Plant Soil

https://doi.org/10.1007/s11104-022-05432-2

RESEARCH ARTICLE

Surface liming triggers improvements in subsoil fertility

and root distribution to boost maize crop physiology, yield
and revenue
João William Bossolani · Carlos Alexandre Costa Crusciol · Letusa Momesso · José Roberto Portugal ·
Luiz Gustavo Moretti · Ariani Garcia · Mariley de Cássia da Fonseca · Vitor Alves Rodrigues ·
Juliano Carlos Calonego · André Rodrigues dos Reis

Received: 1 October 2021 / Accepted: 6 April 2022

© The Author(s), under exclusive licence to Springer Nature Switzerland AG 2022

Abstract  Methods  The treatments consisted of four dolo-

Background and aims  Liming is widely used to alle-
viate soil acidity worldwide. However, the vast major-
ity of studies with liming are restricted to agricultural
systems that incorporate lime into the soil, not con-
sidering its effects as surface applications. Although
liming effects on soil fertility and crop yield are well
understood, there are few studies that elucidate the
role of soil improvements in the established crop
physiology and the revenue in lime-amended soils,
especially when cultivated in regions prone to agro-
climatic risks. Here, we address the effects of surface
liming, for three growing seasons (2017–2019) sub-
sequent to the lime treatment (2016), on soil fertility,
root growth, crop nutrition, photosynthetic pigment
concentrations, gas exchange parameters and produc-
tion costs of maize cultivated in a tropical region on
an acidic soil with low water regime.
Responsible Editor: N. Jim Barrow.
J. W. Bossolani · C. A. C. Crusciol (*) · L. Momesso ·
J. R. Portugal · L. G. Moretti · A. Garcia ·
M. de Cássia da Fonseca · V. A. Rodrigues · J. C. Calonego  in response to lime supply. Additionally, root develop-
College of Agricultural Sciences, Department of Crop
Science, São Paulo State University (UNESP), Botucatu,
São Paulo 18610‑034, Brazil
e-mail: carlos.crusciol@unesp.br
A. R. dos Reis  resulted in greater net profit over the 3 years studied.
School of Sciences and Engineering, Department
of Biosystems Engineering, São Paulo State University
(UNESP), Tupã, São Paulo 17602‑496, Brazil
mitic lime doses applied to the soil surface as follows:
i) control (untreated soil), ii) half the recommended
dose (½ RD), iii) full recommended dose (1 RD) and
iv) twice the recommended dose (2 RD).
Results  Surface liming increased soil fertility, and
higher doses provided better results. ­Ca2+ and ­Mg2+
concentrations increased at greater depths under
higher lime doses, directly influencing maize root
growth. Even under low water availability, also in the
driest year of 2018, this liming induced improvement
of growing conditions and increased growth was
observed’. Maize grown under lime at 2 RD exhibited
better nutrition, improved chlorophylls concentration,
photosynthetic parameters and water use efficiency.
As a result, both shoot growth and grain yield also
increased. Net profit in the first growing season was
higher in 1 RD, whereas in the two following grow-
ing seasons the application of 2 RD resulted in higher
revenue.
Conclusions  Increased water use efficiency, chloro-
phyll and photosynthesis were the main physiological
traits regulating the growth and yield of maize plants
ment was favoured in the entire soil profile, mainly in
deeper layers, after improvements on soil fertility by
cascading effects of liming. These results were more
prominent in 2RD lime-amended soil, which also

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Keywords  Field experiment · Acidic soils · Root
growth · Photosynthesis · Water use efficiency
Introduction
Maize (Zea mays L.) is one of the most important
cereal crops worldwide, and demand for this cereal
has increased in recent years (International Grains
Council 2019). Compared with the productivity of
subtropical regions, the average maize yield in tropi-
cal regions is low, especially when maize is grown
during autumn/winter season (International Grains
Council 2019; USDA 2020). The low maize yield
in tropical regions occurs in detriment of unfavour-
able climatic conditions during cultivation, which are
aggravated by lower soil quality (Kiani et al. 2017).
One of the main factors that reduce the productive
potential of crops most, is the soil acidity (Fageria
and Nascente 2014). Acidic soils represent approxi-
mately 80% of potentially cultivable soils globally
(von Uexküll and Mutert 1995; Gibbs and Salmon
2015) and are mostly located in tropical regions.
Tropical soils are characterized by low nutrient con-
centrations and high concentrations of toxic elements
such as aluminum (­Al3+) and manganese ­ (Mn2+)
(Caires et al. 2011; Crusciol et al. 2019). These toxic
elements inhibit crop root growth, magnify the nega-
tive effects of environmental stresses, and limit the
acquisition of water and nutrients from the subsoil
(Carmeis Filho et  al. 2017a). Therefore, to guaran-
tee food security, agricultural management prac-
tices, such as lime application, are indispensable for
achieving an adequate soil quality and health neces-
sary for crop production. Liming has a potential ben-
efit of increasing soil structure aggregation as well as
the crop’s resilience against agroclimatic variability
in long-term land management systems (Kiani et  al.
2017; Bossolani et al. 2020a, 2021b).
Liming as surface applications can improve the
chemical (Caires et al. 2006, 2011; Joris et al. 2016;
Carmeis Filho et  al. 2017a; Rheinheimer et  al.
2018; Crusciol et  al. 2019), physical (Carmeis Filho
et  al. 2017b, 2018) and biological properties of soil
(Bossolani et al. 2020a; Bossolani et al. 2021b; Hol-
land et al. 2018), and is an appropriate soil manage-
ment practice to reverse the decline of soil quality by
acidification processes, resulting in gains in produc-
tivity. Lime is used extensively in surface applications
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Plant Soil
in no-till systems (Auler et  al. 2019; Nunes et  al.
2019), but its poor solubility raises doubts about
its efficiency in subsurface layers (Bossolani et  al.
2020b). Among the liming materials used in agricul-
ture are dolomite [CaMg(CO3)2], calcium carbonate
­(CaCO3), calcium oxide (CaO), and calcium hydrox-
ide [Ca(OH)2], although dolomite lime is undoubt-
edly the most widely used soil amendment due to
its ability to also supply ­Mg2+ in addition to ­Ca2+
(Caires et  al. 2006; Bossolani et  al. 2018, 2020a; Li
et al. 2019).
Although beneficial impacts of liming on soil qual-
ity have been described, the relationship between soil
quality and the physiological performance of crops
grown in chemically corrected soils has not received
as much attention, especially in long-term field
experiments that reduce contradictory outcomes from
short-term time series and possible misleading inter-
pretations (Cusser et  al. 2020). Therefore, our study
brings an approach of how the improved soil fertility
with liming affects the root growth to deeper layers
and the physiology of maize, culminating in increases
in yield responses and revenue to farmers. In detail,
our study was based on physiological processes that
were indirectly improved by liming in chemically
corrected soil, such as improvements in production
of photosynthetic pigments, net photosynthetic rates,
stomatal conductance, and water use efficiency by
maize grown under low water regime as well as rev-
enue gains over years. The hypotheses of this study
were as follows:
(i) a pluri-annual surface liming improves soil fer-
tility up to 1 m depth;
(ii) in this modified soil profile, root growth is pro-
moted and roots are more uniformly distributed
among soil layers;
(iii) plants established in these lime-amended soils
have greater photosynthetic capacity, even
under limited water availability;
(iv) the combined effect of improvements in soil
and plant properties results in higher crop
yields and revenue of maize production over
years.
These hypotheses were tested in a long-term
experiment started in 2002 under a no-tillage system
on a tropical soil in Brazil during three growing sea-
sons (2017–2019), which evaluated the benefits of
Plant Soil
surface lime application at different doses on soil fer-
tility and the root growth, physiological parameters,
grain yield, as well as costs and net profit of maize
production.
Material and methods
Site description
A long-term field experiment (registered on the
GLTEN Metadata Portal; https://​www.​glten.​org/​
exper​iments/​62) is carried out in Botucatu, São Paulo
State, Southeastern Brazil (48°25′37″ W, 22° 49′50″
S, 765 m above sea level) under no tillage since 2002.
The climate is Cwa, humic subtropical zone, with dry
winters and hot summers, according to the Köppen-
Geiger climate classification system (Alvares et  al.
2013). The long-term (1956–2020) annual maximum
and minimum temperatures registered in this region
were 26.1 and 15.3  °C, respectively. The soil is a
sandy clay loam kaolinitic thermic Typic Haplorthox
(USDA 2014), corresponding to Oxisols (Jahn et  al.
2006), with 347, 108 and 545 g ­kg−1 of clay, silt and
sand, respectively. The chemical properties of the soil
prior to establishment of the study were determined
at 0.0–0.2  m depth according to the methodology
described by van Raij et al. (2001): soil pH (­CaCl2):
4.2; soil organic matter (SOM): 21 g ­kg−1; phospho-
rus ­(Presin): 9.2 mg ­kg−1; potassium ­(K+): 1.2 ­mmolc
­kg−1; exchangeable calcium ­(Ca2+): 14 m ­ molc ­kg−1;
2+
exchangeable magnesium ­ (Mg ): 5 ­ mmolc ­kg−1;
−1
total acidity (H  + 
Al): 37 ­ mmolc ­kg , CEC: 57
­mmolc ­kg−1; BS: 35%.
Experimental design and establishment of treatments
The experimental design is randomized complete
blocks with four replications. Each experimental unit
(plots) covered an area of 56.7  ­m2 (6.3  m × 9.0  m).
The treatments consisted of four sedimentary dolo-
mitic lime doses surface-applied (without incorpora-
tion): i) control (lime-untreated soil), ii) half the rec-
ommended dose (½ RD), iii) full recommended dose
(1 RD) and iv) twice the recommended dose (2 RD).
Over 17 experiment years in the field, the treat-
ments of lime doses were applied four times in 2002,
2004, 2010 and 2016. From 2002 to 2010, liming
doses calculation considered the 0.0–0.2 m soil depth
and are described in detail in (Costa et  al. 2018).
From 2016, the fourth lime application was calcu-
lated considering the lime recommended dose at the
layer of 0.0–0.4  m depth to increase the soil fertil-
ity in deeper layers, and based on previous results
obtained in this experimental area by Carmeis Filho
et  al. (2017a). These authors found that the classic
liming recommendation based on the 0.0–0.2 m layer
represents an underestimate for stable cropping sys-
tems under no-till with crop rotation and high input
of crop residues throughout the agricultural year.
Thus, the new calculation resulted in doses of 0 (no
lime), 3250, 6500 and 13,000  kg  ­ha−1, which were
applied in October 2016. Reapplications were per-
formed following the criterion of new lime dose must
be applied when the soil (recommended dose treat-
ment) reached base saturation (BS) ≤ 50%,. To fulfil
this premise, soil fertility was verified annually. For
all applications, dolomitic lime was used as the lim-
ing agent; lime composition was 233 g ­kg−1 of CaO
(calcium oxide) and 175 g ­kg−1 of MgO (magnesium
oxide). The recommended dose (RD) was calculated
to increase the BS in the topsoil (0.0–0.4 m depth) to
70% adapted from van Raij et al. (1997), as shown in
Eq. (1):
(( ) )
( −1
) BS2 − BS1 × CEC 0.0−0.4 m
RD Mg ha =
ECCE × 10
(1)
where ECCE is the effective calcium carbonate
equivalent of the dolomitic lime, ­BS2 is the stimated
base saturation (70%) and ­BS1 is the base saturation
measured in the soil analysis as follows in Eq. (2):
( 2+
Ca + Mg2+ + K + ×100
)
BS1 (%) = (2)
CEC
where ­Ca2+, ­Mg2+, and ­K+ are basic exchangeable
cations ­(mmolc ­kg−1) [sodium ­ (Na+) is negligible
in these soils], and CEC is the total cation exchange
capacity, calculated as shown in Eq. (3):
CEC mmolc kg−1 = Ca2+ + Mg2+ + K+ + total acidity pH 7 (H + Al)
( ) ( )
(3)
This study corresponds to the first (2017), sec-
ond (2018) and third (2019) year after the fourth
lime reapplication (2016), characterized as long-
term residual effect. Several crops were cultivated
during the agricultural years from 2002 to 2019.
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 Plant Soil

