Journal of Applied Phycology (2019) 31:465–470

https://doi.org/10.1007/s10811-018-1518-y

Novel bioprocess for the cultivation of microalgae in hydroponic

growing system of tomato plants
Valeria Barone 1 & Ivana Puglisi 1 & Ferdinando Fragalà 1 & Angela Roberta Lo Piero 1 & Francesco Giuffrida 1 &
Andrea Baglieri 1

Received: 19 April 2018 / Revised and accepted: 15 May 2018 / Published online: 26 May 2018
# Springer Science+Business Media B.V., part of Springer Nature 2018

Abstract
Hydroponic growing systems and the application of natural biostimulant substances are becoming very attractive option for crop
cultivation due to their economic relevance since they allow reduction in the use of fertilizer and increase the yield. In order to
perform a hydroponic co-cultivation system of microalgae (Chlorella vulgaris or Scenedesmus quadricauda) and tomato plants,
grown in Hoagland nutrient solution, their mutual effect and the influence of a natural biostimulant obtained by alkaline
extraction from a digestate of agro-livestock residues (DHL) were evaluated. The results showed that the co-cultivation system
positively affected the growth of both tomato plants and microalgae. The best option, aimed to a mutual benefit for both cultures,
resulted to be the co-cultivation system of tomato plants and S. quadricauda in the presence of DHL, positively affecting the
growth of tomato plants along with a great increase in microalgal biomass.

Keywords Alkaline extract from digestate . Biostimulant . Chlorella vulgaris . Hoagland solution . Scenedesmus quadricauda .
Solanum lycopersicum

