Asian Journal of Advances in Agricultural Research

10(4): 1-14, 2019; Article no.AJAAR.49116

ISSN: 2456-8864

The Use of Biostimulants in High-density Olive

Growing: Quality and Production
Gilberto Hernández-Hernandez1*, Domingo M. Salazar1,2,
Juan Martínez-Tomé3 and Isabel López-Cortés1,2
1
Department of Plant Production, Universitat Politècnica de València, Spain.
2
Institute for the Conservation and Improvement of Valencian Agrobiodiversity (COMAV), Spain.
3
Department of Plant Production and Microbiology, Universidad Miguel Hernández, Spain.

Authors’ contributions

This work was carried out in collaboration among all authors. Authors GHH and ILC designed the
study and performed the statistical analysis, authors DMS and ILC wrote the protocol and wrote the
first draft of the manuscript. Authors GHH and JMT managed the analyses of the study. Authors JMT
and DMS managed the literature searches. All authors read and approved the final manuscript.

Article Information

DOI: 10.9734/AJAAR/2019/v10i430034
Editor(s):
(1) Dr. Wuyi Liu, Professor, Department of Science and Technology Research, Fuyang Normal College (West), Anhui Province,
China.
Reviewers:
(1) Miriam De La Caridad Núñez Vázquez, National Institute of Agricultural Sciences, Cuba.
(2) Raúl Leonel Grijalva-Contreras, Instituto Nacional de Investigaciones Forestales Agrícolas y Pecuarias, Mexico.
Complete Peer review History: http://www.sdiarticle3.com/review-history/49116

Received 10 May 2019

Accepted 22 July 2019
Original Research Article
Published 29 July 2019

ABSTRACT

Due to the increase of high-density holdings, especially of olive trees, the nutritional requirements
of the plants are higher per unit area, which implies that a greater contribution of fertilizers to the
soil is needed. Opting for fertilizers of inorganic origin will produce an increase in the pollution of
the soil.
In the face of this possible soil contamination, our aim is to analyze the effect of biostimulants as
an alternative to chemical fertilizers, to steadily produce and maintain high quality standards during
the life of the crop. Our objective is using more environmentally friendly products in order to satisfy
one of the most important demands from both consumers and the authorities.
In this study, we carried out five different treatments in addition to a control treatment with a supply
of NPK, from inorganic products, which are used to control fertilization with a solution obtained
from seaweed extracts. These treatments were applied in two crop cycles for two of the most
important varieties in the current olive tree growing scenario: Arbequina and Koroneiki.
_____________________________________________________________________________________________________

*Corresponding author: Email: [email protected], [email protected];

 Hernández-Hernandez et al.; AJAAR, 10(4): 1-14, 2019; Article no.AJAAR.49116

This study was developed in the farm Pozohondo, which is located in a crop zone by the Palancia
river (Castellón, Valencia, Spain), in the southeast of the Iberian Peninsula, where the olive trees
were established in a high-density system with a planting framework of 4 x 1.5 m. We ensured an
exhaustive control of the nutritional needs of the holding by using a fertigation system.
We could notice differences in the productions of each applied treatment, avoiding any possible
biases through the additional control of 100 randomly selected olives from each of the samples.
There is an improvement in the set of physical characteristics of the olives with the treatment that
provides amino acids and extra potassium based on amni acids. We analyzed the quality of the
olive oil obtained from the production of each treatment by measuring the fatty acids, tocopherols
and polyphenols contents. We also carried out an organoleptic tasting analysis following the rules
of the International Olive Committee (IOC).
We observed an improvement with regard to the rest of treatments in the pomological parameters
of the olives when applying the potassium and amino acid biostimulant, while the quality of the oils
was not affected by the type of fertilization applied in each treatment.

Keywords: Biostimulant; NPK; olive growing; chemical fertilizers.

