Journal of Soil Science and Plant Nutrition (2022) 22:743–764

https://doi.org/10.1007/s42729-021-00684-w

ORIGINAL PAPER

Assessing the Potential Role of Compost, PGPR, and AMF in Improving

Tomato Plant Growth, Yield, Fruit Quality, and Water Stress Tolerance
Abdel‑ilah Tahiri1,2 · Abdelilah Meddich2 · Anas Raklami1   · Abdelrahman Alahmad3 · Noura Bechtaoui4 ·
Mohamed Anli2 · Michael Göttfert5 · Thierry Heulin3 · Wafa Achouak3 · Khalid Oufdou1,4

Received: 25 June 2021 / Accepted: 3 November 2021 / Published online: 29 November 2021
© The Author(s) under exclusive licence to Sociedad Chilena de la Ciencia del Suelo 2021

Abstract
Among abiotic stresses, drought is considered the most important growth-limiting factor, particularly in arid and semiarid
regions. Therefore, new management strategies are needed to resolve and mitigate these negative consequences, improve soil
quality and plant growth, and rationalize water use. In this context, we investigated the role of beneficial plant growth–pro-
moting rhizobacteria (PGPR), arbuscular mycorrhizal fungi (AMF) consortium, and compost (Comp) in improving tomato
growth and yield, and drought tolerance. A completely randomized design was used in this experiment with the water stress
as the main factor consisting of two treatments: (1) control well-watered (WW) plants (75% field capacity (FC)) and (2)
water-stressed (WS) plants (35% FC), and the fertilization as a subfactor consisting of eight treatments. Growth parameters
(shoot and root dry weight, leaf number, and area), productivity (fruit number and weight), mineral content ­(Ca2+, ­Na+, ­K+,
and P), biochemical parameters (sugar, protein, and polyphenols), and antioxidant enzyme activities (polyphenoloxidase,
peroxidase, catalase, and superoxide dismutase) were evaluated to investigate the effect of both factor. Soil physicochemical
and microbial properties were examined after the experiment to assess the impact of water stress and applied biofertilizers
on these parameters. Water stress affected negatively plant growth traits and yield and unbalanced the antioxidant enzymes.
However, application of biofertilizers attenuated the negative effect of drought stress. For instance, a significant increase in
shoot biomass of 160%, 120%, and 156% was obtained by Comp, PGPR + Comp, and AMF + Comp treatments compared
to the control, respectively. Indeed, all treatments (except AMF under both conditions and PGPR + AMF + Comp under
WS conditions) significantly increased the fruit number per plant. Concerning fruit yield, Comp, PGPR + Comp, AMF +
Comp, PGPR, and PGPR + AMF treatments were the most effective treatments resulting in 179%, 149%, 111%, 203%, and
181% of the increment, respectively. Concerning the fruit quality, our finding showed a positive effect on sugar content and
a significant decrease in the amount of polyphenol content was recorded by the different applied treatments compared to
control plants under WW conditions. Under WS conditions, the PGPR significantly enhanced sugar and protein by 24% and
177%, respectively. However, they significantly decreased the polyphenol content under WS conditions by 42%. According
to the antioxidant enzymes, a significant decrease in polyphenoloxidase, peroxidase, catalase, and superoxide dismutase
activities in roots was recorded by the different applied treatment plants than WS control plants. In the shoot part, treatments
with PGPR increased the catalase activity under WS conditions. PGPR + Comp, AMF + Comp, PGPR, and PGPR + AMF
treatments significantly decreased the polyphenoloxidase activity and increased the peroxidase activity under WS condi-
tions. In light of these findings, the use of compost alone or in combinations with the beneficial microorganisms (PGPR
and AMF) enhanced the water stress tolerance of tomato plants by improving plant growth, osmolyte accumulation, and
mineral accumulation and by decreasing the amount of antioxidant enzyme activity. This strategy could be vital to resolve
and mitigate the negative consequences of drought stress and rationalize water use.

Keywords  Water stress · PGPR · AMF · Compost · Fruit quality · Antioxidant enzymes · Solanum lycopersicum L

* Anas Raklami
anas.raklami@gmail.com
Extended author information available on the last page of the article

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1 Introduction
Tomato (Solanum lycopersicum L.) is the second most
important vegetable crop after potatoes, and it is widely
cultivated for its commercial interest worldwide and
widely consumed for its edible fruits. It is rich in nutrients
and contains vitamins, lycopene, antioxidants, and essen-
tial minerals (Khanna et al. 2019a, c). Global tomato pro-
duction was 182 million tons in 2018 (FAOSTAT 2018).
Tomato production systems ranged from open-field pro-
duction systems to greenhouse production systems (Ntinas
et al. 2017). In Morocco, tomato production was 1,409,437
tons in 2018 (FAOSTAT 2018). The tomato sector is a real
success story with international influence, and its export
reached 451,000 tons during the 2013–2014 season, with
a value of 2451 million Moroccan Dirhams, 85.2% of them
were exported to the European Union (MAPM 2015). Nev-
ertheless, tomato production is extremely decreased by
abiotic constraints as well as drought (Nahar and Ullah
2018).
Recently, the severity and frequency of drought have
increased significantly, causing severe damage, losses, and
harm worldwide. Climate change is contributing to this
growing trend, presenting a greater threat to society, the
environment, and sectors that rely on rainfall and water
(Parsons et al. 2019). Lack of water is considered as the
most abiotic stress in arid and semiarid areas, reducing
plant development and productivity (Ullah et al. 2016).
Plants respond to drought by a range of multipartite
mechanisms from gene expression, biochemical responses
through individual plant physiological processes to eco-
system levels. Drought limits plant production, and it also
affects morphological, physiological, and biochemical
progress involved in plant growth and development (Kaur
et al. 2021).
Drought is a critical limiting factor in plant growth and
development that affects its elongation, expansive growth,
and cell division (Ansari et al. 2019). Symptoms like wilt-
ing, yellowing, etiolation, premature leaf fall, bark crack-
ing, stunted growth, and canopy thinning are primary vis-
ible responses to water deficit conditions. If the drought
severity is extreme, the plant usually dies (Salehi-Lisar
and Bakhshayeshan-Agdam 2016). A significant decrease
in growth parameters such as plant height, leaf area, bio-
mass, and yield has been reported under water stress in
tomato plants (Nahar and Ullah 2018).
Exposure to drought stress conditions disturbs the
integrity and stability of the cell membranes (Abobatta
2019). Inducing cell injury via the production of reactive
oxygen species (ROS) and increment in the cell tempera-
ture ultimately results in increased cellular viscosity, pro-
tein denaturation, and aggregation. Dehydration provokes
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Journal of Soil Science and Plant Nutrition (2022) 22:743–764
cell shrinkage along with a reduction in cellular volume.
Moreover, solute accumulation leads to cellular toxicity
and enzyme dysfunction that result in reduced photosyn-
thetic rate and water use efficiency (Joshi et al. 2016).
Drought stress alters several metabolic and biochemical
processes within plants besides inducing secondary meta-
bolic molecules to combat the stress (Kaur et al. 2021).
A plant can initiate these events in its tissues by inducing
drought tolerance gene expression, producing antioxidants
or osmolytes, and activating certain antioxidant enzymes
(Feki and Brini 2016). Plants enhance osmolyte/osmopro-
tectant production besides regulating nutrient homeostasis,
as a defense mechanism towards water deficit conditions.
These osmolytes such as proline, polyols, trehalose, and
glycine betaine regulate the osmotic adjustment, provide
protection from ROS, prevent cell membrane injury, and
stabilize various enzymes and proteins (Singh et al. 2015;
Khanna et al. 2019d). Reduction in soil moisture raises also
sugar levels in plants to counteract the drought limiting plant
growth and yield (Kaur et al. 2021).
Plants have evolved an array of cellular, molecular, mor-
phological, and physiological processes in response to water
deficit conditions (Mahmood et al. 2019). The reduction in
net carbon uptake of leaves facing drought is followed by
changes in photoassimilate partitioning at the plant level
that corresponds to an enhanced root/shoot ratio. In gen-
eral, drought tolerance is defined as the ability of plants to
conserve vegetative growth and maintain crop yield during
drought (Abobatta 2019). Abscisic acid (ABA)-mediated
stomatal closure is among the earliest processes induced
under drought conditions. Prolonged exposure to drought
and higher stress intensity leads to further acclimation reac-
tions. Such responses include decreased shoot/root ratio,
osmotic adjustment, reprogrammed metabolism, cell wall
modifications, and activation of the antioxidant system. Most
of the modifications in plants under stressed conditions can
be measured and help characterize the drought stress sever-
ity and include traits like net photosynthesis, mesophyll
conductance, photorespiration, osmoprotectant abundance,
ABA content, tissue water potential, and membrane integrity
(Chen et al. 2015; Laxa et al. 2019).
One way to optimize plant production would be to valor-
ize certain biological components of the soil, including plant
growth–promoting rhizobacteria (PGPR), as well as arbus-
cular mycorrhizal fungi (AMF). The application of PGPR
has been reported to improve tolerance to water stress (Rolli
et al. 2014; Vurukonda et al. 2016). Among the main species
widely used to improve plant productivity, we found Acine-
tobacter sp. and Rahnella aquatilis (Bechtaoui et al. 2019;
Raklami et al. 2019). Both species are known to be major
phosphate solubilizers and indole acetic acid (IAA) produc-
ers and have ACC deaminase activity (Indiragandhi et al.
2008; Mehnaz et al. 2010; Rokhbakhsh-Zamin et al. 2011;
Journal of Soil Science and Plant Nutrition (2022) 22:743–764
Ahemad and Kibret 2014; Kang et al. 2014). In addition,
Acinetobacter sp. produces exopolysaccharides (EPSs) and
siderophores (Rolli et al. 2014). Moreover, Acinetobacter sp.
was able to reduce the activity of plant antioxidant enzymes
under stressful conditions and enhance the amount of gib-
berellic acids (GAs), and these GAs are plant hormones
specialized in plant growth and development (Kang et al.
2009). AMF are beneficial to soil microorganisms with the
ability to colonize the root system of up to 80% of plants to
form a symbiotic association. Such symbiosis benefits host
plants by enhancing water and nutrient uptake from the sur-
rounding soil (Smith and Read 2008; Raklami et al. 2019).
Symbiosis of plants with AMF significantly increases the
explored soil surface area, resulting in greater water and
nutrient uptake (Calvo-Polanco et al. 2016). In our study, we
used an autochthonous AMF consortium isolated from the
Tafilalet palm grove located 500 km southeast of Marrakech.
It is composed of a mixture of native species: (i) Glomus
sp. (15 spores ­g−1 of soil), (ii) Sclerocystis sp. (9 spores
­g−1 of soil), and (iii) Acaulospora sp. (1 spore g­ −1 of soil)
(Meddich et al. 2015). Previous studies have shown that the
application of this AMF consortium significantly enhanced
plant growth in the field (Raklami et al. 2019) and the green-
house (Meddich et al. 2015; Ben-Laouane et al. 2019, 2020;
Anli et al. 2020; Toubali et al. 2020). The AMF consor-
tium improved the plant resistance to abiotic stresses such
as drought (Meddich et al. 2015; Anli et al. 2020), salin-
ity (Ben-Laouane et al. 2019, 2020; Toubali et al. 2020),
and heavy metal stress (Raklami et al. 2020a, b). Numerous
studies have demonstrated that AMF symbiosis and PGPR
can improve host plant stress resistance, including drought
(Armada et al. 2014; Bharti et al. 2016; Calvo-Polanco et al.
2016; Aalipour et al. 2020; Anli et al. 2020). The joint use of
these two groups of microorganisms together could increase
plant growth and yield under water stress conditions (Dodd
and Ruiz-Lozano 2012).
In arid and semiarid regions, soils generally have poor
structure, low organic matter (OM) content, and low water
holding capacity. Under these conditions, limited plant
growth and root exudation can result in poor survival of ben-
eficial microorganisms. The decline in microbial properties
of dry soils is partly due to their gradual decrease in OM con-
tent (Armada et al. 2014). Many studies have reported that
organic amendments improve soil quality (nutrients, OM,
and water retention). Moreover, they increased soil microbial
activity (Trejo et al. 2012; Lopez et al. 2013). In this context,
the application of OM like compost, prior to microbial inocu-
lation, has been suggested (Trejo et al. 2012).
To our knowledge, there are no reports on the combined
use of PGPR and AMF along with the application of com-
post on tomato plants under water stress. Therefore, it is
important to apprehend and evaluate the associated effects of
PGPR and AMF inoculations in combination with compost
745
on tomato plant development and water stress tolerance. In
this context, this study was conducted to answer the fol-
lowing question: Does the combined use of AMF, bacte-
rial strains, and composted green waste promote growth,
yield, and resistance of tomato (S. lycopersicum L.) plants
submitted to drought conditions and alleviate the negative
effect of water deficiency? For that, a greenhouse experiment
was carried out to assess the efficiency of PGPR, AMF, and
compost in alleviating the water stress effect.
2 Materials and Methods
2.1 Experimental Site, Design, Treatment
Applications, and Cultural Practices
Tomato seeds (S. lycopersicum L.) var. Roma VF (Shal,
Spain) were disinfected using diluted sodium hypochlorite
(1:3; v/v). After several washes using distilled water, the seeds
were transferred to polypropylene trays containing sterile
commercial peat. After 20 days of germination, the homog-
enous young seedlings having the same height and showing
the first true leaves were transferred to the 4.5-l pot (15 cm
bottom diameter, 23 cm upper diameter, and 20 cm height)
filled with 5 kg of agriculture soil, sampled in the commune
Saada, prefecture of Marrakech (coordinates 3°37′38.7″N,
8°07′48.4″W, 450 m). Plants were transferred at the rate of
one plant per pot with ten repetitions for each treatment. The
soil used was characterized by a pH of 8.38, soil available P
of 30.45 mg ­kg−1, organic carbon of 0.98%, total N of 0.112%,
and electrical conductivity (EC) of 785 μS ­cm−2 (Table 1).
A completely randomized design was used in this experi-
ment with the water stress as the main factor consisting of
two treatments: (1) control (well-watered (WW) plants [75%
field capacity (FC)]) and (2) water-stressed (WS) plants
(35% FC) after 1 month of seedling transfer, and the fertili-
zation as the subfactor consisted of eight treatments:
1. Control plants without inoculation or amendment (control)
2. Amended with compost (Comp)
3. Inoculated with AMF (AMF)
4. Inoculated with AMF and amended with compost (AMF
+ Comp)
5. Inoculated with PGPR (PGPR)
6. Inoculated with PGPR and amended with compost
(PGPR + Comp)
7. Inoculated with PGPR in combination with the AMF
(PGPR + AMF)
8. Inoculated with PGPR and AMF along with compost
(PGPR + AMF + Comp)
The experimental design was factorial, with 10 replicates
per treatment (160 plants in total).
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Table 1  Physicochemical Parameters Soil Soil + 10% compost Compost