Previous cultivated crops and details about lime Crop management

applications are shown in Table 1.
Maize (simple hybrid Pioneer - P3707VYH) was
sown in March 2017, 2018 and 2019. For all grow-
ing seasons, the fertilization was performed at sowing

Table 1  Scheme of crops and treatments during the experimental period (from 2002 to 2019)
Growing season Crops Treatmentsa

2002/2003 Nov. Oriza sativa Liming

Apr. Avena strigosa (71% ­ECCEb)
2003/2004 Jan. Phaseolus vulgaris –
Apr. A. strigosa
2004/2005 Nov. Arachis hypogaea Liming
Apr. Avena sativa (71% ECCE)
2005/2006 Nov. A. hypogaea –
May A. sativa
2006/2007 Feb. Zea mays intercropped with Urochloa brizantha (cv. Marandu) –
– U. brizantha (cv. Marandu)
2007/2008 Dec. Z. mays intercropped with U. brizantha (cv. Marandu) –
– U. brizantha (cv. Marandu)
2008/2009 Dec. Glycine max –
Jun. A. strigosa
2009/2010 Oct. G. max –
Mar. Sorghum vulgare
2010/2011 Nov. Z. mays Liming
Apr. Crambe abyssinica (88% ECCE)
Sept. Vigna unguiculata
2011/2012 Nov. Z. mays –
May C. abyssinica
Sept. V. unguiculata
2012/2013 Jan. Pennisetum glaucum –
Apr. Triticum aestivum
2013/2014 Nov. P. vulgaris –
Mar. T. aestivum
2014/2015 Dec. P. vulgaris –
Mar. U. brizantha (cv. Marandu)
2015/2016 – U. brizantha (cv. Marandu) –
– U. brizantha (cv. Marandu)
2016/2017c Oct. G. max Liming
Mar. Z. mays intercropped with Urochloa ruziziensis (69% ECCE)
2017/2018 Oct. G. max –
Mar. Z. mays intercropped with U. ruziziensis
2018/2019 Oct. G. max –
Mar. Z. mays intercropped with U. ruziziensis
a
 The reapplications in October 2004, 2010 and 2016 were performed when base saturation reached ≤50%
b
 ECCE: effective calcium carbonate equivalents
c
 Micronutrients applied in total area

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time with 350  kg  ­ha−1 of 08–28–16 (N–P2O5–K2O)
and 90 kg N ­ha−1 as ammonium sulfate in topdress-
ing (Cantarella et al. 1997) at maize V ­ 4 phenological
stage (Ritchie et al. 1993) in all treatments. Phytosan-
itary treatments were carried out according to the
needs of maize. To avoid micronutrient unavailability
to the plants due to the increase in pH by higher lime
doses, a micronutrient based fertilizer was applied
over a total area at rates of 3  kg B ­ha−1 + 1  kg Cu
­ha−1 + 1  kg Mn ­ha−1 + 10  kg Zn ­ha−1 + 0.2  kg Mo
­ha−1.
Meteorological measurements
During the experimental period, meteorological
data (rainfall, solar radiation, wind speed, relative
humidity and maximum and minimum temperatures)
were measured by an automatic meteorology station
Fig. 1  Climatological water balance at Botucatu-SP, Brazil,
from 2002 to 2016 (A), and during the maize crop cycle in
2017 (B), 2018 (C), and 2019 (D). ETc, crop evapotranspira-
installed near to the experiment (~50  m). Evapo-
transpiration reference (­ET0) was calculated by the
Penman-Monteith method (Allen et  al. 1998). Crop
evapotranspiration (ETc) was calculated using the
crop coefficient (Kc) for each phenological stage
(Allen et  al. 1998) of the maize. Using the rainfall
data, climatological water balance was monitored and
calculated using electronic spreadsheet, as described
by Rolim et  al. (1998), following the procedure of
Thornthwaite and Mather (1955) to obtain the real
evapotranspiration (ETr). The climatological water
balance from experimental data (three growing sea-
sons) is shown in Fig. 1.
Soil chemical properties analysis
At 36  months after the last reapplied lime (2016),
eight individual soil subsamples were randomly
tion; ETr, real evapotranspiration. The arrows indicate the
managing and sampling time
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◂Fig. 2  Changes in pH (A), soil organic carbon (SOC) (B),
exchangeable calcium ­(Ca2+) (C) and magnesium ­(Mg2+) (D),
base saturation (BS) (E) and aluminum saturation (AS) (F) in
response to liming doses [control (lime-untreated soil), half
the recommended rate (½ RD), recommended rate (1 RD) and
twice the recommended rate (2 RD)] in 2019, three years after
liming. Different lower-case letters indicate significant dif-
ferences between treatments by Student’s t test at p ≤ 0.05 for
each soil depth. Error bars express the standard error of the
mean (n = 4)
collected from 0.0–0.1, 0.1–0.2, 0.2–0.4, 0.4–0.6,
0.6–0.8, and 0.8–1.0  m depths of each plot, with
the exception of the micronutrients (Fe, Mn, Cu and
Zn) that were sampled and analysed at the depth of
0.0–0.2  m, and were combined into one compos-
ite sample. The samples were dried, sieved (2-mm
sieves), and analysed to determine their chemical
properties according to Cantarella et  al. (1998). The
soil pH was determined in a 0.01 M C­ aCl2 suspension
(1:2.5 soil/solution) (Quaggio and van Raij 2001).
The soil organic carbon (SOC) was determined
according to the Walkley–Black method (Walkley
and Black 1934). Exchangeable basic cations ­(Ca2+,
­Mg2+, and K­ +) were extracted using an ionic resin
(van Raij et  al. 2001) and were determined using a
Shimadzu AA-6300 atomic absorption/flame-emis-
sion spectrophotometer. The total acidity at pH  7.0
(H + Al) was extracted by 0.5  M Ca acetate at pH  7
and determined by titration with 0.025 M NaOH solu-
tion (van Raij et  al. 2001). Exchangeable A ­ l3+ was
extracted with a neutral 1 M KCl solution (1:10 soil/
solution) and determined by titration with a 0.025 M
NaOH solution (van Raij et al. 2001). The micronutri-
ents (Fe, Mn, Cu and Zn) were extracted by solution
containing 0.005  M diethylenetriaminepentaacetic
acid pH  7.3, 0.1  M triethanolamine and of 0.01  M
­CaCl2 and then determined by atomic absorption
spectrophotometry (van Raij et  al. 2001). The base
saturation (BS) and aluminium saturation (AS) val-
ues were calculated as described by Cantarella et al.
(1998) and expressed as percentage.
Root development
At maize full flowering (­R2 phenological stage;
Ritchie et al. 1993), eight root subsamples were col-
lected randomly from each plot and combined to
compose the sample. The root samplings were per-
formed from plant rows and in the middle of the
inter-rows of each sub-plot. A galvanized-steel
probe with a 82-mm diameter cutting tip was used
at 0.0–0.1, 0.1–0.2, 0.2–0.4, 0.4–0.6, 0.6–0.8, and
0.8–1.0  m depths. Roots were carefully separated
from soil and other residues by washing under flow
of swirling water over a 0.5-mm mesh sieve. Root
dry matter was evaluated after drying in a forced-air
oven at 60 °C for 72 h. The results were expresssed in
g ­m−3 and subsequently stimated to Mg ­ha−1 for the
layer of 0.0–1.0 m depth. The root dry matter distri-
bution was calculated by the ratio between root dry
matter in each layer and total root dry matter, with the
result multiplied by 100.
Chemical element analysis
The analyses were performed on the diagnostic leaves
of maize when the plants were at full flowering stage
­(R2 phenological stage; Ritchie et al. 1993). The diag-
nostic leaves of maize used were the fully expanded
sunlit leaves in the top third of the maize canopy.
Except N, all nutrients (P, K, Ca, Mg S, Fe, Mn, Cu
and Zn) were digested via nitroperchloric digestion
and determined via atomic absorption spectropho-
tometry. Nitrogen was digested using a H ­ 2SO4 (sulfu-
ric acid) extraction solution and determined through
the Kjeldahl distillation method (AOAC 2016).
Photosynthetic pigments
Photosynthetic pigments (chlorophylls and carot-
enoids) were performed based on the description of
Lichtenthaler (1987) using N, N-dimethylformamide
(DMF), in diagnostic maize leaves (­R2 phenological
stage; Ritchie et  al. 1993). The extraction was car-
ried out using five discs cut between the edge and the
central rib of the maize leaves, using a paper punch
(0.5 cm in diameter) and 2 mL of DMF. The concen-
trations of chlorophyll a, chlorophyll b, total chloro-
phyll and carotenoids were expressed in μg ­cm−2.
Gas exchange parameters
Evaluations of gas exchange consisted of non-destruc-
tive analyses in diagnostics leaves ­(R2 phenological
stage; Ritchie et  al. 1993) using a Portable Infrared
Gas Analyzer CIRAS-3 Portable Photosynthesis
System (PP Systems Inc., Amesbury, MA, USA).
The following parameters were determined: ­ CO2
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Fig. 3  Changes in iron (Fe)

