Scientia Horticulturae 166 (2014) 9–16

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Scientia Horticulturae
journal homepage: www.elsevier.com/locate/scihorti

Assessment of biofortification with iodine and selenium of lettuce

cultivated in the NFT hydroponic system
Sylwester Smoleń ∗ , Iwona Kowalska, Włodzimierz Sady
University of Agriculture in Krakow, Faculty of Horticulture, Institute of Plant Biology and Biotechnology, Unit of Plant Nutrition, Al. 29 Listopada 54,
31-425 Krakow, Poland

a r t i c l e i n f o a b s t r a c t

Article history: Iodine and selenium are not nutrients for plants but both play important roles in human and animal
Received 8 July 2013 organisms. Diet of many populations around the world contains insufficient amount of these elements.
Received in revised form 4 November 2013 Iodine and selenium biofortification of crop plants can increase its transfer into the food chain. A limiting
Accepted 8 November 2013
factor for the development of agro-technical methods of I and Se application is poor recognition of its
interaction with respect to plant growth and metabolism. The aim of the study was to determine the
Keywords:
possibility of simultaneous biofortification of lettuce with iodine and selenium, applied foliarly or through
Biofortification
the nutrient medium in the hydroponic system of nutrient film technique (NFT). Two-factor experiment
Selenium
Iodine
with greenhouse cultivation of lettuce ‘Melodion’ cv. was conducted for two years. The following five
Nutrient film technique sub-blocks with iodine and selenium introduction into the nutrient medium were distinguished: (1)
Lettuce control, (2) 0.5 mg Se dm−3 , (3) 1 mg I dm−3 , (4) 0.5 mg Se dm−3 + 1 mg I dm−3 , (5) 1.5 mg Se dm−3 + 1 mg
I dm−3 – the respective molar concentration were as follows: (2) 6.33 ␮M Se, (3) 7.88 ␮M I, (4) 6.33 ␮M
Se + 7.88 ␮M I, (5) 19.00 ␮M Se + 7.88 ␮M I. Each sub-block included four combinations with five-time
foliar treatment with: (A) distilled water, (B) 0.005% Se (0.633 mM Se), (C) 0.05% I (3.94 mM I) and (D)
0.005% Se + 0.05% (0.633 mM Se + 3.94 mM I). Iodine and selenium were applied in the form of KIO3 and
Na2 SeO4 , respectively. There were three replicates for each treatment with eleven plants per one replicate.
Tested factors did not negatively affect lettuce yield (average head weight) and nutritional status of leaves
and roots. Only in lettuce from the sub-blocks with the nutrient medium containing 1.0 mg I dm−3 , 0.5 mg
Se + 1.0 mg I dm−3 and 1.5 mg Se + 1.0 mg I dm−3 , foliar application of KIO3 and Na2 SeO4 + KIO3 decreased
the level of Ca, Mg and Fe in roots, when compared to respective plants from the control sub-block.
Introduction of IO3 − or SeO4 2− into the nutrient medium (in a dose of 0.5 mg Se and 1.0 mg I dm−3 ) had
no negative impact on root uptake of SeO4 2− and IO3 − , respectively and its further transport to leaves.
Higher efficiency of iodine and selenium biofortification of lettuce plants was noted after foliar application
of tested compounds rather than through its introduction into the nutrient medium. Foliar spraying with
IO3 − and SeO4 2 did not affect root uptake of iodine and selenium present in the nutrient medium. Foliar
application of iodine together with selenium improved SeO4 2 absorption by leaves when compared to
plants sprayed only with Se. Results obtained in the control sub-block may indirectly suggest that the
transport of iodine and selenium in plants may occur from leaves to roots through phloem.
© 2013 Elsevier B.V. All rights reserved.

1. Introduction
Selenium and iodine are not essential mineral nutrients for
plants (Kopsell and Kopsell, 2007; Kabata-Pendias, 2011) though
Se, together with Al, Co, Si, Na and V, is included to the group of
“beneficial elements” (Kopsell and Kopsell, 2007). It has been fre-
quently noted that iodine applied in low concentrations can have
a positive effect on plants (Lehr et al., 1958; Borst-Pauwels, 1961,
1962; Kabata-Pendias, 2011). The physiological, biochemical and
∗ Corresponding author. Tel.: +48 12 6625239; fax: +48 12 6625240.
E-mail addresses: Sylwester.Smolen@interia.pl, s.smolen@ogr.ur.krakow.pl
(S. Smoleń).
molecular nature of this influence has not yet been recognized
what makes iodine still not being considered “a beneficial element”
(Kabata-Pendias, 2011).
Both elements are crucial for proper functioning of human
and other animal organisms. There is a close relationship – on
the physiological and molecular basis – between iodine and sele-
nium resulting mainly from the fact that three of iodothyronine
deiodinases (D1, D2 and D3) contain selenium in the form of seleno-
cysteine (Arthur et al., 1992; Bobek, 2006).
Approximately two-third of the whole human population in the
world suffer from illnesses and health problems caused by insuffi-
cient supply of iodine and selenium in the daily diet. This situation
is noted not only in the areas with endemic deficiency of these ele-
ments and mainly results from low content of mobile forms of I and

