Journal of Plant Nutrition

ISSN: 0190-4167 (Print) 1532-4087 (Online) Journal homepage: https://www.tandfonline.com/loi/lpla20

Enrichment of lithium in lettuce plants through

agronomic biofortification

Rubens Ribeiro da Silva, Alvaro José Gomes de Faria, Gilson do Carmo

Alexandrino, Evandro Alves Ribeiro, Antônio Carlos Martins dos Santos, Tulio
Teixeira Deusdara, Ildon Rodrigues do Nascimento & Vitor L. Nascimento

To cite this article: Rubens Ribeiro da Silva, Alvaro José Gomes de Faria, Gilson do Carmo
Alexandrino, Evandro Alves Ribeiro, Antônio Carlos Martins dos Santos, Tulio Teixeira
Deusdara, Ildon Rodrigues do Nascimento & Vitor L. Nascimento (2019): Enrichment of
lithium in lettuce plants through agronomic biofortification, Journal of Plant Nutrition, DOI:
10.1080/01904167.2019.1648671

To link to this article: https://doi.org/10.1080/01904167.2019.1648671

Published online: 12 Aug 2019.

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JOURNAL OF PLANT NUTRITION
https://doi.org/10.1080/01904167.2019.1648671

Enrichment of lithium in lettuce plants through agronomic

biofortification
Rubens Ribeiro da Silva, Alvaro Jos
e Gomes de Faria, Gilson do Carmo Alexandrino,
Evandro Alves Ribeiro, Anto^nio Carlos Martins dos Santos, Tulio Teixeira Deusdara,
Ildon Rodrigues do Nascimento, and Vitor L. Nascimento
Plant Production Graduate Program, Universidade Federal do Tocantins – Campus Gurupi, Gurupi,
Tocantins, Brazil

ABSTRACT ARTICLE HISTORY

Biofortification of crops with lithium (Li) is a new trend in agriculture and Received 15 September 2018
researches on this subject still scarce, therefore, this work aims to evaluate Accepted 27 February 2019
the accumulation potential and the effects in agronomic characteristics of
KEYWORDS
lettuce (Lactuca sativa L.) submitted to foliar application of two Li chemical
foliar application; Lactuca
sources and different concentrations. The experiment was conducted in a sativa L.; Li accumulation;
greenhouse and the applications of lithium hydroxide (LiOH) and lithium trace element
sulfate (Li2SO4) were done via foliar spray at concentrations ranging from 0
to 40 mg dm
3. The applications were divided into three applications dis-
tributed at 15, 25, and 35 days after transplanting. Positive and promising
results were observed in agronomic characteristics and Li in accumulation.
High concentrations of Li (40 mg dm
3) may compromise the development
of lettuce plants, however, lower concentrations applied (16 to 26 mg
dm
3 of Li) promoted an increase in dry weight of roots and shoot/root
ratio, both chemical forms, and stem diameter and specific leaf area,
Li2SO4, suggesting beneficial effect of this element. Li accumulation in
shoots depend on the concentrations of this element in the applied solu-
tion of Li but not on its chemical form.

Introduction
The food production maintains the same growth rate that world population and it is expected to
reach 9.6 billion people by 2050 (Gerland et al. 2014; Reis et al. 2014; Crist, Mora, and Engelman
2017). However, with the increase of food production, problems such as hidden hunger—defi-
ciencies in essential vitamins and minerals—have increased in recent years and it has reached
more than 2 billion people worldwide, especially pregnant women, adolescents and children
(Graham et al. 2007; Welch 2001).
Hidden hunger is a condition of starvation characterized by the consumption of enough food,
mainly basic crops, but these foods do not bring adequate levels of nutrients and vitamins to the
health and well-being of people (Ruel-Bergeron et al. 2015). In part, this is due to plant breeding
focused in productivity without the appropriate concern as to the nutritional quality of food
(Garvin, Welch, and Finley 2006; Murphy, Reeves, and Jones 2008; White and Broadley 2009).
Currently, the deficiencies caused by the absence of zinc (Zn), iron (Fe), selenium (Se), iodine (I)
and vitamin A are the most worrying in relation to human health, especially in developing coun-
tries such as Brazil (Allen et al. 2006; Moraes 2008). Moreover, lithium (Li) deficiency is linked