(A), manganese (Mn) (B),
copper (Cu) (C) and zinc
(Zn) (D) concentrations in
soil at 0.0–0.2 m depth, in
response to liming doses
[control (lime-untreated
soil), half the recommended
rate (½ RD), recommended
rate (1 RD) and twice the
recommended rate (2 RD)]
in 2019, three years after
liming. Different lower-case
letters indicate significant
differences between treat-
ments by Student’s t test at
p ≤ 0.05. Error bars express
the standard error of the
mean (n = 4)

assimilation rate expressed by area (A − μmol ­CO2
­m−2 ­s−1), stomatal conductance (gs − mol ­H2O
­m−2 ­s−1), leaf transpiration (E − mmol ­H2O ­m−2 ­s−1),
internal ­CO2 concentration in the substomatal cham-
ber (ic − mmol ­CO2 mol ­air−1) and water use effi-
ciency (WUE; A/E ratio − μmol ­CO2 (mmol ­H2O)−1).
The initial conditions imposed for measurements
were 1000 μmol ­m−2 ­s−1 of photosynthetically active
radiation (PAR), provided by LED lamps, 380  ppm
of ­CO2, and a chamber temperature of 28  °C. The
measurements were performed between 8:00 and
10:00 am.
Agronomic parameters and grain yield
At the end of the experiment, the maize plants were
contained along 5 m of the 4 central rows from each
plot and were harvested. Subsequently, the shoot
and grains were separeted. The shoot was dried in
a forced air circulation oven, weighed using an ana-
lytical balance and the results were expressed in
Mg ­ha−1. The grain yield was calculated based on the
relationship between the grain mass of each plot and
their respective water content (adjusted at 130 g ­kg−1
of water). The root:shoot ratio was calculated by the
ratio between root dry matter (for the 0.0–1.0 m layer)
and shoot dry matter. Harvest index was calculated by
the ratio between grain yield and total shoot dry mat-
ter (shoot dry matter + grain yield).
Economic evaluations
Determination of liming cost was by lime dose
applied (Mg  ­ha−1) plus operational cost per Mg
applied, both in US$ ­ha−1, in each plot only in the first
growing season. Cost per hectare (US$ ­ha−1) of maize
production was calculated according to CONAB
(2021) and was equal for all treatments. Mean total
costs were determined by liming plus maize produc-
tion costs. Gross is the revenue per ha calculated
using the formula: maize grain yield (Mg ­ha−1) × US$
0.21 (i.e., in dollars per unit of production). Net is the
return per ha calculated using the formula: gross ­ha−1
– total cost ­ha−1. Costs of lime application and maize
production, and net profit were calculated using mean
prices of US$ 4.3 of the last three years (2017–2019).

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Analysis of the results
In all of the datasets considered, the normality of
the data was analyzed using the Anderson-Darling
test, and homoscedasticity was analyzed by Levene’s
test. After that, the data were considered normal, and
subsequently, subjected to the analysis of individual
variance ANOVA by the F test (p ≤ 0.05), and when
significant, the means were analysed using the Fish-
er’s protected least significant difference (LSD) at
p ≤ 0.05. Comparisons of the means of soil chemi-
cal properties were conducted for each soil layer to
assess treatment effects. Comparisons were not made
between different soil depths. The heatmap was per-
formed calculating the Pearson correlation coef-
ficients (p ≤ 0.05) and only significant correlations
were shown. Hierarchical clustering was constructed
based on Gower distance of similarity with 1000
bootstraps.
Results
Soil chemical analysis and root development in the
soil profile
At 36  months after the last reapplication of surface
liming, the primary chemical attributes of the acidic
tropical soil differed significantly (p < 0.01), but the
magnitudes of the differences varied according to soil
chemical properties, lime dose and soil depth (Fig. 2).
Soil pH was strongly altered by lime dose com-
pared with the control (untreated soil), mainly in the
uppermost soil layers. Remarkable effects occurred
for the application of twice the recommended rate
(2 RD), which altered soil pH more efficiently than
the other doses down to a depth of 0.4 m (Fig. 2A).
At layers deeper than 0.4  m, the effects of the lime
doses were similar and superior to those of the con-
trol treatment. Notably, applying lime effectively
increased SOC content (Fig.  2B), concentration of
exchangeable ­Ca2+ (Fig. 2C) and ­Mg2+ (Fig. 2D), and
base saturation (Fig. 2E) in all soil layers, and these
effects were proportional to the lime rate: 2 RD > 1
RD > ½ RD > control. In addition, aluminum satura-
tion (AS) (Fig. 2F) was greatly reduced by the appli-
cation of lime at any dose, mainly in the 0.0–0.1 and
0.1–0.2 m layers; effects proportional to the lime dose
were obtained in intermediate layers (0.2–0.4  m). In
the deepest soil layers (0.6–0.8 and 0.8–1.0 m), 2 RD
lime reduced aluminum saturation more efficiently
than the other doses compared with the control. The
concentration of micronutrients in the 0.0–0.2 m lay-
ers was significantly (p < 0.01) changed by the lime
doses (Fig.  3); Fe, Mn, Cu and Zn concentrations
(Fig. 3A−D) were lowest at the highest lime dose.
Lime doses significantly affected root dry matter
production along the soil profile and the root distri-
bution (p < 0.01) (Fig. 4). The production of root dry
matter increased proportionally with the lime dose in
all soil layers up to 1.0 m depth, regardless of grow-
ing season (Fig.  4A, C and E). The root dry matter
distribution revealed that the proportion of roots in
the 0.0–0.1  m and 0.1–0.2  m layers was highest in
the control treatment, followed by ½ RD, whereas in
the soils amended with 1 RD and, in particular, 2 RD,
roots were better distributed in the soil profile, includ-
ing at deeper depths (Fig. 4B, D and F).
Nutritional status of plants
Uptake of N, Mg and S was higher in plants estab-
lished in 2 RD lime-amended soil, regardless of grow-
ing season (Table 2). Interestingly, in 2017 and 2018,
leaf concentration of P and Ca did not differ between
½ RD, 1 RD and 2 RD treatments, but were higher
than the control. On the other hand, in 2019, maize
plants grown under 2 RD-amended soil presented
the highest values of P and Ca, when compared with
other treatments. By contrast, the leaf concentrations
of Fe, Mn, Cu and Zn were highest in plants grown in
untreated and ½ RD lime-amended soil. In the three
growing seasons, higher doses of lime provided lower
micronutrient concentrations in maize leaves.
Photosynthetic pigments and gas exchange
measurements
Lime application led to increased (p < 0.01) chloro-
phyll and carotenoid concentrations in all growing
seasons, mainly at 2 RD treatment (Fig. 5). Leaf gas
exchange also changed by liming doses (Fig. 6). Net
photosynthetic rate (A) (Fig.  6A), stomatal conduct-
ance (gs) (Fig.  6B) and water use efficiency (WUE)
(Fig. 6E) increased proportionally with the lime dose,
whereas the opposite occurred for internal C ­ O2 (ic)
and leaf transpiration (E) (Fig. 6C, D).
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◂Fig. 4  Root dry matter (A) and root dry matter distribution
(B) of maize in response to lime doses [control (lime-untreated
soil), half the recommended rate (½ RD), recommended rate
(1 RD) and twice the recommended rate (2 RD)] in the three
growing seasons. Different lower-case letters indicate sig-
nificant differences between treatments by Student’s t test at
p ≤ 0.05 for each soil depth. Error bars express the standard
error of the mean (n = 4)
Agronomic parameters and grain yield
In the three growing seasons, all agronomic param-
eters of maize strongly differed (p < 0.01) by the lime
doses, except for root:shoot ratio (Fig. 7B). Untreated
and ½ RD lime-amended soils provide the lowest
shoot dry matter (Fig. 7A) and grain yield (Fig. 7C),
as well as the lowest harvest index (Fig.  7D). Con-
sidering the average of the three growing seasons,
the two highest lime doses produced ~46% more
shoot dry matter than control and ½ RD. On the other
hand, shoot dry matter production of maize grown in
2 RD lime-amended soil was 15% higher than 1 RD.
In addition, also considering the average of the three
growing seasons, liming increased the grain yield by
~58, 134 and 171%, respectively for ½ RD, 1 RD and
2 RD, compared with the control treatment. In the
second growing season, root:shoot ratios clearly indi-
cated an increase in the proportion of root production
to the detriment of low shoot dry matter production
(Fig. 7B). Higher lime doses (1 RD and, in particular,
2 RD) provided visually more vigorous maize plants
at 45 days after emergence, and higher ear growth at
harvest time than those established in ½ RD lime-
treated and untreated soil (Fig. 7E) .
Pearson’s correlation analysis of soil chemical and
maize plant parameters and hierarchical clustering of
treatments
The changes in soil pH correlated positively with
SOC and nutrients concentration in the soil, with
direct effects on root growth, leaf pigment concen-
trations, gas exchange, and shoot and grain yield
(Fig.  8A). Negative correlations occurred only for
­Al3+ saturation (AS) and substomatal internal ­CO2
concentration, which were negatively correlated with
the other evaluated parameters but positively corre-
lated with each other. Hierarchical clustering analysis
involving all evaluated soil and plant parameters pri-
marily revealed (based on threshold line) a clustering
of untreated soil (control) apart from lime-amended
soils (½ RD, 1 RD and 2 RD); whereas, a second-
ary segregation occurred with ½ RD and 1 RD lime-
amended soils into one group, and 2 RD-amended
soil in another one.
Revenue
Economic evaluation revealed that liming strongly
increased maize production’s net profit (Table  3). In
2017, liming with ½ RD, 1 RD and 2 RD promoted an
increase in lime costs and thus, total costs compared
to untreated soil. On the other hand, even with the
highest total production cost, the two highest doses of
lime resulted in the highest net profits (1 RD = US$
886 and 2 RD = 796 ­ha−1, respectively). Thus, in the
first growing season after lime application, the profit
from the 1 RD was about 11% greater than the 2 RD,
and about 2 and 6.6 times greater than the ½ RD and
control. Interestingly, in 2018 (from this year there is
no cost with liming), only the two largest lime doses
resulted in positive net profits (1 RD = US$ 25 and 2
RD = 265  ­ha−1), where 2 RD presented a profit ~11
times higher than 1 RD. Untreated soil and ½ RD of
lime presented losses of US$ -400 and − 267  ­ha−1,
respectively. In the third growing season (2019), lim-
ing with 2 RD resulted in the highest net profit (US$
1001  ­ha−1), values ​​ ~ 1.25, 2.11, and 8 times greater
than the profit obtained by treatments 1 RD, ½ RD
and control, respectively. At the end of 3  years of
maize cultivated in a soil managed with different lime
doses, the accumulated net profit was: control −141
US$ ­ha−1; ½ RD = 653 US$ h­ a−1; 1 RD = 1709 US$
­ha−1; and 2 RD = 2062 US$ h­ a−1.
Discussion
Weather conditions
Maize received 306, 200 and 294 mm of pluvial pre-
cipitation from first to third growing season, respec-
tively. From early-March (maize sowing) until mid-
July (maize physiological maturity) of each year, due
to low amounts of rain and high evapotranspiration,
negative hydric balance occurred during the entire
maize crop cycle. According to Fancelli (2015),
the maize needs between 350 and 600  mm of rain
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Table 2  Nutritional status of maize plants for each growing season (2017, 2018 and 2019) in response to lime doses [control (lime-
untreated soil), half the recommended rate (½ RD), recommended rate (1 RD) and twice the recommended rate (2 RD)]
Liming N P K Ca Mg S Fe Mn Cu Zn
(g ­kg−1) (mg ­kg−1)