parameters of the used soil, the
compost, and the mixture of soil pH 8.38 (0.08) 8.06 (0.01) 7.74 (0.01)
and compost (9:1 w/w)
EC (μS c­ m−2) 785 (0.45) 1395 (0.25) 5460 (0.2)
Soil TN (%) 0.112 (0.01) 0.252 (0.01) 1.316 (0.01)
NH4+ (%) – – 0.088 (0.002)
NO3− (%) – – 0.307 (0.002)
TOC (%) 0.98 (0.12) 1.68 (0.04) 5.72 (0.45)
OM (%) 1.68 (0.2) 2.89 (0.07) 9.86 (0.78)
C/N ratio – – 7.49
NH4+/NO3− – – 0.29
Soil available P (mg k­ g−1) 30.45 (5.22) 196.83 (2.89) 489.95 (20.29)
Na+ (mg ­kg−1) 990 (30) 1240 (80) 2110 (50)
K+ (mg ­kg−1) 890 (50) 1430 (10) 5590 (150)
Ca2+ (mg ­kg−1) 11,480 (480) 17,770 (2030) 37,380 (1840)
Bacteria (UFC g­ −1) 4.95 × ­105 (4.5 × ­104) 5.9 × 1­ 06 (3.95 × ­105) 1.23 × 1­ 08 (7 × ­106)
Fungi (UFC g­ −1) 1.95 × 1­ 05 (1.5 × ­104) 3.3 × 1­ 05 (0) 1.6 × ­106 (1.5 × ­104)

Data presented are means ± SD

EC electrical conductivity, TN total nitrogen, TOC total organic carbon, OM organic matter, C/N carbon-to-
nitrogen, UFC units forming colony

The experiment was conducted under greenhouse condi-
tions. Plants were grown under natural light, 16 h/8 h (day/
night). The average temperature was 33 °C/25 °C, and the
relative humidity was 55%/86%. Throughout the 4 first
weeks of the experiment, soil water content was maintained
close to 75% of FC for both groups of plants (WW and WS).
Thereafter, the WS pots were left to dry out until the soil
water content reached 35% of FC and maintained under
these conditions for the additional 12 weeks. To reach this,
the pots were weighted twice daily and water was added to
achieve 35% of FC. The methodology for the application of
water stress is described by Meddich et al. (2015). P1 is the
weight of the pot filled with 5 kg of dry soil. Then, the soil
was watered to saturation and allowed to drain under grav-
ity, to a constant weight. The soil was hence at FC. P2 was
the weight of the pot after the flow of excess water. The dif-
ference (P2 − P1) corresponded to the volume required for
obtaining the FC of the used soil (100% FC). For equivalent
humidity of the soil, of 75%, and 35% of FC, we added, to
the series of pots containing dry soil, water volumes corre-
sponding to 0.75 × (P2 − P1) and 0.35 × (P2 − P1), respec-
tively. P3 and P4 weights of the pots at 75% and 35% of FC,
respectively, were then presented.
material was dried at 80 °C to constant weight for dry weight
and nutrient determination. The yield was determined by
measuring the total number of fruits and their weight.
The nutrients as C­ a2+, ­Na+, and K
­ + contents in shoots and
roots were measured by the flame spectrometer as described
by Raklami et al. (2019). Total P in the compartments was
determined by the colorimetric method (Olsen et al. 1954).
Gas exchange (gs) was determined by a portable porometer
(Leaf Porometer, Decagon Device, Inc.) between 11 and 13 h
on four leaves per treatment.
Chlorophyll fluorescence (Fv/Fm) was measured by a fluo-
rometer (Opti-Sciences, OS30p). The eclipses were placed
on the upper side of the leaves for 20 min to keep the leaves
in the dark. This parameter was measured in well-developed
leaves of the same rank (four repetitions per treatment) and is
expressed in Fv/Fm, with Fv as the variable fluorescence (Fm
− F0) and Fm as the maximum fluorescence, and F0 being the
fluorescence in the initial state when all photosystem II reac-
tion centers are open (oxidized quinones).
The leaf water potential (leaf Ψ) was recorded using the
pressure chamber method described by Scholander et  al.
(1965) by placing a leaf in a pressure chamber instrument
(PMS Instrument Company, USA).