CONTACT Rubens Ribeiro da Silva rrs2002@uft.edu.br Universidade Federal do Tocantins, Campus Gurupi, 77402-970
Gurupi, Tocantins, Brazil
ß 2019 Taylor & Francis Group, LLC
2 R. R. DA SILVA ET AL.

to the occurrence of mood disorders, including bipolar disorder (Mleczek et al. 2017). In add-
ition, at optimal levels in the body Li has a positive role in the prophylaxis of diseases such as
Alzheimer’s (Young 2011; Forlenza et al. 2011), amyotrophic sclerosis, schizophrenia, phobic anx-
iety (Martinsson et al. 2013), reduction of diabetes, heart disease and alcohol dependence
(Schrauzer, Shrestha, and Flores-Arce 1992; Schrauzer 2002; Norra et al. 2008).
Li is not an essential nutrient for plants, however, in relative low concentrations there is
stimulation in the growth and accumulation of this element in interesting parts for human con-
sumption (Hawrylak-Nowak, Kalinowska, and Szyma
nska 2012). There is still little information
about the application of Li in plants; however, there is great evidence of deficiency of this element
in the population. In order to maintain adequate levels of Li in the human body, a daily intake of
food is required which contains sufficient levels of this element and the use of biofortified crop
products becomes a low-cost strategy to increase food sustainability (Gonçalves et al. 2015).
Biofortification consists of the agronomic practice of adding, during cultivation, one or more ele-
ments, to obtain a food enriched with essential nutrients to the human being (White and
Broadley 2009). The biofortification should not alter the basic characteristics of the products or
interfere in the existing logistics between producers and consumers, in addition, there is aggrega-
tion of quality and value to the product of interest (Martinez 2013).
Foliar fertilization allows a balanced complementation of soil fertilization and experiments
based on this application are widely found in the literature, especially when the aim is the appli-
cation of micronutrients, in the order of g ha
1, seeking a rapid response of the crop, to improve
the distribution of the applied product and at the same time reducing the costs of production.
Positive results are reported in several crops, as lettuce (Luz et al. 2010), cabbage (Silva et al.
2014), common bean (Lopes et al. 2014) and soybean (Barbaro et al. 2009).
Thus, the practice of agronomic biofortification via leaves stands out for conducting this
experiment, because the present study involves a trace element that may have reduced availability
through application via soil. In addition, the Li content in the soil decreases with the advance
of the weathering process and that this element is strongly incorporated into the clay minerals,
presenting a strong adsorption by the organic matter and oxides of manganese interfering in
the absorption of this element by the cultures of agronomic interest (Kabata-Pendias and
Mukherjee 2007).
Due to importance of studies with biofortification of crops with Li, associated to the scarcity
of information on this subject and the benefits that this element at optimal levels brings to
human health, this work aims to evaluate the potential of Li bioaccumulation and the agronomic
characteristics of lettuce submitted to foliar application of different sources and concentrations of
this element.

Material and methods

The experiment was carried out in a greenhouse at the Universidade Federal do Tocantins
(UFT) - Campus Gurupi, located in the southern region of the Tocantins State, 11 430 4500 S and
49 040 0700 W and altitude of 280 m. The climate of the region is Aw type (tropical with dry
season) with annual rainfall superior than 900 mm and 24.3  C average temperature (Alvares
et al. 2013).
The Li biofortification was carried out via foliar application, in five concentrations of Li (0, 10,
20, 30, and 40 mg dm
3), comparing two mineral sources of Li (lithium sulfate - Li2SO4 P.A.-
A.C.S. and lithium hydroxide - LiOH P.A.-A.C.S.). The experiment was implanted following
a completely randomized design with four repetition. The treatments were arranged in a 2  5
factorial scheme in which the first factor was the chemical source and the second was the concen-
trations of Li, of respective sources. Each experimental plot was composed of 20 pots of 5.0 dm
3
of capacity with only one plant per pot. The lettuce (Lactuca sativa L.) cultivar Elba was chosen
 JOURNAL OF PLANT NUTRITION 3

Table 1. Physical and chemical parameters of substrate used in the experiment.