0304-4238/$ – see front matter © 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.scienta.2013.11.011
10 S. Smoleń et al. / Scientia Horticulturae 166 (2014) 9–16
Se in soils. The consequence is the low transfer of both elements to
the food chain in which plants are the first level. One of the most
promising approaches to combat this problem may be biofortifi-
cation (enrichment) of crop plants with these mineral nutrients.
This strategy may be realized through agronomic or biotechnolo-
gical methods (Lyons et al., 2004; White and Broadley, 2005, 2009;
Hirschi, 2009; Bouis et al., 2011).
Numerous studies report the issue of selenium biofortification
of crop pants. In the case of lettuce, the influence of Se form (SeO3 2− ,
SeO4 2− ) and dose on: biomass, selenium accumulation, antioxida-
tive activity, the content of secondary metabolites (Cary, 1980;
Hartikainen et al., 1997; Pezzarossa et al., 2007; Ramos et al., 2010;
Ríos et al., 2008, 2010) or nitrogen metabolism (Ríos et al., 2010)
has been documented.
Physiological and biochemical reactions of plants to iodine still
remain unclear. It comes from the fact that for the last few decades
studies in this aspect have been rarely conducted. What is more,
proposed conclusions regarding plant fertilization with iodine have
been inconsistent (Marschner, 1995). One of the main causes is
analytical difficulty related to iodine determination and interpre-
tation of obtained results (Smoleń et al., 2011). In the last decade a
growing popularity on studies of iodine biogeochemistry has been
observed. In the case of lettuce cultivation in perlite/vermiculite (in
growth chambers or greenhouse) the influence of various doses of
iodide (I− ) and iodate (IO3 − ) form of this element on plant toxicity,
biomass, biofortification efficiency, physiological and biochemical
processes, the content of primary or secondary metabolites and
mineral nutrition of plants has been documented (Blasco et al.,
2008, 2010a,b, 2011a,b, 2012, 2013; Voogt and Jackson, 2010; Voogt
et al., 2010).
Basically, little information is available on iodine or selenium
influence on plants cultivated in recirculating hydroponic systems
– conditions comparable to those used in large-scale horticulture
production. In the studies conducted in the NFT system, a diverse
effect of foliar nutrition with iodine was found with reference to
the content of: dry matter, soluble sugars, ascorbic acid and free
amino acids in lettuce heads depending on KIO3 concentration in
the nutrient solution (Ledwożyw-Smoleń et al., 2011).
In above context it seems reasonable to carry out the study
with simultaneous application of iodine and selenium in plant cul-
tivation. Despite its importance, studies focused on this subject
are scarce. Only Zhu et al. (2004) conducted the experiment with
hydroponic cultivation of spinach with the application of SeO4 2−
and IO3 − into the nutrient medium. There is no available infor-
mation assessing the effect of simultaneous application of iodine
and selenium through foliar spraying and introduction of these ele-
ments into the rhizosphere. Development of agro-technical rules of
biofortification of crop plants with iodine and selenium requires the
recognition of all aspects of its interaction with respect to I and Se
uptake and distribution as well as possible side-effects. An impor-
tant issue is to select the most efficient methods of increasing the
content of I and Se in yield. It seems that the first step should be to
conduct studies in the hydroponic system to eliminate the influence
of soil factors on the uptake of mineral nutrients by plants.
The aim of the study was to determine the possibility of simul-
taneous application biofortification of lettuce with iodine and
selenium applied foliarly or through the nutrient medium in the
hydroponic system of NFT.
2. Materials and methods
2.1. Plant culture
The experiment was conducted in the years 2010–2011 in a
greenhouse of the Faculty of Horticulture, University of Agriculture
Table 1
The approximate amount of iodine and selenium applied to plants through foliar
spraying.
Foliar application Total for five foliar treatments
in each sub-block
cm3 water/ mg mg
solution plant−1 Se plant−1 I plant−1
H2 O – control 19.09 0.000 0.000
Se 19.09 0.955 0.000
I 19.09 0.000 9.545
Se + I 19.09 0.955 9.545
in Krakow. Lettuce ‘Melodion’ cv. was cultivated in autumn sea-
son in the hydroponic system of NFT. Each year seeds were sown
into rockwool plugs in the first half of September (10.09.10 and
15.09.11). In a two-leaf phase seedlings were placed in the holes
(spaced 25 cm apart) of styrofoam slabs filling NFT beds (“dry-
hydroponic” method).
After plant seedling, day and night temperature was set at
15 and 10 ◦ C, respectively. From the second decade of November
(10.11.2010 and 2011) to the end of the experiment light supple-
mentation was applied for the following hours: 6.00–7.00 and
15.00–17.00 with the use of 600 W high-pressure sodium lamps
in order to extend day-length.
Studies were conducted according to a two-factorial design
in which five sub-blocks (units) with the introduction of sele-
nium and iodine into the nutrient medium (first factor) were
distinguished: (1) control, (2) 0.5 mg Se dm−3 , (3) 1 mg I dm−3 , (4)
0.5 mg Se dm−3 + 1 mg I dm−3 , (5) 1.5 mg Se dm−3 + 1 mg I dm−3 –
the respective molar concentration were as follows: (2) 6.33 ␮M Se,
(3) 7.88 ␮M I, (4) 6.33 ␮M Se + 7.88 ␮M I, (5) 19.00 ␮M Se + 7.88 ␮M
I. Within each sub-block four combinations with five-time foliar
application of Se and I were introduced (second factor): (A) dis-
tilled water – control for foliar application, (B) 0.005% Se (0.633 mM
Se), (C) 0.05% I (3.94 mM I) and (D) 0.005% Se + 0.05% (0.633 mM
Se + 3.94 mM I). The chemical forms of selenium and iodine used
for both methods were: Na2 SeO4 and KIO3 , respectively. Both
compounds were introduced into the nutrient medium after trans-
planting seedlings into NFT system. Foliar spraying with iodine and
selenium was conducted every 7 days beginning with the six-leaf
phase and ending 18 days before the harvest. There were three
replicates with eleven plants per one replicate in each combination
– the total number of plants in the experiment was 660 each year.
Totally, for five treatments 210 cm3 of water or tested solutions of
iodine and/or selenium salts were used in each repetition, which is
approximately 19.09 cm3 plan−1 . Estimated amounts of iodine and
selenium applied foliarly to lettuce plants are presented in Table 1.
In each sub-block plants were growing in the nutrient medium
of pH 6.0, EC 1.8 mS cm−1 and the following content of macro- and
micronutrients (mg dm−3 ): 120 N, 40 P, 170 K, 35 Mg, 150 Ca, 1.5 Fe,
0.55 Mn, 0.25 Zn, 0.2 B, 0.09 Cu, 0.04 Mo [the respective molar con-
centrations were as follows: 8.6 mM N, 1.3 mM P, 4.3 mM K, 1.4 mM
Mg, 3.7 mM Ca, 26.86 ␮M Fe, 10.01 ␮M Mn, 3.82 ␮M Zn, 18.50 ␮M B,
1.42 ␮M Cu, 0.42 ␮M Mo]. For each sub-block, 1000 dm3 of nutri-
ent medium were stored in separate containers and periodically
administered into cultivation slabs. The frequency of watering was
adjusted to the growth stage of lettuce and weather conditions.
Plants were cultivated in the recirculating system. Water losses
caused by evaporation and uptake by plants were being adjusted to
the set volume automatically, while the content of mineral nutri-
ents, iodine and selenium (in the respective sub-blocks) was being
supplemented to the initial concentration basing on the chemical
analysis of the nutrient medium. In both years of the study one
time during the cultivation (10.11.10 and 15.11.11) the whole vol-
ume of the nutrient medium was replaced with the fresh one while
maintaining the set chemical composition.
 S. Smoleń et al. / Scientia Horticulturae 166 (2014) 9–16 11