1. INTRODUCTION and to improve nutrient use efficiency, tolerance

to abiotic stress, and crop quality when applied to
The olive tree is a traditional growing throughout plants or the rhizosphere. Council [7]. According
the Mediterranean Basin and it plays a key role to Chen et al. [8] this type of compounds
in the so-called Mediterranean Diet [1]. At enhance soil microbial activity, thereby improving
present, the surface cultivated in Spain of olive the fungal and bacterial activity in the long term,
grove is 2.697.445 hectares, which supposes as well as improving the crop itself.
more than 50 % of the surface devoted to the
cultivation of woody species in this country. Its oil Algae extracts are one of the most important
is said to have nutraceutical properties, mainly components in the composition of biostimulants.
due to its monounsaturated fatty acids, They enhance plant development and are
polyphenols and tocopherols contents, which beneficial for both human and animal health [9].
provide antioxidant, antimicrobial and The major components of commercial SWE are
carcinogenic activities, among others [2]. polysaccharides, followed by phenolics, vitamins
precursors, osmolytes (mannitol),
There is a clear tendency nowadays towards the phytohormones, and hormone-like compounds
use of environmentally friendly cropping [10]. Furthermore, they improve plant resistance
techniques, there is a special interest in the to both biotic and abiotic stress [11], activating
practice of organic fertilization with products different metabolites that provide the plant with a
coming from extracts of algae and/or crops, better defense against pathogens [12].
which provide a high organic matter content that
delivers the necessary nutrients to the plant. The market for biostimulants has not stopped
growing since the year 2013, at the rate of about
It is well documented that a suitable irrigation 12% per year, and their main destinations are
regime increases the size and weight of the European holdings, which meant more than 6
olives, in addition to improving the pulp/endocarp million hectares in our continent that year [13].
relation [3], the difference is greater when a
custom fertilization is applied [4]. The use of In addition to improving plant development,
fertilizers exceeds 100 billion kilograms per year. biostimulants increase the biomass in different
This value has increased steadily in recent years, crops such as the almond tree [14]. Another type
along with the introduction of growings in high- of products that falls within the definition of
density systems, which increase fertilizer biostimulants, such as compost, improves the
consumption and can lead to overuse development and growth of peach trees [15]. On
contamination [5] producing salinization and the other hand, it is important to point out that
sodification of soils [6]. there is not only an increase in the productions,
but also an organoleptic improvement of
In general, biostimulants have been described as production in the case of fruit trees [16].
products that contain substances and/or
microorganisms whose function is to stimulate Some studies consider that fertilization has no
natural processes, to enhance nutrient uptake, effect on the organoleptic characteristics of the

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product obtained, however, it can alter the friendly fertilization. Our study focuses on the
composition of compounds such as polyphenols search for an environmentally friendly fertilization
in olive oils [17]. as well as the achievement of an optimum
production while maintaining the highest
It has also been written that products grown in standards of both chemical and organoleptic
more environment-friendly conditions are tastier qualities.
[4], on the other hand, oils show a higher content
of monounsaturated fatty acids [18]. People have 2. MATERIALS AND METHODS
been proven to have a greater interest in
pesticide-free products that present some type of The study was carried out in an olive tree
certification, such as ecological or organic exploitation located in the province of Castellón
products [19], so it is interesting to carry out (Spain) (39°53'50.1" N 0°31'28.0" W) in the
studies in this area. southeast of the Iberian Peninsula, in an area
with an average temperature of 14.2°C and an
Each cultivar possesses singular characteristics average annual rainfall of 384 mm per year. The
in composition of virgin olive oil [20], but the planting pattern was 1.50 x 4.00 meters, for trees
location and climatic conditions also influence in of two of the main varieties that are used in this
this characteristics [21]. type of growings, the Arbequina and the
The Arbequina variety is known for adapting to Koroneiki varietyies, that are 2.50 meters high,
high density cultivation, it is a Spanish origin 20 years of age, and in full production. The plot
cultivar whose fruits are small and round and its had a fertigation system with which the
oils are smooth only slightly bitter and peppery. contributions of irrigation and fertilization were
made, irrigation was 3500 m3 per hectare per
The Koroneiki is a Greek origin variety, it is very year, distributed throughout the periods when the
important in the production of oils. It provides an cultivation needed the most water, from June,
intense green color which is very much when olives are in BBCH 69 state (end of the
appreciated by consumers. Its fruits are large flowering and ripening of the fruit), until mid-
and oval, the oils obtained from its olives have a September, when the trees are in BBCH 89 state
bitter are peppery taste, as opposed to the (the fruits acquire the characteristic color of their
Arbequina cultivar. variety, they remain turgid. Fruits are suitable for
the extraction of the oil).
Despite all the benefits involved in the use of this
type of products (biostimulant fertilizing The biostimulants were tested in an Arbequina
treatments), it is necessary to understand that and Koroneiki cultivar tree holding, given their
carrying out a fertilization process of this type is a importance in the current olive growing, and
complex activity that requires meeting the more specifically in high-density cultivation
nutritional needs of the plant as well as ensuring systems.
soil fertility [22]. That is why this study aims to
evaluate the possible production and quality Each of the cultivars had 5 different fertilizing
differences in an intensive cultivation of olive treatments, in addition to a control treatment with
trees by comparing biostimulant fertilizing fertilizer NPK (T0). The composition of each of
treatments in order to prove if it is possible to the products applied in each treatment can be
maintain the productive performance of an seen in Table 1. The treatments applied were T1
intensive system holding, using environmentally (potassium fertilization), T2 (fertilization with