2.2 Determination of Plant Growth, Yield, 2.3 Molecular Identification of Bacterial Strains

and Physiological Performance
After 4 months, plants were harvested. The number of leaves
and leaf area were determined to assess plant growth. Sam-
ples of the harvested plants were frozen in liquid nitrogen
and stored at − 20 °C for enzyme assays. The rest of the plant
2.3.1 DNA Extraction
The bacterial strains BS14 and BS36 were isolated from
the rhizosphere of faba bean roots sampled in the Ait Ourir
region, Morocco (31°46′01″N, 07°67′65″W). Their genomic

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Journal of Soil Science and Plant Nutrition (2022) 22:743–764
DNA was extracted as described by Bechtaoui et al. (2019).
The bacterial strains were cultivated in Trypticase Soy Agar
medium at 28 °C for 48 h. Then, 4 ml of the obtained bacte-
rial suspension was centrifuged. The pellet was washed once
using Tris-EDTA (TE) buffer (10 mM Tris, 1 mM EDTA,
pH 8), and bacterial cells were resuspended in 300 μl TE
buffer. To the bacterial suspension, 100 μl SDS (5%) and
100 μl pronase E (2.5 mg ­ml−1 in TE buffer) were added.
Then, the mixture was incubated overnight. Then, the DNA
was carefully cropped using a syringe. DNA purification
was performed by two extractions with 300 μl Tris-buffered
phenol and one extraction with methylene chloride. DNA
was collected by precipitation using 0.1 volumes of 3 M
sodium acetate and 2.5 volumes of ethanol.
2.3.2 16S Ribosomal DNA Amplification of Sequencing
The 16S ribosomal DNA (rDNA) gene was multiplied using
16Sa (5′-CGCT ​ GGC​ GGC ​ AGG ​ CTT ​ AAC
​ A-3′) and 16Sb (5′-
CCAG ​ CCG​ CAG ​ GTT​ CCC ​ CT-​ 3′) primers (Van Berkum and
Fuhrmann 2000). The reaction mixture consisting of 100 ng
bacterial DNA, DreamTaq buffer, 100 pmol dNTPs, 1.25 U
DreamTaq polymerase, and Milli-Q water in a total volume
of 50 μl. PCR was carried out under the following condi-
tions: 5 min of initial denaturation at 95 °C, followed by
30 cycles (30 s of denaturation at 95 °C, 30 s of annealing
at 58 °C, and 1.5 min of extension at 72 °C), then 10 min
of a final elongation at 72 °C. Horizontal gel electrophore-
sis (1% agarose) in Tris-acetate-EDTA (TAE) buffer was
used to check PCR products. The PCR product was purified
using “MEGAquick-spin™ Total Fragment DNA Purifica-
tion Kit.” Nucleotide sequencing was carried out using the
same primers by GATC Biotech (Konstanz) on both strands.
Phylogenetic analysis was conducted with MEGA, version
6 (Tamura et al. 2013).
2.4 PGPR Activity Characterization and Inoculum
The isolates were tested for their capacity to display different
PGPR activities. Phosphate and potassium solubilization was
performed in NRBIP and Alexandrov solid medium (Alikhani
et al. 2006). Positive results were visualized by the presence
of a clear zone around the bacterial cells. EPS production was
assessed using the methods described by Lee et al. (2007).
Siderophore production was tested on CAS medium (Schwyn
and Neilands 1987). A change in the color of the medium
from blue to orange in the zone surrounding the colony was
considered a positive result. For IAA production, strains were
cultivated in Luria-Bertani (LB) medium supplemented with
−1
l-tryptophan (1 g ­l ). After 96 h of incubation at 28 °C and
140 rpm, 1.5-ml aliquots of cultures were centrifuged and 1 ml
of the supernatants was added to 2 ml Salkowski reagent and
100 μl of 0.5 M phosphoric acid. The absorbance was measured
747
at 530 nm after 30 min of darkness (Raklami et al. 2019). The
ability of isolates to produce hydrocyanic acid (HCN) is deter-
mined by the method described by Lorck (1948).
Tomato plants were inoculated by BS14 and BS36 strains.
The inoculum was prepared by strain multiplication in Tryp-
ticase Soy Broth medium and shaken for 24 to 48 h at 28 °C
until an absorbance of 1 at 600 nm was obtained (~ ­109 CFU
­ml−1). Plant inoculation was carried out by spraying 50 ml
of the bacterial suspension formed from the two abovemen-
tioned strains in equal volumes as close as possible to the roots.
Boosters were carried out after 15 days by inoculating 50 ml of
the bacterial suspension onto the plant roots to enhance the rate
of PGPR in the soil and infect the newly formed small roots.
2.5 Arbuscular Mycorrhizal Fungus
The AMF complex used in this study was isolated from the
Tafilalet palm grove located 500 km southeast of Marrakech.
It is composed of a mixture of native species: (i) Glomus sp.
(15 spores g­ −1 of soil), (ii) Sclerocystis sp. (9 spores g­ −1 of
soil), and (iii) Acaulospora sp. (1 spore ­g−1 of soil) (Meddich
et al. 2015). Corn plants (Zea mays L.) were cultivated in the
sampled soil to trap and multiply the autochthonous mycorrhi-
zal consortium, which naturally forms a symbiotic association
with palms.
Upon transplantation, seedlings were inoculated with 10
g of sand containing spores and corn roots pre-mycorrhized
by the above AMF. Non-AMF plants were inoculated by 10
g of autoclaved inoculum. After harvest, the AMF infection
frequency (F) and infection intensity (M) of tomato roots by
AMF were determined by the technique described by Phillips
and Hayman (1970). Briefly, root fragments were cleaned with
10% KOH and stained with 0.05% trypan blue in lactic acid
(v/v). The rate of AMF colonization and the intensity of AMF
infection were determined using the following formula:
( )
Infected root segments
AMF infection frequency (F) (%) =
Total root segments
× 100
(95n5 + 70n4 + 30n3 + 5n2 + n1)
AMF infection intensity (M) (%) =
Total root segments
where n5 is the number of roots with colonization level
5 (colonization rate between 90 and 100%), n4 is the root
number at level 4 (colonization rate between 50 and 90%),
n3 is the root number at level 3 (colonization rate between
10 and 50%), n2 is the root number at level 2 (colonization
rate between 1 and 10%), and n1 is the root number at level
1 (colonization rate between 0 and 1%).
2.6 Compost
The compost used in this work was obtained from the
composting unit of the “Faculty of Sciences Semlalia
13
748
Marrakech” (FSSM) (Marrakech, Morocco). The compost
produced from the green waste was collected selectively dur-
ing the pruning of gardens. Plant debris were fragmented
and aggregated in heap then moistened. Each week, the plant
debris was turned up and humidified until stable and fine
compost was obtained. The compost was amended at a rate
of 10%. Their main characteristics are reported in Table 1.
2.7 Parameters Measured
2.7.1 Proline Content
The proline content was determined following the method of
Bates et al. (1973); 0.5 g of plant material was homogenized
in 10 ml of 3% aqueous sulfosalicylic acid, and the homoge-
nate was filtered through Whatman #2 filter paper. Two mil-
liliters of filtrate was reacted with 2 ml acid ninhydrin and 2
ml of glacial acetic acid in a test tube for 1 h at 100 °C, and
the reaction stopped in an ice bath. The reaction mixture was
extracted with 4 ml toluene and mixed vigorously with a test
tube stirrer for 15–20 s. The chromophore containing toluene
was aspirated from the aqueous phase and warmed to room
temperature, and the absorbance was read at 520 nm using
toluene as blank. The proline concentration was determined
from a proline standard curve.
2.7.2 Sugar, Protein, and Polyphenol Concentration
For the determination of sugar content, 0.1 g of leaves, root,
or fruit tissues (to determine fruit quality) was homogenized
with 8 ml of ethanol (80%) and centrifuged for 10 min at
5000 rpm. To 0.2 ml of supernatant, 200 μl of phenol (5%)
and 1 ml H ­ 2SO4 were added and stirred. After cooling, the
absorbance at 485 nm was read (Dubois et al. 1956).
Soluble proteins in the fruits as fruit quality marker were
measured following the method of Bradford (1976). Briefly,
5 ml of Bradford reagent was supplemented with 0.1 ml
of the ethanolic extract. After homogenization, the reaction
mixture was placed for 30 min at 30 °C. Then, the absorb-
ance was read at 595 nm.
Polyphenol content was measured according to the
method described by Yamamoto et al. (1977) with slight
modifications. Leaves, roots, or fruit tissues (1 g fresh
weight (FW)) were ground in 8 ml of methanol (80%). To
ensure a maximum extraction, two supplementary extrac-
tions were made by washing the residues with methanol
(80%). The mixture of the filtrate and additional filtration
was centrifuged at 1000 g for 5 min. Then, 0.2 ml of the
supernatant was supplemented with 0.4 ml of Folin-Denis
reagent and distilled water in a total reaction mixture of 3
ml. After 3 min, 1 ml of saturated aqueous ­Na2CO3 solution
was added at ambient temperature for 1 h to complete the
13
Journal of Soil Science and Plant Nutrition (2022) 22:743–764
reaction. After that, the absorbance was determined at 765
nm using gallic acid as standard.
2.7.3 Antioxidant Enzyme Activities and Protein Content
The extraction of antioxidant enzymes was performed at 4
°C. For this, 0.1 g of previously frozen leaves and roots parts
was crushed in 4 ml of 0.1 mM sodium phosphate buffer
(pH 6.0) with 5% (w/v) insoluble polyvinylpolypyrrolidone
(PVPP) and 0.1 mM Na-EDTA on ice. The homogenate was
centrifuged at 18,000 g for 10 min at 4 °C. The supernatant
was used for enzymatic dosages and the determination of
protein content.
Soluble proteins were measured following the method of
Bradford (1976). Briefly, 5 ml of Bradford reagent was sup-
plemented with 0.1 ml of the extract. After homogenization,
the reaction mixture was placed for 30 min at 30 °C. Then,
the absorbance was read at 595 nm.
Polyphenoloxidase (PPO) activity was determined at 410
nm following the catechol oxidation according to the method
of Moore and Flurkey (1990). The enzyme reaction was per-
formed in a 2.1 ml reaction mixture at 23 °C. The mixture
consisted of 0.1 mM sodium phosphate buffer (pH 6.0) and
10 mM catechol. The evolution of absorbance was followed
at 410 nm for 3 min. Total peroxidase (POD) activity was
determined according to Bergmeyer et al. (1974), and the
assay solution contained 100 μl of enzymatic extract, 2 ml
(0.1 mM) of phosphate buffer (pH 6.0), and 1 ml (20 mM)
of guaiacol. The reaction started by adding 0.5 ml (10 mM)
of ­H2O2. Enzyme activity was monitored by increasing the
optical density at 470 nm. For the catalase (CAT) assay, the
assay contained 890 μl of phosphate buffer and 100 μl of
extract. The reaction was launched by adding 10 μl of ­H2O2
(15%). The activity was assessed by monitoring the decrease
in optical density at 240 nm (Duarte et al. 2015). Superoxide
dismutase (SOD) was monitored using the reduction of nitro
blue tetrazolium (NBT) on formazan blue at 560 nm after
30 min under blue light (Beyer and Fridovich 1987). The
assay contained 2550 μl sodium phosphate buffer (0.1 mM,
pH 6.0), 75 μl of methionine (55 mM), 300 μl of NBT (0.75
mM), and 50 μl of extract. The reaction was started when 60
μl of riboflavin (0.1 mM) was added. Blanks were carried
out in the absence of extract to assess the auto-oxidation of
the reagents. Antioxidant activities were expressed in U mg
­protein−1 ­min−1.
2.7.4 Physicochemical and Microbial Properties of the Soil
The physicochemical characteristics of the soil were deter-
mined before and after the greenhouse experiment, as fol-
lows: the pH and EC of soil (soil:distilled water; 1:5; w/v)
were estimated using a digital pH meter (pH 21, Hanna
Journal of Soil Science and Plant Nutrition (2022) 22:743–764
Instruments, Romania) and conductivity meter (LF92,
WTW, France), respectively. The determination of the soil
texture consisted of estimating the percentage of the vari-
ous mineral fractions constituting the aggregates contained
in a fine soil sample (2 mm). After digestion of the OM
with 20% hydrogen peroxide and deflocculation of the clays
with sodium hexametaphosphate (50 g ­l−1), the sand frac-
tion was determined by passing the soil solution through a
50-μm sieve. The clay and silty fractions were determined
using Robinson’s pipette (Baize 2000). Total organic car-
bon (TOC) was assessed after oxidation with 1 N potassium
dichromate (Baize 2000). Soil total nitrogen (TN) was deter-
mined by the Kjeldahl method (Baize 2000), using a nitro-
gen estimation apparatus (PRO-NITRO S; Selecta, Spain).
The available form of P was determined with 1 M acidic
ammonium fluoride (Olsen et al. 1954) using a colorimetric
method (VR-2000 Spectrophotometer, Selecta, Spain).
Determination of minerals ­(Na+, ­K+, and ­Ca2+) was per-
formed after soil mineralization. Briefly, 0.5 g of soil was
mineralized for 6 h at 550 °C in a muffle furnace. Then, the
samples were soaked for 1 h in 3 ml hydrochloric acid (5N),
filtered, and diluted with distilled water to 100 ml. ­Na+, ­K+,
and ­Ca2+ elements were measured using a flame photometer
(AFP 100 flame photometer).
In addition, the estimation of the number of bacteria and
fungi in the soil before and after the experiment was carried
out by the suspension-dilution technique, and 0.1 ml of each
dilution was spread on TSA medium for bacteria and PDA
medium supplemented with 100 mg l­ −1 of chloramphenicol
for fungi. Colony counts were performed after 24 h of incu-
bation at 28 °C for bacteria and after 96 h of incubation at
25 °C for fungi.
2.8 Statistical Analyses
The results are the means ± standard deviation (SD) of
eight determinations for growth parameters: four for
physiological parameters and three for mineral nutrients,
biochemical parameters, enzymatic activity, and soil
parameters. After importing the data, analysis of vari-
ance (ANOVA) was applied using the aov function of the
package (“stats”) (R Core Team 2013). Then, multiple
comparisons of treatments were achieved using the least
significant difference (LSD) test with the grouping of treat-
ments using the LSD .test() function of the agricolae pack-
age (de Mendiburu and Yaseen 2020). For this, we used
the default value for the alpha argument that represents
the 5% significance level (p ≤ 0.05) and the Bonferroni
method for adjusting the probability value. In addition,
data integrity was checked and normalized by data scaling:
auto-scaling (mean-centered and divided by the standard
deviation of each variable) using the SanityCheckData()
and Normalization() functions, respectively, from the
749
“MetaboAnalystR” package (Chong and Xia 2018). The
analysis was proceeded, starting with two-factor (treat-
ments vs. water stress) independent samples univariate
analysis considering their interaction. Two-way (between-
subjects) ANOVA type I (the means are weighted) was
calculated using the ANOVA2.Anal() function with the
adjusted p value cutoff of 0.05 and the false discovery rate
(FDR) method for multiple testing correction. Hierarchi-
cal cluster analysis was conducted, and the results were
then displayed using the two-way heatmap visualization by
the PlotHeatMap2() function with Euclidean distance as a
similarity measure and Average as a clustering algorithm
(clustering uses the centroids of the observations).
3 Results
3.1 PGPR Strain Identification and Characterization
The 16S rDNA gene sequences showed that BS14 strain is
closely related to Acinetobacter sp. (97.46% of similarity to
Acinetobacter sp. 10169 (HE651913.1)) and strain BS36 is
very similar to Rahnella aquatilis with 100% of similarity
with Rahnella aquatilis PGP30 (MN006387.1) (Fig. 1).
Both tested strains showed significant phytobeneficial
traits (Table 2). They can both solubilize tricalcium phos-
phate, while BS14 can also solubilize potassium. Further-
more, the rhizobacterial strains are able to produce EPS
up to 152.31 mg of CR/OD600 (BS36) and IAA arranged
from 12.69 μg m ­ l−1 for BS36 to 195.64 μg m­ l−1 for BS14.
However, no strain was able to produce siderophores and
HCN.
3.2 Effect of Water Stress on Tomato Root
Colonization by AMF
Both studied factors (water stress and biological treat-
ments) had a significant (p < 0.001) effect on the fre-
quency and the intensity of AMF infection (Table  3).
Under WW conditions, AMF inoculation resulted in a
significant (p < 0.05) increase in the AMF frequency in
all four AMF-inoculated treatments (Fig. A1a). Moreo-
ver, PGPR inoculation (PGPR treatment) significantly
enhanced the AMF frequency compared to the control
plants (Fig. A1a). Under WS conditions, AMF, PGPR,
PGPR + AMF, and PGPR + AMF + Comp treatments
showed a significant increase (p < 0.05) in the AMF fre-
quency (Fig. A1a). For instance, AMF alone or in combi-
nation with PGPR increased the AMF frequency by 166%
and 125%, respectively. AMF intensity was improved by
PGPR + AMF treatment under both water levels, while
AMF alone increased these parameters only under WW
conditions.
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750 Journal of Soil Science and Plant Nutrition (2022) 22:743–764