Lettuce harvest followed by the assessment of average head

weight and the collection of leaf and root samples was conducted
on 30.11.10 and 10.12.11.

2.2. Plant analysis

Whole root systems as well as one-fourth of each head harvested

in the experiment were collected for further analysis. Root and leaf
samples were dried at 70 ◦ C in a laboratory dryer with forced air
circulation. Dried samples were ground in variable speed rotor mill
Pulverisette 14, FRITSCH using 0.5 mm sieve.
In plant samples iodine and selenium concentration was deter-
mined after sample incubation with 1 ml of 25% TMAH and 10 ml of
double-distilled water. Incubation of 0.5 g plant material was con-
ducted for 3 h at 90 ◦ C in closed falcon tubes (PN-EN 15111, 2008).
In root and leaf samples N content was assayed by the Kjeldahl
method with the use of VELP Scientifica UDK 193 distillation unit
(Persson and Wennerholm, 1999). The level of P, K, Ca, Mg, S, B, Cu,
Fe, Mn, Mo and Zn in plant samples was analyzed after mineraliza-
tion in 65% super pure HNO3 in microwave system CEM MARS-5
Xpress (Pasławski and Migaszewski, 2006). The content of iodine,
selenium and mineral nutrients was determined with the use of
high-dispersion spectrometer ICP-OES Prodigy Teledyne Leeman
Labs.

2.3. Data analysis

Since the effect of year was not significant the results are means
of two years (2010 and 2011). Than the data were subjected to
analysis of variance using ANOVA module of Statistica 10.0 PL. To
determine the significance between means the Bonferroni test was Fig. 2. Selenium content in leaves (A) and roots (B) of lettuce depending on the
used. The significance was declared at P < 0.05. interaction between foliar application and introduction to the nutrient medium of
iodine and selenium (n = 6). Means followed by the same letters are not significantly
different at P < 0.05; bars indicate standard error.
3. Results

3.1. Biomass and mineral nutrition
No statistically significant interaction was found between tested
factors (introduction of SeO4 2− /IO3 − into the nutrient medium and
foliar treatment) on yield quantity expressed as a mean weight of
a lettuce head (Fig. 1). The measurement of the influence of tested
factors on plants (apart form the yield assessment) bases on its
effect on nutritional status of plants. In the conducted studies no
such interaction was statistically significant with respect to the
content of macro- and micronutrients in leaves and roots of let-
tuce (Tables 2–5), with the exception of the content of Ca, Mg and
Fig. 1. Influence of foliar application and introduction to the nutrient medium of
iodine and selenium on lettuce yield (n = 6). n.s., differences are not significant at
P < 0.05; bars indicate standard.
Fe in roots (Tables 4 and 5). In lettuce from the sub-blocks with
the nutrient medium containing 1.0 mg I dm−3 , 0.5 mg Se + 1.0 mg
I dm−3 and 1.5 mg Se + 1.0 mg I dm−3 , foliar application of KIO3
and Na2 SeO4 + KIO3 decreased the level of Ca, Mg and Fe in roots,
when compared to respective plants from the control sub-block.
The highest concentration of these three elements was determined
in roots of plants from the control sub-block foliarly treated with
Na2 SeO4 + KIO3 .
3.2. Selenium and iodine
A significant interaction between tested methods of iodine and
selenium application (into the nutrient medium or foliarly) was
noted with respect to the content of these elements in lettuce leaves
and roots (Figs. 2A and B and 3A). Along with increasing dose of sele-
nium introduced into the nutrient medium (0 – control, 0.5, 1.5 mg
Se dm−3 – the latter one applied together with iodine) a signifi-
cant increase in Se level in roots and leaves of lettuce was observed
(Fig. 2A and B). In comparison to the control, introduction of iodine
into the nutrient medium caused a significant increase in its level
in lettuce leaves and roots (Fig. 3A and B). It should be underlined
that Se content in roots and leaves of plants grown in the nutrient
medium supplemented with 1.5 mg Se + 1.0 mg I dm−3 was almost
3-fold higher than in plants from the sub-block with proportion-
ally lower Se content in the nutrient medium, i.e. 0.5 mg Se dm−3 .
It is interesting that in plants sprayed with distilled water and cul-
tivated in the nutrient medium containing iodine and/or selenium
the content of Se in roots was higher than in leaves while for iodine
– comparable levels of this element were noted in both organs
(Figs. 2 and 3A and B). Basically, foliar application of Na2 SeO4 and
12 S. Smoleń et al. / Scientia Horticulturae 166 (2014) 9–16

Table 2
The content of N, P, K, Ca, Mg and S in lettuce leaves depending on the interaction between foliar application and introduction of I and Se into the nutrient medium.