Ca2þ Mg2þ Al3þ H þ Al S.B.a C.E.C.b O.M.c
cmolcdm
3 g dm
3
3.4 2.2 0 2.2 5.6 7.8 5.5
K P pH CaCl2 Sand Silt Clay V%d
mg dm
3 (%)
5.22 103.5 5.4 57.5 10 32.5 80
a
sum of bases; bcation exchangeable capacity in pH 7.0; corganic matter content; dbase saturation.

to conduct the experiment because it is much appreciated in the region and because it allowed
successive crops in the same year.
The Li application was subdivided into three applications of equal proportions (1/3 of each
concentration per application) during plant development, being applied at 15, 25, and 35 days after
transplanting. The applications were carried out by means of a hand sprayer, adopting a fixed volume
of 05 ml of solution per plant.
The soil used was characterized as a dystrophic red-yellow oxisol (EMBRAPA 2013), collected
at the UFT’s experimental area, in the 0 to 0.2 m deep layer, and then was enriched with 1:1:1
organic compound (soil, composted ruminal content, and charred rice husk). The aim of this
formulation was to improve the soil physical and chemical conditions and to allow a better devel-
opment for the lettuce root system during the experiment. After the formulation of the substrate,
the physical and chemical characterization was performed in the soil laboratory of UFT according
to methodology recommended by EMBRAPA (1997). From the chemical analysis of the substrate
and nutrient availability levels, it was verified that it would not be necessary to carry out the
chemical fertilization (Table 1).
The seedlings were produced in trays of expanded polystyrene (IsoporV) wherein each cell in the
R

tray has 40 cm3 and were sown three seeds per cell. The thinning occurred eight days after sowing,
leaving only the most vigorous seedlings. The seedlings were irrigated manually twice a day and the
transplanting to the pots was carried out when the seedlings presented between 4 and 6 well-formed
leaves, choosing the seedlings of greater vigor and homogeneity. After transplanting, the fertilization
followed the recommendations proposed by Ribeiro, Guimar~aes, and Alvarez (1999) to lettuce plants,
with urea (45% N) at the dose of 150 kg ha
1 of N, with an application at the planting (20% of the
recommendation) and three cover fertilizations (1st with 20% and the 2nd and 3rd with 30%, each).
The application of the insecticide Actara 250 WGV (200 g ha
1) was carried out weekly, and the
R

application of the fungicide Amistar TopV (400 ml ha
1) after transplanting.

R

The lettuce harvest was performed 40 days after transplanting and the traits evaluated were:
plant height (cm); stem diameter (mm); stem length (cm); number of leaves; dry weight of leaves
(g); dry weight of stem (g); dry weight of shoot (g); dry weight of roots (g); shoot/root ratio;
plant area (cm2); total leaf area (cm2); specific leaf area (cm2 g
1); and Li accumulation in leaves
(mg kg
1 of dry weight).
Plant area (cm2) was determined by measuring each plant diameter and subsequently
converted into cm2 according to Equation 1:
ðpD2 Þ
PA ¼ (1)
4
wherein: PA: plant area, in cm2; p: 3.1416; D: plant diameter, in cm.
Total leaf area (cm2) was calculated using the disc method, according to studies done by
Huerta and Alvim (1962), in this way, the leaf area of each plant was calculated according to
Equation 2:
ðND  DA  DWLÞ
TLA ¼ (2)
DWD
4 R. R. DA SILVA ET AL.