Introduction about possible hydroponic systems for cultivating tomato va-

Microalgae are photosynthetic organisms which can be auto-
trophic, heterotrophic, or mixotrophic. The production of
microalgae and their cultivation systems have attracted the
attention of researchers for their great economic and commer-
cial relevance (Mata et al. 2010; Hultberg et al. 2013). In fact,
their biomass can potentially be used to produce biofuels and
biomaterials such as oligopeptides, proteins, oligosaccharides,
and carbohydrates for the food and feed sectors (Maurya et al.
2016). Given the versatility of employing microalgae, several
studies have focused on their production and economic yield.
A successful strategy for increasing the cost-effectiveness of a
microalgal culture system requires the optimal utilization of
substrates, the application of low-cost resources as well as the
use of cheap fertilizers (Lee 2001; Mata et al. 2010; Gong and
Jiang 2011). Recently, Zhang et al. (2017) carried out a study
* Ivana Puglisi
ipuglisi@unict.it
1
Dipartimento di Agricoltura, Alimentazione e Ambiente (Di3A),
Università di Catania, Via S. Sofia 98, 95123 Catania, Italy
rieties in association with the microalga Chlorella infusionum.
They showed that algal photosynthesis enhanced the crop bio-
mass, while crop root respiration and exudation which are
sources of carbon, increased microalgal biomass.
Hydroponic cultures for crop production have significantly
increased worldwide in the last few years (Hickman 2011;
Kumar and Cho 2014). In these soilless systems, water manage-
ment is cost effective and efficient so that it is possible to reduce
water use in crop cultivation. Therefore, numerous studies have
been performed to improve production both in quantitative and
qualitative terms. The development of a sustainable and environ-
mental friendly agricultural systems can include the use of plant
biostimulants. These compounds are a wide range of molecules
that promote plant growth when applied in small quantities in the
soil or directly onto the leaf surface. The application of
biostimulants in hydroponic cultures has been proven to enhance
crop performance and, when applied in these cultivation systems,
can also reduce the use of fertilizer and the nutrient solution
concentration (Vernieri et al. 2006; Povero et al. 2016).
Biostimulants have shown a beneficial effect on specific aspects
of plant growth such as increased root and shoot growth, increas-
ing both the tolerance to abiotic stress and the water uptake
(Parrado et al. 2008; Alam et al. 2014; Bulgari et al. 2015).
466
Ertani et al. (2013) demonstrated that humic-like substances
(HLs) extracted from agro-industrial residues coming from
Brassica napus, Ricinus communis, Linum usitatissimum, and
from a digestate increased corn seedling protein content and the
activity of those enzymes connected to the assimilation of ni-
trogen and photosynthesis. Similarly, it has been shown that
extracts obtained by alkaline hydrolysis from tomato plant res-
idues produce a biostimulating effect on bean plants leading
mainly to an increase in the quantity of assimilated nitrogen
(Baglieri et al. 2014).
Interestingly, it was also observed that HLs also exert a
biostimulant effect on maize and bean plants as well as on
the microalgae Chlorella vulgaris and Scenedesmus
quadricauda (Ertani et al. 2013; Baglieri et al. 2014; Puglisi
et al. 2018). In particular, among the extracts applied to the
growth medium, HLs extracted by alkaline hydrolysis from a
digestate of agro-livestock residues (D-HL) seems to have a
great effect on microalgal biomass and their lipid production
(Puglisi et al. 2018). Furthermore, Barone et al. (2018) found
that extracts from C. vulgaris and S. quadricauda are promis-
ing biostimulants in the early stages of plant growth in sugar
beet. The putative biostimulation mechanism induced by
microalgae is a very complex mechanism and not completely
understood. Most likely, it could be due to the interaction of
several molecules and not necessarily by only one of them. It
is well known that microalgae secrete a large number of bio-
logically active extracellular molecules. These active mole-
cules are generally secondary metabolites and are known as
allelochemicals (Bacellar Mendes and Vermelho 2013).
The aim of this work was to carry out a hydroponic co-
cultivation of microalgae and plants to evaluate a possible