Nutrient medium Foliar application % d.w. (n = 6, ±SE)

N P K Ca Mg S

Control H2 O – control 5.59 ± 0.08 0.94 ± 0.01 8.62 ± 0.01 1.00 ± 0.01 0.34 ± 0.004 0.29 ± 0.013
Se 5.44 ± 0.07 0.94 ± 0.02 8.57 ± 0.02 0.95 ± 0.02 0.32 ± 0.009 0.27 ± 0.006
I 5.47 ± 0.03 0.92 ± 0.02 8.71 ± 0.03 0.98 ± 0.03 0.32 ± 0.009 0.26 ± 0.010
Se + I 5.39 ± 0.11 0.92 ± 0.01 8.47 ± 0.01 0.96 ± 0.01 0.33 ± 0.002 0.26 ± 0.001

0.5 mg Se dm−3 H2 O – control 5.38 ± 0.03 0.86 ± 0.02 8.69 ± 0.02 0.91 ± 0.02 0.31 ± 0.012 0.25 ± 0.003
Se 5.47 ± 0.01 0.84 ± 0.01 8.25 ± 0.00 0.87 ± 0.00 0.29 ± 0.004 0.25 ± 0.002
I 5.54 ± 0.03 0.88 ± 0.03 9.10 ± 0.01 0.95 ± 0.01 0.30 ± 0.002 0.26 ± 0.006
Se + I 5.60 ± 0.03 0.90 ± 0.01 8.45 ± 0.01 0.91 ± 0.01 0.32 ± 0.006 0.26 ± 0.007

1.0 mg I dm−3 H2 O – control 5.23 ± 0.04 0.95 ± 0.02 8.82 ± 0.02 1.11 ± 0.02 0.35 ± 0.008 0.26 ± 0.005
Se 5.26 ± 0.01 0.96 ± 0.02 8.65 ± 0.01 1.05 ± 0.01 0.32 ± 0.006 0.27 ± 0.007
I 5.32 ± 0.01 0.91 ± 0.02 8.73 ± 0.02 1.11 ± 0.02 0.33 ± 0.005 0.26 ± 0.003
Se + I 4.65 ± 0.44 0.89 ± 0.01 8.29 ± 0.03 1.02 ± 0.03 0.33 ± 0.010 0.26 ± 0.010

0.5 mg Se + 1.0 mg I dm−3 H2 O – control 5.29 ± 0.05 0.93 ± 0.04 8.94 ± 0.03 1.05 ± 0.03 0.34 ± 0.014 0.27 ± 0.009
Se 5.42 ± 0.04 0.93 ± 0.03 8.47 ± 0.02 0.98 ± 0.02 0.32 ± 0.005 0.27 ± 0.006
I 5.25 ± 0.11 0.94 ± 0.03 9.01 ± 0.01 1.06 ± 0.01 0.33 ± 0.006 0.26 ± 0.007
Se + I 5.26 ± 0.04 0.92 ± 0.01 8.70 ± 0.01 1.00 ± 0.01 0.33 ± 0.006 0.27 ± 0.002

1.5 mg Se + 1.0 mg I dm−3 H2 O – control 5.39 ± 0.04 0.91 ± 0.03 9.01 ± 0.00 1.02 ± 0.00 0.34 ± 0.006 0.27 ± 0.006
Se 5.54 ± 0.07 0.90 ± 0.02 8.71 ± 0.01 0.95 ± 0.01 0.31 ± 0.003 0.27 ± 0.003
I 5.47 ± 0.04 0.90 ± 0.01 8.89 ± 0.02 1.01 ± 0.02 0.33 ± 0.006 0.28 ± 0.002
Se + I 5.47 ± 0.04 1.04 ± 0.09 10.05 ± 0.17 1.14 ± 0.17 0.37 ± 0.045 0.31 ± 0.024

Test F for nutrient medium × foliar application n.s. n.s. n.s. n.s. n.s. n.s.

SE, standard error; n.s., differences are not significant at P < 0.05.

KIO3 turned out more effective for iodine and selenium biofortifi-
cation of lettuce leaves than the introduction of these compounds
into the nutrient medium.
3.2.1. Nutrient medium and iodine/selenium interaction
Interaction between iodine and selenium introduced into the
nutrient medium requires in-depth analysis particularly with
respect to its influence on the accumulation of these two ele-
ments in plants. In substance, the description of these relations
was limited only to plants sprayed with distilled water, I and Se
– respectively in sub-blocks with the introduction of only Se or I
into the nutrient medium.
Simultaneous application of Se and I (0.5 mg Se + 1.0 mg I dm−3 )
into the nutrient medium did not significantly change the content
of selenium in roots and leaves of lettuce sprayed with distilled
water when compared to the sub-block with 0.5 mg Se dm−3 con-
centration in the medium (Fig. 2A and B). Interesting is also the
observation that within these two sub-blocks (0.5 mg Se dm−3 and
0.5 mg Se + 1.0 mg I dm−3 ) foliar application of KIO3 did not affect
Se uptake from the nutrient medium by plants – as compared to
spraying with distilled water.
In plants sprayed with distilled water or Na2 SeO4 simulta-
neous introduction of Se and I into the nutrient medium (in a
dose of 0.5 mg Se + 1.0 mg I dm−3 ) did not affect iodine content in

Table 3
The content of B, Cu, Fe, Mn, Mo and Zn in lettuce leaves depending on the interaction between foliar application and introduction of I and Se into the nutrient medium.