Table 2. Analysis of variance of the traits plant height (PH), stem diameter (SD), stem length (SL), number of leaves (NL), dry
weight of leaves (DWL), dry weight of stem (DWSt), dry weight of shoot (DWSh), dry weight of roots (DWR), shoot/root
ratio (SRR), plant area (PA), total leaf area (TLA), specific leaf area (SLA), and Li accumulation in leaves (LAL) of lettuce plants,
as a function of foliar application of chemical forms (LiOH and Li2SO4) and concentrations (0, 10, 20, 30, and 40 mg dm
3).
Source of variation
Degrees of freedom
Li forms (F) Li concentration (C) FC Residual
Traits 1 4 4 30 Average C.V. (%)
PH 2.13ns 14.29 0.16ns 0.7 17.93 4.67
SD 2.27ns 3.34 4.78 0.8 15.49 5.79
SL 8.28ns 4.98ns 1.20ns 2.24 9.88 15.18
NL 0.02ns 1.96ns 0.46ns 1.5 26.02 4.72
DWL 0.07 ns
12.30 1.83 ns
4.48 11.15 18.99
DWSt 0.05ns 0.23ns 0.08ns 0.18 2.22 19.46
DWSh 2.37ns 15.82 2.25ns 1.35 13.6 8.54
DWR 5.06 1.77 1.20ns 0.56 5.38 13.91
SRR 0.024ns 0.035 0.021ns 0.0097 0.4 24.18
PA 12.95ns 14637.2ns 4330.92ns 7592.97 1204.28 7.24
TLA 81.13ns 2091.48 333.81ns 401.32 196.95 10.17
SLA 0.98ns 9.67 6.12 1.47 17.65 6.88
LAL 0.01 2970.34 115.13 21.24 20.06 22.97
Significant by t-test and regression analysis (p < 0.01); Significant by t-test and regression analysis (p < 0.05); ns
non-signifi-
cant by t-test and regression analysis; CV: Coefficient of Variation.

wherein: TLA: total leaf area, in cm2; ND: number of disks; DA: disc’s area, in cm2; DWL: dry
weight of leaves, in g
1; and DWD: dry weight of the discs, in g
1.
Specific leaf area (cm2 g
1) was determined according to Equation 3.
TLA
SLA ¼ (3)
DWL
wherein: SLA: specific leaf area, in cm2 g
1; TLA: total leaf area, in cm2; and DWL: dry weight of
leaves, in g
1.
In relation to Li content in leaves (mg kg
1 MS
1), particular attention was paid to cleaning
the leaves with water to remove the excess of Li held on the outer surface of the leaves, after the
drying process of the leaves, the samples were ground in a Willey type mill with a 1.27 mm sieve.
Then the material was subjected to wet digestion using H2SO4 and catalytic mixture, consisting
of Na2SO4 and CuSO4  5H2O  Li content was determined according to the methodology
described by Malavolta, Vitti, and Oliveira (1997). The reading was carried out in a QuimisV
R

flame photometer, for which the digested was diluted in the ratio of one part to five of deionized
water (5 ml þ 25 ml) and transferred to a 50 ml container before reading.
The data were submitted to analysis of variance and their averages were compared by the
Tukey test, adopting 1 and 5% probability using Assistat, 7.7V. The regressions were plotted using
R

Sigma PlotV 10.0.

R

Results
The analysis of variance for all traits is shown in Table 2. It is observed that the sources of Li
applied had slight influence on the evaluated characteristics, and only dry weight of roots and Li
accumulation in leaves presents a significant response. On the other hand, increasing concentra-
tions of Li presented a significant effect on the plant height, stem diameter, dry weight of leaves,
dry weight of shoot, dry weight of roots, shoot/root ratio, total leaf area, specific leaf area, and
Li accumulation in leaves. It was also verified that the interaction between sources and levels
of Li resulted in a significant effect on the following characteristics: stem diameter, specific leaf
 JOURNAL OF PLANT NUTRITION 5