2017
Control 27.9 c 2.21 b 20.6 a 1.81 b 2.21 c 1.32 c 276 a 30.6 a 13.8 a 78.5 a
½ RD 31.7 bc 2.36 a 21.0 a 2.66 ab 3.33 b 1.94 bc 204 b 24.5 b 12.1 a 53.3 b
1 RD 35.3 ab 2.36 a 20.9 a 3.05 a 3.68 b 2.08 b 196 b 23.4 b 12.7 a 45.0 b
2 RD 36.6 a 2.33 a 20.8 a 2.94 a 5.31 a 3.07 a 150 c 21.5 b 12.1 a 42.4 b
2018
Control 22.9 c 1.82 b 16.2 a 1.49 b 1.81 c 1.08 c 227 a 25.1 a 11.7 a 64.3 a
½ RD 26.0 bc 1.93 a 16.7 a 2.18 a 2.73 b 1.59 bc 167 b 20.1 b 10.6 a 43.7 b
1 RD 29.0 ab 1.93 a 17.2 a 2.50 a 3.02 b 1.71 b 161 b 19.2 b 10.4 a 36.9 b
2 RD 30.0 a 1.91 a 17.3 a 2.41 a 4.35 a 2.52 a 150 b 17.6 b 9.9 a 34.7 b
2019
Control 30.2 c 2.15 c 18.2 c 1.84 c 2.14 c 1.28 c 279 a 76.0 a 12.5 a 29.7 a
½ RD 34.0 b 2.30 b 20.1 b 2.02 bc 2.35 c 1.41 c 262 a 77.9 a 11.8 a 30.4 a
1 RD 34.9 b 2.34 b 22.0 a 3.08 ab 3.21 b 1.82 b 171 b 39.3 b 10.6 ab 20.4 b
2 RD 38.6 a 2.89 a 22.9 a 3.85 a 4.02 a 2.27 a 153 b 37.0 b 9.37 b 18.8 b

Different lowercase letters indicate significant difference between treatments by Student’s t test at p ≤ 0.05

Fig. 5  Chlorophyll a
(A), chlorophyll b (B),
total chlorophyll (C) and
carotenoids (D) contents in
maize leaves in response to
lime doses [control (lime-
untreated soil), half the
recommended rate (½ RD),
recommended rate (1 RD)
and twice the recommended
rate (2 RD)] in the three
growing seasons. Different
lower-case letters indicate
significant differences
between treatments by Stu-
dent’s t test at p ≤ 0.05 for
each growing season. Error
bars express the standard
error of the mean (n = 4)