Nutrient medium Foliar application mg kg−1 d.w. (n = 6, ±SE)

B Cu Fe Mn Mo Zn

Control H2 O – control 31.87 ± 0.26 6.32 ± 0.19 98.99 ± 4.67 137.79 ± 1.60 0.53 ± 0.02 54.65 ± 1.84
Se 31.73 ± 0.56 6.32 ± 0.05 75.11 ± 0.65 127.97 ± 4.38 0.51 ± 0.05 52.79 ± 2.31
I 30.78 ± 1.53 6.62 ± 0.39 76.15 ± 1.55 132.05 ± 5.82 0.50 ± 0.02 52.56 ± 2.88
Se + I 29.67 ± 0.85 6.36 ± 0.04 76.55 ± 4.66 128.42 ± 3.08 0.46 ± 0.04 49.78 ± 0.31

0.5 mg Se dm−3 H2 O – control 30.73 ± 0.83 5.66 ± 0.34 80.36 ± 0.94 116.14 ± 7.76 0.46 ± 0.04 45.01 ± 2.27
Se 29.98 ± 0.98 5.88 ± 0.28 75.48 ± 3.34 117.19 ± 2.22 0.31 ± 0.06 43.36 ± 1.75
I 30.71 ± 0.52 6.04 ± 0.12 84.28 ± 1.87 122.08 ± 2.22 0.43 ± 0.11 47.84 ± 1.01
Se + I 28.87 ± 0.54 5.70 ± 0.08 82.00 ± 1.66 118.01 ± 2.56 0.44 ± 0.03 48.96 ± 1.80

1.0 mg I dm−3 H2 O – control 33.99 ± 1.54 5.86 ± 0.20 96.32 ± 7.42 114.55 ± 3.46 0.52 ± 0.03 48.56 ± 0.18
Se 30.05 ± 0.13 6.93 ± 0.87 80.63 ± 0.78 111.89 ± 2.48 0.47 ± 0.04 53.90 ± 1.17
I 31.94 ± 0.59 6.76 ± 0.48 86.69 ± 1.14 121.88 ± 4.21 0.38 ± 0.03 50.28 ± 1.35
Se + I 30.14 ± 1.09 6.78 ± 0.97 86.92 ± 3.83 106.97 ± 0.98 0.55 ± 0.02 45.38 ± 1.24

0.5 mg Se + 1.0 mg I dm−3 H2 O – control 31.90 ± 0.28 5.95 ± 0.24 78.11 ± 4.05 110.00 ± 3.35 0.59 ± 0.04 49.07 ± 2.04
Se 30.11 ± 0.35 6.21 ± 0.41 78.84 ± 1.16 106.39 ± 2.33 0.52 ± 0.05 51.46 ± 3.49
I 32.54 ± 0.11 6.68 ± 0.43 79.93 ± 0.52 116.41 ± 1.12 0.48 ± 0.06 49.91 ± 1.00
Se + I 30.71 ± 0.70 5.90 ± 0.14 85.31 ± 4.73 108.46 ± 3.31 0.61 ± 0.03 49.25 ± 2.21

1.5 mg Se + 1.0 mg I dm−3 H2 O – control 31.39 ± 0.20 5.62 ± 0.07 92.17 ± 2.61 109.71 ± 1.02 0.58 ± 0.01 48.05 ± 1.64
Se 30.05 ± 1.12 6.23 ± 0.35 80.55 ± 2.30 105.22 ± 2.11 0.43 ± 0.02 51.27 ± 2.16
I 30.72 ± 0.61 6.14 ± 0.16 83.95 ± 2.95 108.31 ± 2.89 0.52 ± 0.05 49.18 ± 1.53
Se + I 34.97 ± 4.51 6.90 ± 1.06 93.80 ± 0.42 125.01 ± 11.93 0.51 ± 0.03 55.89 ± 4.93

Test F for nutrient medium × foliar application n.s. n.s. n.s. n.s. n.s. n.s.

SE, standard error; n.s., differences are not significant at P < 0.05.
 S. Smoleń et al. / Scientia Horticulturae 166 (2014) 9–16 13

Table 4
The content of N, P, K, Ca, Mg and S in lettuce roots depending on the interaction between foliar application and introduction of I and Se into the nutrient medium.

Nutrient medium Foliar application % d.w. (n = 6, ±SE)

N P K Ca Mg S

Control H2 O – control 4.34 ± 0.16 1.08 ± 0.06 5.93 ± 0.28 1.34 ± 0.03abc 0.50 ± 0.01abc 0.67 ± 0.03
Se 4.53 ± 0.09 1.15 ± 0.03 6.54 ± 0.13 1.49 ± 0.27abc 0.50 ± 0.07abc 0.69 ± 0.01
I 4.50 ± 0.16 1.30 ± 0.06 6.43 ± 0.17 1.74 ± 0.16bc 0.55 ± 0.04bc 0.71 ± 0.02
Se + I 4.43 ± 0.09 1.25 ± 0.04 6.15 ± 0.30 1.83 ± 0.13c 0.60 ± 0.02c 0.66 ± 0.02

0.5 mg Se dm−3 H2 O – control 4.45 ± 0.06 1.09 ± 0.02 5.97 ± 0.13 1.49 ± 0.05abc 0.52 ± 0.02bc 0.70 ± 0.02
Se 4.46 ± 0.06 1.11 ± 0.06 6.15 ± 0.24 1.27 ± 0.11abc 0.42 ± 0.02ab 0.71 ± 0.02
I 4.43 ± 0.07 1.18 ± 0.04 6.11 ± 0.12 1.51 ± 0.09abc 0.48 ± 0.03abc 0.71 ± 0.02
Se + I 4.43 ± 0.02 1.12 ± 0.02 5.90 ± 0.18 1.56 ± 0.09abc 0.54 ± 0.02bc 0.68 ± 0.01

1.0 mg I dm−3 H2 O – control 4.35 ± 0.09 1.00 ± 0.02 5.75 ± 0.26 1.32 ± 0.08abc 0.44 ± 0.02ab 0.66 ± 0.04
Se 4.45 ± 0.05 1.07 ± 0.03 6.21 ± 0.06 1.42 ± 0.08abc 0.44 ± 0.02ab 0.71 ± 0.01
I 3.81 ± 0.80 1.08 ± 0.03 5.85 ± 0.18 1.10 ± 0.08a 0.35 ± 0.03a 0.67 ± 0.02
Se + I 4.44 ± 0.13 1.05 ± 0.01 5.74 ± 0.03 1.39 ± 0.12ab 0.44 ± 0.03ab 0.65 ± 0.01