Table 1. Composition of applied treatmentsº

Treatment Composition
T0 NPK-based fertilization (130 UF N, 35 UF P2O5, 180 UF K2O)
T1 Potassium fertilization (60 % K2O)
T2 Fertilization with seaweed-based biostimulant (2,08 % Bo, 0,02 % Mo and GA142
seaweed filtrate)
T3 Potassium nitrate based fertilization (60 % NO3 + 38 % K2O)
T4 Potassium and algae-based biostimulant fertilization (60 % K2O) + (2,08 % Bo,
0,02 % Mo and GA142 seaweed filtrate)
T5 Potassium fertilization and amino acid based biostimulant (60 % K2O) + (12 %
Aminoácidos libres + 8,5 % N + 2,5 % MgO)

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seaweed-based biostimulant, whose main through tasting was carried out on the rest
ingredients are Boron and Molybdenum), of the sample, in accordance with the rules
T3 (potassium nitrate based fertilization), of the International Olive Oil Council (IOOC)
T4 (potassium and algae-based [23].
biostimulant fertilization) and T5 (potassium
fertilization and amino acid based In order to determine the fatty acid composition
biostimulant. Amino acids were composed mainly of the olive oil a sample was subjected to
of free amino acids, nitrogen and manganese transesterification with methanolic
oxide). potassium hydroxide and n-heptane. The
following fatty acids were determined: palmitic
In order to calculate the production of the trees, acid (C16:0), palmitoleic acid (C16:1),
fruit from 4 randomly selected trees per heptadecanoic acid (C17:0), stearic acid
treatment and cultivate was collected manually. (C18:0), oleic acid (C18:1), linoleic acid
To this effect, 2 trees of each of the rows treated (C18:2), linolenic acid (C18:3), eicosanoic–
with each treatment were selected, avoiding the arachidic acid (C20:0), docosanoic–behemic acid
trees at the beginning and the end of (C22:0), and tetracosanoic–lignoceric acid
the rows that might be affected by passing (C24:0).
vehicles.
Three sterols were examined: β-sitosterol,
The first step taken to analyze the olives was stigmasterol and campesterol. The oil sample
characterizing them pomologically following norm was saponified with an ethanolic potassium
UPOV-CPVO (Union for the Protection of Variety hydroxide solution. The unsaponifiable fraction
Obtention) of the olive tree TG/99/4, as a was removed with an ethyl ether. The
system to establish a pomological unsaponifiable sterol fraction was separated by
characterization of the olive material to be used silica gel plate chromatography. Separation and
in the study. quantification of the silanized sterol fraction was
carried out by means of a capillary column in a
Once the pomological analysis was carried out, gas chromatograph, Hewlett-Packard model HP
we conducted a pomometric analysis of the 5840 gas chromatograph, equipped with an FID-
olives by measuring the weight, length, width A 300, which worked at 290°C. The sample was
and width B of each of them, after which we injected at 280°C, following an isothermal
proceeded to the study of the endocarps, and at process at 265°C for 45 min using a HP-5MS
the same time obtained the pulp/endocarp capillary column (30 m × 0.25 mm × 0.22 μm).
relation. This column was filled with film OB5 Tracer-
Tecnocroma. The working conditions were as
In a pilot plant installation, we proceeded to follows: Helium flow was 1 mL/min; the injector
obtain the oil production from each of the temperature was 300°C; and the detector
samples. These olives were crushed in a temperature was 290°C. The injection volume
hammer mill in order to obtain the olive mass that was 0.2 mL at a flow rate of 1.1 mL/min
was then poured into a blender in a bath to keep (Commission Regulation (EEC) No. 2568/91,
the temperature below 21°C and thus extract the corresponding to AOCS method Ch 6–91). The
individual oil in each of the fertigation trials. After compounds were quantified by addition of an
this process was completed, the mass was then internal pattern (5-α-cholestanol). The sterol
centrifuged to separate the oil from the solid and concentration was expressed as mg/100 g of
aqueous phase obtained after the blending fatty matter. The area of peaks generated by the
phase. sterols was carried out by an automatic
integrator.
Once the oils were separated, a sample of each
of them was taken to be analyzed in the α-Tocopherol was evaluated following AOCS
laboratory, in order to get the parameters that method Ce 8–89. A solution of oil in hexane was
indicate their quality from a chemical point of analyzed on an Agilent Technologies HPLC
view by analyzing the polyphenols, tocopherols system (1100 series) on a silica gel Lichrosorb
and fatty acids contents. This process was aimed Si-60 column (particle size 5 μm × 250 mm × 4
at verifying that they were extra virgin olive oils mm i.d. of Sugerlabor, Madrid, Spain) using n-
(EVOO), complying with the highest standards of hexane/2-propanol (98.5/1.5, vol/vol) at a flow
quality as well as obtaining a complete chemical rate of 1 mL/min. A fluorescence detector
characterization. An organoleptic analysis (Thermo-Finnigan FL3000) was used, with