positive effect on their growth. Tomato seedlings were culti-
vated hydroponically in association with living microalgal
species (C. vulgaris or S. quadricauda) in order to estimate
simultaneously microalgal biomass growth in the presence of
plant roots in the optimal medium for crops as well as the
effects of microalgae on tomato plants. At the same time, the
alkaline extract from a digestate of agro-livestock residues (D-
HL), showing activity as a biostimulant of plants as well as
microalgae, was added to the nutrient solution to evaluate its
effect in the co-cultivation system.
Materials and methods
Humic-like substances
The humic-like substances (HLs) were extracted from a
digestate from wastes of an agro-livestock farm (DHL).
Alkaline-hydrolyzed extracts were obtained as described in
Ertani et al. (2013). Briefly, 200 g of finely ground waste was
treated with aqueous 0.1 mol L−1 KOH at a 1:5 w/v ratio (waste/
solution). The suspension was shaken under N2 for 20 h,
J Appl Phycol (2019) 31:465–470
allowed to settle overnight, and centrifuged at 3000 rpm for
20 min. In order to solubilize and totally remove the HLs, three
treatments with 0.1 mol L−1 KOH were carried out on solid
residue. The supernatant collected was dialyzed and freeze
dried for further analysis. This fraction has been previously
characterized (Ertani et al. 2013; Baglieri et al. 2014).
Microalgae culture
Microalgae employed in this study were Chlorella vulgaris
(CCAP 211/11C) and Scenedesmus quadricauda (isolated
from algal company raceway pond, placed in Borculo,
Gelderland, Netherland, in 2011) obtained and maintained in
algal collection of Swansea University (Wales, UK).
Microalgae were cultivated in a growth chamber using stan-
dard BG11 algae culture medium (Stanier et al. 1971), bub-
bled with air and an average photon flux (PPF) of
100 μmol photons m−2 s−1 (Philips SON-T AGRO 400) with
a 12-h photoperiod (Baglieri et al. 2016).
Experimental conditions
The experimental apparatus included a transparent container
(40 × 20 × 10 cm), covered with wire net (mesh size 0.5 cm),
used for crop fixation with an average temperature of around
25 °C. Co-cultivation was conducted in 4 L of modified
Hoagland solution: Ca(NO3)2·4H20, 1250 mg L−1; KNO3,
410 mg L −1 ; NH 4 H 2 PO 4 , 280 mg L −1 ; MgCl 2· 6H 2 0,
624 mg L −1 ; FeSO 4· 7H 2 O, 60 mg L −1 ; EDTA-Na 2 ,
80 mg L−1; H3BO3, 6 mg L−1; MnCl2·4H2O, 4 mg L−1;
ZnSO4·7H2O, 0.04 mg L−1; and CuSO4·5H2O, 0.04 mg L−1
(Zhang et al. 2017). Nutrient solution was aerated for 12 h per
day by bubbling with air through an aeration pump. Exactly
50 mL of microalgae suspension was harvested at OD680 = 1.0
and centrifuged at 2500 rpm for 10 min; the pellet was recov-
ered and washed several times until reached an electric con-
ductivity lower than 200 μS cm−1 electric conductivity, and
used to inoculate Hoagland solution. Moreover, the experi-
mental scheme provided two additional treatments of a
50 mL aliquot of each microalgae suspension and DHL
(100 mg L −1 ). Corresponding controls received only
50 mL of distilled water. All cultures were characterized
by constant electric conductivity, measured weekly by a
conductometer.
Treatments are hereafter referred to as (1) untreated = con-
trol plants; (2) Cv = containing C. vulgaris suspension; (3) Sq
= containing S. quadricauda suspension; (4) DHL+ Cv =
containing C. vulgaris suspension and DHL; (5) DHL+ Sq =
containing S. quadricauda suspension and DHL; (6) DHL =
containing only DHL.
Tomato seedlings (Solanum lycopersicum cultivar
Missouri) were provided by Ecofaber (Noto, Siracusa, Italy).
After germination in a vermiculite-peat substrate, five
J Appl Phycol (2019) 31:465–470
seedlings of tomato per container were transplanted at the
appearance of the first true leaf. A completely random design
with five replications for each treatment was used.
The experiment was conducted for 46 days in a growth
chamber at 25 ± 2 °C, and an average photon flux (PPF) of
100 μmol photons m−2 s−1 (PHILIPS SON-T AGRO 400)
with a 12-h photoperiod.
Microalgae recovery
Microalgae were harvested from the experimental plots
46 days after transplant. Microalgal suspensions were centri-
fuged at 1500×g for 10 min and the pellets were washed three
times with distilled water and lyophilized, then biomass was
weighed.
Physiological parameters in tomato plants
Tomato plants were harvested at 46 days of cultivation in
Hoagland solution. For dry weight measurement, the samples