Fig. 1  Maximum likelihood
phylogenetic tree of the strains
BS14 and BS36 based on 16S
rDNA gene sequences, show-
ing the position of the strains
with regard to related species.
Bootstrap values based on 1500
replications are given at branch
points. Numbers in parentheses
represent the sequence acces-
sion numbers in GenBank.
Scale bar: substitutions per
nucleotide position

Table 2  Plant growth–promoting rhizobacterial properties of the tested rhizobacteria

Strain Phosphate solu- Potassium solu- Exopolysaccharide produc- Siderophore IAA production (μg ­ml−1) HCN
bilization bilization tion (μg eq CR/OD600) production produc-
tion

BS36 *** – 152.31 (0.58) – 12.69 (0.30) –

BS14 *** *** 77.49 (3.65) – 195.64 (15.05) –

Data presented are means ± SD; –: absence

CR Congo red, IAA indole acetic acid, HCN hydrocyanic acid
***High

3.3 Effect of Biofertilizers and Water Stress
on Growth, Yield, and Mineral Nutrition
of Tomato
We have demonstrated the significant effect (p < 0.01) of
both factors (water stress and biological treatments) and
the significant interaction (p < 0.01) between treatments
and water stress, indicating that water stress had different
effects on growth parameters depending on the treatments
(Table 3).
enhanced the shoot biomass by 160%, 120%, and 156%,
respectively (Fig.  2a). This result might be due to
increased water availability to the plants by compost.
All treatments under water stress conditions had signifi-
cantly higher (p < 0.05) shoot biomass or equivalent to
the control treatment under WW conditions, suggesting
that all treatments had the potential to offset the negative
effect of water stress.
3.3.2 Root Biomass

3.3.1 Shoot Biomass Under WW conditions, compost application and its com-

All treatments (except AMF treatment) resulted in a
significant increase (p < 0.05) in shoot biomass under
WW conditions (Fig. 2a). Under WS conditions, all the
applied treatments, except PGPR and AMF treatments,
resulted in a significant (p < 0.05) increase over control.
Comp, PGPR + Comp, and AMF + Comp treatments
bination with PGPR and/or AMF and the PGPR + AMF
treatments had a significant (p < 0.05) and positive effect
on root biomass up to 252% for the compost treatment com-
pared to control (Fig. 2b). Under WS conditions, Comp,
PGPR + Comp, and Comp + AMF treatments significantly
increased (p < 0.05) root biomass (Fig. 2b) by 196%, 59%,
and 156%, respectively. All treatments under WS conditions