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excitation and emission wavelengths set at 290 biostimulants, similar to our T2 treatment, while
and 330 nm, respectively. those who had lower values for the pomometric
parameters in study were those in the control
We used the program Statgraphics Centurion treatment, with a fruit weight between 0.45 and
XVII for the statistical analysis, performing 0.54 grams, which is reflected in Table 3, where
variance analysis (ANOVAs) with a 95% the pulp/endocarp relation of the fruits under the
significance to analyze each of the parameters T5 treatment (extra supply of potassium and
individually. amino acid based biostimulant) presented an
average value in both campaigns of 73%,
3. RESULTS AND DISCUSSION whereas in the control treatment, they varied
between 61% and 64%.
In the pomometric characterization of the
Arbequina variety, we observed differences With regard to the productions per tree, the same
between the studied treatments in the size and applies for the pomometry, trees that showed a
weight of the fruits and their endocarps, as better performance, and therefore increased
reflected in Table 2. In the two studied production during the two campaigns of
campaigns, we could observe that the heaviest cultivation under study, were those belonging to
fruits were the ones who had received an extra the crop lines treated with an extra supply of
intake of potassium and amino acids biostimulant potassium and amino acid biostimulant (T5) for
(T5), with an average weight between 1.30 and both varieties. There was an average production
1.38 grams in each campaign, while the lighter of 6.35 kg per tree in the trees of the Arberquina
fruits were the control treatment with a weight variety in which this treatment was applied, while
between 0.92 and 0.93 grams in each campaign, the trees in the control treatment barely achieved
this has an impact on the pulp endocarp that an average production of 4.87 kg per tree. On
usually marks the performance of the fruits, so it the other hand, in the Koroneiki variety,
is one of the most relevant values that are production was 7.45 kg per tree in the lines
generally studied. Thus, the treatments that treated with an extra supply of potassium and
represented the maximum and minimum values amino acid biostimulant (T5), while the trees of
for this parameter were repeated, and the fruits the control treatment lines obtained an average
with a higher pulp/endocarp relation, ranging production of 4.8 kg per tree.
between 76 % and 78 %, came from trees
treated with an extra supply of amino acids and After analyzing the composition of the obtained
potassium (T5), while the fruits of the control oils, as shown in Tables 4 and 5, we found that
treatment that received conventional NPK the fatty acids, polyphenols and tocopherols
fertilization, recorded a lower pulp/endocarp contents were not significantly affected in any of
relation of between 65% and 70%, just like the various combinations variety-treatment, there
Laila et al. [24], in our study, we improved the were only small variations in some of them.
caliber of the olives with biofertilizer However, other authors such as Tekaya et al.
contributions. [26] have seen significant variations in the
content of tocopherols. This may be due to the
In the case of the Koroneiki variety fruits, the fact that our study was conducted in a high
differences between treatments were lower than density growing which was not the case in the
in the Arbequina variety, even so, in the two studies of these authors.
campaigns in study, we observed an
improvement in the size and the pulp/endocarp When carrying out the organoleptic
relation in the fruits treated with an extra supply characterization of the oils obtained for each
of potassium and amino acid biostimulant (T5) variety-treatment combination, we proved that
with respect to the rest of the treatments. The none of the treatments applied had altered the
average weight of the fruits collected in the trees characteristics of the monovarietal oils of the
that received this treatment was between 0.75 varieties under study. So there has been no
and 0.80 grams. differences between the values obtained from
each of the flavours appreciated by this method.
On the other hand, the treatment with lighter This allows to establish that, in the use of
fruits and less pulp/endocarp relation was the biostimulants, organoleptic conditions remain
control treatment. Chouliaras et al. [25] obtained unchanged and will continue to be of interest to
an improvement in the pomometry of the fruits of consumers who are used to these varietal
this variety when applying algae extract features.