were placed in a drying oven until constant weight at 105 °C
was reached, allowed to cool for 2 h inside a closed bell jar,
then leaves and roots of each plot were weighted.
Root length was measured with a flexible ruler to the
nearest 0.5 mm.
The measurement of the SPAD index was performed using
the SPAD-502 Leaf Chlorophyll Meter (Minolta Camera Co.,
Ltd., Japan) on three points of the last fully expanded leaf of
tomato plants.
Statistical analysis
Data were analyzed by one-way ANOVA (P < 0.05) followed
by Tukey’s test for multiple comparison procedures.
Results and discussion
Table 1 shows the dry biomass of microalgae grown in the co-
cultivation system with tomato plants, harvested after 46 days.
The co-cultivation of C. vulgaris with tomato plants yielded a
biomass of 0.77 g L−1 corresponding to a volumetric produc-
tivity of biomass of 0.019 g L−1 day−1. The presence of DHL
in the hydroponic culture seems to have a positive effect on
final C. vulgaris biomass, yielding an increase of 32% com-
pared to the biomass obtained without DHL and a volumetric
productivity of 0.022 g L−1 day−1. These values were compa-
rable to those reported by Mata et al. (2010) for C. vulgaris
grown in different optimal substrate and conditions, ranging
from 0.02 to 0.20 g L−1 day−1. Similarly, Zhang et al. (2017)
found, after 76 days, for C. infusionum, a microalgal biomass
productivity of 0.032 g L−1 day−1 in a simple eco-hydroponic
system with tomato.
467
Table 1 Dry biomass of microalgae and productivity of biomass grown
in co-cultivation with tomato plants after 46 days (Cv, Chlorella vulgaris;
Sq, Scenedesmus quadricauda; DHL, digestate humic-like substance).
Data are means ± SD. The values are means of data from five
replications. Values of the same microalgae species followed by
different letters are significantly different (P < 0.05)
Treatment Biomass (g −1) Productivity of biomass (g L−1 day−1)
Cv 0.77 ± 0.07 b 0.019 ± 0.004 a
DHL + Cv 1.02 ± 0.09 a 0.022 ± 0.003 a
Sq 1.02 ± 0.06 b 0.022 ± 0.002 b
DHL + Sq 1.33 ± 0.1 a 0.030 ± 0.004 a
Similar results were obtained for S. quadricauda co-
cultivated with tomato. In this case, the volumetric productiv-
ity of biomass was of 0.022 g L−1 day−1, a value slightly lower
than that reported by Mata et al. (2010) for Scenedesmus sp. of
about 0.03–0.26 g L−1 day−1. The biomass increment caused
by DHL in the hydroponic culture for S. quadricauda was
30% higher than the biomass obtained without DHL. In this
latter case, the volumetric productivity of biomass increased to
0.030 g L −1 day −1 , similar to the values reported for
Scenedesmus sp. cultivated in an optimal substrate by Mata
et al. (2010).
In a successful co-cultivation system, both cultures need to
have benefit; therefore, tomato plant growth during the exper-
imental period was also estimated.
All the treatments positively affected fresh whole plant
weight as well as dry weight, except DHL + Cv which showed
fresh and dry weight values comparable to the untreated plants
(Fig. 1). In particular, the fresh and dry weight of whole plants
grown with C. vulgaris (11.95 and 0.90 g, respectively), in-
creased about 2.5-fold compared with the control (4.54 and
0.40 g, respectively) (Fig. 1).
The highest values of shoot fresh and dry weight were
observed in plants grown in co-cultivation with C. vulgaris
(9.88 and 0.94 g, respectively), recording an increase (3.70
and 0.31 g, respectively) of 167 and 139% compared to the
control, respectively (Fig. 1). Similarly to whole plants, shoot
fresh and dry weight increased in all treatments, except that
for DHL + Cv, which was comparable to that of the control
(Fig. 1). Nevertheless, as shown in Table 1, the percentage
of dry matter was higher in DHL + Cv, both in the whole
plants and in the shoots (9.8 and 9.1%, respectively), with
respect to the percentage calculated in samples grown only
with C. vulgaris (7.6% in whole plants and 7.4% in shoots)
as well as in the control (8.8% in whole plants and 8.3% in
shoots). It is noteworthy that the treatments with S.
quadricauda with and without DHL (0.55 and 0.53 g, respec-
tively) increased similarly shoot dry weight with respect to the
control (0.31 g), as well as shoot fresh weight (7.31 g for DHL
+ Sq, 5.15 g for Sq, and 3.70 for control) recording a signifi-
cant increase with respect to the control. In plants grown with
468 J Appl Phycol (2019) 31:465–470