13
Journal of Soil Science and Plant Nutrition (2022) 22:743–764 751

Table 3  Result of two-way Parameters Treatments Water stress Treatments × water

ANOVA test for treatment stress
(compost, PGPR, and AMF)
effect, water stress effect, and F value p value F value p value F value p value
their interactions
AMF frequency 23.683 *** 31.148 *** 2.2593 ns
AMF intensity 28.669 *** 30.205 *** 8.755 ***
Shoot dry weight 89.832 *** 552.61 *** 19.872 ***
Root dry weight 33.342 *** 29.177 *** 7.6822 ***
Leaf number 52.437 *** 118.68 *** 5.7731 ***
Leaf area 83.31 *** 413.28 *** 14.958 ***
Fruit number 38.486 *** 97.808 *** 10.855 ***
Fruit weight (yield) 19.297 *** 435.35 *** 12.549 ***
Na+ (shoot) 996.83 *** 1340.5 *** 52.502 ***
Na+ (root) 145.12 *** 621.73 *** 152.16 ***
K+ (shoot) 796.55 *** 3901.5 *** 806.52 ***
K+ (root) 143.93 *** 271.19 *** 14.333 ***
Ca2+ (shoot) 79.34 *** 413.98 *** 90.776 ***
Ca2+ (root) 88.533 *** 34.828 *** 85.636 ***
P (plant) 658.92 *** 4577.7 *** 294.02 ***
Fv/Fm 4.9921 *** 59.333 *** 1.5763 ***
Gas exchange 12.743 *** 2634.6 *** 7.4662 ***
Water potential (Ψ) 40.441 *** 929.78 *** 19.899 ns
Proline (shoot) 30.554 *** 107.55 *** 13.874 ***
Proline (root) 11.997 *** 117.59 *** 14.629 ***
Sugar (shoot) 106.94 *** 120.29 *** 122.67 ***
Sugar (root) 299.52 *** 1042.3 *** 26.977 ***
Sugar (fruit) 14.093 *** 9.2035 ** 26.776 ***
Protein (shoot) 64.469 *** 320.93 *** 23.272 ***
Protein (root) 80.202 *** 866.93 *** 24.32 ***
Protein (fruit) 51.209 *** 3.9401 ns 89.599 ***
Polyphenol (shoot) 18.119 *** 1.4663 ns 1.4795 ns
Polyphenol (root) 27.097 *** 5.3445 * 9.0613 ***
Polyphenol (fruit) 16.512 *** 80.878 *** 7.1735 ***
PPO (shoot) 19.032 *** 327.39 *** 14.74 ***
PPO (root) 81.802 *** 416.79 *** 23.679 ***
POD (shoot) 177.15 *** 439.06 *** 108.91 ***
POD (root) 408.13 *** 3324.5 *** 373.05 ***
CAT (shoot) 19.998 *** 418.04 *** 19.985 ***
CAT (root) 91.959 *** 49.305 *** 47.644 ***
SOD (shoot) 5.4514 *** 50.586 *** 5.2 ***
SOD (root) 2.8748 * 31.835 *** 3.7622 **
pH 121.95 *** 369.12 *** 116.92 ***
EC 1516.8 *** 2299.1 *** 1136 ***
Soil available P 1358.1 *** 825.28 *** 248.67 ***
TOC (%) 186.2 *** 34.727 *** 40.441 ***
OM (%) 186.48 *** 34.583 *** 40.731 ***
Na+ (soil) 4.6695 *** 31.872 *** 3.4376 **
K+ (soil) 8.7249 *** 0.86889 ns 8.9306 ***
Ca2+ (soil) 90.445 *** 26.241 *** 27.414 ***
Bacterial count 68.849 *** 72.902 *** 3.2193 *
Fungal count 67.603 *** 72.902 *** 3.2193 ***

ns not significant, AMF arbuscular mycorrhizal fungi, PPO polyphenoloxidase, POD peroxidase, CAT​cat-
alase, SOD superoxide dismutase, EC electrical conductivity, TOC total organic carbon, OM organic matter
*Significant at p < 0.05; **significant at p < 0.005; ***significant at p < 0.001

13
752 Journal of Soil Science and Plant Nutrition (2022) 22:743–764

Fig. 2  Effects of water stress

and biofertilizers on a shoot
dry weight (SDW), b root dry
weight, c leaf number, d leaf
area, e fruit number, and f
yield measured by the weight
of fruits/plant of S. lycopersi-
cum plants. Comp compost;
AMF arbuscular mycorrhizal
fungal consortium; PGPR plant
growth–promoting rhizobacte-
rial strains (BS14 and BS36
strains); WW well-watered
(75% field capacity (FC)); WS
water-stressed (35% FC). Data
presented are means ± SD. Bars
sharing the same letters in each
graphic are not significantly
different (p < 0.05)

had significantly (p < 0.05) higher root biomass or equiva-
lent to the control treatment under WW conditions, suggest-
ing that all treatments had the potential to compensate for
the negative effect of water stress.
3.3.3 Leaf Number and Leaf Area
For these two leaf parameters, the results revealed that
the application of compost alone or in combination with
PGRP and/or AMF resulted in a significant (p < 0.05)
and positive effect on leaf number regardless of water
supply conditions (Fig. 2c). For leaf area, all treatments,
except AMF treatment under WW conditions and PGPR
+ AMF under WS conditions, had significantly (p < 0.05)
increased this parameter compared to control plants. All
treatments under WS conditions had significantly (p <
0.05) higher leaf number and leaf surface or equivalent to
the control treatment under WW conditions, suggesting
that all treatments had the potential to offset the effect of
water stress.
3.3.4 Fruit Number and Fruit Weight
Under WW conditions, almost all treatments (except AMF)
significantly (p < 0.05) increased the fruit number per plant.
There was an improvement of 395% and 278% for the Comp
and PGPR + Comp treatments, respectively (Fig. 2e). In
WS conditions, the treatments Comp, PGPR + Comp, AMF
+ Comp, PGPR, and PGPR + Comp had significantly (p
< 0.05) increased the fruit numbers in comparison to the
control treatment, suggesting that these treatments had the
potential to compensate for the effect of water stress. Con-
cerning the yield under WW conditions, PGPR, PGPR +
AMF, and Comp treatments were the most effective treat-
ments resulting in 203%, 181%, and 176% of the increment,
respectively (Fig. 2f). None of the seven treatments has
enhanced fruit yield under WS conditions (Fig. 2f).
3.3.5 Mineral Nutrition of Tomato
The concentration of three ions ­(Na+, ­K+, and ­Ca2+) was
measured in both plant compartments (roots and shoots).

13
Journal of Soil Science and Plant Nutrition (2022) 22:743–764 753

The findings displayed a significant effect (p < 0.01) of the
two factors (water stress and biological treatments) and a
significant interaction (p < 0.01) between treatments and
water stress, indicating that water stress had different effects
depending on the treatments (Table 3).
The concentration of N ­ a+ and K­ + was increased in the
control treatment under WS conditions (Fig. 3a–d). ­Na+
concentration in roots and shoots increased significantly
(p < 0.05) in almost all treatments (Fig. 3a, b). The situ-
ation is significantly different for ­K+ as its concentration
increased significantly (p < 0.05) under the both condi-
tions in Comp, PGPR + Comp, AMF + Comp, PGPR +
AMF + Comp, and AMF treatments, except AMF + Comp
under WW conditions, and decreased significantly (p <
0.05) for the treatments with PGPR and PGPR + AMF
in shoot compartments (Fig. 3c). The ­K+ in the root part
was enhanced only by AMF + Comp treatment under WW
conditions. The ­Ca2+ concentration in the shoot and root
decreased significantly (p < 0.05) in the control treatment
under WS conditions. Biological treatments significantly
increased (p < 0.05) the ­Ca2+ concentration in the root
tissues, especially under WS conditions, suggesting that
these biological treatments could counteract the effect of
water stress (Fig. 3f). In the shoot compartment, the pic-
ture is contrasted according to the water supply conditions:
­ a2+
most treatments significantly decreased (p < 0.05) the C
concentration under WW conditions while four treatments
with PGPR inoculation (PGPR + Comp, PGPR + AMF +
Comp, PGPR, and PGPR + AMF) significantly increased
(p < 0.05) its concentration, counteracting the effect of
water stress (Fig. 3e).
P in tomato plants was decreased by water stress
(Fig. A2). Under WW conditions, Comp ± PGPR ± AMF
treatments increased significantly (p < 0.05) the amount
of P in plants. The highest amount was recorded in plants
amended with compost with an increment of 280% over
the control plants (Fig. A2). Under WS conditions, plants
amended with compost alone or in combination with the
AMF (Comp ± AMF) presented a significant increment (p
< 0.05) in P content of 84% and 75%, respectively (Fig. A2).

Fig. 3  Effects of water stress

and biofertilizers on a, b ­Na+
content, c, d ­K+ content, and
e, f ­Ca2+ content of the shoot
(a, c, e) and root (b, d, f) of
S. lycopersicum plants. Comp
compost; AMF arbuscular
mycorrhizal fungal consortium;
PGPR plant growth–promoting
rhizobacteria (BS14 and BS36
strains); WW well-watered
(75% field capacity (FC)); WS
water-stressed (35% FC). Data
presented are means ± SD. Bars
sharing the same letters in each
graphic are not significantly
different (p < 0.05)