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Table 2. Fruit pomometric characterization of cultivar Arbequina

Cultivar arbequina first year

T0 T1 T2 T3 T4 T5
e c d d b a
Fruit weight (g) 0.93 ± 0.19 1.14 ± 0.22 0.99 ± 0.29 1.03 ± 0.27 1.21 ± 0.23 1.38 ± 0.24
Fruit length (mm) 12.41 ± 1.14d 13.49 ± 1.12b 12.66 ± 1.25 c
13.36 ± 1.16b 13.46 ± 1.02b 14.56 ± 1.11a
Fruit width A (mm) 10.60 ± 0.93d 11.51 ± 0.86b 10.67 ± 1.09d 11.13 ± 1.02c 11.51 ± 0.93b 12.80 ± 0.88a
d b d c b a
Fruit width B (mm) 10.20 ± 0.84 11.13 ± 0.83 10.19 ± 1.08 10.77 ± 1.02 11.09 ± 0.86 12.39 ± 0.87
d b cd b a bc
Endocarp weight (g) 0.27 ± 0.06 0.30 ± 0.06 0.29 ± 0.07 0.30 ± 0.06 0.33 ± 0.06 0.30 ± 0.05
d b c bc a ab
Endocarp length 9.33 ± 0.81 10.05 ± 0.85 9.76 ± 0.94 9.92 ± 0.84 10.36 ± 0.82 10.14 ± 0.80
(mm)
d b bc cd a cd
Endocarp width A 6.63 ± 0.48 6.82 ± 0.48 6.75 ± 0.50 6.70 ± 0.47 7.03 ± 0.46 6.66 ± 0.35
(mm)
Endocarp width B 6.45 ± 0.46d 6.64 ± 0.43b 6.57 ± 0.52bc 6.50 ± 0.44cd 6.82 ± 0.42a 6.52 ± 0.36cd
(mm)
Pulp/endocarp 0.70 ± 0.04c 0.74 ± 0.04b 0.70 ± 0.05c 0.70 ± 0.04c 0.73 ± 0.03b 0.78 ± 0.03a
relation
Cultivar Arbequina second year
T0 T1 T2 T3 T4 T5
Fruit weight (g) 0.92 ± 0.21d 0.96 ± 0.32d 1.06 ± 0.20b 1.00 ± 0.21c 1.03 ± 0.23bc 1.30 ± 0.30a
Fruit length (mm) 12.93 ± 0.97d 12.50 ± 1.30d 13.03 ± 1.13 b
13.41 ± 1.27bc 12.89 ± 1.18c 13.91 ± 1.15a
c d c b c a
Fruit width A (mm) 10.44 ± 0.86 10.32 ± 1.18 11.12 ± 0.85 10.96 ± 0.83 10.84 ± 0.97 12.01 ± 1.06
Fruit width B (mm) 10.11 ± 0.82d 10.00 ± 1.15d 10.72 ± 0.82 b
10.79 ± 0.91b 10.49 ± 0.92c 11.75 ± 1.05a
b c a b b b
Endocarp weight (g) 0.31 ± 0.06 0.28 ± 0.06 0.34 ± 0.08 0.32 ± 0.07 0.31 ± 0.07 0.31 ± 0.06
Endocarp length 10.27 ± 0.97b 9.92 ± 0.92c 10.49 ± 1.09a 10.17 ± 1.04b 10.19 ± 0.86b 10.16 ± 0.88b
(mm)
bc e a b de cd
Endocarp width A 6.87 ± 0.50 6.72 ± 0.51 7.31 ± 0.71 6.93 ± 0.65 6.77 ± 0.51 6.83 ± 0.51
(mm)
bc d a b cd bc
Endocarp width B 6.66 ± 0.44 6.53 ± 0.48 6.97 ± 0.63 6.69 ± 0.58 6.60 ± 0.48 6.64 ± 0.48
(mm)
d b c d b a
Pulp/endocarp 0.65 ± 0.08 0.69 ± 0.08 0.67 ± 0.06 0.65 ± 0.06 0.69 ± 0.08 0.76 ± 0.03
relation
T0 (NPK), T1 (potassium fertilization), T2 (fertilization with seaweed-based biostimulant), T3 (potassium nitrate based fertilization), T4 (potassium and algae-based biostimulant
fertilization) and T5 (potassium fertilization and amino acid based biostimulant).Different letters indicate statistical significant differences in a 95%