14
Plant a A fresh weight and 0.17 g for dry weight) when seedlings were
12 Shoot co-cultivated with S. quadricauda (107% for fresh weight and
Root a 88% for dry weight). Root fresh weight obtained for other
10 b
treatments showed values similar to the control (P > 0.05)
b
(Fig. 1A). The treatments DHL + Sq (0.11 g) and DHL
Fresh weight (g)

8 b b (0.11 g) had similar increments (22%) in dry weight of roots

b with respect to the control (0.09 g) (Fig. 1B). Conversely,
b Ertani et al. (2013) observed that, after DHL treatment of
6
c
c maize plants, root dry weight decreased (− 18%) with respect
c c
4 to the control. Probably, at this concentration, the DHL stim-
ulating effect on roots was higher in tomato than in maize
a a
2 b plants. The different behavior could also be attributed to a
b b b
different duration of the DHL treatment. However, the dry
0 root/shoot ratio (0.25) confirms that DHL positively affected
1.2
the plant weights mostly at shoot level (Table 2), as observed
Plant B
by Ertani et al. (2013). As well as for shoot dry weight, in
Shoot a
1 the co-cultivation system C. vulgaris and tomato seedlings,
Root
the addition of DHL to the nutrient solution did not affect
a
0.8 b root dry weight, showing similar value to the control plants
b
(Fig. 1B). Noteworthy, dry matter (%) was significantly
Dry weight (g)

b b b decreased in plants co-cultivated with C. vulgaris (8.15)

0.6
c c
0.4 c c
a a
0.2
c b
c greater amount of water inside the vacuoles (Taiz et al.
0 2015). Whereas, the treatment with DHL + Cv showed a
Untreated Cv Sq DHL+Cv
Fig. 1 Fresh (A) and dry (B) weight (g) of tomato seedlings (whole plant,
shoot, and root) grown in co-cultivation with microalgae after 46 days
(Cv, Chlorella vulgaris; Sq, Scenedesmus quadricauda; DHL, digestate
humic-like substance). Error bars indicate standard deviation. The values
are means of data from five replicates. Values for root, shoot, and plant
followed by different letters are significantly different (P < 0.05)
S. quadricauda, a greater percentage of dry matter (10% for
whole plant and for shoot) was observed, whereas the presence
of DHL keeps dry matter at values similar to that of the control
both in the whole plants and shoots (Table 2). Finally, a sig-
nificant increase in fresh and dry weights (58 and 33%, respec-
tively) with respect to the control (3.70 and 0.31 g, respective-
ly) was achieved in shoots of plants treated with DHL (5.86
and 0.45 g, respectively). Similarly, it was found that 14-day-
old maize plants treated with the same concentration of DHL
(100 mg L−1) augmented shoot dry weight by 14% with re-
spect to the control (Ertani et al. 2013). These differences
might be attributed to either the different crop or to the longer
DHL treatment period.
As shown in Fig. 1, the highest value of root fresh and dry
weight was obtained in co-cultivation with C. vulgaris (2.07
and 0.17 g, respectively), resulting the best treatment in order
to increase (144 and 88%, respectively) total plant biomass.
Moreover, root weights were positively influenced (1.76 g for
b when compared with the control (10.74) (Table 2). A ra-
tional explanation for the higher water content in roots of
plants treated with both microalgae may be due to the sharp
increase of root length (Fig. 2), being closely related to a
b
distension of root cells achieved by the accumulation of a
DHL+Sq DHL percentage of dry matter (13.3) significantly similar to the
value calculated for control plants (10.7).
Normally, the root system and epigeous part grow at the
same rate; however, the root and shoot ratio (both on the basis
of fresh and dry weight) can be heavily affected by stress
conditions (Erice et al. 2010). The co-cultivation with
microalgae or DHL did not greatly affect the fresh and dry
root/shoot ratio, ranging from 0.13 to 0.34 for fresh weight
and ranging from 0.21 to 0.32 for dry weight, thus confirming
that plants were not affected by stress conditions.
Interestingly, the lowest values for fresh (0.13) and dry
(0.21) shoot/root ratios were obtained in tomato plants co-
cultivated with S. quadricauda + DHL (Table 2).
Considering that both fresh and dry root weights of plants in
DHL + Sq increased compared with the control, the lower
root/shoot ratio may be attributed to a general wellness of
plants enhancing growth of the epigeous part. These results
are in agreement with Ertani et al. (2013), who found that in
14-day-old maize plants, the treatment with DHL induced
decrements in fresh and dry root/shoot ratio.
Figure 2 shows root length of tomato plants at the end of
experimental period. The overall trend is that all the treatments
positively affect the root length, as already observed for the
root dry weight (Fig. 1B). Interestingly, the co-cultivation of
plants with microalgae had an increase in root length by about
J Appl Phycol (2019) 31:465–470 469