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throughout its development to achieve maximum pro-
ductivity. In our study, the amount of accumulated
rain was below the amount needed, resulting in a
severely negative hydric balance, a condition hardly
appropriate for maize cultivation.
Soil chemical analysis and root development in the
soil profile
The long-term liming enhanced all chemical param-
eters compared with initial soil fertility (2002; see
Material and Methods section). Soil pH from con-
trol treatment at surface layers remained close to that
obtained in 2002, however, even in the control treat-
ment, the SOC increased and was changed in soil
with liming. Exchangeable C ­ a2+ and ­Mg2+ presented
the highest increase through liming. Our results con-
firm that the long-term management and maintenance
with lime is crucial to raise the productive potential
of these tropical soils.
Soil acidity was strongly reduced by liming, and
these effects were most pronounced at depths of up
to 0.4  m when 2 RD lime was applied. Lime cre-
ates a correction front (migration of lime particles)
towards deep layers (Caires et al. 2011; Rheinheimer
et  al. 2018). Lime reactivity is lower when higher
doses are applied to the soil surface (higher pH) (Tiri-
tan et  al. 2016; Rheinheimer et  al. 2018; Bossolani
et  al. 2020b). Intensive tropical agricultural systems
promote strong acidification of the soil through N
fertilization and decomposition of organic mate-
rial (Crusciol et  al. 2019). No tillage-management
resulted in the formation of biopores (channels from
root decomposition) and soil fracture points (Tiritan
et al. 2016); as a result, the particles react in the soil
and can migrate to deep layers. In addition, lime-
amended soils improve physical properties and thus
enable crop plant growth and its functional relation-
ships with soil characteristics (Hernandez-Ramirez
et al. 2014). These features increase lime percolation,
increasing the efficiency of acidity correction in deep
layers even when lime is applied to the surface.
Increasing pH by liming reduced the concentration
of all micronutrients in the soil at 0.0–0.2  m depth.
Micronutrient concentrations were measured only in
the arable layer since SOC is the largest contributor
of these elements in tropical soils (Stevenson 1991)
and may have helped sustain micronutrient levels
within the appropriate range for crop development
(Stevenson 1991; Dhaliwal et  al. 2019). In addition,
fertile soils induce higher biomass production (above-
and belowground) by crops, which in turn, increases
organic C content over time(Briedis et  al. 2012).
Thus, long-term surface liming can provide essential
benefits in highly weathered tropical soils by increas-
ing SOC accumulation and soil fertility.
The maximum rate of lime application (2 RD)
provided the largest increase in ­Ca2+ and M ­ g2+ con-
centration and base saturation in deeper layers. Long-
term surface liming efficiently increased the concen-
tration of nutrients in the soil profile, even at depths
greater than 0.6  m. In tropical regions, which are
characterized by dry spells in autumn/winter, surface
liming has become an important strategy for improv-
ing subsoil fertility and providing favourable condi-
tions for root growth (Bossolani et al. 2021a, 2021b)
and, consequently, water and nutrient uptake. Another
important factor for improving soil chemical condi-
tions and root growth is the reduction of aluminum
saturation, mainly in deep layers in the soil. Alu-
minum saturation significantly down to depths of
0.2 m when liming was performed, regardless of the
applied rate; however, 2 RD lime was more efficient
in reducing aluminum saturation down to depths of
up to 0.8 m.
Through cascading effects by liming benefits on
soil fertility, maize root dry matter increased through-
out the soil profile in all three years. This effect may
be attributable to the higher concentration of C ­ a2+
and lower aluminum saturation in the uppermost soil
layers, particularly at 2 RD of lime. The improve-
ment of soil fertility throughout the entire soil pro-
file provided favourable conditions for root growth,
particularly during drought periods when the crops
quickly exhaust the available soil water in the sur-
face layers (Barbosa et al. 2014). One way of reduc-
ing the effects of the dry spells is increasing the root
growth to deeper layers through improving soil fer-
tility (Bossolani et  al. 2021a, 2021b). Calcium plays
a fundamental role in root growth, participating in
the cell division and the growth of apical meristems
(Wilkins et  al. 2016). On the other hand, Ca mobil-
ity in soil is low, and the same occurs in plant tissues
(Hepler 2005). Almost all Ca uptake by the roots
takes place in the root tip (Millaway and Wiersholm
1979; Hepler 2005), for this reason, it is essential
to have adequate Ca concentration in deeper layers
(Ritchey et al. 1982; Wilkins et al. 2016), and in this
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◂Fig. 6  Net photosynthesis rate–A (A), stomatal conductance–
gs (B), internal C
­ O2 concentration–ic (C), leaf transpiration–
E (D), and water use efficiency–WUE (E) of maize plants in
response to lime doses [control (lime-untreated soil), half the
recommended rate (½ RD), recommended rate (1 RD) and
twice the recommended rate (2 RD)] in the three growing
seasons. Different lower-case letters indicate significant dif-
ferences between treatments by Student’s t test at p ≤ 0.05 for
each growing season. Error bars express the standard error of
the mean (n = 4)
scenario, long-term application of higher lime doses
is the way to go, especially in high weathered soils,
such as those from tropical regions.
In addition to the root growth, the distribution of
these roots is of general importance because the dis-
tribution of nutrients and water is not uniform in the
soil (Fageria and Moreira 2011). The root dry matter
distribution along the soil profile was shallowest in
untreated soil. Although root production is an excel-
lent indicator of root development, the distribution
of roots along the soil profile is essential to ensure
that plants can acquire soil resources under low
water regime (Kiani et  al. 2017; Costa et  al. 2018).
Both root dry weight at different soil depths, and the
extent and distribution of rooting were linked to dif-
ferences in yield and the ability of plants to escape
drought (Fageria and Moreira 2011; Basu et al. 2016).
Therefore, improvements in soil chemical conditions
in lime-amended soils ensured a better distribution of
maize roots along the soil profile, particularly at the
highest doses of lime.
Maize nutritional status, photosynthetic pigments and
gas exchange measurements
The concentrations of all macronutrients increased
in maize leaves as the lime dose increased, which
were within or above the range considered sufficient
for crop development when 1 RD and 2 RD lime
were applied (Cantarella et  al. 1997). In the 2018
growing season, the concentrations of N and P were
lower than considered appropriate for maize (Canta-
rella et  al. 1997); this growing season corresponded
with lower rainfall during maize development. The
higher N concentrations in maize leaves may be due
to increases in soil pH resulting in higher biological
N processes responsible for mineralization and nitri-
fication in bulk soils and maize rhizosphere, further
leading to an increase in N bioavailability (Rosolem
et  al. 2003; Bossolani et  al. 2020a). Therefore, the
greater root growth in lime-amended soils, especially
at higher doses of lime, may have further enabled
greater uptake of this nutrient. Regarding to Ca and
Mg concentrations, lime is the main source of these
nutrients in soil, making this practice essential for the
adequate supply of these nutrients to plants (Caires
et al. 2008; Carmeis Filho et al. 2017a), as occurred
in our study. In addition, changes in soil pH by lime
doses also changed soil nutrient availability and
uptake by maize plants. Higher concentrations of P,
K, Ca, Mg and S in maize leaves occurred in lime-
amended soil as a result of improved soil fertility. In
contrast to macronutrients, maize leaf micronutrient
concentrations were lower in soil amended with the
highest lime doses; however, they were within the
range considered adequate for maize development
(Cantarella et al. 1997).
In this study, liming improved soil fertility, maize
root development and, consequently, nutrient uptake,
with sharp increases in C­ a2+ and ­Mg2+ concentrations
in the soil and leaves, converging to a better leaf pho-
tosynthetic pigment production and corroborating the
positive correlations among these factors. Calcium
and Mg play important functions in the structure and
biochemical processes of chloroplasts (Farhat et  al.
2016; Wang et al. 2019). Calcium is involved in pre-
serving thylakoid morphology and chloroplast ultra-
structure, promoting increased chloroplast width and
area (Liu et  al. 2015), and synthesizing many chlo-
roplast proteins (Wang et  al. 2019). Magnesium is
a central element of chlorophyll structure (Liu et  al.
2008) and, as part of Mg-chelatase, catalyzes the
insertion of Mg into protoporphyrin IX, which is the
first step in the synthesis of chlorophyll (Walker and
Weinstein 1991). Thus, the higher concentrations of
these nutrients underlie the increased pigment pro-
duction as observed in our study.
In addition, the Mg concentrations in maize leaves
in all treatments are within the sufficient range for
crop development (1.5–5.0  g  ­kg−1; Cantarella et  al.
1997); however, maize plants established in the con-
trol treatment presented concentrations close to the
lower limit of Mg sufficiency (average of three grow-
ing seasons = 2.05 g ­kg−1). In field conditions, we did
not observe classic visual symptoms of Mg deficiency
in maize plants, but control plants were visually less
green. This finding corroborates our results of chloro-
phyll concentrations.
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◂Fig. 7  Shoot dry matter (A), root:shoot ratio (B), grain yield
(C) and harvest index of maize in response to lime doses [con-
trol (lime-untreated soil), half the recommended rate (½ RD),
recommended rate (1 RD) and twice the recommended rate
(2 RD)] in the three growing seasons. Different lower-case
letters indicate significant differences between treatments by
Student’s t test at p ≤ 0.05 for each growing season. Error bars
express the standard error of the mean (n = 4). The photos rep-
resents the effect of liming doses on maize development (2018)
at 45  days after emergence, and ear development at harvest
time (E)
Modern maize cultivars are more productive and
therefore more nutrient-demanding (such as Mg) to
express full metabolism and yield capacity (Bender
et  al. 2013). Magnesium levels in plant tissue are
highly important for chlorophylls synthesis (Ceylan
et al. 2016; Jaghdani et al. 2021). The low availability
of Mg in the cytosol (even without visible symptoms)
can result in poor protein synthesis, degradation
of chlorophylls, and low photosynthetic efficiency
(Farhat et al. 2016).
Although high Mg concentrations in leaf tis-
sue are of great importance for the functioning of
plant metabolism, Hauer-Jákli and Tränkner (2019)
demonstrated that plant growth and photosynthetic
efficiency increase up to a certain threshold of Mg
concentrations. These results suggest that Mg con-
centration is perhaps not the only reason for the low
chlorophyll concentrations. The low N and S concen-
trations in leaf tissue may be the cause of chloroses in
the leaves in the absence of liming. In the control and
in the lowest lime dose (½ RD), the concentrations of
both nutrients are below the sufficient range for crop
development (N = 27–35  g  ­kg−1; S = 1.5–3.0  g  ­kg−1;
Cantarella et al. 1997). SOM mineralization depends
on soil pH (Gharmakher et al. 2009), and under low
soil pH SOM mineralization rates decrease (Holland
et al. 2018). It is important to note that the largest S
source in the soil comes from the mineralization of
organic compounds (Vicensi et  al. 2020). Thus, the
increase in soil pH caused by liming can increase
the mineralization of ­SO42− from SOM (Bolan et al.
2003), as well as increasing sulphate desorption (Bar-
row and Debnath 2015), making it available for plant
uptake.
The A rate, gs and WUE of maize leaves increased
with the lime dose, whereas ic reduced. These results
suggest that the maize plants in lime-amended soil
exhibited a greater photosynthetic rate and higher
­CO2 carboxylation efficiency. The greater root devel-
opment along the soil profile with increasing lime
rate due to improved soil chemical conditions likely
increased the ability of the maize plants to access
water and nutrients at greater depths, as supported
by the higher concentrations of maize roots in the
uppermost soil layers in the control and ½ RD lime
treatments and the better distribution of roots in all
layers of the soil in the 2 RD lime treatment. Even
in 2018, when rainfall during maize plant develop-
ment was lower than in the other two growing sea-
sons, photosynthetic parameters increased as the
lime rate increased. These observations indicate that
even under stress, the ability of deeper roots to access
­Ca2+ and ­Mg2+ can mitigate the effect of lower water
availability, as indicated by the better WUE of these
plants. As a result, less water was lost due to leaf tran-
spiration during ­CO2 assimilation via photosynthesis.
In addition, the low S availability to the plants (as
occurred in control and ½ RD treatments) can reduce
the biosynthesis of chlorophyll, which induces chlo-
rosis in young leaves, in addition to altering the effi-
ciency of the photosynthetic apparatus (Shafiq et  al.
2021).
Flexas et al. (2004) showed that the stress caused
by reduced water availability can be quantified by
stomatal conductance, where values ≥0.2  mol ­H2O
­m−2 ­s−1 represent no drought stress; 0.1–0.2  mol
­H2O ­m−2 ­s−1 moderate drought stress, and ≤ 0.1  mol
­H2O ­m−2 ­s−1 severe drought stress. The maize plants
grown in the control treatment and under a lower
rate of liming exhibited stomatal conductance val-
ues below 0.1 and 0.2 mol H ­ 2O ­m−2 ­s−1, respectively,
suggesting that plants with lower root development
have lower water accessibility, which causes a severe
decline in photosynthesis.
According to Sousa et  al. (2020), photosynthesis
is very sensitive to environmental stress. Under pro-
longed drought conditions, plants initiate stomatal
limitations that directly affect photosynthetic activ-
ity, including lower stomatal conductance and C ­ O2
assimilation. Greater stomatal conductance and, con-
sequently, the photosynthetic ­CO2 assimilation rate
are directly related to the ability of plants to transport
water from roots to transpiring tissues (Basu et  al.
2016; Gupta et al. 2020). Consequently, the improve-
ments in root development obtained by lime appli-
cation can effectively ensure higher photosynthetic
rates by plants. In addition, stomatal movement is
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Fig. 8  Heatmap correlation (Pearson) (A) and the hierarchi-
cal clustering (B) based on soil chemical properties and physi-
ological and agronomic parameters of maize. Only significant
correlations at p ≤ 0.05 are shown. Hierarchical clustering was
constructed based on Gower distance of similarity with 1000
bootstraps. Treatments of lime doses as follow: control: lime-
untreated soil; ½ RD: half the recommended rate; 1 RD recom-
mended rate and; 2 RD: twice the recommended rate. ­R1, ­R2,
co-regulated by a signalling network that includes the
­Ca2+ signal transduction pathway (Wang et al. 2019);
thus plants with better Ca nutrition can regulate the
water status in plant cells and maintain photosyn-
thetic activity.
Agronomic parameters, grain yield and revenue
Shoot dry matter production, grain yield and rev-
enue were always greatest at high doses of lime in all
years. However, large differences were observed in
2018. In this growing season, rainfall during maize
development was lowest, and shoot dry matter and
grain yield were sharply reduced. Under adverse cli-
matic conditions such as long dry spells, which are
common in tropical regions, corrective soil manage-
ment by means of surface liming is of paramount
importance to ensure that crops are able to withstand
stresses and produce grains (Carmeis Filho et  al.
2017a; Costa et al. 2018), as well as enhance revenue
to farmers. Accordingly, in 2018, maize established
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­R3 and ­R4 represents the replicates of all parameters evaluated.
Soil organic carbon (SOC), base saturation (BS), aluminum
saturation (AS), root dry matter (RDM), chlorophyll a (Chl a),
Chlorophyll b (Chl b), total chlorophyll (T. chl), carotenoids
(Carot), Net photosynthesis rate (A), stomatal conductance
(gs), leaf transpiration (E), internal ­ CO2 concentration (ic),
water use efficiency (WUE), shoot dry matter (SDM) and grain
yield (GY)
in 2 RD lime-amended soil provided shoot dry mat-
ter and grain yields approximately 5.2 and 2.1 times
higher than the control treatment, respectively. These
results are highly dependent on greater root growth.
Rooting depth and distribution are important traits
related to absorption of water and nutrients from the
soil profile, which make the plants able to overcome
periods of low water availability, and ensure higher
yields (Bossolani et al. 2021a).
Hierarchical clustering analysis clearly showed
that the limed soils were superior to the control
treatment; moreover, there was strong segregation
between application doses, demonstrating that higher
lime doses (mainly 2 RD) provided superior results.
Interestingly, the lower shoot dry matter productivity
in 2018 was associated with an increase in the maize
root:shoot ratio under low water availability, although
this ratio did not differ between lime doses. The root
system is the first plant compartment to sense low
water availability (Basu et  al. 2016), but it is shoot
growth that is inhibited (Liu et  al. 2004). Although
Plant Soil