Table 3. Number of leaves, stem length (cm), dry weight of stem (g) and plant area (cm2) as a function of foliar application
of LiOH and Li2SO4 at increasing concentrations (0, 10, 20, 30, and 40 mg dm
3) in lettuce plants.
Number of leaves Stem length (cm)
Chemical forms Chemical forms
Levels
(mg dm
3) LiOH Li2SO4 Average LiOH Li2SO4 Average
0 26.25 26.25 26.25 a 10.8 10.8 10.80 a
10 25.75 25.5 25.62 a 9.62 8.87 9.25 a
20 26.25 26.75 26.50 a 9.75 9.12 9.43 a
30 26.66 26.25 26.45 a 10.66 8.87 9.77 a
40 26.25 25.25 25.75 a 10.75 10.25 10.5 a
Average 26.25 a 26.00 a 10.36 a 9.58 a
Dry weight of stem (g) Plant area (cm
2)
Levels Chemical forms Chemical forms
(mg dm
3) LiOH Li2SO4 Average LiOH Li2SO4 Average
0 2.42 2.42 2.42 a 1272.28 1272.28 1272.28 a
10 2.22 2.08 2.15 a 1163.76 1164.54 1164.15 a
20 2.08 2.09 2.08 a 1182.03 1240.88 1211.45 a
30 1.97 2.12 2.04 a 1236.37 1166.11 1201.24 a
40 2.22 2.26 2.24 a 1164.15 1180.44 1172.29 a
Average 2.18 a 2.19 a 1203.72 a 1204.85 a

area, and Li accumulation in leaves. No significant effect was observed for number of leaves, stem
length, dry weight of stem, and plant area (Table 3). For the traits that present a significant effect
on sources of variation, a linear or quadratic response was calculated as a function of the increas-
ing levels of LiOH and Li2SO4 applied in lettuce plants (Figures 1 and 2). The maximum applied
Li dose (40 mg dm
3), for both chemical sources, provided a significant reduction in all the char-
acteristics, excepting specific leaf area after using LiOH. In contrast, the lettuce accumulated the
most Li in relation to control plants, reaching up to 84 and 61 times of superiority for the
hydroxide and sulfate form of Li, respectively.
The increase of the Li concentration in the applied solution promoted a reduction in the let-
tuce growth independent of the chemical form used (Figure 1A). For both sources, the level of
40 mg dm
3 presented about of 15% reduction in plant height in relation to the control.
However, it is observed that regardless of the Li chemical form the initial concentration of 10 mg
dm
3 occurred an expressive reduction in the plant height, up to 12%, in relation to the control.
In relation to the stem diameter (Figure 1B), LiOH promoted a negative response at the highest
level applied, 40 mg dm
3 of Li, reaching a reduction of 12% in relation to the control plants. In
contrast, a positive result was observed with Li2SO4, and to this source the most efficient level
through the quadratic function derivation was 19.7 mg dm
3, this dose provided 16.2 mm of stem
diameter, corresponding to an increase of 6% in comparison to control plants. As well as plant
height, the increase of Li concentrations for both sources also promoted a reduction in the dry
weight of leaves and shoots of lettuce (Figures 1C) and 1D. At the concentration of 40 mg dm
3
the dry weight of leaves was reduced by 24%, for both chemical forms, and for the dry weight of
stem this reduction was 20 and 15%, for LiOH and Li2SO4, respectively.
For dry weight of roots, similar behavior was observed among the Li forms tested (Figure 2A).
For the LiOH and Li2SO4 the highest growth-stimulation efficiency estimated concentrations was
22 and 16.8 mg dm
3 of Li, in these levels the lettuce plants presented 6.36 g and 5.65 g of dry
weight of roots, respectively. In comparison with the control plants these concentrations caused
an increase in dry weight of roots by 31%, after application of LiOH, and 18%, after application
of Li2SO4. At 40 mg dm
3 for both Li forms there was a reduction of 22% in the dry weight of
lettuce roots, in relation to the higher response found for each source, however, was no decrease
in root biomass in relation to the control plants. The application of increasing concentrations of
Li affected the shoot/root ratio of lettuce, with a quadratic effect (Figure 2B). This parameter was
6 R. R. DA SILVA ET AL.