0.5 mg Se + 1.0 mg I dm−3 H2 O – control 4.42 ± 0.13 0.88 ± 0.02 5.87 ± 0.09 1.26 ± 0.05abc 0.43 ± 0.01ab 0.66 ± 0.01
Se 4.35 ± 0.06 0.90 ± 0.02 5.80 ± 0.11 1.21 ± 0.06ab 0.41 ± 0.01ab 0.64 ± 0.01
I 4.29 ± 0.04 0.95 ± 0.03 5.93 ± 0.14 1.02 ± 0.03a 0.35 ± 0.01a 0.64 ± 0.03
Se + I 4.53 ± 0.04 0.95 ± 0.02 5.93 ± 0.08 1.10 ± 0.04a 0.39 ± 0.01ab 0.66 ± 0.02

1.5 mg Se + 1.0 mg I dm−3 H2 O – control 4.29 ± 0.03 0.92 ± 0.03 5.71 ± 0.15 1.54 ± 0.08abc 0.51 ± 0.01abc 0.69 ± 0.01
Se 4.19 ± 0.08 0.98 ± 0.05 5.77 ± 0.21 1.47 ± 0.12abc 0.46 ± 0.03abc 0.70 ± 0.02
I 4.46 ± 0.06 1.03 ± 0.02 5.97 ± 0.10 1.29 ± 0.15ab 0.42 ± 0.05ab 0.70 ± 0.01
Se + I 4.44 ± 0.03 0.97 ± 0.02 5.94 ± 0.02 1.18 ± 0.08ab 0.40 ± 0.03ab 0.69 ± 0.01

Test F for nutrient medium × foliar application n.s. n.s. n.s. * *

n.s.

SE, standard error; n.s., differences are not significant at P < 0.05. Means followed by the same letters are not significantly different at P < 0.05.
*
Means are significantly different.

lettuce leaves and roots when compared to plants from the sub-
block with the nutrient medium supplemented only with iodine
(Fig. 3A and B). In plants cultivated in the nutrient medium with
1.5 mg Se + 1.0 mg I dm−3 and sprayed with distilled water a ten-
dency of decreasing iodine content in roots, and to a lesser extent
in leaves, was observed when compared to respective objects from
the sub-blocks with the nutrient medium enriched with I alone or
0.5 mg Se + 1.0 mg I dm−3 .
3.2.2. Foliar treatment and iodine/selenium interaction
Interesting were the results regarding iodine and selenium con-
tent in lettuce leaves after foliar application of Na2 SeO4 or KIO3
in sub-blocks with the nutrient media lacking iodine or selenium,
respectively (Figs. 2 and 3A and B). During lettuce cultivation in the
control medium or with the presence of 1 mg I dm−3 a significantly
higher Se content in leaves was noted after simultaneous spraying
with I and Se when compared to the application of Na2 SeO4 only

Table 5
The content of B, Cu, Fe, Mn, Mo and Zn in lettuce roots depending on the interaction between foliar application and introduction of I and Se into the nutrient medium.

Nutrient medium Foliar application mg kg−1 d.w. (n = 6, ±SE)

B Cu Fe Mn Mo Zn

Control H2 O – control 20.07 ± 0.34 31.27 ± 4.75 3959.2 ± 130.0abc 584.89 ± 10.96 9.59 ± 0.25 109.33 ± 5.83
Se 21.21 ± 0.54 29.61 ± 1.34 4802.3 ± 734.7abc 631.43 ± 29.09 8.66 ± 0.25 113.22 ± 3.59
I 22.48 ± 0.99 31.86 ± 0.87 5552.7 ± 427.2b 684.56 ± 25.51 8.60 ± 0.36 127.22 ± 2.08
Se + I 21.78 ± 0.65 30.70 ± 0.72 5638.8 ± 226.6c 650.73 ± 5.52 9.19 ± 0.38 116.93 ± 0.62

0.5 mg Se dm−3 H2 O – control 21.90 ± 0.47 30.18 ± 1.05 4296.0 ± 195.1abc 793.49 ± 25.51 9.66 ± 0.36 130.88 ± 6.02
Se 22.93 ± 0.40 28.70 ± 2.18 3707.5 ± 614.8abc 692.38 ± 21.89 9.18 ± 0.14 136.45 ± 7.97
I 22.14 ± 0.52 30.96 ± 1.06 4607.2 ± 228.3abc 737.65 ± 31.55 9.53 ± 0.35 143.43 ± 5.17
Se + I 22.63 ± 0.21 29.67 ± 0.21 4430.9 ± 302.1abc 753.91 ± 32.54 9.76 ± 0.08 135.22 ± 6.71

1.0 mg I dm−3 H2 O – control 21.50 ± 0.16 33.02 ± 0.68 4045.3 ± 317.1ab 525.85 ± 25.59 8.93 ± 0.21 131.09 ± 4.39
Se 23.69 ± 0.94 36.04 ± 1.59 4594.4 ± 255.1abc 545.72 ± 17.87 8.80 ± 0.21 128.59 ± 4.61
I 22.03 ± 0.44 31.60 ± 0.80 3162.7 ± 321.2a 458.73 ± 18.26 8.33 ± 0.15 122.99 ± 5.17
Se + I 22.04 ± 0.21 33.77 ± 1.56 3912.8 ± 316.3ab 542.60 ± 12.36 8.90 ± 0.24 131.15 ± 2.08