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Table 3. Fruit pomometric characterization of cultivar Koroneiki

Cultivar Koroneiki first year

T0 T1 T2 T3 T4 T5
d b c bc a a
Fruit weight (g) 0.54 ± 0.18 0.73 ± 0.13 0.69 ± 0.18 0.70 ± 0.13 0.79 ± 0.18 0.80 ± 0.18
Fruit length (mm) 13.61 ± 1.11d 14.79 ± 1.11b 14.01 ± 1.64 c
15.04 ± 1.21ab 15.12 ± 1.35a 15.19 ± 1.44a
Fruit width A (mm) 8.11 ± 0.89e 9.01 ± 0.65c 8.85 ± 0.83d 9.27 ± 0.72b 9.49 ± 0.73a 9.52 ± 0.89a
d bc c b a a
Fruit width B (mm) 7.81 ± 0.95 8.65 ± 0.64 8.59 ± 0.81 8.78 ± 0.73 9.19 ± 0.75 9.13 ± 0.87
c
Endocarp weight (g) 0.18 ± 0.04 0.21 ± 0.04b 0.19 ± 0.05c
0.19 ± 0.04 c
0.22 ± 0.05a 0.21 ± 0.04b
c a c b a a
Endocarp length 10.98 ± 1.05 11.66 ± 0.91 11.07 ± 1.24 11.37 ± 0.87 11.82 ± 1.06 11.75 ± 1.09
(mm)
c a bc b a a
Endocarp width A 5.40 ± 0.36 5.62 ± 0.38 5.44 ± 0.38 5.47 ± 0.32 5.58 ± 0.42 5.58 ± 0.35
(mm)
Endocarp width B 5.29 ± 0.36d 5.46 ± 0.37a 5.33 ± 0.38cd 5.37 ± 0.33bc 5.43 ± 0.39ab 5.47 ± 0.35a
(mm)
Pulp/endocarp 0.64 ± 0.07d 0.71 ± 0.04c 0.72 ± 0.04bc 0.73 ± 0.03ab 0.72 ± 0.04c 0.73 ± 0.05a
relation
Cultivar Koroneiki second year
Control T1 T2 T3 T4 T5
Fruit weight (g) 0.45 ± 0.12e 0.56 ± 0.09d 0.55 ± 0.13d 0.64 ± 0.11c 0.67 ± 0.19b 0.75 ± 0.13a
Fruit length (mm) 13.23 ± 1.02de 13.19 ± 0.89e 13.34 ± 1.16 d
13.99 ± 1.03c 14.43 ± 1.39b 15.00 ± 1.29a
e c d b b a
Fruit width A (mm) 7.74 ± 0.60 8.35 ± 0.42 8.08 ± 0.75 8.73 ± 0.49 8.66 ± 0.91 9.21 ± 0.61
Fruit width B (mm) 7.48 ± 0.55e 8.00 ± 0.42c 7.77 ± 0.72 d
8.38 ± 0.49b 8.41 ± 0.88b 8.86 ± 0.61a
d d c c a b
Endocarp weight (g) 0.17 ± 0.03 0.17 ± 0.03 0.18 ± 0.03 0.18 ± 0.03 0.20 ± 0.04 0.20 ± 0.04
Endocarp length 10.69 ± 0.76c 10.59 ± 0.75c 10.86 ± 0.73b 10.99 ± 0.86b 11.40 ± 0.93a 11.46 ± 1.03a
(mm)
d d c c a b
Endocarp width A 5.23 ± 0.25 5.24 ± 0.29 5.30 ± 0.27 5.32 ± 0.32 5.49 ± 0.32 5.37 ± 0.36
(mm)
e de c cd a b
Endocarp width B 5.11 ± 0.24 5.12 ± 0.28 5.17 ± 0.27 5.16 ± 0.30 5.37 ± 0.31 5.25 ± 0.36
(mm)
e c d b c a
Pulp/endocarp 0.61 ± 0.08 0.69 ± 0.06 0.66 ± 0.07 0.72 ± 0.03 0.69 ± 0.06 0.73 ± 0.05
relation
T0 (NPK), T1 (potassium fertilization), T2 (fertilization with seaweed-based biostimulant), T3 (potassium nitrate based fertilization), T4 (potassium and algae-based biostimulant
fertilization) and T5 (potassium fertilization and amino acid based biostimulant).Different letters indicate statistical significant differences in a 95%