Table 3  Economic evaluation of maize production for each growing season (2017, 2018 and 2019) in response to lime doses [con-
trol (lime–untreated soil), half the recommended rate (½ RD), recommended rate (1 RD) and twice the recommended rate (2 RD)]
Treatment Cost GYa Grossb Netc
Liming Maize production Total
–1
US$ ­ha kg ­ha−1 US$ ­ha–1

2017
Control 0 560 560 3250 694 134
½ RD 108 560 668 5230 1114 446
1 RD 216 560 776 7800 1662 886
2 RD 432 560 992 8350 1788 796
2018
Control – 560 560 750 160 −400
½ RD – 560 560 1380 293 −267
1 RD – 560 560 2740 585 25
2 RD – 560 560 3870 825 265
2019
Control – 560 560 3210 685 125
½ RD – 560 560 4850 1034 474
1 RD – 560 560 6370 1358 798
2 RD – 560 560 7320 1561 1001
3–years accumulated
Control – – 1680 7210 1539 −141
½ RD – – 1788 11,460 2441 653
1 RD – – 1896 16,910 3605 1709
2 RD – – 2112 19,540 4174 2062
a
 GY is the maize grain yield
b
 Gross is the revenue h­ a−1
c
 Net is the return ­ha−1

drought stress has little effect on primary root
growth, lateral root growth is strongly limited (Xu
et  al. 2015). This relationship highlights the relative
biomass allocation between roots and shoots. Root
growth is guided by source-to-sink sugar partition-
ing, such that roots become stronger sinks than shoots
(Xu et  al. 2015; Basu et  al. 2016). Consequently,
improving photosynthesis is critical to increase car-
bohydrate production by maize for use in root growth,
as observed when lime was applied. In addition, the
harvest index and net profit clearly demonstrated
that liming is essential to ensure a better relationship
between grain and shoot productivity. It is remarkable
that under severe water shortages, as in 2018, the ½
RD treatment resulted in a lower index than the 1 RD
and 2 RD treatments. The differences between the
effects of the liming doses were always more accen-
tuated under stress conditions, especially for revenue.
Lime amended soil benefited the net profit in maize
production over the years since the high dose of
lime generated revenue gains even, and especially, in
growing seasons highly stressful due to drought. Soil
managed with lasting lime effect of 2 RD ensured
that maize was able to produce grain, whereas in the
treatment with 1 RD, although positive, the net profit
was low, almost in line with the cost of production of
maize. In contrast, under high water restriction, the
use of ½ RD of lime, and the absence of liming (con-
trol treatment) caused strong economic losses, mak-
ing maize production unfeasible for the farmers.