Figure 1. Plant height (cm) (A), stem diameter (mm) (B), dry weight of leaves (g) (C), and dry weight of shoot (g) (D) of lettuce
plants, as a function of foliar application of chemical forms (LiOH and Li2SO4) and concentrations (0, 10, 20, 30, and
40 mg dm
3).

found to be 0.51 for LiOH and 0.42 for Li2SO4. The study of the dry weight of shoot demonstrate
the reduction of this trait due to the increase of the Li concentration applied, which corresponds
to the increase of the shoot/root ratio, therefore, the effect of Li on the roots is less than on the
above-ground parts of the lettuce. There were no impacts in total leaf area of lettuce plants at
lower Li concentrations (Figure 2C). Both sources provided similar results and were not very
expressive, and at 7.5 mg dm
3 lettuce plants had a total leaf area of approximately 210 cm2 and
an increase of 3% (non-significant) for both sources in relation to the control plants was found.
However, at 40 mg dm
3, the LiOH and Li2SO4 reduced leaf area by 10% and 19% in relation to
control plants. In relation to the specific leaf area (Figure 2D), it was found that the
 JOURNAL OF PLANT NUTRITION 7

Figure 2. Dry weight of roots (g) (A), shoot/root ratio (B), total leaf area (cm2) (C), and specific leaf area (cm2 g
1) (D) of lettuce
plants, as a function of foliar application of chemical forms (LiOH and Li2SO4) and concentrations (0, 10, 20, 30, and
40 mg dm
3).

concentrations causing the largest increase of this parameter were 40 mg dm
3 (LiOH) and 21 mg
dm
3 (Li2SO4). Under these concentrations the lettuce plants get 19.75 cm2 g
1 and 18.32 cm2
g
1, corresponding to 18 and 13% of increase in relation to the control plants.
The analysis of Li accumulation in leaves of the lettuce plants demonstrates that it was directly
proportional to the concentrations of this element applied through leaves and was not dependent
to chemical form of Li (Figure 3). The Li concentration, which was low in the control plants,
increased significantly after the foliar application and both forms induced maximum concentra-
tion of Li at 40 mg dm
3. After using LiOH the leaves accumulated 51 mg Li kg
1 dry matter,
8 R. R. DA SILVA ET AL.

Figure 3. Li accumulation in leaves (mg Kg
1 of dry matter) of lettuce plants, as a function of foliar application of chemical
forms (LiOH and Li2SO4) and concentrations (0, 10, 20, 30, and 40 mg dm
3).

representing an increase of 84 times compared to the control plants which present 0.61 mg kg
1
of dry matter. For the Li2SO4, application of 40 mg dm
3 caused accumulation of 37.7 mg
Li kg
1 dry matter, this result represents an increase of 61 times in relation to the control plants.
However, at this concentration there is an expressive reduction of approximately 24% in the dry
weight of shoot when the different sources were used.
Taking the dry weight of lettuce leaves as the main trait of crop yield, the foliar application of
Li at increasing concentrations presented a negative result. However, the tested Li concentration
that provided the lowest reduction of this parameter was 10 mg dm
3 for both sources
(Figures 1C and 1D). Under this concentration the lettuce plants accumulated 11.5 and 7.0 mg
Li kg
1 with an increase of 15 (LiOH) and 9 (Li2SO4) times in relation to the control.

Discussion
A better understanding of the Li effects on plant growth and its accumulation in tissues remain
of great importance for improving knowledge about the biological activity of Li compounds in
crops. In our study, the increase of Li concentrations through the foliar application of LiOH and
Li2SO4 did not influenced only the follow characteristics: number of leaves, stem length, dry
weight of stem, and plant area (Table 3).
It is also worth mentioning that visually there was no formation of necrotic or chlorotic spots
on the new or old leaves, regardless of the Li chemical form or concentration used. Possibly, this
result can be explained by the splitting of the Li concentrations in three applications, of equal
proportion, during the development of lettuce plants. This method of application may have led to
a reduction in the toxic effects of the Li, even after the application of the maximum concentration
(40 mg dm
3). However, negative response on agronomic characteristics and formation
of necrotic spots in lettuce plants, cv. Justyna, grown in nutrient solution containing increasing
concentrations of Li has already been documented by Kalinowska, Hawrylak-Nowak, and
Szyma
nska (2013). The same negative effect of the Li ions has also been described in other
 JOURNAL OF PLANT NUTRITION 9

