016.

https://doi.org/10.1007/s00726-016-2179-2

THOMAS, A. L.; GUERREIRO, S. M. C.; SODEK, L.

Aerenchyma formation and recovery from hypoxia of the

flooded root system of nodulated soybean. Annals of

Botany, v. 96, n. 7, p. 1191-1198, 2005.

https://doi.org/10.1093/aob/mci272

TIAN, L.; LI, J.; BI, W.; ZUO, S.; LI, L.; LI, W.; SUN, L.

Effects of waterlogging stress at different growth stages

on the photosynthetic characteristics and grain yield of

spring maize (Zea mays L.) under field conditions.

Agricultural Water Management, v. 218, n. 8, p. 250-

258, 2019. https://doi.org/10.1016/j.agwat.2019.03.054

VELASCO, N. F.; LIGARRETO, G. A.; DÍAZ, H. R.;

FONSECA, L. P. M. Photosynthetic responses and

tolerance to root-zone hypoxia stress of five bean

cultivars (Phaseolus vulgaris L.). South African Journal of

Botany, v. 123, n. 4, p. 200-207, 2019.

https://doi.org/10.1016/j.sajb.2019.02.010

VOESENEK, L. A. C. J.; COLMER, T. D.; PIERIK, R.;

MILLENAAR, F. F.; PEETERS, A. J. M. How plants

cope with complete submergence. New phytologist, v.

170, n. 2, p. 213-226, 2006.

https://doi.org/10.1111/j.1469-8137.2006.01692.x

WANG, S., ZHOU, H., FENG, N., XIANG, H., LIU, Y.,

WANG, F., LI, W.; FENG, S.; LIU M.; ZHENG, D.

Physiological response of soybean leaves to uniconazole

under waterlogging stress at R1 stage. Journal of Plant

Physiology, v. 268, n. 1, p. 153579, 2022.

https://doi.org/10.1016/j.jplph.2021.153579

WIENTJES, E.; PHILIPPI, J.; BORST, J. W.; VAN

AMERONGEN, H. Imaging the Photosystem

I/Photosystem II chlorophyll ratio inside the leaf.

Biochimica et Biophysica Acta (BBA)-

Bioenergetics, v. 1858, n. 3, p. 259-265, 2017.

https://doi.org/10.1016/j.bbabio.2017.01.008

XING, W.; LI, D.; LIU, G. Antioxidative responses of Elodea

nuttallii (Planch.) H. St. John to short-term iron

exposure. Plant Physiology and Biochemistry, v. 48,

n. 10-11, p. 873-878, 2010.

https://doi.org/10.1016/j.plaphy.2010.08.006

XU, S.; LIN, D.; SUN, H.; YANG, X.; ZHANG, X. Excess

iron alters the fatty acid composition of chloroplast

membrane and decreases the photosynthesis rate: a study

Lapaz et al.

Nativa, Sinop, v. 10, n. 3, p. 319-327, 2022.

327

in hydroponic pea seedlings. Acta Physiologiae

Plantarum, v. 37, n. 10, p. 1-9, 2015.

https://doi.org/10.1007/s11738-015-1969-6

XU, Y.; SUN, X.; ZHANG, Q.; LI, X.; YAN, Z. Iron plaque

formation and heavy metal uptake in Spartina alterniflora at

different tidal concentrations and waterlogging

conditions. Ecotoxicology and environmental safety,

v. 153, n. 7, p. 91-100, 2018.

https://doi.org/10.1016/j.ecoenv.2018.02.008

YAN, K.; ZHAO, S.; CUI, M.; HAN, G.; WEN, P.

Vulnerability of photosynthesis and photosystem I in

Jerusalem artichoke (Helianthus tuberosus L.) exposed to

waterlogging. Plant Physiology and Biochemistry, v.

125, n. 4, p. 239-246, 2018.

https://doi.org/10.1016/j.plaphy.2018.02.017

ZHANG, F.; ZHU, K.; WANG, Y. Q.; ZHANG, Z. P.; LU,

F.; YU, H. Q.; ZOU, J. Q. Changes in photosynthetic and

chlorophyll fluorescence characteristics of sorghum

under drought and waterlogging stress.

Photosynthetica, v. 57, n. 4, p. 1156-1164, 2019.

https://doi.org/10.32615/ps.2019.136

ZHANG, Y.; CHEN, Y.; LU, H.; KONG, X.; DAI, J.; LI,

Z.; DONG, H. Growth, lint yield and changes in

physiological attributes of cotton under temporal

waterlogging. Field Crops Research, v. 194, p. 83-93,

2016. https://doi.org/10.1016/j.fcr.2016.05.006

ZHENG, X. D.; ZHOU, J. Z.; TAN, D. X.; WANG, N.;

WANG, L.; SHAN, D. Q.; KONG, J. Melatonin

improves waterlogging tolerance of Malus baccata (Linn.)

Borkh. seedlings by maintaining aerobic respiration,

photosynthesis and ROS migration. Frontiers in Plant

Science, v. 8, p. 483, 2017.

https://doi.org/10.3389/fpls.2017.00483