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Table 4. Olive oils fatty acids composition of the studied cultivars

Cultivar Treatment Fatty acids composition

Miristic Palmitic Palmitoleic Margaric Margaroleic Estearic Oleic Linoleic Linollenic Araquidic Gadoleic Behenic Lignoceric
Arbequina T0 ˂0,01 11,01 1,19 0,06 0,08 2,27 78,60 5,31 0,58 0,42 0,31 0,15 0,02
Arbequina T1 ˂0,01 10,97 1,12 0,08 0,10 2,38 78,53 5,38 0,56 0,41 0,34 0,11 0,02
Arbequina T2 ˂0,01 10,88 1,10 0,05 0,08 2,35 78,72 5,32 0,57 0,44 0,30 0,15 0,04
Arbequina T3 ˂0,01 10,98 1,15 0,05 0,09 2,34 78,55 5,37 0,54 0,44 0,33 0,14 0,02
Arbequina T4 ˂0,01 11,12 1,18 0,04 0,10 2,36 78,69 4,96 0,60 0,43 0,35 0,14 0,03
Arbequina T5 ˂0,01 11,02 1,16 0,04 0,11 2,35 78,94 4,86 0,57 0,42 0,35 0,15 0,03
Koroneiki T0 ˂0,01 9,86 0,58 0,04 0,08 2,32 81,02 4,50 0,59 0,46 0,34 0,16 0,05
Koroneiki T1 ˂0,01 9,88 0,59 0,04 0,07 2,35 81,01 4,51 0,63 0,44 0,30 0,14 0,04
Koroneiki T2 ˂0,01 9,87 0,63 0,04 0,08 2,25 81,20 4,41 0,63 0,41 0,31 0,14 0,03
Koroneiki T3 ˂0,01 9,88 0,61 0,04 0,07 2,38 80,81 4,68 0,65 0,42 0,29 0,13 0,04
Koroneiki T4 ˂0,01 9,85 0,64 0,04 0,07 2,34 80,88 4,61 0,64 0,42 0,33 0,14 0,04
Koroneiki T5 ˂0,01 9,82 0,59 0,05 0,07 2,36 80,97 4,59 0,62 0,44 0,31 0,14 0,04
T0 (NPK), T1 (potassium fertilization), T2 (fertilization with seaweed-based biostimulant), T3 (potassium nitrate based fertilization), T4 (potassium and algae-based biostimulant fertilization) and T5 (potassium
fertilization and amino acid based biostimulant)

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Table 5. Tocopherols and Poliphenols content in olive oils of the studied cultivars