crops such as corn and sunflower (Hawrylak-Nowak, Kalinowska, and Szyma
nska, 2012), as well
as in Brassica carinata seedlings (Li et al. 2009).
The results of the present study indicate that the sources of Li used here did not differ consid-
erably in their phytotoxicity for lettuce plants and that a concentration of 40 mg dm
3 caused a
great reduction in the stem diameter, dry weight of roots, total leaf area, and specific leaf area
(only for Li2SO4) but affected mainly plant height, dry weight of leaves and dry weight of shoot
(Figures 1 and 2). Similar result was also obtained by Li et al. (2009) and these authors concluded
that high concentrations of Li (about 200 mg dm
3) promoted a reduction in the growth of
Brassica carinata seedlings. Also, Stolarz, Kr
ol, and Dziubi

nska (2015) observed that concentra-
tions of 60- and 80-mM Li can reduce the hypocotyl height of sunflower seedlings by 34 and
55%, respectively, in relation to the control plants. Recently, Kalinowska, Hawrylak-Nowak, and
Szyma
nska (2013) found that the maximum concentration of Li used (100 mg dm
3) in their
experiment inhibited lettuce growth, since the plants reached only 5–9% of the dry weight of the
control plants. Moreover, Hawrylak-Nowak, Kalinowska, and Szyma
nska (2012) concluded that
concentrations of 50 mg dm
3 of Li (their maximum applied concentration) reduced the biomass
of sunflower and maize by 27 and 32%, respectively. Negative effects that cause reduction in
height and production of dry weight by plants depend mainly on the concentration of Li in plant
tissue and on the plant species. In the experiments carried out by Magalh~aes et al. (1990), radish
and watercress cultivated under hydroponic conditions under Li stress showed a reduction in the
dry weight of roots and leaves, which resulted from the reduction of water retention in
the tissues.
Li at relatively low concentration may stimulate plant growth, whereas at higher concentrations
it may completely inhibit growth and may lead to plant death (Shahzad et al. 2016). Thus, at low
concentrations, between 7.5 and 22 mg dm
3, Li2SO4 and LiOH provided significant increase in
some parameters determined. For dry weight of roots (Figure 2A) this increase was 31% for
LiOH and 16.8% for Li2SO4. Furthermore, there was noted an increase in specific leaf area
(Figure 2D) by 18% after using LiOH and 13% after using Li2SO4. The stems diameter
(Figure 1B) presented a 6% increase in plants submitted to 19.7 mg dm
3 of Li for Li2SO4. These
results are in agreement with Kalinowska, Hawrylak-Nowak, and Szyma
nska (2013), that observed
a 62% increase in dry weight of roots of lettuce plants grown in nutrient solution containing
2.5 mg dm
3 of Li, using LiOH as a Li source. Hawrylak-Nowak, Kalinowska, and Szyma
nska
(2012) observed different biological effects of Li in maize and sunflower, both grown in nutrient
solution. In maize plants under 5 mg dm
3, there was a 15% increase in biomass and concentra-
tions of 5 and 25 mg dm
3 promoted a 22 and 29% increase in leaf area, respectively. On the
other hand, sunflower plants submitted to increasing concentrations of Li showed an expressive
reduction in these characteristics. In spinach and mustard, the addition of Li to the soil also
increased plant biomass, but only under conditions of reduced light levels (Makus, Zibilske, and
Lester 2006). Mcstay, Rogers, and Anderson (1980) demonstrated that 4 mg dm
3 of Li caused an
increase in the plant height, and leaf area of common bean.
The determination of the relationship between root and shoot dry weight is important because
it shows in which organs the plant prioritizes the distribution of photoassimilates reflecting the
accumulation of dry matter. In the present work, increasing levels of Li applied via leaves caused
a change in this characteristic (Figure 2B), wherein at the maximum Li level, it was observed that
the dry weight of shoot presents a great reduction while the dry weight of root demonstrate
increase to the detriment of the photoassimilates accumulation in this region, possibly in the
sense of mitigating the consequences of the application of the element.
Li is absorbed by all plant species, although not essential for plant growth and development.
As already discussed, low-level supplementation of this element can give to plants an increase
in their biomass, growth stimulation, and there is evidence of its positive effects in the photo-
synthesis, synthesis and translocation of sugars, many enzymatic processes and N metabolism
10 R. R. DA SILVA ET AL.