Table 2 Morphological traits of tomato plants grown in co-cultivation fresh weight; DW, dry weight. Data are means ± SD. The values are
with microalgae and DHL after 46 days (Cv, Chlorella vulgaris; Sq, means of data from five replications. Values followed by different
Scenedesmus quadricauda; DHL, digestate humic-like substance). FW, letters are significantly different (P < 0.05)

Treatment Root dry matter % Shoot dry matter % Plant dry matter % FW root/shoot DW root/shoot

Control 10.7 ± 0.8 a 8.3 ± 0.3 b 8.8 ± 0.4 b 0.23 ± 0.04 b 0.30 ± 0.02 a
Cv 8.1 ± 0.5 c 7.4 ± 0.6 b 7.6 ± 0.6 b 0.21 ± 0.02 b 0.23 ± 0.02 c
Sq 9.5 ± 0.3 b 10.2 ± 1.1 a 10.0 ± 0.6 a 0.34 ± 0.04 a 0.32 ± 0.04 a
DHL+ Cv 13.3 ± 2.3 a 9.1 ± 0.6 a 9.8 ± 0.5 a 0.18 ± 0.02 b 0.27 ± 0.01 b
DHL+ Sq 12.0 ± 2.1 a 7.5 ± 0.5 b 8.0 ± 0.6 b 0.13 ± 0.02 c 0.21 ± 0.01 c
DHL 9.4 ± 0.3 b 7.6 ± 0.7 b 7.9 ± 0.5 b 0.20 ± 0.03 b 0.25 ± 0.02 b

130% for C. vulgaris (18.5 cm) and S. quadricauda (15.5 cm).
The addition of DHL to the nutrient solution resulted in an
increase of root length (12.26 cm) of 71%, with respect to the
untreated plant. Furthermore, in co-cultivation with S.
quadricauda, the presence of DHL (15.66 cm) produced an
increase by 119% with respect to the control (7.15 cm).
Conversely, in DHL + Cv treatment, root length was similar
to the control (Fig. 2).
Finally, the SPAD index was measured to verify the treatment
effect on chlorophyll content (Fig. 3). The obtained results indi-
cated that chlorophyll content was rather constant in all treatments.
These results suggest that the co-cultivation of tomato
plants with C. vulgaris or S. quadricauda has a clear benefi-
cial effect on crop culture in terms of fresh and dry weight of
plants, similar to the results obtained by Zhang et al. (2017)
using C. infusionum. As suggested by Zhang et al. (2017), the
positive effect on the physiological parameters measured on
tomato plants may be mainly attributed to the continuous algal
photosynthesis constantly supplying oxygen in the
hydroponic solution. Moreover, the enhanced growth of
plants may be also due to the presence of biostimulating sub-
stances or allelochemicals slowly released by microalgae in
the nutrient solution. Formally, allelopathy could be positive
or negative, depending on the beneficial or detrimental effect
on the target organisms, respectively. A deep investigation
convinced Rice (1984) that most organic compounds,
resulting inhibitors at certain concentrations, may act as
stimulant for the same processes at lower concentration.
Such evidences suggest that the strong competition with
the young growing tomato roots could induce microalgae
to produce allelochemical substances in order to inhibit the
growth of competitors. However, according to Rice (1984),
the low concentration of these substances played a stimu-
latory function.
The presence of DHL in the co-cultivation system with C.
vulgaris did not affect growth of tomato plants, whereas in co-
cultivation with S. quadricauda enhanced shoot growth with
respect to the control. These results suggested that the

25.0

30 a
a
a a a
20.0 a
25 a
a a

ab
Root lenght (cm)