13
754
3.4 Effect of Water Stress Under Different
Biofertilizer Treatments on Physiological
Parameters
Water stress affected negatively the gs, leaf Ψ, and Fv/Fm (p
< 0.01, Table 3). Under WS conditions, we noted that the
PGPR, PGPR + AMF, and AMF treatments significantly
increased the Fv/Fm (p < 0.05). Under WW conditions,
PGPR and AMF alone or in combination with compost
increased the gs in tomato plants than that in the control
plants (95.08 mmol ­H2O ­m−2 ­s−1) (Table  A1). Moreo-
ver, plants inoculated with PGPR alone or in combina-
tion with AMF significantly (p < 0.05) enhanced the leaf
Ψ (− 8 bar and − 8.83 bar) than WW control plants (− 10
bar, Table A1). According to the leaf Ψ, the applied biofer-
tilizers significantly improved the leaf Ψ than the control
plants under WS conditions, except for “AMF + Comp” and
“PGPR + AMF” treatments that had no significant effect
compared to the control plants (Table A1).
3.5 Effect of Water Stress Under Different
Biofertilizer Treatments on Proline Content
The effect of water stress on proline concentration in shoots of
the control treatment was very important, and a significant (p
< 0.05) increase reaching 471% was observed (Tables 3 and
A1). PGPR, AMF, and PGPR + AMF significantly decreased
(p < 0.05) the proline concentration in shoot under WS condi-
tions, counteracting the effect of water stress (Table A1). In the
root compartment, the observation was completely different as
there was no effect of water stress on proline concentration in
control plants. Under WW conditions, treatments with com-
post significantly (p < 0.05) decreased the root proline content.
The significant decrease (p < 0.05) in proline concentration
was only observed under WW conditions (Table A1).
3.6 Effect of Water Stress Under Different
Biofertilizer Treatments on Sugar, Protein,
and Polyphenol Content
Sugar, protein, and polyphenol concentrations were meas-
ured in three plant compartments (root, shoot, and fruit).
We recorded a significant effect (p < 0.01) of both factors
(water stress and biological treatments) and a significant
interaction (p < 0.01) between treatments and water stress
(except the concentration of polyphenol in roots), indicat-
ing that water stress had different effects depending on the
treatments (Table 3).
3.6.1 Sugar Content
In shoots, water stress significantly increased (p < 0.05)
the sugar concentration of the control treatment (Fig. 4a),
13
Journal of Soil Science and Plant Nutrition (2022) 22:743–764
although some treatments (Comp, Comp + AMF, and PGPR)
significantly increased (p < 0.05) shoot sugar content under
WW conditions. Under WS conditions, all treatments sig-
nificantly decreased (p < 0.05) shoot sugar concentration
(Fig. 4a). Water stress significantly decreased (p < 0.05)
the sugar concentration in the roots of the control treatment
(Fig. 4b). Under WW conditions, most of the treatments had
no effect or significantly decreased (p < 0.05) sugar concen-
tration, except for the Comp and AMF treatments for which
a positive and significant (p < 0.05) effect was observed
(Fig. 4b). However, PGPR and AMF treatments had signifi-
cantly (p < 0.05) increased root sugar content (Fig. 4b). For
sugar content in tomato fruits as fruit quality marker, water
stress significantly increased (p < 0.05) the sugar concentra-
tion of the control treatment (Fig. 4c). Sugar content increased
significantly (p < 0.05) in all treatments under WW conditions
(Fig. 4c). Under WS conditions, plants inoculated with PGPR
presented a significantly (p < 0.05) higher fruit sugar content.
3.6.2 Protein Content
The significant (p < 0.05) and positive effect on shoot pro-
tein content was observed in plants treated by Comp, AMF
+ Comp, PGPR + AMF + Comp, and PGPR + AMF under
both water regimes (Fig. 4d). Almost all the treatments had
a significant increase (p < 0.05) in protein concentration
of the root part under both conditions. Under WW condi-
tions, AMF + Comp, PGPR, and PGPR + AMF + Comp
treatments induced a high increment of 187%, 210%, and
223%, respectively, over WW control (Fig. 4e). Under WS
conditions, PGPR and AMF + Comp treatments resulted in
the high increment in the root protein content of 171% and
130%, respectively. Concerning the protein concentrations in
tomato fruits, no significant difference was recorded between
treatments and water supply conditions (Fig. 4f), except for
PGPR + AMF + Comp which enhanced the fruit quality
under WW conditions and PGPR alone which enhanced the
fruit quality under WS conditions.
3.6.3 Polyphenol Content
The general trend was the absence of water stress effects on
polyphenol content in root, shoot, and fruit compartments
(Fig. 4g–i). Under WW conditions, only PGPR + AMF treat-
ment had increased the shoot polyphenol content (Fig. 4g).
Under WS conditions, plants treated by PGPR + AMF +
Comp, PGPR, PGPR + AMF, and AMF presented signifi-
cantly higher polyphenol content (Fig. 4g). In the root part,
a decrease in the polyphenol content was recorded in plant
treated with Comp, PGPR + Comp, AMF + Comp, PGPR +
AMF + Comp, and PGPR treatments (Fig. 4h). On the other
hand, fruits’ polyphenol content decreased significantly (p <
0.05) in all treatments compared to the control under WW
Journal of Soil Science and Plant Nutrition (2022) 22:743–764
Fig. 4  Effects of water stress and biofertilizers on a–c sugar content,
d–f protein content, and g–i polyphenol contents of the shoot (a, d,
g), root (b, e, h), and fruits (c, f, i) of S. lycopersicum plants. Comp
compost; AMF arbuscular mycorrhizal fungal consortium; PGPR
conditions (Fig. 4i). Under WS conditions, the triple combina-
tion increased the polyphenol content. However, PGPR and
PGPR + AMF treatments decreased their amount (Fig. 4i).
3.7 Effect of Water Stress Under Different
Biofertilizer Treatments on Antioxidant Enzyme
Activities
The activities of the four antioxidant enzymes (PPO, POD,
CAT, and SOD) were measured in both plant compartments
(roots and shoots). We found a significant effect (p < 0.01)
of water stress considering all treatments (Compost, PGPR,
and AMF) and a significant interaction (p < 0.01) between
treatments and water stress, indicating that water stress had
different effects according to treatments (Table 3).
3.7.1 PPO and POD Activities
Both antioxidant enzyme activities were significantly
increased (p < 0.05) in the control treatment under WS
755
plant growth–promoting rhizobacteria (BS14 and BS36 strains); WW
well-watered (75% field capacity (FC)); WS water-stressed (35% FC).
Data presented are means ± SD. Bars sharing the same letters in each
graphic are not significantly different (p < 0.05)
conditions (Fig. 5a, b). In the root compartment, the gen-
eral trend was a significant decrease (p < 0.05) in PPO
and POD activities in all treatments, especially under WS
conditions (Fig. 5a, b), suggesting that the effect of the
biological treatments counterbalances that of water stress
in roots. In the shoot compartment, the effects were dif-
ferent (increase or decrease) depending on the treatments
(Fig. 5a, b). The PPO activity in the shoot was increased
by PGPR + Comp, PGPR, and AMF treatments under
WW conditions. Under WS conditions, it was decreased
by PGPR + Comp, AMF + Comp, PGPR + AMF + Comp,
and PGPR + AMF treatments (Fig. 5a). The shoot POD
activity was increased by treatments that contain PGPR
in addition to the plant inoculated with AMF alone and
decreased by AMF + Comp treatment. Under WS condi-
tions, the POD activity was increased by PGPR + Comp,
AMF + Comp, PGPR, and PGPR + AMF treatments and
decreased by Comp, PGPR + AMF + Comp, and AMF
treatments (Fig. 5b).
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756 Journal of Soil Science and Plant Nutrition (2022) 22:743–764

Fig. 5  Effects of water stress

and biofertilizers on a polyphe-
noloxidase (PPO), b peroxidase
(POD), c catalase (CAT), and
d superoxide dismutase (SOD)
activities in the root and shoot
part of tomato plants. Comp
compost; AMF arbuscular
mycorrhizal fungal consortium;
PGPR plant growth–promoting
rhizobacteria (BS14 and BS36
strains); WW well-watered
(75% field capacity (FC)); WS
water-stressed (35% FC). Data
presented are means ± SD.
Bars sharing the same letters in
each graphic and each part of
the plant (shoot or root) are not
significantly different (p < 0.05)

3.7.2 CAT and SOD Activities
As PPO and POD activities, CAT and SOD activities were sig-
nificantly increased (p < 0.05) in the control treatment under
WS conditions (Fig. 5c, d). In the root compartment, the over-
all trend was a significant decrease (p < 0.05) in CAT activities
in almost all treatments and regardless of the water supply sta-
tus, especially under WS conditions (Fig. 5c), and a significant
decrease of SOD activities under WS conditions (Fig. 5d). The
effect of biological treatments seems to counteract the effect of
water stress in the root compartment. In the shoot compartment,
no significant effect of the application of the biofertilizers under
WW conditions (Fig. 5c, d). Under WS conditions, CAT activity
in the shoot was increased by the application of the PGPR alone
or in combination with compost and/or AMF (Fig. 5c). The shoot
SOD activity was increased by the PGPR + Comp and PGPR +
AMF treatments. However, it was decreased by the application
of Comp, AMF + Comp, and AMF treatments (Fig. 5d).
3.8 Effect of Water Stress on Soil Physicochemical
and Microbial Properties
­ a+, ­K+,
Several soil parameters (pH, EC, soil available P, N
2+
­Ca , and TOC) were measured in soil fractions associated