Cultivar Treatment Isomers trans Tocopherols/tocotrienols Total poliphenols

Trans oleics Tr L+Tr Ln Total tocopherols α-Tocopherol β-Tocopherol γ-Tocopherol δ-Tocopherol Poliphenoles (Cafeic)
Arbequina T0 ˂0,03 ˂0,03 288,4 284,0 1,4 1,1 ˂1 155
Arbequina T1 ˂0,03 ˂0,03 290,5 286,3 1,4 1,1 ˂1 152
Arbequina T2 ˂0,03 ˂0,03 279,1 274,6 1,8 1,3 ˂1 149
Arbequina T3 ˂0,03 ˂0,03 282,1 275,9 1,6 1,0 ˂1 160
Arbequina T4 ˂0,03 ˂0,03 276,0 273,6 1,3 1,2 ˂1 153
Arbequina T5 ˂0,03 ˂0,03 291,9 289,1 1,6 1,1 ˂1 152
Koroneiki T0 ˂0,03 ˂0,03 239,8 228,8 2,0 3,6 ˂1 174
Koroneiki T1 ˂0,03 ˂0,03 242,2 236,7 2,2 3,3 ˂1 185
Koroneiki T2 ˂0,03 ˂0,03 236,4 231,3 2,1 3,1 ˂1 175
Koroneiki T3 ˂0,03 ˂0,03 228,5 223 2,3 3,2 ˂1 172
Koroneiki T4 ˂0,03 ˂0,03 249,5 243,3 2,3 4 ˂1 165
Koroneiki T5 ˂0,03 ˂0,03 247,6 239,6 2,4 3,4 ˂1 182
T0 (NPK), T1 (potassium fertilization), T2 (fertilization with seaweed-based biostimulant), T3 (potassium nitrate based fertilization), T4 (potassium and algae-based biostimulant fertilization) and T5 (potassium
fertilization and amino acid based biostimulant)

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4. CONCLUSIONS 4. Rosati A, Cafiero C, Paoletti A, Alfei B,

Caporali S, Casciani L. Effect of
We achieved an improvement in production by agronomical practices on carpology, fruit
making different extra biostimulant contributions, and oil composition, and oil sensory
which can be said to replace, at least under our properties in olive (Olea europea L.). Food
working conditions, fertilizers of an inorganic Chemistry. 2014;159:236-243.
origin. This means it is possible to maintain or 5. Neilsen D, Millard P, Neilsen GH, Hogue
even enhance yields in this type of growing given EJ. Sources of N for leaf growth in a high-
that we slightly increased production in our study. density appel (Malus domestica) orchard
At the same time, we cultivated in a more irrigated with ammonium nitrate solution.
environmentally friendly way, highlighting the Tree Physiology. 1997;17(11):733-739.
extra supply of potassium and amino acid 6. Ramos TB, Darouich H, Simunek J,
biostimulant among the applied treatments. On Gonçalves MC, Martins JC. Soil
the other hand, none of the treatments altered salinization in very high-density olive
the chemical composition nor the organoleptic orchards grown in southern Portugal:
quality of the oils, so the specific characteristics Current risks and posible trends.
of the oils from the studied varieties were Agricultural Water Management.
maintained in the implementation of the different 2019;217:265-281.
fertilizing treatments. 7. EBIC. Definition of Biostimulants by
European Bioestimulant Industry Council.
ACKNOWLEDGEMENTS (Accessed 25 April 2019)
Available: www.biostimulants.eu.
8. Chen S, Subler S, Edwards CA.
This work was funded by Project AICO/2017/047.
Effects of agricultural biostimulants on
Development of methods of quantification of
soil microbial activity and nitrogen
riparian vegetation biomass for the management
dynamics. Applied Soil Ecology.
of channels of the Comunitat Valenciana.
2002;19:249-259.
Dirección General de Universidades. Generalitat
9. Khan W, Rayirath UP, Subramanian S,
Valenciana (Spain).
Jithesh MN, Rayorath P, Hodges DM,
Critchley AT, Craigie JS, Norrie J,
COMPETING INTERESTS Prithiviraj B. Seaweed extracts as
Biostimulants of plant growth and
Authors have declared that no competing development. J. Plant Growth Regul.
interests exist. 2009;28:386-399.
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