0.5 mg Se + 1.0 mg I dm−3 H2 O – control 21.01 ± 0.32 30.36 ± 0.56 3758.0 ± 263.0abc 480.23 ± 22.87 12.38 ± 0.34 99.79 ± 5.31
Se 21.66 ± 0.61 29.89 ± 0.85 3724.7 ± 369.4abc 427.18 ± 29.07 10.91 ± 0.53 94.82 ± 3.98
I 21.04 ± 0.38 29.49 ± 1.38 3270.8 ± 342.2a 419.58 ± 16.23 11.40 ± 0.43 91.99 ± 3.25
Se + I 21.81 ± 0.60 35.73 ± 4.01 3349.4 ± 266.3a 445.79 ± 25.20 12.72 ± 0.24 122.39 ± 13.12

1.5 mg Se + 1.0 mg I dm−3 H2 O – control 21.65 ± 0.19 31.75 ± 1.18 4812.5 ± 386.1abc 701.58 ± 23.70 12.80 ± 0.36 123.60 ± 5.37
Se 22.34 ± 0.49 34.44 ± 1.78 5067.7 ± 590.2abc 707.10 ± 24.26 13.21 ± 0.53 133.88 ± 7.07
I 22.44 ± 0.81 31.45 ± 1.42 4071.9 ± 497.0ab 644.45 ± 31.81 12.05 ± 0.44 133.26 ± 7.40
Se + I 21.82 ± 0.23 32.65 ± 1.72 3476.0 ± 171.8ab 648.28 ± 29.05 12.14 ± 0.44 134.54 ± 5.66

Test F for nutrient medium × foliar application n.s. n.s. *

n.s. n.s. n.s.