13
Journal of Soil Science and Plant Nutrition (2022) 22:743–764
with the roots of all treatments. We found a significant effect
(p < 0.01) of both factors (water stress and biological treat-
ments) and a significant interaction (p < 0.01) between treat-
ments and water stress, indicating that water stress had dif-
ferent effects depending on the treatments (Table 3).
3.8.1 pH
The pH variations ranged between 7.94 and 8.69. The effect
of water stress in the control treatment was a significant
decrease (p < 0.05) of pH (0.42 unit, Table A2). Indeed,
compost treatment resulted in a significant decrease (p <
0.05) of soil pH under WS conditions, while PGPR and/or
AMF inoculation resulted in a significant increase (p < 0.05)
in soil pH under both water supply conditions (Table A2).
3.8.2 Soil EC
The effect of water stress in the control treatment was a
significant increase (p < 0.05) in EC (+ 148%, Table A2).
Under WW conditions, treatments with compost alone or in
combination with PGPR and/or AMF significantly increased
(p < 0.05) the EC (Table A3). Under WS conditions, there
was a significant increase (p < 0.05) of EC by Comp and
PGPR + AMF + Comp treatments (Table A2).
­ a2+, K
3.8.3 Mineral Elements (Soil Available P, C ­ +, and ­Na+)
The treatments combining compost and PGPR/AMF sig-
nificantly increased (p < 0.05) the concentration of avail-
able P under both water supply conditions (WW and WS)
(Table A2). Under WS conditions, inoculation of PGPR
also significantly increased the concentration of available
P (Table A2). The effect of water stress in the control treat-
ment was a significant increase (p < 0.05) in ­Ca2+ concen-
tration (Table A2). Under WW conditions, Comp, PGPR,
PGPR + AMF, PGPR + Comp, and PGPR + AMF + Comp
treatments significantly increased the soil ­Ca2+ concentra-
tion (p < 0.05, Table A2). Under WS conditions, Comp and
PGPR + Comp treatments significantly increased the C ­ a2+
content (Table A2). As for the concentrations of ­K+ and ­Na+
in soil, Comp, AMF + Comp, and PGPR + AMF treatments
increased the amount of ­Na+ under WW conditions and the
­K+ under WS conditions (Table A2).
3.8.4 TOC and the OM Percentage
Water stresses significantly decreased (p < 0.05) TOC and
OM. Almost all treatments carried out under WW condi-
tions (except PGPR + AMF) and under WS conditions
significantly increased (p < 0.05) the TOC and OM in soil
(Table A2). The biological treatments with PGPR and AMF
had a compensatory effect of water stress (no significant
757
difference of these treatments under WS conditions and the
control treatment under WW conditions) (Table A2). The
effect of the compost treatments was even stronger since
the values of these treatments under WS conditions were
significantly higher than those in the control treatment under
WW conditions (Table A2).
After the enumeration of bacteria and fungi in the soil
after the experiment, we found a significant (p < 0.05) effect
of water stress on the bacterial and fungal counts in most
treatments including the control treatment (Table A2). The
inoculation of PGPR strains resulted in a significant increase
(p < 0.01) in the bacterial counts in the four treatments
inoculated by the two PGPR strains, regardless of the water
supply condition (WW vs. WS, Table A3). Fungal counts
showed a significant increase (p < 0.05) for 4 out of the 6
treatments (Table A2).
4 Discussion
The drought is considered as one of the serious environ-
mental constraints that affect plant development, yield, and
mineral nutrition (Armada et al. 2014; Meddich et al. 2015;
Ortiz et al. 2015; Anli et al. 2020). Accordingly, the present
study was conducted to assess the efficiency of AMF and/
or PGPR inoculation and/or compost amendment to allevi-
ate water stress effect on the growth and development of S.
lycopersicum.
Our results confirmed that the WS conditions exerted a
negative impact on the growth, development, and productiv-
ity of tomato plants compared to the WW conditions. Con-
sequently, a significant decrease in the case of shoot dry
weight (SDW) and the root dry weight (RDW) was observed.
It is well documented that water stress negatively affects
plant growth and yield (Armada et al. 2014; Calvo-Polanco
et al. 2016; Aalipour et al. 2020). In harmony with these
results, Ullah et al. (2016) and Nahar and Ullah (2018) dem-
onstrated that growing S. lycopersicum under drought stress
reduced plant elongation, SDW, RDW, and fruit yield. Simi-
lar results were found by Hashem et al. (2019) who recorded
a significant decrease in plant growth under WS conditions.
Drought influences the transport and availability of soil
nutrients (Vurukonda et al. 2016) and affects the morpholog-
ical, physiological, and nutritional traits of plants especially
water content, leaf water potential, photosynthetic pigment,
stomatal conductance, and P and N absorption (Jaleel et al.
2009; Augé et al. 2014; Baslam et al. 2014; Symanczik et al.
2018).
The major consequence of water deficit in plants is the
decrease or suppression of photosynthesis. Water deficit
occurs in plants in part due to the loss of water from the
leaves when the stomata open to allow the entry of C ­ O2
from the atmosphere to be used in photosynthesis. The first
13
758
response of plants during drought stress is to close the sto-
mata to avoid water loss through transpiration (Kaur et al.
2021). Consequently, stomatal closure during low water
supply minimizes transpirational water loss. Because of the
ongoing photosynthetic process in the light, the enhanced
gas diffusion barrier leads to depletion of intercellular
­CO2. Lower ­CO2 diffusion from the environment to the
carboxylation site is considered to be the major reason
behind decreased photosynthetic rate during mild to mod-
erate drought stress (Pinheiro and Chaves 2011). Besides, it
stimulates oxygenation of ribulose-1,5-bisphosphate (RuBP)
thus leading to a photorespiratory burst of hydrogen perox-
ide ­(H2O2) in the peroxisomes (Laxa et al. 2019) disturbing
the cellular ionic balance in plants (Abobatta 2019). Water-
deprived conditions also alter the electron transport chain
(ETC), which further enhances ROS within cell organelles,
thereby reducing the pigment levels and causing disruption
of thylakoid structures. The impairment of ETC is mainly
attributed to P700 oxidation under drought conditions that
not only decrease photosystem II functioning, but also
reduce the quantum efficiency of photosystem I. This is also
accompanied by elevated non-photochemical quenching and
reduced plastoquinone pools (Tikhonov 2013; Wada et al.
2019). Consequently, the declining pigment levels collapse
the entire chloroplast membrane to nullify photosynthesis by
affecting ribulose-1,-5-bisphosphate carboxylase/oxygenase
(Rubisco), carboxylation, amino acid levels, and RuBP for-
mation (Faraloni et al. 2011).
The lower intracellular ­CO2 levels reduce the reactions
of the Calvin cycle, which leads to lower consumption of
NADPH and ATP. This response results in a lack of regener-
ation of electron acceptors ­(NADP+, ­NAD+, FAD), facilitat-
ing the transference of electrons from the electron transport
chain to oxygen and leading to greater production of ROS
such as H­ 2O2 (hydrogen peroxide), O ­ 2− (superoxide), and
−
­OH (hydroxyl) radicals (Chiappero et al. 2019).
To counteract ROS overproduction under water stress
conditions, plants produce and accumulate non-enzymatic
metabolites (proline and polyphenols) and antioxidant
enzymes (CAT, SOD, PPO, and POD). It is well reported
that drought promotes overproduction of ROS in plant cells,
inducing oxidative stress (Ruiz-Lozano 2003; Armada et al.
2014; Ortiz et al. 2015; Khanna et al. 2019a, c). In this study,
treated tomato plants recorded high soluble protein content
in the leaves and roots of WS plants. Compost application
and inoculation with AMF and PGPR allayed or reduced
RNA disassembly and could increase the capacity of the
non-enzymatic antioxidant defense system using soluble
proteins (Abbaspour et  al. 2012). In our study, tomato
plants inoculated with PGPR increased the level of antioxi-
dant enzymes, especially in the shoot part. SOD scavenges
­O2− by converting it into ­H2O2. Indeed, H­ 2O2 is originated
in plants by two possible pathways, with the involvement of
13
Journal of Soil Science and Plant Nutrition (2022) 22:743–764
SOD or via oxidases. In addition, it is known that an excess
of ­H2O2 can be reduced into H ­ 2O by POD, CAT, ascorbate
peroxidase, glutathione reductase, monodehydroascorbate
reductase, and dehydroascorbate reductase at different cel-
lular compartments (Chiappero et al. 2019; Khanna et al.
2019b, 2020; Singh et al. 2021). Many reports have revealed
that inoculation of PGPR under water deficiency was related
to ROS scavenging enzymes, resulting in an increased accu-
mulation of several antioxidant enzymes diminishing oxida-
tive damage (Gururani et al. 2013; Ortiz et al. 2015; Sara-
vanakumar et al. 2011). In accordance, Ortiz et al. (2015)
have reported that plants inoculated with autochthonous bac-
teria and/or AMF increased antioxidant enzyme activities
resulting in better protection against ROS. Moreover, they
found that the application of autochthonous microorganisms
(bacterial and mycorrhizal inoculum) enhanced mineral ele-
ments: P by 89%, Ca by 131%, Mg by 79%, and Zn by 62%
under water stress. Besides, the dual inoculation of these
two microorganisms significantly enhanced the K content.
K is an important element; it is implicated in plant resistance
in response to water stress. This nutrient is reported to be
capable to keep a high photosynthesis rate and atmospheric
carbon fixation and to preserve the photosynthetic system
against oxidative injury (Ortiz et al. 2015).
The application of compost, AMF, and PGPR in our study
alleviated water stress. PGPR application has been reported
to improve water stress tolerance (Rolli et al. 2014; Vuru-
konda et al. 2016). For instance, Rolli et al. (2014) have
demonstrated that the application of Pseudomonas, Acine-
tobacter, and Bacillus has improved the growth of pepper
plants under water stress. This improvement could be attrib-
uted to the ability of these species to solubilize phosphates,
IAA production, and their ACC deaminase activity (Indira-
gandhi et al. 2008; Kang et al. 2009; Mehnaz et al. 2010;
Rokhbakhsh-Zamin et al. 2011; Ahemad and Kibret 2014;
Kang et al. 2014) and/or to the ability of Acinetobacter sp.
to produce EPS and siderophores (Rolli et al. 2014). EPS
production by bacteria is an important trait for plant root
colonization (Santaella et al. 2008) and for structuring the
soil surrounding the roots (Amellal et al. 1998; Alami et al.
2000; Bezzate et al. 2000). The used PGPR strains were able
to reduce the activity of plant antioxidant enzymes under
stressful conditions and increase the amount of GAs (Kang
et al. 2014). AMF are known to improve plant uptake of
water and nutrients from the surrounding soil (Smith and
Read 2008; Raklami et al. 2019), by increasing the explored
soil surface area, resulting in more water and nutrient uptake
(Calvo-Polanco et al. 2016). Previous studies have shown
that the application of the autochthonous AMF consortium
significantly improved plant growth under osmotic stress
conditions (Meddich et al. 2015; Ben-Laouane et al. 2019,