Vol.: (0123456789)
13

Finally, the long-term results confirmed the fea-
sibility of surface liming for improving soil fertility,
root growth, physiological responses and crop yield,
as well as the positive correlations among these fac-
tors. The highest lime rate (2 RD) proved to be the
most efficient for guaranteeing maize productive
potential and profit gains in a tropical soil with dry
winters and frequent dry spells during maize develop-
ment (autumn/winter). Our results suggest that liming
dose recommendations for tropical soils need to be
revised, as the emergence of no-tillage systems and
the accompanying soil benefits permit the application
of higher lime doses, which would prolong the ben-
efits of acidity management over time, in addition to
increase the farm profit.
Conclusions
Long-term lime surface application under no-tillage
cropping efficiently alleviated soil acidity through-
out the evaluated soil profile (1.0  m), especially at
dose of 2 RD, in addition to increasing nutrient con-
centration and mitigating the toxic effects of A ­ l3+.
This practice proved to be strongly feasible and is
therefore recommended for tropical soils. The mag-
nitude of the effect of liming depends on the dose
and the reaction time. The analysis of the soil pro-
file at depths of up to 1 m provided insights on soil
chemical stratification as well as root production
and distribution, revealing differences between the
top and bottom depths that would not have been
recognized in typical studies at shallower depths.
The combined effect of improving soil fertility and
root development in deep soil layers was reflected
in improvements in photosynthetic responses and,
consequently, maize plant development and grain
yield, even under limited water availability during
the crop cycle, which is common in tropical regions.
We also demonstrate that soils managed with 2 RD
of lime provide greater net profit for maize cultiva-
tion, especially when climatic conditions are unfa-
vourable. This study precisely details how liming at
different doses changes soil chemistry and how the
response of maize plants to these changes results in
higher grain yield and revenue to farmers.
Acknowledgements  CACC, JCC and ARR would like to
thank the National Council for Scientific and Technological
Vol:. (1234567890)
13
Plant Soil
Development (CNPq) for an award for excellence in research.
We also thank the Calcário Guapirama Company (www.​calca​
riogu​apira​ma.​com.​br) for providing the sedimentary lime used
in this experiment.
Author contributions  J.W.B. and C.A.C.C. worked on the
research designing and conduction, data analysis, writing and
formatting the manuscript. L.M., J.R.P., L.G.M., A.G., M.C.F.,
V.A.R., J.C.C., and A.R.R. revised this draft by rewriting, dis-
cussing and commenting. All authors contributed significantly
on this manuscript.
Funding  This study was funded by the São Paulo Research
Foundation (FAPESP) (Grant 2018/11063–7 and 2019/12764–
1) and the National Council for Scientific and Techno-
logical Development (CNPq) (Universal Research Project:
421637/2018–8).
Data availability  Data can be provided upon request.
Code availability  Not Applicable.
Declarations 
Conflict of interest  The authors declare that they have no
competing interests.
References
Allen RG, Pereira LS, Raes D, Smith M (1998) Crop evapo-
transpiration-guidelines for computing crop water require-
ments-FAO irrigation and drainage paper 56. Fao, Rome
300:D05109
Alvares CA, Stape JL, Sentelhas PC et al (2013) Köppen’s cli-
mate classification map for Brazil. Meteorol Zeitschrift
22:711–728. https://​doi.​org/​10.​1127/​0941-​2948/​2013/​
0507
AOAC (2016) Official methods of analysis of AOAC inter-
national. Rockville, MD: AOAC International, ISBN:
978-0-935584-87-5
Auler AC, Caires EF, Pires LF et  al (2019) Lime effects in a
no-tillage system on Inceptisols in southern Brazil. Geo-
derma Reg 16:e00206. https://​doi.​org/​10.​1016/j.​geodrs.​
2019.​e00206
Barbosa FDS, Coelho RD, Maschio R et  al (2014) Drought
resistance of sugar-cane crop for different levels of water
availability in the soil. Eng Agric 34:203–210. https://​doi.​
org/​10.​1590/​S0100-​69162​01400​02000​02
Barrow NJ, Debnath A (2015) Effect of phosphate status and
pH on sulphate sorption and desorption. Eur J Soil Sci
66:286–297. https://​doi.​org/​10.​1111/​ejss.​12223
Basu S, Ramegowda V, Kumar A, Pereira A (2016) Plant adap-
tation to drought stress. F1000Research 5:1–10. https://​
doi.​org/​10.​12688/​F1000​RESEA​RCH.​7678.1
Bender RR, Haegele JW, Ruffo ML, Below FE (2013) Nutrient
uptake, partitioning, and remobilization in modern, trans-
genic insect-protected maize hybrids. Agron J 105:161–
170. https://​doi.​org/​10.​2134/​agron​j2012.​0352
Plant Soil
Bolan NS, Adriano DC, Curtin D (2003) Soil acidification and
liming interactions with nutrient and heavy metal transfor-
mation and bioavailability. Adv Agron 78:5–272
Bossolani JW, Lazarini E, Santos FL et al (2018) Surface reap-
plication of lime and gypsum on maize cultivated sole and
intercropped with Urochloa. Commun Soil Sci Plant Anal
49:1855–1868. https://​doi.​org/​10.​1080/​00103​624.​2018.​
14755​65
Bossolani JW, Crusciol CAC, Merloti LF et al (2020a) Long-
term lime and gypsum amendment increase nitrogen fixa-
tion and decrease nitrification and denitrification gene
abundances in the rhizosphere and soil in a tropical no-till
intercropping system. Geoderma 375:114476. https://​doi.​
org/​10.​1016/j.​geode​rma.​2020.​114476
Bossolani JW, Santos FL, Meneghette HHA et al (2020b) Soy-
bean in crop rotation with maize and palisade grass inter-
cropping enhances the long-term effects of surface liming
in no-till system. J Soil Sci Plant Nutr 20:1–12. https://​doi.​
org/​10.​1007/​s42729-​020-​00347-2
Bossolani JW, Crusciol CAC, Garcia A et  al (2021a) Long-
term lime and phosphogypsum amended-soils allevi-
ates the field drought effects on carbon and antioxidative
metabolism of maize by improving soil fertility and root
growth. Front Plant Sci 12:1437. https://​doi.​org/​10.​3389/​
fpls.​2021.​650296
Bossolani JW, Crusciol CAC, Leite MFA et al (2021b) Modu-
lation of the soil microbiome by long-term Ca-based soil
amendments boosts soil organic carbon and physicochem-
ical quality in a tropical no-till crop rotation system. Soil
Biol Biochem 156. https://​doi.​org/​10.​1016/j.​soilb​io.​2021.​
108188
Briedis C, Sá JC d M, Caires EF et al (2012) Soil organic mat-
ter pools and carbon-protection mechanisms in aggregate
classes influenced by surface liming in a no-till system.
Geoderma 170:80–88. https://​doi.​org/​10.​1016/j.​geode​rma.​
2011.​10.​011
Caires EF, Corrêa JCL, Churka S et  al (2006) Surface appli-
cation of lime ameliorates subsoil acidity and improves
root growth and yield of wheat in an acid soil under no-
till system. Sci Agric 63:502–509. https://​doi.​org/​10.​1590/​
S0103-​90162​00600​05000​13
Caires EF, Garbuio FJ, Barth G (2008) Surface application of
lime and cover black oat and corn residues for no-till soy-
bean production. Commun Soil Sci Plant Anal 39:2102–
2118. https://​doi.​org/​10.​1080/​00103​62080​21351​38
Caires EF, Joris HAW, Churka S (2011) Long-term effects of
lime and gypsum additions on no-till corn and soybean
yield and soil chemical properties in southern Brazil. Soil
Use Manag 27:45–53. https://​doi.​org/​10.​1111/j.​1475-​
2743.​2010.​00310.x
Cantarella H, van Raij B, Camargo CEO (1997) Adubação de
cereais. RAIJ, B van al, eds Recom adubação e calagem
para o Estado São Paulo 2:43–50
Cantarella H, van Raij B, Quaggio JA (1998) Soil and plant
analyses for lime and fertilizer recommendations in Bra-
zil. Commun Soil Sci Plant Anal 29:1691–1706. https://​
doi.​org/​10.​1080/​00103​62980​93700​60
Carmeis Filho ACA, Crusciol CAC, Castilhos AM (2017a)
Liming demand and plant growth improvements for an
Oxisol under long-term no-till cropping. J Agric Sci
155:1093–1112. https://​doi.​org/​10.​1017/​S0021​85961​
70002​35
Carmeis Filho ACA, Crusciol CAC, Guimarães TM et  al
(2017b) Impact of amendments on the physical properties
of soil under tropical long-term no till conditions. PLoS
One 12:27959897. https://​doi.​org/​10.​1371/​journ​al.​pone.​
01730​11
Carmeis Filho ACA, Crusciol CAC, Guimarães TM et  al
(2018) Changes in soil physical properties and carbon
protection mechanisms by surface application of lime in
a tropical no-tillage system. Soil Sci Soc Am J 82:56–65.
https://​doi.​org/​10.​2136/​sssaj​2017.​04.​0120
Ceylan Y, Kutman UB, Mengutay M, Cakmak I (2016) Mag-
nesium applications to growth medium and foliage affect
the starch distribution, increase the grain size and improve
the seed germination in wheat. Plant Soil 406:145–156.
https://​doi.​org/​10.​1007/​s11104-​016-​2871-8
CONAB. National supply Company (2021) Cost of Agricul-
tural Production. https://​www.​conab.​gov.​br/​info-​agro/​cus-
tos-​de-​produ​cao/​plani​lhas-​de-​custo-​de-​produ​cao/​iteml​ist/​
categ​ory/​821-​milho. Accessed 8 Aug 2021
Costa CHM, Carmeis Filho ACA, Crusciol CAC et al (2018)
Intensive annual crop production and root development
in a tropical acid soil under long-term no-till and soil-
amendment management. Crop Pasture Sci 69:488–505.
https://​doi.​org/​10.​1071/​CP172​33
Crusciol CAC, Marques RR, Carmeis Filho ACA et  al
(2019) Lime and gypsum combination improves crop
and forage yields and estimated meat production and
revenue in a variable charge tropical soil. Nutr Cycl
Agroecosyst 115:347–372. https://​doi.​org/​10.​1007/​
s10705-​019-​10017-0
Cusser S, Bahlai C, Swinton SM et  al (2020) Long-term
research avoids spurious and misleading trends in sustain-
ability attributes of no-till. Glob Chang Biol 26:3715–
3725. https://​doi.​org/​10.​1111/​gcb.​15080
Dhaliwal SS, Naresh RK, Mandal A et al (2019) Dynamics and
transformations of micronutrients in agricultural soils as
influenced by organic matter build-up: a review. Environ
Sustain Indic 1–2:100007. https://​doi.​org/​10.​1016/j.​indic.​
2019.​100007
Fageria NK, Moreira A (2011) The role of mineral nutrition on
root growth of crop plants, 1st edn. Elsevier Inc.
Fageria NK, Nascente AS (2014) Management of soil acidity
of south American soils for sustainable crop production.
Elsevier
Fancelli AL (2015) Ecofisiologia, fenologia e implicações bási-
cas de manejo. In: BORÉM, A.; GALVÃO, JCC; PIMEN-
TEL, MA Milho: do plantio à colheita. Viçosa: Ed. UFV,
pp 50–76
Farhat N, Elkhouni A, Zorrig W et al (2016) Effects of magne-
sium deficiency on photosynthesis and carbohydrate par-
titioning. Acta Physiol Plant 38. https://​doi.​org/​10.​1007/​
s11738-​016-​2165-z
Flexas J, Bota J, Loreto F et al (2004) Diffusive and metabolic
limitations to photosynthesis under drought and salin-
ity in C3 plants. Plant Biol 6:269–279. https://​doi.​org/​10.​
1055/s-​2004-​820867
Gharmakher HN, Machet J-M, Beaudoin N, Recous S (2009)
Estimation of sulfur mineralization and relationships with
nitrogen and carbon in soils. Biol Fertil Soils 45:297–304
Vol.: (0123456789)
13