(Shkolnik 1984; Schrauzer 2002; Aral and Vecchio-Sadus 2008, Jiang et al. 2014). According to
Bradford (1966), low levels of Li also improved plant production in some cases, increasing yield,
accelerating maturation and increasing resistance to disease. This increase in agronomic charac-
teristics that reflected in productivity may be due to the hormonal effect that has been described
in the literature as stimulation of plant growth from lower levels of potentially toxic agents, as
well as the Li (Allender et al. 1997) or some other unknown effects that Liþ ions can exert on the
metabolism of a plant. Kabata-Pendias and Mukherjee (2007) explain that yet no function of Liþ
ions is known as a cofactor for any enzymatic activity, it is known only that this element has an
affinity for enzymes activated by Ca and/or Mg. Therefore, the beneficial effect of Li compounds
on plant growth still needs further research.
In relation to the accumulation of Li in the shoot of the lettuce plants, the increase in the con-
centration of this element in the applied solution influences positively on foliar Li concentration
(Figure 3). Li concentrations in leaves showed that the accumulation was proportional to the con-
centrations of this element in a solution and was not related to its chemical form. Similar results
were found by Kalinowska, Hawrylak-Nowak, and Szyma
nska (2013). These authors obtained a
linear response in Li accumulation in shoot and root of lettuce plants independent of the sources
used (LiOH and LiCl). Accumulation of this element did not occur only with the application of
high concentrations, at initial concentrations (10 mg dm
3), lettuce plants accumulated about 15
times more Li compared to the control plants for the both forms. Thus, it is possible to perform
an accumulation of this element in quite expressive quantities without great damage in the agro-
nomic characteristics of lettuce plants. These results demonstrate that the lettuce culture has a
great capacity to perform Li bioaccumulation independent of the chosen source.
According to Jurkowska and Rog
oz_ (1993), Li content is considered higher in dicots plants
than in monocots, and among dicots, lettuce may contain relatively high levels of this element
(Schafer 2004). However, according to Magalh~aes et al. (1990), lettuce can present half of Li con-
tent in their leaves as radish and watercress cultivated under hydroponic conditions and suggests
that lettuce is able to accumulate less Li among the studied dicot plants. Hawrylak-Nowak,
Kalinowska, and Szyma
nska (2012) explains that the observed differences in Li accumulation by
different plant species may also be partially related to the influence of some experimental condi-
tions, including different plant ages, when Li compounds were added to the nutrient solution and
several exposure times to the element.
It is important to recall that Li levels in the human body may be beneficial mainly in mood
stabilization (bipolar disorder) and it could indirectly reduce suicide rates the population,
Alzheimer’s disease prevention (Young 2011; Forlenza et al. 2011), reduction of amyotrophic
sclerosis, schizophrenia, phobic anxiety (Martinsson et al. 2013), reduction of diabetes, heart dis-
ease and alcohol dependence (Schrauzer, Shrestha, and Flores-Arce 1992; Schrauzer 2002; Norra
et al. 2008). Thus, adequate consumption of biofortified foods with Li may be considered as a
new aspect in agriculture to help combat diseases and disorders caused by deficiency of this elem-
ent in the human body (Vita, De Peri, and Sacchetti 2015).
In this work, promising results were obtained after the application of two chemical forms of Li
at different concentration in lettuce plants. The foliar applications, as well as the splitting in equal
proportions of the Li concentrations during the development of the lettuce plants, provided great
accumulation of Li in the shoot independent of the Li source used. Relatively low concentrations
of this element also provide bioaccumulation, however, with minimal losses in agronomic charac-
teristics, showing that lettuce plants have great potential for biofortification with this element.

ORCID
Vitor L. Nascimento http://orcid.org/0000-0001-5276-2638
 JOURNAL OF PLANT NUTRITION 11

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