2020; Anli et al. 2020; Toubali et al. 2020). The AMF con-
sortium improved plant tolerance to water stress (Meddich
Journal of Soil Science and Plant Nutrition (2022) 22:743–764
et al. 2015; Anli et al. 2020) and salinity (Ben-Laouane et al.
2019, 2020; Toubali et al. 2020). The beneficial effect of
mycorrhizal and PGPR associations and compost amend-
ment on the growth of tomato plants, under water deficit,
could be explained by the greater uptake of nutrients with
low mobility, such as P and N contained in the soil (Jaleel
et al. 2009; Augé et al. 2014; Baslam et al. 2014; Symanczik
et al. 2018; Raklami et al. 2019). In this study, we showed
that inoculated and amended plants had considerably higher
mineral nutrient content (P and N) as compared to controls
under water deficit conditions allowing higher plant perfor-
mance. This resulted from the better absorption of the sur-
face area provided by extensive AMF hyphae and/or a direct
uptake from compost to plant roots and/or the mobilization
and absorption of various nutrients from the soil to plants
by PGPR.
Comparable results were recorded by Armada et  al.
(2014) who evaluated the impact of the combined applica-
tion of beneficial soil microorganisms (bacteria and AMF)
and agro-waste residue to increase plant growth under water
deficiency in a semiarid area. They found that the plant bio-
mass of compost-amended plants was higher than plants co-
inoculated with bacteria (Bacillus megaterium) and AMF
(autochthonous consortium composed of Septoglomus
constrictum, Diversispora aunantia, Archaeospora trap-
pei, Glomus versiforme, and Paraglomus occultum) and not
amended with compost. The highest total yielded biomass,
after three harvests, was recorded in plants treated with bac-
teria and amended with compost. Tartoura (2010) showed
that the positive effect of compost on soil was related to
the improvement of water retention and enhancing the FC
of the soil providing more water to the stressed plants. The
association with AMF amends the plants’ water regulation
by triggering hormonal signaling such as ABA mediating
stomatal conductance or by stimulating osmolytes. Other
studies showed, under drought stress, the development of
microorganism-mediated mechanisms, including modifi-
cations in the content of plant hormones (strigolactones,
jasmonic acid, and ABA) and improvement in plant water
status by increasing hydraulic conductivity (Chaumont and
Tyerman 2014; Fernández-Lizarazo and Moreno-Fonseca
2016). The increase in root hydraulic conductivity can be
related to an enhanced expression in AMF or plant aquapor-
ins (Sánchez-Romera et al. 2016).
Many studies have reported that the application of
AMF and PGPR and compost amendments improved plant
growth and tolerance to water stress (Meddich et al. 2015;
Armada et al. 2014; Bowles et al. 2016; Calvo-Polanco et al.
2016; Razavipour et al. 2018; Chiappero et al. 2019). This
effectiveness might be attributed to multiple mechanisms
like nutritional and physiological ameliorations through
increased water and nutrient acquisition. The application of
PGPR has been reported to enhance water stress tolerance
759
(Vurukonda et al. 2016), and they are able to mitigate water
stress by the synthesis of EPS, phytohormone, ACC deami-
nase, and volatile compounds. Moreover, they increased
osmolytes and antioxidant enzymes, regulating stress-
responsive genes and inducing changes in the root system
(Vurukonda et al. 2016; Khan et al. 2019). On the other
hand, AMF are attractive for increasing water supply by
improving soil characteristics and enhancing P acquisition
(Augé 2004; Amiri et al. 2015). AMF can positively influ-
ence the growth of 80% of their host plants by improving
the mineral and water nutrition of plants (Smith and Read
2008). Furthermore, compost beneficially influences plant
growth and biological yield advantageously and increases
OM, thereby improving nutrient availability (Weber et al.
2014). Consequently, inoculated and amended plants could
be less affected by water stress (Armada et al. 2014).
Accumulation of osmolytes is important for plant protec-
tion and osmotic adjustment under water stress. Water stress
induced an important accumulation of proline in the shoot
part of stressed plants. Proline, a non-protein amino acid
that accumulates in various parts of the plant exposed to
water deficiency, is considered one of the best compounds
that regulate osmosis in plants (Kishor and Sreenivasulu
2014) and could be easily degraded when the normal water
regime comes back (Singh et al. 2015). In addition to the
role of osmotic adjustments, proline stabilizes cellular struc-
tures like proteins and membranes, decreased the amount
of ROS, and adjusts redox potential (Hayat et  al. 2012;
Ortiz et al. 2015). The accumulation of proline in plant tis-
sues is directly related to the capacity of plants to mitigate
drought (Ortiz et al. 2015). Furthermore, they reported that
high accumulation of proline is an indicator of improved
osmoprotective ability and the capacity of inoculated plants
to ensure high water uptake under water stress (Ortiz et al.
2015).
In our study, AMF and PGPR increased sugar content in
WS plants compared to non-inoculated plants. This could be
a strategy of plants to mitigate the negative effect of water
stress by accumulating soluble sugar as osmolytes (Khaleghi
et al. 2019). Previous researchers have reported that envi-
ronmental stresses (such as drought and salinity) affect fruit
quality (Ozbahce and Tari 2010; Patanè et al. 2011; Zhang
et al. 2016). In this study, we have demonstrated that water
stress enhanced sugar accumulation. In harmony with this
outcome, Ozbahce and Tari (2010) reported that increasing
water stress levels (50% FC, and 25% FC) have proportion-
ally enhanced the total soluble sugar content. This finding
could be explained by a fall in water accumulation by the
fruit without any significant modification in the quantity of
accumulated sugars (Patanè et al. 2011). Concerning the
protein concentrations in tomato fruits, no significant dif-
ference was recorded between treatments and water supply
conditions. However, Wakchaure et al. (2020) have found
13
760
that water stress greatly increased the protein concentrations
in developing fruit of eggplant (Solanum melongena L.).
Acinetobacter sp. and Rahnella aquatilis applications have
increased the fruit protein content under WS conditions.
This phenomenon could be explained by the ability of PGPR
to produce iron-binding protein and siderophores that pro-
vide easy absorption of nutrients during drought stress. For
instance, Flores-Félix et al. (2015) have demonstrated that
inoculation with a strain of Phyllobacterium, able to produce
siderophores and biofilms and to colonize strawberry roots,
is able to increase the yield and quality of strawberry plants
under drought stress.
The biofertilizers tested in our study showed an
enhancement of soil properties after the experiment. We
noted a significant increment in OM and available P, espe-
cially with the application of compost alone or in combi-
nation with the PGPR or the AMF. This result could be
attributed to the high content of OM and available P in
the compost. Moreover, a high amount of available P was
recorded in stressed plants and amended with PGPR that
solubilize tricalcium phosphate and potassium. A decrease
in soil pH was recorded after compost supplementation.
This result could be assigned to the transformation of OM,
resulting in a ­CO2 release (Ben Achiba et al. 2009). Simi-
lar changes in pH were reported after compost applica-
tion by Lakhdar et al. (2011). Composts usually contain
soluble salts in great quantities, and their amendment on
soils raises EC values. It is commonly documented that the
use of compost enhances soil characteristics by improving
the amount of organic carbon and EC (Weber et al. 2007,
2014). The increase of OM and EC had a great impact
on soils that are low in mineral and organic compounds,
with poor fertility and mineral nutrient scarcity (Weber
et al. 2007). Moreover, compost application enhanced soil
fertility by improving the amounts of exchangeable micro-
nutrients and cations (Fagnano et al. 2011; Lakhdar et al.
2011; Weber et al. 2014).
5 Conclusion
Compost application alone or in combination with plant
growth–promoting rhizobacteria (PGPR) or arbuscular
mycorrhizal fungi (AMF) alleviated the negative effect
of water stress and, thus, a significant increase in plant
growth, development, and nutrient acquisition. Applica-
tion of all biofertilizers improved fruit quality by enhanc-
ing the sugar and protein content under well-watered
conditions. Indeed, PGPR boosted the fruit quality by
increasing sugar and protein content under stressful con-
ditions. Drought induction in tomato triggered several
13
Journal of Soil Science and Plant Nutrition (2022) 22:743–764
physiological, biochemical, and enzymatic responses. It
is further observed that the application of PGPR alone or
in combinations increased chlorophyll fluorescence and
boosted the catalase, superoxide dismutase, and peroxi-
dase activities in the shoot part of the tomato plants. The
overall results revealed the promising effect of compost,
PGPR, and AMF in water stress tolerance of tomato plants.
Our results provide new opportunities to harness the
potential of biofertilizer combinations to mitigate abiotic
stress in agricultural crops. Further research is still needed
to uncover the underlying mechanism of improved plant
drought tolerance by compost, PGPR, and AMF. Finally,
the authors suggest an open-field experiment to assess
the effects of these tested biofertilizers on the growth and
yield of tomato under non-controlled conditions before
their commercialization as a biofertilizer for drought man-
agement in tomato.
Supplementary Information  The online version contains supplemen-
tary material available at https://d​ oi.o​ rg/1​ 0.1​ 007/s​ 42729-0​ 21-0​ 0684-w.
Acknowledgements  This work was partially supported by the project
PPR2/2016/42, CNRST Morocco, and the Alexander von Humboldt
foundation.
Author Contribution  Conceptualization: O.K. and M.A.; methodology:
T.A., R.A., B.N., and A.M.; data curation and formal analysis: T.A.,
R.A., and A.A.; investigation: T.A., R.A., and A.A.; writing of the
original draft: T.A.; writing (review and editing): O.K., M.A., G.M.,
R.A., A.W., and H.T.; funding acquisition: O.K. and M.A.; resources:
O.K., M.A., and G.M.; supervision: O.K. and M.A.
Declarations 
Conflict of Interest  The authors declare no competing interests.
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Authors and Affiliations
Abdel‑ilah Tahiri1,2 · Abdelilah Meddich2 · Anas Raklami1   · Abdelrahman Alahmad3 · Noura Bechtaoui4 ·
Mohamed Anli2 · Michael Göttfert5 · Thierry Heulin3 · Wafa Achouak3 · Khalid Oufdou1,4
1 3
Laboratory of Microbial Biotechnologies, Agrosciences
and Environment (BioMAgE), Labeled Research
Unit‑CNRST N 4, Faculty of Sciences Semlalia, Cadi Ayyad
University, PO Box 2390, Marrakech, Morocco
2 4
Laboratory of Agro‑Food, Biotechnologies and Valorization
of Plant Bioresources (Agrobioval), Faculty of Sciences
Semlalia, Cadi Ayyad University, PO Box 2390, Marrakech,
Morocco
13
Journal of Soil Science and Plant Nutrition (2022) 22:743–764
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