Gibbs HK, Salmon JM (2015) Mapping the world’s degraded
lands. Appl Geogr 57:12–21. https://​doi.​org/​10.​1016/j.​
apgeog.​2014.​11.​024
Gupta A, Rico-Medina A, Caño-Delgado AI (2020) The
physiology of plant responses to drought. Science (80- )
368:266–269. https://​doi.​org/​10.​1126/​scien​ce.​aaz76​14
Hauer-Jákli M, Tränkner M (2019) Critical leaf magnesium
thresholds and the impact of magnesium on plant growth
and photo-oxidative defense: a systematic review and
meta-analysis from 70 years of research. Front Plant Sci
10:1–15. https://​doi.​org/​10.​3389/​fpls.​2019.​00766
Hepler PK (2005) Calcium: a central regulator of plant growth
and development. Plant Cell 17:2142–2155. https://​doi.​
org/​10.​1105/​tpc.​105.​032508
Hernandez-Ramirez G, Lawrence-Smith EJ, Sinton SM et al
(2014) Root responses to alterations in macroporosity
and penetrability in a silt loam soil. Soil Sci Soc Am
J 78:1392–1403. https://​doi.​org/​10.​2136/​sssaj​2014.​01.​
0005
Holland JE, Bennett AE, Newton AC et  al (2018) Liming
impacts on soils, crops and biodiversity in the UK: a
review. Sci Total Environ 610–611:316–332. https://​doi.​
org/​10.​1016/j.​scito​tenv.​2017.​08.​020
International Grains Council (2019) Five-year baseline projec-
tions of supply and demand for wheat, maize (corn), rice
and soyabeans to 2023/24. 44:1–32
Jaghdani SJ, Jahns P, Tränkner M (2021) Mg deficiency
induces photo-oxidative stress primarily by limiting CO2
assimilation and not by limiting photosynthetic light uti-
lization. Plant Sci 302. https://​doi.​org/​10.​1016/j.​plant​sci.​
2020.​110751
Jahn R, Blume HP, Asio VB et  al (2006) Guidelines for soil
description. FAO
Joris HAW, Caires EF, Scharr DA et  al (2016) Liming in the
conversion from degraded pastureland to a no-till crop-
ping system in southern Brazil. Soil Tillage Res 162:68–
77. https://​doi.​org/​10.​1016/j.​still.​2016.​04.​009
Kiani M, Hernandez-Ramirez G, Quideau S et al (2017) Quan-
tifying sensitive soil quality indicators across contrasting
long-term land management systems: crop rotations and
nutrient regimes. Agric Ecosyst Environ 248:123–135.
https://​doi.​org/​10.​1016/j.​agee.​2017.​07.​018
Li Y, Cui S, Chang SX, Zhang Q (2019) Liming effects on soil
pH and crop yield depend on lime material type, appli-
cation method and rate, and crop species: a global meta-
analysis. J Soils Sediments 19:1393–1406. https://​doi.​org/​
10.​1007/​s11368-​018-​2120-2
Lichtenthaler HK (1987) Chlorophylls and carotenoids: pig-
ments of photosynthetic biomembranes. In: Methods in
enzymology. Elsevier, pp 350–382
Liu HS, Li FM, Xu H (2004) Deficiency of water can enhance
root respiration rate of drought-sensitive but not drought-
tolerant spring wheat. Agric Water Manag 64:41–48.
https://​doi.​org/​10.​1016/​S0378-​3774(03)​00143-4
Liu H, Chen X, Chen R et al (2008) Effects of magnesium defi-
ciency on growth and photosynthesis of flowering Chi-
nese cabbage. Acta Hortic 767:169–176
Liu YF, Zhang GX, Qi MF, Li TL (2015) Effects of calcium on
photosynthesis, antioxidant system, and chloroplast ultra-
structure in tomato leaves under low night temperature
Vol:. (1234567890)
13
Plant Soil
stress. J Plant Growth Regul 34:263–273. https://​doi.​org/​
10.​1007/​s00344-​014-​9462-9
Millaway RM, Wiersholm L (1979) Calcium and metabolic
disorders. Commun Soil Sci Plant Anal 10:1–28. https://​
doi.​org/​10.​1080/​00103​62790​93668​75
Nunes MR, Denardin JE, Vaz CMP et  al (2019) Lime move-
ment through highly weathered soil profiles. Environ Res
Commun 1:115002. https://​doi.​org/​10.​1088/​2515-​7620/​
ab4eba
Quaggio JA, Raij B van (2001) Determinação do pH em cloreto
de cálcio e da acidez total. Análise química para avaliação
da Fertil solos Trop 181–188
Rheinheimer DS, Tiecher T, Gonzatto R et al (2018) Residual
effect of surface-applied lime on soil acidity properties in
a long-term experiment under no-till in a southern Brazil-
ian sandy Ultisol. Geoderma 313:7–16. https://​doi.​org/​10.​
1016/j.​geode​rma.​2017.​10.​024
Ritchey KD, Silva JE, Costa UF (1982) Calcium defi-
ciency in clayey B horizons of savanna oxisols. Soil Sci
133:378–382
Ritchie SW, Hanway JJ, Benson GO (1993) How a corn plant
develops. Special report n. 48. Iowa State University of
Science and Technology. Coop Ext Serv Ames, IA, USA
Rolim G d S, Sentelhas PC, Barbieri V (1998) Planilhas no
ambiente EXCEL TM para os cálculos de balanços hídri-
cos: normal, sequencial, de cultura e de produtividade real
e potencial. Rev Bras Agrometeorol 6:133–137
Rosolem CA, Foloni JSS, De Oliveira RH (2003) Dinâmica do
nitrogênio no solo em razão da calagem e adubação nitro-
genada, com palha na superficie. Pesqui Agropecu Bras
38:301–309. https://​doi.​org/​10.​1590/​s0100-​204x2​00300​
02000​18
Shafiq BA, Nawaz F, Majeed S et  al (2021) Sulfate-based
fertilizers regulate nutrient uptake, photosynthetic gas
exchange, and enzymatic antioxidants to increase sun-
flower growth and yield under drought stress. J Soil
Sci Plant Nutr 21:2229–2241. https://​doi.​org/​10.​1007/​
s42729-​021-​00516-x
Sousa LF, de Menezes-Silva PE, Lourenço LL et  al (2020)
Improving water use efficiency by changing hydraulic and
stomatal characteristics in soybean exposed to drought:
the involvement of nitric oxide. Physiol Plant 168:576–
589. https://​doi.​org/​10.​1111/​ppl.​12983
Stevenson FJ (1991) Organic matter-micronutrient reactions in
soil, 2nd edn. Wiley Online Library
Thornthwaite CW, Mather JR (1955) The water balance cli-
matology. Laboratory of climatology, Philadelphia, PA:
Drexel Institute of Technology
Tiritan CS, Büll LT, Crusciol CAC et al (2016) Tillage system
and lime application in a tropical region: soil chemical
fertility and corn yield in succession to degraded pastures.
Soil Tillage Res 155:437–447. https://​doi.​org/​10.​1016/j.​
still.​2015.​06.​012
USDA (2014) Keys to soil taxonomy. US Gov. Print. Office,
Washington, DC, pp 327–328
USDA (2020) World agricultural supply and demand esti-
mates. In: USDA, Washington, DC. https://wwwusdagov/
oce/commodity/wasde/. Accessed 27 Jul 2020
van Raij B, Cantarella H, Quaggio JA, Furlani AMC (1997)
Recomendações de adubação e calagem para o Estado
Plant Soil
de São Paulo. Instituto Agronômico/Fundação IAC
Campinas
van Raij B, Quaggio JA, Cantarella H, Abreu CA (2001) Os
métodos de análise química do sistema IAC de análise de
solo no contexto nacional. Análise química para avaliação
da Fertil solos Trop 5–39
Vicensi M, Lopes C, Koszalka V et  al (2020) Gypsum rates
and splitting under no-till: soil fertility, corn performance,
accumulated yield and profits. J Soil Sci Plant Nutr
20:690–702
von Uexküll HR, Mutert E (1995) Global extent, development
and economic impact of acid soils. Plant Soil 171:1–15.
https://​doi.​org/​10.​1007/​BF000​09558
Walker CJ, Weinstein JD (1991) Further characterization of
the magnesium chelatase in isolated developing cucum-
ber chloroplasts: substrate specificity, regulation, intact-
ness, and ATP requirements. Plant Physiol 95:1189–1196.
https://​doi.​org/​10.​1104/​pp.​95.4.​1189
Walkley A, Black IA (1934) An examination of the Degt-
jareff method for determining soil organic matter, and
a proposed modification of the chromic acid titration
method. Soil Sci 37:29–38
Wang Q, Yang S, Wan S, Li X (2019) The significance of cal-
cium in photosynthesis. Int J Mol Sci 20:1–14. https://​doi.​
org/​10.​3390/​ijms2​00613​53
Wilkins KA, Matthus E, Swarbreck SM, Davies JM (2016)
Calcium-mediated abiotic stress signaling in roots. Front
Plant Sci 7:1–17. https://​doi.​org/​10.​3389/​fpls.​2016.​01296
Xu W, Cui K, Xu A et  al (2015) Drought stress condition
increases root to shoot ratio via alteration of carbohy-
drate partitioning and enzymatic activity in rice seed-
lings. Acta Physiol Plant 37. https://​doi.​org/​10.​1007/​
s11738-​014-​1760-0
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