SE, standard error; n.s., differences are not significant at P < 0.05. Means followed by the same letters are not significantly different at P < 0.05.
*
Means are significantly different.
14 S. Smoleń et al. / Scientia Horticulturae 166 (2014) 9–16
Fig. 3. Iodine content in leaves (A) and roots (B) of lettuce depending on the inter-
action between foliar application and introduction to the nutrient medium of iodine
and selenium (n = 6). Means followed by the same letters are not significantly dif-
ferent at P < 0.05; bars indicate standard error.
(Fig. 2A). Such relations were not found with respect to selenium
content in roots (Fig. 2B). Diverse observations were noted for Se
and I interaction (when applied foliarly) on iodine content in lettuce
plants (Fig. 3A and B). Application of KIO3 alone or together with
Na2 SeO4 did not significantly change iodine content in leaves of
lettuce plants cultivated in the control medium as well as with the
addition of 0.5 mg Se, 1.0 mg I or 0.5 mg Se + 1.0 mg I dm−3 into the
nutrient medium (Fig. 3A). Only in the sub-block with the highest
selenium content in the medium (1.5 mg Se + 1.0 mg I dm−3 ) simul-
taneous application of Na2 SeO4 + KIO3 caused a little decrease of
I level in lettuce leaves when compared to KIO3 treatment. Basi-
cally, iodine content in roots of plants treated foliarly with KIO3
or Na2 SeO4 + KIO3 was at a similar level in both combinations
(Fig. 3B).
4. Discussion
4.1. Biomass and mineral nutrition
Zhu et al. (2004) noted a significant decrease of shoot and root
biomass of spinach grown in hydroponic cultivation due to simul-
taneous application of SeO4 2− (in the concentration of 0, 10 and
20 ␮M Se) and IO3 − (in a dose of 0.10 and 50 ␮M I). Lack of signif-
icant differences in terms of biomass values that was noted in our
studies suggests that double biofortification of lettuce with iodine
and selenium did not negatively affect lettuce growth. However,
it is worth to mention that few days before the last foliar spray-
ing with Na2 SeO4 (alone or simultaneously with KIO3 ) chlorotic
spots turning into necrosis occurred on the oldest leaves with no
such damages noted in the younger ones. Most likely, it may have
been caused by the retention of the droplets of the solution due to
its flowing down on the surface of the lowest (oldest) leaves. As an
effect the longer contact of Na2 SeO4 solution with the surface of the
oldest leaves (when compared to younger ones) could have directly
contributed to the occurrence of the damages that was limited to
these leaves. Only in this aspect our studies confirm previous infor-
mation on a relatively high SeO4 2− toxicity on plants (Kopsell and
Kopsell, 2007).
With a comparable level of lettuce head biomass in all com-
binations, understandable is the lack of major differences in the
content of macro- and micronutrients in leaves and roots. Only a
decrease in the level of Ca, Mg and Fe (in roots of plants grown
in the nutrient medium containing 0 mg I dm−3 , 0.5 mg Se + 1.0 mg
I dm−3 or 1.5 mg Se + 1.0 mg I dm−3 and foliarly treated with KIO3
or Na2 SeO4 + KIO3 ) indirectly indicates that simultaneous applica-
tion of selenium and iodine may impair the functioning of mineral
nutrition in plants. It seems that obtained differences in the con-
tent of these three elements in roots may have resulted from foliar
spraying with iodine and selenium rather than its addition into the
nutrient medium. Reasons for these relations need to be studied in
further experiments.
4.2. Interaction between iodine and selenium
In the above-mentioned studies conducted by Zhu et al. (2004)
no significant influence of IO3 − treatment on selenium content
as well as the application of SeO4 2− on iodine accumulation in
spinach plants was noted. In our experiment, lettuce plants foliarly
treated with distilled water (in the sub-blocks with Se, I and Se + I
content in the nutrient medium) were characterized by a higher
selenium content in roots than in leaves. In the case of iodine,
accumulation of this element in roots and leaves remained at a com-
parable level in these combinations (Figs. 2 and 3A and B). Obtained
results indirectly suggest that iodine and selenium undergo sepa-
rate mechanisms of uptake and transport from roots to leaves. It
is stated that iodine transport in plants is analogous to chloride
(Cl− ) movement occurring on the basis of: symport (H+ /anion),
antiport (Na:K/Cl), through ion channels conducting Cl− /I− (White
and Broadley, 2001; Roberts, 2006; Colmenero-Flores et al., 2007)
or with the use of carriers – most probably halide-specific (White
and Broadley, 2009). SeO4 2− ions may be transported by the same
carriers as SO4 2− what leads to the antagonism between these
two types of anions (Kopsell and Kopsell, 2007). It is generally
considered that SeO4 2− anions (in contrast to SeO3 2− ) are easily
transported from roots to upper parts of the plant in which they
are accumulated (White and Broadley, 2009). No such relations
were however found in our studies. The obtained differences in
I and Se accumulation in roots and leaves (in plants foliarly treated
with distilled water) may have been caused by various detoxifica-
tion mechanisms for both of these elements in plants. It is known
that SeO4 2− form of selenium is more readily taken up but is also
more toxic for plants than SeO3 2− (Kopsell and Kopsell, 2007).
After the uptake, selenium can be accumulated in roots due to
intensive biosynthesis of phytochelatines – proteins responsible for
detoxification of heavy metals and metalloids, including selenium.
Those proteins contain significant amounts of sulphur amino acids:
cysteine, homocysteine and methionine (Spain and Rabenstein,
2004). It has been noted that after the application of selenium
compounds accumulation of non-protein SH groups (present in
cysteine) in spinach and tomato plants was observed (Hawrylak
and Szymańska, 2004). Simojoki et al. (2003) revealed that after the
treatment with low SeO4 2− doses (0 and 1 ␮g Se·3.5 dm−3 pot−1 ) Se
accumulation was observed mainly in roots, while with increasing
Se dose (10, 100, 500 and 1,000 ␮g Se·3.5 dm−3 pot−1 ) the content
of this element in lettuce roots and leaves tended to be balanced.
It is widely considered that one of the most efficient mechanisms
 S. Smoleń et al. / Scientia Horticulturae 166 (2014) 9–16
of iodine detoxification in plants is based on its transportation to
leaves where iodine methylation takes place. As an effect iodine
is volatilized into the atmosphere in the form of CH3 I (Muramatsu
and Yoshida, 1995; Sain et al., 1995; Landini et al., 2012).
Results obtained in our studies indicate that foliar application
of KIO3 resulted in a synergistic effect on SeO4 2− absorption in
leaves. Foliar spraying with Na2 SeO4 2 basically did not influence
leaf uptake of IO3 − . It is generally known that, when applied foliarly,
apolar substances (such as urea) are the most quickly absorbed
molecules by leaves, followed by cations and anions (Franke, 1986;
Michałojć and Szewczuk, 2003). Through the cuticular layer that
covers leaf surface, water-soluble polar compounds (cations and
anions) are transported by the “hydrophilic path” (Franke, 1967).
For both cations and anions, its penetration from the outer to the
inner part of the leaf prevails over the transport in the oppo-
site direction – efflux (Smoleń, 2012). It seems that the causes
for increased SeO4 2− absorption by leaves observed with simul-
taneous foliar application of IO3 − can be ascribed to: a specific
influence of iodine on the cuticle structure (Shaw et al., 2007) or
synergistic effect of IO3 − towards SeO4 2− uptake by leaf meso-
phyll cells. Studies conducted by Shaw et al. (2007) revealed dozens
of times faster sorption of iodine than sulphate SO4 2− by adaxial
leaf surface of broad bean – it was also found that the absorp-
tion of Cs2+ is five times lower than iodine. Observed results of
significant level of iodine absorption by leaves were explained
by: (a) organophilic nature of iodine – and its ability to interact
with cuticular waxes or (b) a possible oxidation of I− to I2 on
leaf surface significantly facilitating iodine penetration through the
cuticle.
Another interesting issue directly related to our studies is iodine
and selenium movement in plants. These two elements are effi-
ciently transported through xylem (White and Broadley, 2009).
Additionally, selenium in the form of SeO4 2− and/or organosele-
nium compounds can be redistributed within the plant through
phloem (White et al., 2007). Studies conducted by numerous
authors indicate that the process of iodine redistribution to fruits
and seed through phloem is limited (Muramatsu et al., 1993, 1995;
Salau et al., 2008; Shinonaga et al., 2000). However, results obtained
by Landini et al. (2011) provided evidence for relatively high level
of iodine redistribution from tomato leaves to fruits after foliar
application of iodides (I− ). In our experiment indirect evidence
for an efficient transport of SeO4 2− and IO3 − from leaves to roots
by phloem was obtained in the sub-block with the control nutri-
ent medium. It is based on the observation of increased content
of iodine and selenium in roots of plants treated foliarly with:
Na2 SeO4 and Na2 SeO4 + KIO3 (for selenium content in roots) or KIO3
and Na2 SeO4 + KIO3 (for iodine content in roots) in relation to other
combinations with foliar application in the control sub-block. The
causes as well as mechanisms underlying this effect are still unclear
and require further studies.
5. Conclusion
An observed lack of the influence of tested combinations with
iodine and selenium application on the biomass yield and min-
eral composition of plants (its nutritional status with respect to
macro- and microelements) suggests the possibility of conducting
safe double biofortification of lettuce with iodine and selenium in
hydroponic cultivation.
Introduction of iodine or selenium alone in the form of IO3 − or
SeO4 2− into the nutrient medium (in a dose of 0.5 mg Se and 1.0 mg
I dm−3 ) did not affect root uptake of SeO4 2− and IO3 − , respectively
as well as its further transport into upper parts of the plant. How-
ever, a significant tendency of a 3-fold increase of selenium dose
lowering iodine uptake from the nutrient medium by lettuce plants
was noted.
15
Conducting biofortification with iodine and selenium through
foliar spraying did not affect root uptake of selenium and iodine
from the nutrient medium, respectively. A synergistic interaction
of IO3 − on SeO4 2− absorption by leaves was noted with foliar appli-
cation of these two compounds. Probably both tested ions were
transported from leaves to roots by phloem tissue.
Acknowledgement
This work was financed by research grants from the Ministry of
Science and Higher Education Republic of Poland.
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