15.0 20
SPAD

b 15
10.0 b

10

5.0
5

0.0 0
Untreated Cv Sq DHL+Cv DHL+Sq DHL Untreated Cv Sq DHL+Cv DHL+Sq DHL
Fig. 2 Length (cm) of root of tomato plants grown in co-cultivation with Fig. 3 SPAD index in leaves of tomato plants grown in co-cultivation
microalgae after 46 days (Cv, Chlorella vulgaris; Sq, Scenedesmus with microalgae after 46 days (Cv, Chlorella vulgaris; Sq, Scenedesmus
quadricauda; DHL, digestate humic-like substance). Error bars indicate quadricauda; DHL, digestate humic-like substance). Error bars indicate
standard deviation. The values are means of data from five replicates. standard deviation. The values are means of data from five replicates.
Values followed by different letters are significantly different (P < 0.05) Values followed by different letters are significantly different (P < 0.05).
470
bioavailability of DHL for plants could be different on the
basis of the microalgae species. A hypothesis might be that,
in the co-cultivation system, C. vulgaris resulted to be more
competitive than S. quadricauda for the utilization of DHL
with respect to tomato plants. Interestingly, as already ob-
served in maize by Ertani et al. (2013), the addition of DHL
positively affected the shoot rather than the root.
The co-cultivation system showed a benefit for microalgal
growth, especially when DHL was added to the Hoagland
solution, considering that the Hoagland solution is not an op-
timal medium for the growth of microalgae. Probably, the
presence of DHL exerted a bioactive effect on microalgae,
leading to a more efficient use of nutrients in the Hoagland
solution, hence an increase in their biomass.
Conclusion
The hydroponic co-cultivation system exerted a great benefit for
the growth of both tomato plants and microalgae. This is the first
work, reporting an associated co-cultivation system between
tomato and S. quadricauda or C. vulgaris, resulting in a
biostimulant effect for tomato plants. Successfully, in the co-
cultivation system with S. quadricauda, the presence of DHL
affected positively the shoot growth of tomato plants, along with
an increase in microalgal biomass. This novel agricultural sys-
tem might lead to a significant cost reduction allowing a double
output to be obtained using inputs only for tomato cultivation.
On the other hand, in the co-cultivation with C. vulgaris, the
addition of DHL may be considered as a regulatory, useful and
cheap step when the final objective is to produce a greater con-
tent of microalgal biomass while tomato plants are cultivated.
Moreover, these results showed that the presence of DHL dif-
ferently affects tomato plant growth depending on the
microalgae species.
Considering that hydroponic crop production increased in
the world, it is important to underline that the co-cultivation of
tomato and microalgae in nutrient solution of hydroponic sys-
tem could be strongly recommended as a new strategy for a
better, cheaper and ecofriendly cultivation of tomato plants.
Funding This work was supported by the grant “PIANO PER LA
RICERCA 2016-2018”—University of Catania (Italy).
References
Alam MZ, Braun G, Norrie J, Hodges DM (2014) Ascophyllum extract
application can promote plant growth and root yield in carrot asso-
ciated with increased root-zone soil microbial activity. Can J Plant
Sci 94:337–348
Bacellar Mendes LB, Vermelho AB (2013) Allelopathy as a potential strategy
to improve microalgae cultivation. Biotechnol Biofuels 6:152–165
J Appl Phycol (2019) 31:465–470
Baglieri A, Cadili V, Monterumici CM, Gennari M, Tabasso S, Montoneri
E, Nardi S, Negre M (2014) Fertilization of bean plants with tomato
plants hydrolysates. Effect on biomass production, chlorophyll con-
tent and N assimilation. Sci Hortic 176:194–199
Baglieri A, Sidella S, Barone V, Fragalà F, Silkina A, Nègre M, Gennari
M (2016) Cultivating Chlorella vulgaris and Scenedesmus
quadricauda microalgae to degrade inorganic compounds and pes-
ticides in water. Environ Sci Pollut Res 23:18165–18174
Barone V, Baglieri A, Stevanato P, Broccanello C, Bertoldo G, Bertaggia
M, Cagnin M, Pizzeghello D, Moliterni VMC, Mandolino G,
Fornasier F, Squartini A, Nardi S, Concheri G (2018) Root morpho-
logical and molecular responses induced by microalgae extracts in
sugar beet (Beta vulgaris L.). J Appl Phycol 30:1061–1071
Bulgari R, Cocetta G, Trivellini A, Vernieri P, Ferrante A (2015)
Biostimulants and crop responses: a review. Biol Agric Hortic 31:
1–17
Erice G, Louahlia S, Irigoyen JJ, Sanchez-Diaz M, Avice JC (2010)
Biomass partitioning, morphology and water status of four alfalfa
genotypes submitted to progressive drought and subsequent recov-
ery. J Plant Physiol 167:114–120
Ertani A, Pizzeghello D, Baglieri A, Cadili V, Tambone F (2013) Humic-
like substances from agro-industrial residues affect growth and ni-
trogen assimilation in maize (Zea mays L.) plantlets. J Geochem
Explor 129:103–111
Gong Y, Jiang M (2011) Biodiesel production with microalgae as feed-
stock: from strains to biodiesel. Biotechnol Lett 33:1269–1284
Hickman GW (2011) Greenhouse vegetable production statistics: a re-
view of current data on the international production of vegetables in
greenhouses. Cuesta Roble Greenhouse Consultants, Mariposa
Hultberg M, Carlsson AS, Gustafsson S (2013) Treatment of drainage
solution from hydroponic greenhouse production with microalgae.
Bioresour Technol 136:401–406
Kumar RR, Cho JY (2014) Reuse of hydroponic waste solution. Environ
Sci Pollut Res 21:9569–9577
Lee YK (2001) Microalgal mass culture systems and methods: their lim-
itation and potential. J Appl Phycol 13:307–315
Mata TM, Martins AA, Caetano NS (2010) Microalgae for biodiesel
production and other applications: a review. Renew Sustain
Energy Rev 14:217–232
Maurya R, Paliwal C, Chokshi K, Pancha I, Ghosh T, Satpati GG, Pal R,
Ghosh A, Mishra S (2016) Hydrolysate of lipid extracted microalgal
biomass residue: an algal growth promoter and enhancer. Bioresour
Technol 207:197–204
Parrado J, Bautista J, Romero EJ, García-Martínez AM, Friaza V, Tejada
M (2008) Production of a carob enzymatic extract: potential use as a
biofertilizer. Bioresour Technol 99:2312–2318
Povero G, Mejia JF, Di Tommaso D, Piaggesi A, Warrior P (2016) A
systematic approach to discover and characterize natural plant
biostimulants. Front Plant Sci 7:435
Puglisi I, Barone V, Sidella S, Coppa M, Broccanello C, Gennari M, Baglieri
A (2018) Biostimulant activity of humic-like substances from agro-
industrial waste on Chlorella vulgaris and Scenedesmus quadricauda.
Eur J Phycol; https://doi.org/10.1080/09670262.2018.1458997
Rice EL (1984) Allelopathy, 2nd edn. Academic Press, Orlando
Stanier RY, Kunisawa R, Mandel M, Cohen-Bazire G (1971) Purification
and properties of unicellular blue-green algae (order
Chroococcales). Bacteriol Rev 35:171–205
Taiz L, Zeiger E, Møller IM, Murphy A (2015) Plant physiology and
development, 6th edn. Sinauer Associates, NY
Vernieri P, Borghesi E, Tognoni F, Serra G, Ferrante A, Piaggesi A (2006)
Use of biostimulants for reducing nutrient solution concentration in
floating system. Acta Hortic 718:477–484
Zhang J, Wang X, Zhou Q (2017) Co-cultivation of Chlorella spp and
tomato in a hydroponic system. Biomass Bioenergy 97:132–138